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Hanitrarimalala V, Prgomet Z, Hedhammar M, Tassidis H, Wingren AG. In vitro 3D modeling of colorectal cancer: the pivotal role of the extracellular matrix, stroma and immune modulation. Front Genet 2025; 16:1545017. [PMID: 40376304 PMCID: PMC12078225 DOI: 10.3389/fgene.2025.1545017] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2024] [Accepted: 04/23/2025] [Indexed: 05/18/2025] Open
Abstract
Colorectal cancer (CRC) is a leading global cancer with high mortality, especially in metastatic cases, with limited therapeutic options. The tumor microenvironment (TME), a network comprising various immune cells, stromal cells and extracellular (ECM) components plays a crucial role in influencing tumor progression and therapy outcome. The genetic heterogeneity of CRC and the complex TME complicates the development of effective, personalized treatment strategies. The prognosis has slowly improved during the past decades, but metastatic CRC (mCRC) is common among patients and is still associated with low survival. The therapeutic options for CRC differ from those for mCRC and include surgery (mostly for CRC), chemotherapy, growth factor receptor signaling pathway targeting, as well as immunotherapy. Malignant CRC cells are established in the TME, which varies depending on the primary or metastatic site. Herein, we review the role and interactions of several ECM components in 3D models of CRC and mCRC tumor cells, with an emphasis on how the TME affects tumor growth and treatment. This comprehensive summary provides support for the development of 3D models that mimic the interactions within the TME, which will be essential for the development of novel anticancer therapies.
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Affiliation(s)
- Veroniaina Hanitrarimalala
- Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden
- Biofilms-Research Center for Biointerfaces, Malmö University, Malmö, Sweden
| | - Zdenka Prgomet
- Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden
- Biofilms-Research Center for Biointerfaces, Malmö University, Malmö, Sweden
| | - My Hedhammar
- KTH Royal Institute of Technology, Division of Protein Technology, Stockholm, Sweden
| | - Helena Tassidis
- Department of Bioanalysis, Faculty of Natural Sciences, Kristianstad University, Kristianstad, Sweden
| | - Anette Gjörloff Wingren
- Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden
- Biofilms-Research Center for Biointerfaces, Malmö University, Malmö, Sweden
- Department of Bioanalysis, Faculty of Natural Sciences, Kristianstad University, Kristianstad, Sweden
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2
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Kataki AD, Gupta PG, Cheema U, Nisbet A, Wang Y, Kocher HM, Pérez-Mancera PA, Velliou EG. Mapping Tumor-Stroma-ECM Interactions in Spatially Advanced 3D Models of Pancreatic Cancer. ACS APPLIED MATERIALS & INTERFACES 2025; 17:16708-16724. [PMID: 40052705 PMCID: PMC11931495 DOI: 10.1021/acsami.5c02296] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 02/03/2025] [Revised: 02/19/2025] [Accepted: 02/20/2025] [Indexed: 03/21/2025]
Abstract
Bioengineering-based in vitro tumor models are increasingly important as tools for studying disease progression and therapy response for many cancers, including the deadly pancreatic ductal adenocarcinoma (PDAC) that exhibits a tumor/tissue microenvironment of high cellular/biochemical complexity. Therefore, it is crucial for in vitro models to capture that complexity and to enable investigation of the interplay between cancer cells and factors such as extracellular matrix (ECM) proteins or stroma cells. Using polyurethane (PU) scaffolds, we performed a systematic study on how different ECM protein scaffold coatings impact the long-term cell evolution in scaffolds containing only cancer or only stroma cells (activated stellate and endothelial cells). To investigate potential further changes in those biomarkers due to cancer-stroma interactions, we mapped their expression in dual/zonal scaffolds consisting of a cancer core and a stroma periphery, spatially mimicking the fibrotic/desmoplastic reaction in PDAC. In our single scaffolds, we observed that the protein coating affected the cancer cell spatial aggregation, matrix deposition, and biomarker upregulation in a cell-line-dependent manner. In single stroma scaffolds, different levels of fibrosis/desmoplasia in terms of ECM composition/quantity were generated depending on the ECM coating. When studying the evolution of cancer and stroma cells in our dual/zonal model, biomarkers linked to cell aggressiveness/invasiveness were further upregulated by both cancer and stroma cells as compared to single scaffold models. Collectively, our study advances the understanding of how different ECM proteins impact the long-term cell evolution in PU scaffolds. Our findings show that within our bioengineered models, we can stimulate the cells of the PDAC microenvironment to develop different levels of aggressiveness/invasiveness, as well as different levels of fibrosis. Furthermore, we highlight the importance of considering spatial complexity to map cell invasion. Our work contributes to the design of in vitro models with variable, yet biomimetic, tissue-like properties for studying the tumor microenvironment's role in cancer progression.
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Affiliation(s)
- Anna-Dimitra Kataki
- Centre
for 3D models of Health and Disease, Division of Surgery and Interventional
Science, University College London, London W1W 7TY, U.K.
| | - Priyanka G. Gupta
- Centre
for 3D models of Health and Disease, Division of Surgery and Interventional
Science, University College London, London W1W 7TY, U.K.
- School
of Life and Health Sciences, Whitelands College, University of Roehampton, London SW15 4JD, U.K.
| | - Umber Cheema
- Centre
for 3D models of Health and Disease, Division of Surgery and Interventional
Science, University College London, London W1W 7TY, U.K.
| | - Andrew Nisbet
- Department
of Medical Physics and Biomedical Engineering, University College London, London WC1E 6BT, U.K.
| | - Yaohe Wang
- Centre
for Cancer Biomarkers and Biotherapeutics, Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, U.K.
| | - Hemant M. Kocher
- Centre
for Tumour Biology and Experimental Cancer Medicine, Barts Cancer
Institute, Queen Mary University of London, London EC1M 6BQ, U.K.
| | - Pedro A. Pérez-Mancera
- Department
of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool L69 3GE, U.K.
| | - Eirini G. Velliou
- Centre
for 3D models of Health and Disease, Division of Surgery and Interventional
Science, University College London, London W1W 7TY, U.K.
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3
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Zambuto SG, Theriault H, Jain I, Crosby CO, Pintescu I, Chiou N, Oyen ML, Zoldan J, Underhill GH, Harley BAC, Clancy KBH. Endometrial decidualization status modulates endometrial microvascular complexity and trophoblast outgrowth in gelatin methacryloyl hydrogels. NPJ WOMEN'S HEALTH 2024; 2:22. [PMID: 39036057 PMCID: PMC11259096 DOI: 10.1038/s44294-024-00020-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/05/2023] [Accepted: 04/20/2024] [Indexed: 07/23/2024]
Abstract
The endometrium undergoes rapid cycles of vascular growth, remodeling, and breakdown during the menstrual cycle and pregnancy. Decidualization is an endometrial differentiation process driven by steroidal sex hormones that is critical for blastocyst-uterine interfacing and blastocyst implantation. Certain pregnancy disorders may be linked to decidualization processes. However, much remains unknown regarding the role of decidualization and reciprocal trophoblast-endometrial interactions on endometrial angiogenesis and trophoblast invasion. Here, we report an engineered endometrial microvascular network embedded in gelatin hydrogels that displays morphological and functional patterns of decidualization. Vessel complexity and biomolecule secretion are sensitive to decidualization and affect trophoblast motility, but that signaling between endometrial and trophoblast cells was not bi-directional. Although endometrial microvascular network decidualization status influences trophoblast cells, trophoblast cells did not induce structural changes in the endometrial microvascular networks. These findings add to a growing literature that the endometrium has biological agency at the uterine-trophoblast interface during implantation. Finally, we form a stratified endometrial tri-culture model, combining engineered microvascular networks with epithelial cells. These endometrial microvascular networks provide a well-characterized platform to investigate dynamic changes in angiogenesis in response to pathological and physiological endometrial states.
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Affiliation(s)
- Samantha G. Zambuto
- Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Hannah Theriault
- Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Ishita Jain
- Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Cody O. Crosby
- Department of Physics, Southwestern University, Georgetown, TX 78626, USA
- Department of Biomedical Engineering, University of Texas at Austin, Austin, TX 78712, USA
| | - Ioana Pintescu
- Department of Molecular and Cellular Biology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Noah Chiou
- Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Michelle L. Oyen
- Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130, USA
- Center for Women’s Health Engineering, Washington University in St. Louis, St. Louis, MO 63130, USA
- Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO 63130, USA
| | - Janet Zoldan
- Department of Biomedical Engineering, University of Texas at Austin, Austin, TX 78712, USA
| | - Gregory H. Underhill
- Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Brendan A. C. Harley
- Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
- Department Chemical and Biomolecular Engineering, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
- Cancer Center at Illinois, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
| | - Kathryn B. H. Clancy
- Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
- Department of Anthropology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
- Beckman Institute for Advanced Science & Technology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA
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4
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Torabizadeh F, Talaei-Khozani T, Yaghobi A, Walker M, Mirzaei E. Enhancing chondrogenic differentiation of mesenchymal stem cells through synergistic effects of cellulose nanocrystals and plastic compression in collagen-based hydrogel for cartilage formation. Int J Biol Macromol 2024; 272:132848. [PMID: 38830491 DOI: 10.1016/j.ijbiomac.2024.132848] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2024] [Revised: 05/22/2024] [Accepted: 05/31/2024] [Indexed: 06/05/2024]
Abstract
Collagen-based (COL) hydrogels could be a promising treatment option for injuries to the articular cartilage (AC) becuase of their similarity to AC native extra extracellular matrix. However, the high hydration of COL hydrogels poses challenges for AC's mechanical properties. To address this, we developed a hydrogel platform that incorporating cellulose nanocrystals (CNCs) within COL and followed by plastic compression (PC) procedure to expel the excessive fluid out. This approach significantly improved the mechanical properties of the hydrogels and enhanced the chondrogenic differentiation of mesenchymal stem cells (MSCs). Radially confined PC resulted in higher collagen fibrillar densities together with reducing fibril-fibril distances. Compressed hydrogels containing CNCs exhibited the highest compressive modulus and toughness. MSCs encapsulated in these hydrogels were initially affected by PC, but their viability improved after 7 days. Furthermore, the morphology of the cells and their secretion of glycosaminoglycans (GAGs) were positively influenced by the compressed COL-CNC hydrogel. Our findings shed light on the combined effects of PC and CNCs in improving the physical and mechanical properties of COL and their role in promoting chondrogenesis.
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Affiliation(s)
- Farid Torabizadeh
- Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Tahereh Talaei-Khozani
- Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Atefeh Yaghobi
- Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
| | - Matthew Walker
- Centre for the Cellular Microenvironment, University of Glasgow, UK
| | - Esmaeil Mirzaei
- Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.
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Balukova A, Bokea K, Barber PR, Ameer-Beg SM, MacRobert AJ, Yaghini E. Cellular Imaging and Time-Domain FLIM Studies of Meso-Tetraphenylporphine Disulfonate as a Photosensitising Agent in 2D and 3D Models. Int J Mol Sci 2024; 25:4222. [PMID: 38673807 PMCID: PMC11050357 DOI: 10.3390/ijms25084222] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/04/2024] [Revised: 03/28/2024] [Accepted: 03/29/2024] [Indexed: 04/28/2024] Open
Abstract
Fluorescence lifetime imaging (FLIM) and confocal fluorescence studies of a porphyrin-based photosensitiser (meso-tetraphenylporphine disulfonate: TPPS2a) were evaluated in 2D monolayer cultures and 3D compressed collagen constructs of a human ovarian cancer cell line (HEY). TPPS2a is known to be an effective model photosensitiser for both Photodynamic Therapy (PDT) and Photochemical Internalisation (PCI). This microspectrofluorimetric study aimed firstly to investigate the uptake and subcellular localisation of TPPS2a, and evaluate the photo-oxidative mechanism using reactive oxygen species (ROS) and lipid peroxidation probes combined with appropriate ROS scavengers. Light-induced intracellular redistribution of TPPS2a was observed, consistent with rupture of endolysosomes where the porphyrin localises. Using the same range of light doses, time-lapse confocal imaging permitted observation of PDT-induced generation of ROS in both 2D and 3D cancer models using fluorescence-based ROS together with specific ROS inhibitors. In addition, the use of red light excitation of the photosensitiser to minimise auto-oxidation of the probes was investigated. In the second part of the study, the photophysical properties of TPPS2a in cells were studied using a time-domain FLIM system with time-correlated single photon counting detection. Owing to the high sensitivity and spatial resolution of this system, we acquired FLIM images that enabled the fluorescence lifetime determination of the porphyrin within the endolysosomal vesicles. Changes in the lifetime dynamics upon prolonged illumination were revealed as the vesicles degraded within the cells.
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Affiliation(s)
- Andrea Balukova
- Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK; (A.B.); (K.B.)
| | - Kalliopi Bokea
- Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK; (A.B.); (K.B.)
| | - Paul R. Barber
- Department of Oncology, UCL Cancer Institute, University College London, London WC1E 6DD, UK;
- Comprehensive Cancer Centre, School of Cancer and Pharmaceutical Sciences, King’s College London, London SE1 9RT, UK;
| | - Simon M. Ameer-Beg
- Comprehensive Cancer Centre, School of Cancer and Pharmaceutical Sciences, King’s College London, London SE1 9RT, UK;
| | - Alexander J. MacRobert
- Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK; (A.B.); (K.B.)
| | - Elnaz Yaghini
- Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London NW3 2QG, UK; (A.B.); (K.B.)
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6
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Bakkalci D, Al-Badri G, Yang W, Nam A, Liang Y, Khurram SA, Heavey S, Fedele S, Cheema U. Spatial transcriptomic interrogation of the tumour-stroma boundary in a 3D engineered model of ameloblastoma. Mater Today Bio 2024; 24:100923. [PMID: 38226014 PMCID: PMC10788620 DOI: 10.1016/j.mtbio.2023.100923] [Citation(s) in RCA: 3] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2023] [Revised: 11/24/2023] [Accepted: 12/17/2023] [Indexed: 01/17/2024] Open
Abstract
Stromal cells are key components of the tumour microenvironment (TME) and their incorporation into 3D engineered tumour-stroma models is essential for tumour mimicry. By engineering tumouroids with distinct tumour and stromal compartments, it has been possible to identify how gene expression of tumour cells is altered and influenced by the presence of different stromal cells. Ameloblastoma is a benign epithelial tumour of the jawbone. In engineered, multi-compartment tumouroids spatial transcriptomics revealed an upregulation of oncogenes in the ameloblastoma transcriptome where osteoblasts were present in the stromal compartment (bone stroma). Where a gingival fibroblast stroma was engineered, the ameloblastoma tumour transcriptome revealed increased matrix remodelling genes. This study provides evidence to show the stromal-specific effect on tumour behaviour and illustrates the importance of engineering biologically relevant stroma for engineered tumour models. Our novel results show that an engineered fibroblast stroma causes the upregulation of matrix remodelling genes in ameloblastoma which directly correlates to measured invasion in the model. In contrast the presence of a bone stroma increases the expression of oncogenes by ameloblastoma cells.
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Affiliation(s)
- Deniz Bakkalci
- UCL Centre for 3D Models of Health and Disease, Division of Surgery and Interventional Science, University College London, Charles Bell House, 43-45 Foley Street, W1W 7TS, London, UK
| | - Georgina Al-Badri
- Department of Mathematics, University College London, 25 Gordon Street, WC1H 0AY, London, UK
| | - Wei Yang
- NanoString Technologies, 530 Fairview Ave N, Seattle, WA 98109, USA
| | - Andy Nam
- NanoString Technologies, 530 Fairview Ave N, Seattle, WA 98109, USA
| | - Yan Liang
- NanoString Technologies, 530 Fairview Ave N, Seattle, WA 98109, USA
| | - Syed Ali Khurram
- Unit of Oral and Maxillofacial Pathology, School of Clinical Dentistry, University of Sheffield, 19 Claremont Crescent, S10 2TA, Sheffield, UK
| | - Susan Heavey
- UCL Centre for 3D Models of Health and Disease, Division of Surgery and Interventional Science, University College London, Charles Bell House, 43-45 Foley Street, W1W 7TS, London, UK
| | - Stefano Fedele
- Eastman Dental Institute, University College London, London, UK
| | - Umber Cheema
- UCL Centre for 3D Models of Health and Disease, Division of Surgery and Interventional Science, University College London, Charles Bell House, 43-45 Foley Street, W1W 7TS, London, UK
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Hassani I, Anbiah B, Moore AL, Abraham PT, Odeniyi IA, Habbit NL, Greene MW, Lipke EA. Establishment of a tissue-engineered colon cancer model for comparative analysis of cancer cell lines. J Biomed Mater Res A 2024; 112:231-249. [PMID: 37927200 PMCID: PMC12083550 DOI: 10.1002/jbm.a.37611] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/10/2022] [Revised: 08/13/2023] [Accepted: 08/30/2023] [Indexed: 11/07/2023]
Abstract
To overcome the limitations of in vitro two-dimensional (2D) cancer models in mimicking the complexities of the native tumor milieu, three-dimensional (3D) engineered cancer models using biomimetic materials have been introduced to more closely recapitulate the key attributes of the tumor microenvironment. Specifically, for colorectal cancer (CRC), a few studies have developed 3D engineered tumor models to investigate cell-cell interactions or efficacy of anti-cancer drugs. However, recapitulation of CRC cell line phenotypic differences within a 3D engineered matrix has not been systematically investigated. Here, we developed an in vitro 3D engineered CRC (3D-eCRC) tissue model using the natural-synthetic hybrid biomaterial PEG-fibrinogen and three CRC cell lines, HCT 116, HT-29, and SW480. To better recapitulate native tumor conditions, our 3D-eCRC model supported higher cell density encapsulation (20 × 106 cells/mL) and enabled longer term maintenance (29 days) as compared to previously reported in vitro CRC models. The 3D-eCRCs formed using each cell line demonstrated line-dependent differences in cellular and tissue properties, including cellular growth and morphology, cell subpopulations, cell size, cell granularity, migration patterns, tissue growth, gene expression, and tissue stiffness. Importantly, these differences were found to be most prominent from Day 22 to Day 29, thereby indicating the importance of long-term culture of engineered CRC tissues for recapitulation and investigation of mechanistic differences and drug response. Our 3D-eCRC tissue model showed high potential for supporting future in vitro comparative studies of disease progression, metastatic mechanisms, and anti-cancer drug candidate response in a CRC cell line-dependent manner.
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Affiliation(s)
- Iman Hassani
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
- Department of Chemical Engineering, Tuskegee University, Tuskegee, AL 36088, USA
| | - Benjamin Anbiah
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Andrew L. Moore
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Peter T. Abraham
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Ifeoluwa A. Odeniyi
- Department of Nutritional Sciences, Auburn University, Auburn, AL 36849, USA
| | - Nicole L. Habbit
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Michael W. Greene
- Department of Nutritional Sciences, Auburn University, Auburn, AL 36849, USA
| | - Elizabeth A. Lipke
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
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Mohammad Hadi L, Stamati K, Yaghini E, MacRobert AJ, Loizidou M. Treatment of 3D In Vitro Tumoroids of Ovarian Cancer Using Photochemical Internalisation as a Drug Delivery Method. Biomedicines 2023; 11:biomedicines11020572. [PMID: 36831108 PMCID: PMC9953023 DOI: 10.3390/biomedicines11020572] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2022] [Revised: 02/05/2023] [Accepted: 02/05/2023] [Indexed: 02/18/2023] Open
Abstract
Photochemical internalisation (PCI) is a means of achieving spatio-temporal control of cytosolic drug delivery using sub-lethal photodynamic therapy (PDT), with a photosensitiser that can be activated by non-ionising visible light. Various 3D models including those developed at our laboratory, where spheroids are grown in a compressed collagen matrix, have been used for studying anti-cancer drug effects. However, the use of a more biomimetic tumouroid model which consists of a relatively hypoxic central cancer mass surrounded by its microenvironment (stroma) has not yet been explored in either toxicity or phototoxicity studies involving PCI. Here, we examined the efficacy of PCI using a porphyrin photosensitiser and a cytotoxin (Saporin) on ovarian cancer tumouroids, with HEY ovarian cancer cells in the central cancer compartment, and HDF fibroblast cells and HUVEC endothelial cells in the surrounding stromal compartment. The efficacy was compared to tumouroids treated with either Saporin or PDT alone, or no treatment. PCI treatment was shown to be effective in the tumouroids (determined through viability assays and imaging) and caused a considerable decrease in the viability of cancer cells both within the central cancer mass and those which had migrated into the stroma, as well as a reduction in the cell density of surrounding HUVEC and HDFs. Post-treatment, the mean distance of stromal invasion by cancer cells from the original cancer mass following treatment with Saporin alone was 730 μm vs. 125 μm for PCI. PDT was also effective at reducing viability in the central cancer mass and stroma but required a higher photosensitiser dose and light dose than PCI. Tumouroids, as tissue mimics, are suitable models for interrogating multicellular events following pharmacological assault.
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Cadamuro F, Marongiu L, Marino M, Tamini N, Nespoli L, Zucchini N, Terzi A, Altamura D, Gao Z, Giannini C, Bindi G, Smith A, Magni F, Bertini S, Granucci F, Nicotra F, Russo L. 3D bioprinted colorectal cancer models based on hyaluronic acid and signalling glycans. Carbohydr Polym 2023; 302:120395. [PMID: 36604073 DOI: 10.1016/j.carbpol.2022.120395] [Citation(s) in RCA: 12] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2022] [Revised: 11/21/2022] [Accepted: 11/22/2022] [Indexed: 12/03/2022]
Abstract
In cancer microenvironment, aberrant glycosylation events of ECM proteins and cell surface receptors occur. We developed a protocol to generate 3D bioprinted models of colorectal cancer (CRC) crosslinking hyaluronic acid and gelatin functionalized with three signalling glycans characterized in CRC, 3'-Sialylgalactose, 6'-Sialylgalactose and 2'-Fucosylgalactose. The crosslinking, performed exploiting azide functionalized gelatin and hyaluronic acid and 4arm-PEG-dibenzocyclooctyne, resulted in biocompatible hydrogels that were 3D bioprinted with commercial CRC cells HT-29 and patient derived CRC tumoroids. The glycosylated hydrogels showed good 3D printability, biocompatibility and stability over the time. SEM and synchrotron radiation SAXS/WAXS analysis revealed the influence of glycosylation in the construct morphology, whereas MALDI-MS imaging showed that protein profiles of tumoroid cells vary with glycosylation, indicating that sialylation and fucosylation of ECM proteins induce diverse alterations to the proteome of the tumoroid and surrounding cells.
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Affiliation(s)
- Francesca Cadamuro
- Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 Milan, Italy.
| | - Laura Marongiu
- Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 Milan, Italy.
| | - Michele Marino
- Department of Civil Engineering and Computer Science, University of Rome Tor Vergata, 00133 Rome, Italy.
| | - Nicolò Tamini
- School of Medicine and Surgery, University of Milano-Bicocca, 20126 Milan, Italy; ASST San Gerardo Hospital, 20900 Monza, Italy
| | - Luca Nespoli
- School of Medicine and Surgery, University of Milano-Bicocca, 20126 Milan, Italy; ASST San Gerardo Hospital, 20900 Monza, Italy.
| | | | - Alberta Terzi
- Institute of Crystallography, National Research Council, v. Amendola 122/O, 70126 Bari, Italy.
| | - Davide Altamura
- Institute of Crystallography, National Research Council, v. Amendola 122/O, 70126 Bari, Italy.
| | - Zirui Gao
- Paul Scherrer Institute, Villigen PSI 5232, Switzerland.
| | - Cinzia Giannini
- Institute of Crystallography, National Research Council, v. Amendola 122/O, 70126 Bari, Italy.
| | - Greta Bindi
- Department of Medicine and Surgery, Proteomics and Metabolomics Unit, University of Milano-Bicocca, 20854 Vedano al Lambro, Italy.
| | - Andrew Smith
- Department of Medicine and Surgery, Proteomics and Metabolomics Unit, University of Milano-Bicocca, 20854 Vedano al Lambro, Italy.
| | - Fulvio Magni
- Department of Medicine and Surgery, Proteomics and Metabolomics Unit, University of Milano-Bicocca, 20854 Vedano al Lambro, Italy.
| | - Sabrina Bertini
- G. Ronzoni Institute for Chemical and Biochemical Research, 20133 Milan, Italy.
| | - Francesca Granucci
- Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 Milan, Italy.
| | - Francesco Nicotra
- Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 Milan, Italy.
| | - Laura Russo
- Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 Milan, Italy; CÚRAM, SFI Research Centre for Medical Devices, National University of Ireland Galway, H91TK33 Galway, Ireland.
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10
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Nyga A, Stamati K, Redondo PA, Azimi T, Feber A, Neves JB, Hamoudi R, Presneau N, El Sheikh S, Tran MGB, Emberton M, Loizidou M, Cheema U. Renal tumouroids: challenges of manufacturing 3D cultures from patient derived primary cells. J Cell Commun Signal 2022; 16:637-648. [PMID: 35102500 PMCID: PMC9733748 DOI: 10.1007/s12079-022-00666-2] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2021] [Accepted: 01/04/2022] [Indexed: 12/14/2022] Open
Abstract
Recent advancements in 3D in vitro culture have allowed for the development of cancer tissue models which accurately recapitulate the tumour microenvironment. Consequently, there has been increased innovation in therapeutic drug screening. While organoid cultures show great potential, they are limited by the time scale of their growth in vitro and the dependence upon commercial matrices, such as Matrigel, which do not allow for manipulations of their composition or mechanical properties. Here, we show a straightforward approach for the isolation and culture of primary human renal carcinoma cells and matched non-affected kidney. This approach does not require any specific selection for cancer cells, and allows for their direct culture in amenable 3D collagen-based matrices, with the preservation of cancer cells as confirmed by NGS sequencing. This method allows for culture of patient-derived cancer cells in 3D microenvironment, which can be used for downstream experimentation such as investigation of cell-matrix interaction or drug screening.
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Affiliation(s)
- Agata Nyga
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK ,Cell Biology Division, MRC Laboratory for Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge, CB2 0QH UK
| | - Katerina Stamati
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK
| | - Patricia A. Redondo
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK
| | - Tayebeh Azimi
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK
| | - Andrew Feber
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK ,Centre for Molecular Pathology, Royal Marsden NHS Trust, London, UK
| | - Joana B. Neves
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK ,Specialist Centre for Kidney Cancer, Royal Free London NHS Foundation Trust, London, UK
| | - Rifat Hamoudi
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK ,Sharjah Institute for Medical Research, College of Medicine, University of Sharjah, Sharjah, United Arab Emirates
| | - Nadège Presneau
- School of Life Sciences, University of Westminster, London, UK
| | - Soha El Sheikh
- Cellular Pathology Department, Royal Free London Foundation Trust, London, UK
| | - Maxine G. B. Tran
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK ,Specialist Centre for Kidney Cancer, Royal Free London NHS Foundation Trust, London, UK
| | - Mark Emberton
- Centre for 3D Models of Health and Disease, Research Department of Targeted Intervention, Division of Surgery & Interventional Science, University College London, Charles Bell House,43-45 Foley Street, London, W1W 7TS UK ,Department of Urology, University College London Hospitals NHS Foundation Trust, London, UK
| | - Marilena Loizidou
- Research Department of Surgical Biotechnology, Division of Surgery & Interventional Science, University College London, London, UK
| | - Umber Cheema
- Centre for 3D Models of Health and Disease, Research Department of Targeted Intervention, Division of Surgery & Interventional Science, University College London, Charles Bell House,43-45 Foley Street, London, W1W 7TS UK
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11
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Iman H, Benjamin A, Peyton K, Habbit NL, Ahmed B, Heslin MJ, Mobley JA, Greene MW, Lipke EA. Engineered colorectal cancer tissue recapitulates key attributes of a patient-derived xenograft tumor line. Biofabrication 2022; 14:10.1088/1758-5090/ac73b6. [PMID: 35617932 PMCID: PMC9822569 DOI: 10.1088/1758-5090/ac73b6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2021] [Accepted: 05/26/2022] [Indexed: 01/11/2023]
Abstract
The development of physiologically relevantin vitrocolorectal cancer (CRC) models is vital for advancing understanding of tumor biology. Although CRC patient-derived xenografts (PDXs) recapitulate key patient tumor characteristics and demonstrate high concordance with clinical outcomes, the use of thisin vivomodel is costly and low-throughput. Here we report the establishment and in-depth characterization of anin vitrotissue-engineered CRC model using PDX cells. To form the 3D engineered CRC-PDX (3D-eCRC-PDX) tissues, CRC PDX tumors were expandedin vivo, dissociated, and the isolated cells encapsulated within PEG-fibrinogen hydrogels. Following PEG-fibrinogen encapsulation, cells remain viable and proliferate within 3D-eCRC-PDX tissues. Tumor cell subpopulations, including human cancer and mouse stromal cells, are maintained in long-term culture (29 days); cellular subpopulations increase ratiometrically over time. The 3D-eCRC-PDX tissues mimic the mechanical stiffness of originating tumors. Extracellular matrix protein production by cells in the 3D-eCRC-PDX tissues resulted in approximately 57% of proteins observed in the CRC-PDX tumors also being present in the 3D-eCRC-PDX tissues on day 22. Furthermore, we show congruence in enriched gene ontology molecular functions and Hallmark gene sets in 3D-eCRC-PDX tissues and CRC-PDX tumors compared to normal colon tissue, while prognostic Kaplan-Meier plots for overall and relapse free survival did not reveal significant differences between CRC-PDX tumors and 3D-eCRC-PDX tissues. Our results demonstrate high batch-to-batch consistency and strong correlation between ourin vitrotissue-engineered PDX-CRC model and the originatingin vivoPDX tumors, providing a foundation for future studies of disease progression and tumorigenic mechanisms.
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Affiliation(s)
- Hassani Iman
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Anbiah Benjamin
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Kuhlers Peyton
- Department of Nutrition, Dietetics, and Hospitality Management, Auburn University, Auburn, AL 36849, USA
| | - Nicole L. Habbit
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
| | - Bulbul Ahmed
- Department of Nutrition, Dietetics, and Hospitality Management, Auburn University, Auburn, AL 36849, USA
| | - Martin J. Heslin
- Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35233, USA
| | - James A. Mobley
- Department of Anesthesiology and Perioperative Medicine, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35205-3703, USA
- Division of Molecular and Translational Biomedicine, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35205-3703, USA
| | - Michael W. Greene
- Department of Nutrition, Dietetics, and Hospitality Management, Auburn University, Auburn, AL 36849, USA
| | - Elizabeth A. Lipke
- Department of Chemical Engineering, Auburn University, Auburn, AL 36849, USA
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12
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Yang H, Kolben T, Meister S, Paul C, van Dorp J, Eren S, Kuhn C, Rahmeh M, Mahner S, Jeschke U, von Schönfeldt V. Factors Influencing the In Vitro Maturation (IVM) of Human Oocyte. Biomedicines 2021; 9:1904. [PMID: 34944731 PMCID: PMC8698296 DOI: 10.3390/biomedicines9121904] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2021] [Revised: 12/08/2021] [Accepted: 12/10/2021] [Indexed: 02/07/2023] Open
Abstract
In vitro maturation (IVM) of oocytes is a promising assisted reproductive technology (ART) deemed as a simple and safe procedure. It is mainly used in patients with impaired oocyte maturation and in fertility preservation for women facing the risk of losing fertility. However, to date, it is still not widely used in clinical practice because of its underperformance. The influencing factors, such as biphasic IVM system, culture medium, and the supplementation, have a marked effect on the outcomes of oocyte IVM. However, the role of different culture media, supplements, and follicular priming regimens in oocyte IVM have yet to be fully clarified and deserve further investigation.
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Affiliation(s)
- Huixia Yang
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Thomas Kolben
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Sarah Meister
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Corinna Paul
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Julia van Dorp
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Sibel Eren
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Christina Kuhn
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
- Department of Obstetrics and Gynecology, University Hospital Augsburg, 86156 Augsburg, Germany
| | - Martina Rahmeh
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Sven Mahner
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
| | - Udo Jeschke
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
- Department of Obstetrics and Gynecology, University Hospital Augsburg, 86156 Augsburg, Germany
| | - Viktoria von Schönfeldt
- Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University, 81377 Munich, Germany; (H.Y.); (T.K.); (S.M.); (C.P.); (J.v.D.); (S.E.); (C.K.); (M.R.); (S.M.); (V.v.S.)
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13
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Habanjar O, Diab-Assaf M, Caldefie-Chezet F, Delort L. 3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages. Int J Mol Sci 2021; 22:12200. [PMID: 34830082 PMCID: PMC8618305 DOI: 10.3390/ijms222212200] [Citation(s) in RCA: 256] [Impact Index Per Article: 64.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2021] [Revised: 11/05/2021] [Accepted: 11/07/2021] [Indexed: 01/09/2023] Open
Abstract
The traditional two-dimensional (2D) in vitro cell culture system (on a flat support) has long been used in cancer research. However, this system cannot be fully translated into clinical trials to ideally represent physiological conditions. This culture cannot mimic the natural tumor microenvironment due to the lack of cellular communication (cell-cell) and interaction (cell-cell and cell-matrix). To overcome these limitations, three-dimensional (3D) culture systems are increasingly developed in research and have become essential for tumor research, tissue engineering, and basic biology research. 3D culture has received much attention in the field of biomedicine due to its ability to mimic tissue structure and function. The 3D matrix presents a highly dynamic framework where its components are deposited, degraded, or modified to delineate functions and provide a platform where cells attach to perform their specific functions, including adhesion, proliferation, communication, and apoptosis. So far, various types of models belong to this culture: either the culture based on natural or synthetic adherent matrices used to design 3D scaffolds as biomaterials to form a 3D matrix or based on non-adherent and/or matrix-free matrices to form the spheroids. In this review, we first summarize a comparison between 2D and 3D cultures. Then, we focus on the different components of the natural extracellular matrix that can be used as supports in 3D culture. Then we detail different types of natural supports such as matrigel, hydrogels, hard supports, and different synthetic strategies of 3D matrices such as lyophilization, electrospiding, stereolithography, microfluid by citing the advantages and disadvantages of each of them. Finally, we summarize the different methods of generating normal and tumor spheroids, citing their respective advantages and disadvantages in order to obtain an ideal 3D model (matrix) that retains the following characteristics: better biocompatibility, good mechanical properties corresponding to the tumor tissue, degradability, controllable microstructure and chemical components like the tumor tissue, favorable nutrient exchange and easy separation of the cells from the matrix.
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Affiliation(s)
- Ola Habanjar
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
| | - Mona Diab-Assaf
- Equipe Tumorigénèse Pharmacologie Moléculaire et Anticancéreuse, Faculté des Sciences II, Université Libanaise Fanar, Beyrouth 1500, Liban;
| | - Florence Caldefie-Chezet
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
| | - Laetitia Delort
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
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14
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da Mata S, Franchi-Mendes T, Abreu S, Filipe B, Morgado S, Mesquita M, Albuquerque C, Fonseca R, Santo VE, Boghaert ER, Rosa I, Brito C. Patient-Derived Explants of Colorectal Cancer: Histopathological and Molecular Analysis of Long-Term Cultures. Cancers (Basel) 2021; 13:cancers13184695. [PMID: 34572922 PMCID: PMC8465429 DOI: 10.3390/cancers13184695] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2021] [Accepted: 09/14/2021] [Indexed: 12/11/2022] Open
Abstract
Simple Summary Colorectal cancer is the third most common cancer type among men and women. Prescription of medical treatments for cancer often relies on a process of trial and potential error, more recently guided by patient stratification based on biomarkers. Nonetheless, available biomarkers do not accurately predict patient response and there is a need for predictive and translational models to provide proper clinical information on treatment guidance. Herein, we developed an ex vivo model of colorectal cancer, using fresh tumour samples to establish explant cultures, taking advantage of agitation-based culture systems. We performed a thorough characterisation over one month in culture and observed preservation of original tumour genetic features and partial preservation of architecture and non-malignant cells that compose the tumour microenvironment. Our findings highlight the importance of detailed model characterisation and support the applicability of our model in pre- and co-clinical settings. Abstract Colorectal cancer (CRC) is one of the most common cancers worldwide. Although short-term cultures of tumour sections and xenotransplants have been used to determine drug efficacy, the results frequently fail to confer clinically useful information. Biomarker discovery has changed the paradigm for advanced CRC, though the presence of a biomarker does not necessarily translate into therapeutic success. To improve clinical outcomes, translational models predictive of drug response are needed. We describe a simple method for the fast establishment of CRC patient-derived explant (CRC-PDE) cultures from different carcinogenesis pathways, employing agitation-based platforms. A total of 26 CRC-PDE were established and a subset was evaluated for viability (n = 23), morphology and genetic key alterations (n = 21). CRC-PDE retained partial tumor glandular architecture and microenvironment features were partially lost over 4 weeks of culture. Key proteins (p53 and Mismatch repair) and oncogenic driver mutations of the original tumours were sustained throughout the culture. Drug challenge (n = 5) revealed differential drug response from distinct CRC-PDE cases. These findings suggest an adequate representation of the original tumour and highlight the importance of detailed model characterisation. The preservation of key aspects of the CRC microenvironment and genetics supports CRC-PDE potential applicability in pre- and co-clinical settings, as long as temporal dynamics are considered.
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Affiliation(s)
- Sara da Mata
- Serviço de Anatomia Patológica, Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (S.d.M.); (S.M.); (M.M.); (R.F.)
- NOVA Medical School, Universidade Nova de Lisboa, Campo dos Mártires da Pátria 130, 1169-056 Lisboa, Portugal
| | - Teresa Franchi-Mendes
- Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; (T.F.-M.); (S.A.); (V.E.S.)
- Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
| | - Sofia Abreu
- Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; (T.F.-M.); (S.A.); (V.E.S.)
- Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
| | - Bruno Filipe
- Unidade de Investigação em Patobiologia Molecular (UIPM), Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (B.F.); (C.A.)
| | - Sónia Morgado
- Serviço de Anatomia Patológica, Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (S.d.M.); (S.M.); (M.M.); (R.F.)
| | - Marta Mesquita
- Serviço de Anatomia Patológica, Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (S.d.M.); (S.M.); (M.M.); (R.F.)
| | - Cristina Albuquerque
- Unidade de Investigação em Patobiologia Molecular (UIPM), Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (B.F.); (C.A.)
| | - Ricardo Fonseca
- Serviço de Anatomia Patológica, Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal; (S.d.M.); (S.M.); (M.M.); (R.F.)
- Faculdade de Medicina da Universidade de Lisboa, Avenida Prof. Egas Moniz MB, 1649-028 Lisboa, Portugal
| | - Vítor E. Santo
- Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; (T.F.-M.); (S.A.); (V.E.S.)
- Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
| | - Erwin R. Boghaert
- Abbvie Inc., 1 North Waukegan Road, North Chicago, IL 60064-6098, USA;
| | - Isadora Rosa
- Serviço de Gastrenterologia, Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG, EPE), Rua Prof. Lima Basto, 1099-023 Lisboa, Portugal
- Correspondence: (I.R.); (C.B.)
| | - Catarina Brito
- Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; (T.F.-M.); (S.A.); (V.E.S.)
- Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
- The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus, Av. da República, 2780-157 Oeiras, Portugal
- Correspondence: (I.R.); (C.B.)
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15
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The Role of Biomimetic Hypoxia on Cancer Cell Behaviour in 3D Models: A Systematic Review. Cancers (Basel) 2021; 13:cancers13061334. [PMID: 33809554 PMCID: PMC7999912 DOI: 10.3390/cancers13061334] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/27/2021] [Revised: 03/05/2021] [Accepted: 03/13/2021] [Indexed: 12/18/2022] Open
Abstract
Simple Summary Cancer remains one of the leading causes of death worldwide. The advancements in 3D tumour models provide in vitro test-beds to study cancer growth, metastasis and response to therapy. We conducted this systematic review on existing experimental studies in order to identify and summarize key biomimetic tumour microenvironmental features which affect aspects of cancer biology. The review noted the significance of in vitro hypoxia and 3D tumour models on epithelial to mesenchymal transition, drug resistance, invasion and migration of cancer cells. We highlight the importance of various experimental parameters used in these studies and their subsequent effects on cancer cell behaviour. Abstract The development of biomimetic, human tissue models is recognized as being an important step for transitioning in vitro research findings to the native in vivo response. Oftentimes, 2D models lack the necessary complexity to truly recapitulate cellular responses. The introduction of physiological features into 3D models informs us of how each component feature alters specific cellular response. We conducted a systematic review of research papers where the focus was the introduction of key biomimetic features into in vitro models of cancer, including 3D culture and hypoxia. We analysed outcomes from these and compiled our findings into distinct groupings to ascertain which biomimetic parameters correlated with specific responses. We found a number of biomimetic features which primed cancer cells to respond in a manner which matched in vivo response.
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16
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Redmond J, McCarthy H, Buchanan P, Levingstone TJ, Dunne NJ. Advances in biofabrication techniques for collagen-based 3D in vitro culture models for breast cancer research. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 122:111944. [PMID: 33641930 DOI: 10.1016/j.msec.2021.111944] [Citation(s) in RCA: 38] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/16/2020] [Revised: 01/26/2021] [Accepted: 01/29/2021] [Indexed: 12/19/2022]
Abstract
Collagen is the most abundant component of the extracellular matrix (ECM), therefore it represents an ideal biomaterial for the culture of a variety of cell types. Recently, collagen-based scaffolds have shown promise as 3D culture platforms for breast cancer-based research. Two-dimensional (2D) in vitro culture models, while useful for gaining preliminary insights, are ultimately flawed as they do not adequately replicate the tumour microenvironment. As a result, they do not facilitate proper 3D cell-cell/cell-matrix interactions and often an exaggerated response to therapeutic agents occurs. The ECM plays a crucial role in the development and spread of cancer. Alterations within the ECM have a significant impact on the pathogenesis of cancer, the initiation of metastasis and ultimate progression of the disease. 3D in vitro culture models that aim to replicate the tumour microenvironment have the potential to offer a new frontier for cancer research with cell growth, morphology and genetic properties that more closely match in vivo cancers. While initial 3D in vitro culture models used in breast cancer research consisted of simple hydrogel platforms, recent advances in biofabrication techniques, including freeze-drying, electrospinning and 3D bioprinting, have enabled the fabrication of biomimetic collagen-based platforms that more closely replicate the breast cancer ECM. This review highlights the current application of collagen-based scaffolds as 3D in vitro culture models for breast cancer research, specifically for adherence-based scaffolds (i.e. matrix-assisted). Finally, the future perspectives of 3D in vitro breast cancer models and their potential to lead to an improved understanding of breast cancer diagnosis and treatment are discussed.
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Affiliation(s)
- John Redmond
- School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin 9, Ireland; Centre for Medical Engineering Research, Dublin City University, Dublin 9, Ireland
| | - Helen McCarthy
- School of Pharmacy, Queen's University, Belfast BT9 7BL, United Kingdom; School of Chemical Sciences, Dublin City University, Dublin 9, Ireland
| | - Paul Buchanan
- School of Nursing and Human Science, Dublin City University, Dublin 9, Ireland; National Institute of Cellular Biotechnology, Dublin City University, Dublin 9, Ireland
| | - Tanya J Levingstone
- School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin 9, Ireland; Centre for Medical Engineering Research, Dublin City University, Dublin 9, Ireland; Tissue Engineering Research Group, Department of Anatomy and Regenerative Medicine, Royal College of Surgeons in Ireland, Dublin 2, Ireland; Advanced Manufacturing Research Centre (I-Form), School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin 9, Ireland; Advanced Processing Technology Research Centre, Dublin City University, Dublin 9, Ireland; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland
| | - Nicholas J Dunne
- School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin 9, Ireland; Centre for Medical Engineering Research, Dublin City University, Dublin 9, Ireland; Advanced Manufacturing Research Centre (I-Form), School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin 9, Ireland; Advanced Processing Technology Research Centre, Dublin City University, Dublin 9, Ireland; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland; Advanced Materials and Bioengineering Research Centre (AMBER), Trinity College Dublin, Dublin 2, Ireland; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Dublin 2, Ireland.
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17
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Fernando K, Kwang LG, Lim JTC, Fong ELS. Hydrogels to engineer tumor microenvironments in vitro. Biomater Sci 2021; 9:2362-2383. [DOI: 10.1039/d0bm01943g] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Illustration of engineered hydrogel to recapitulate aspects of the tumor microenvironment.
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Affiliation(s)
- Kanishka Fernando
- Department of Biomedical Engineering
- National University of Singapore
- Singapore
| | - Leng Gek Kwang
- Department of Biomedical Engineering
- National University of Singapore
- Singapore
| | - Joanne Tze Chin Lim
- Department of Biomedical Engineering
- National University of Singapore
- Singapore
| | - Eliza Li Shan Fong
- Department of Biomedical Engineering
- National University of Singapore
- Singapore
- The N.1 Institute for Health
- National University of Singapore
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Fitzgerald AA, Li E, Weiner LM. 3D Culture Systems for Exploring Cancer Immunology. Cancers (Basel) 2020; 13:cancers13010056. [PMID: 33379189 PMCID: PMC7795162 DOI: 10.3390/cancers13010056] [Citation(s) in RCA: 53] [Impact Index Per Article: 10.6] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2020] [Revised: 12/20/2020] [Accepted: 12/23/2020] [Indexed: 12/12/2022] Open
Abstract
Simple Summary To study any disease, researchers need convenient and relevant disease models. In cancer, the most commonly used models are two-dimensional (2D) culture models, which grow cells on hard, rigid, plastic surfaces, and mouse models. Cancer immunology is especially difficult to model because the immune system is exceedingly complex; it contains multiple types of cells, and each cell type has several subtypes and a spectrum of activation states. These many immune cell types interact with cancer cells and other components of the tumor, ultimately influencing disease outcomes. 2D culture methods fail to recapitulate these complex cellular interactions. Mouse models also suffer because the murine and human immune systems vary significantly. Three-dimensional (3D) culture systems therefore provide an alternative method to study cancer immunology and can fill the current gaps in available models. This review will describe common 3D culture models and how those models have been used to advance our understanding of cancer immunology. Abstract Cancer immunotherapy has revolutionized cancer treatment, spurring extensive investigation into cancer immunology and how to exploit this biology for therapeutic benefit. Current methods to investigate cancer-immune cell interactions and develop novel drug therapies rely on either two-dimensional (2D) culture systems or murine models. However, three-dimensional (3D) culture systems provide a potentially superior alternative model to both 2D and murine approaches. As opposed to 2D models, 3D models are more physiologically relevant and better replicate tumor complexities. Compared to murine models, 3D models are cheaper, faster, and can study the human immune system. In this review, we discuss the most common 3D culture systems—spheroids, organoids, and microfluidic chips—and detail how these systems have advanced our understanding of cancer immunology.
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Gregory E, Dugan R, David G, Song YH. The biology and engineered modeling strategies of cancer-nerve crosstalk. Biochim Biophys Acta Rev Cancer 2020; 1874:188406. [PMID: 32827578 DOI: 10.1016/j.bbcan.2020.188406] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2020] [Revised: 07/23/2020] [Accepted: 07/26/2020] [Indexed: 02/06/2023]
Abstract
A recent finding critical to cancer aggravation is the interaction between cancer cells and nerves. There exist two main modes of cancer-nerve interaction: perineural invasion (PNI) and tumor innervation. PNI occurs when cancer cells infiltrate the adjacent nerves, and its relative opposite, tumor innervation, occurs when axons extend into tumor bodies. Like most cancer studies, these crosstalk interactions have mostly been observed in patient samples and animal models at this point, making it difficult to understand the mechanisms in a controlled manner. As such, in recent years in vitro studies have emerged that have helped identify various microenvironmental factors responsible for cancer-nerve crosstalk, including but not limited to neurotrophic factors, neurotransmitters, chemokines, cancer-derived exosomes, and Schwann cells. The versatility of in vitro systems warrants continuous development to increase physiological relevance to study PNI and tumor innervation, for example by utilizing biomimetic three-dimensional (3D) culture systems. Despite the wealth of 3D in vitro cancer models, comparatively there exists a lack of 3D in vitro models of nerve, PNI, and tumor innervation. Native-like 3D in vitro models of cancer-nerve interactions may further help develop therapeutic strategies to curb nerve-mediated cancer aggravation. As such, we provide an overview of the key players of cancer-nerve crosstalk and current in vitro models of the crosstalk, as well as cancer and nerve models. We also discuss a few future directions in cancer-nerve crosstalk research.
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Affiliation(s)
- Emory Gregory
- Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR 72701, United States of America.
| | - Reagan Dugan
- Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR 72701, United States of America.
| | - Gabriel David
- Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR 72701, United States of America.
| | - Young Hye Song
- Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR 72701, United States of America.
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Azimi T, Loizidou M, Dwek MV. Cancer cells grown in 3D under fluid flow exhibit an aggressive phenotype and reduced responsiveness to the anti-cancer treatment doxorubicin. Sci Rep 2020; 10:12020. [PMID: 32694700 PMCID: PMC7374750 DOI: 10.1038/s41598-020-68999-9] [Citation(s) in RCA: 27] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/09/2019] [Accepted: 07/02/2020] [Indexed: 12/12/2022] Open
Abstract
3D laboratory models of cancer are designed to recapitulate the biochemical and biophysical characteristics of the tumour microenvironment and aim to enable studies of cancer, and new therapeutic modalities, in a physiologically-relevant manner. We have developed an in vitro 3D model comprising a central high-density mass of breast cancer cells surrounded by collagen type-1 and we incorporated fluid flow and pressure. We noted significant changes in cancer cell behaviour using this system. MDA-MB231 and SKBR3 breast cancer cells grown in 3D downregulated the proliferative marker Ki67 (P < 0.05) and exhibited decreased response to the chemotherapeutic agent doxorubicin (DOX) (P < 0.01). Mesenchymal markers snail and MMP14 were upregulated in cancer cells maintained in 3D (P < 0.001), cadherin-11 was downregulated (P < 0.001) and HER2 increased (P < 0.05). Cells maintained in 3D under fluid flow exhibited a further reduction in response to DOX (P < 0.05); HER2 and Ki67 levels were also attenuated. Fluid flow and pressure was associated with reduced cell viability and decreased expression levels of vimentin. In summary, aggressive cancer cell behaviour and reduced drug responsiveness was observed when breast cancer cells were maintained in 3D under fluid flow and pressure. These observations are relevant for future developments of 3D in vitro cancer models and organ-on-a-chip initiatives.
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Affiliation(s)
- Tayebeh Azimi
- School of Life Sciences, University of Westminster, 115 New Cavendish St, London, W1W 6UW, UK
| | - Marilena Loizidou
- Division of Surgery and Interventional Science, Department of Surgical Biotechnology, UCL Medical School Royal Free Campus, Rowland Hill Street, London, NW3 2PF, UK
| | - Miriam V Dwek
- School of Life Sciences, University of Westminster, 115 New Cavendish St, London, W1W 6UW, UK.
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21
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Demir Duman F, Sebek M, Thanh NTK, Loizidou M, Shakib K, MacRobert AJ. Enhanced photodynamic therapy and fluorescence imaging using gold nanorods for porphyrin delivery in a novel in vitro squamous cell carcinoma 3D model. J Mater Chem B 2020; 8:5131-5142. [PMID: 32420578 DOI: 10.1039/d0tb00810a] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
Nanocomposites of gold nanorods (Au NRs) with the cationic porphyrin TMPyP (5,10,15,20-tetrakis(1- methyl 4-pyridinio)porphyrin tetra(p-toluenesulfonate)) were investigated as a nanocarrier system for photodynamic therapy (PDT) and fluorescence imaging. To confer biocompatibility and facilitate the cellular uptake, the NRs were encapsulated with polyacrylic acid (PAA) and efficiently loaded with the cationic porphyrin by electrostatic interaction. The nanocomposites were tested with and without light exposure following incubation in 2D monolayer cultures and a 3D compressed collagen construct of head and neck squamous cell carcinoma (HNSCC). The results showed that Au NRs enhance the absorption and emission intensity of TMPyP and improve its photodynamic efficiency and fluorescence imaging capability in both 2D cultures and 3D cancer constructs. Au NRs are promising theranostic agents for delivery of photosensitisers for HNSCC treatment and imaging.
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Affiliation(s)
- Fatma Demir Duman
- Division of Surgery and Interventional Science, Centre for Nanomedicine and Surgical Theranostics, University College London, Royal Free Campus, Rowland Hill St, London, NW3 2PE, UK.
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Mohammad Hadi L, Yaghini E, MacRobert AJ, Loizidou M. Synergy between Photodynamic Therapy and Dactinomycin Chemotherapy in 2D and 3D Ovarian Cancer Cell Cultures. Int J Mol Sci 2020; 21:E3203. [PMID: 32366058 PMCID: PMC7247344 DOI: 10.3390/ijms21093203] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2020] [Revised: 04/27/2020] [Accepted: 04/28/2020] [Indexed: 01/05/2023] Open
Abstract
In this study we explored the efficacy of combining low dose photodynamic therapy using a porphyrin photosensitiser and dactinomycin, a commonly used chemotherapeutic agent. The studies were carried out on compressed collagen 3D constructs of two human ovarian cancer cell lines (SKOV3 and HEY) versus their monolayer counterparts. An amphiphilc photosensitiser was employed, disulfonated tetraphenylporphine, which is not a substrate for ABC efflux transporters that can mediate drug resistance. The combination treatment was shown to be effective in both monolayer and 3D constructs of both cell lines, causing a significant and synergistic reduction in cell viability. Compared to dactinomycin alone or PDT alone, higher cell kill was found using 2D monolayer culture vs. 3D culture for the same doses. In 3D culture, the combination therapy resulted in 10 and 22 times higher cell kill in SKOV3 and HEY cells at the highest light dose compared to dactinomycin monotherapy, and 2.2 and 5.5 times higher cell kill than PDT alone. The combination of low dose PDT and dactinomycin appears to be a promising way to repurpose dactinomycin and widen its therapeutic applications.
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Affiliation(s)
- Layla Mohammad Hadi
- Division of Surgery & Interventional Science, Faculty of Medical Sciences, University College London, London NW3 2QG, UK; (E.Y.); (A.J.M.)
| | | | | | - Marilena Loizidou
- Division of Surgery & Interventional Science, Faculty of Medical Sciences, University College London, London NW3 2QG, UK; (E.Y.); (A.J.M.)
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23
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Li ZL, Wang ZJ, Wei GH, Yang Y, Wang XW. Changes in extracellular matrix in different stages of colorectal cancer and their effects on proliferation of cancer cells. World J Gastrointest Oncol 2020; 12:267-275. [PMID: 32206177 PMCID: PMC7081112 DOI: 10.4251/wjgo.v12.i3.267] [Citation(s) in RCA: 43] [Impact Index Per Article: 8.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/01/2019] [Revised: 01/12/2020] [Accepted: 02/07/2020] [Indexed: 02/05/2023] Open
Abstract
BACKGROUND The extracellular matrix is the main component of the tumor microenvironment. Extracellular matrix remodels with the oncogenesis and development of tumors. Previous studies usually focused on the changes of proteins in normal colorectal tissues and colorectal cancers. Little is known about the changes in the extracellular matrix in different stages of colorectal cancer and the effects of these changes on the development of this cancer. AIM To test the changes of type I collagen, type IV collagen, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-3 (TIMP-3) in different stages of colorectal cancer and the effects of these changes on the proliferation of cancer cells. METHODS The extracellular matrix from various stages of colorectal cancer and normal colon tissue was obtained by using acellular technology. We used proteomics to detect the differential expression of proteins between normal colon tissues and colorectal cancer tissues, and then we used Western blot to observe their expression in each stage of colorectal cancer and in normal colon tissue. By co-culturing the extracellular matrix and HT29 colon cancer cells in vivo and in vitro, we tested the cancer cell proliferation rate in vitro by methyl thiazolyl tetrazolium (MTT) assay and in vivo by measuring the tumor volume. RESULTS The expression of type I collagen and MMP-2 increased with increased tumor stage. The expression of MMP-9 was higher in colorectal cancer tissues and was highest in stage III cancer. The expression of type IV collagen and TIMP-3 decreased with increased tumor stage. The proliferation rate of cancer cells in the extracellular matrix of colorectal cancer was higher than that in the extracellular matrix of the normal colon. CONCLUSION These data suggest that the extracellular matrix structure and composition become disorganized during the development of tumors, which is more conducive for the growth of cancer cells.
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Affiliation(s)
- Zhu-Lin Li
- Department of General Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
| | - Zhen-Jun Wang
- Department of General Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
| | - Guang-Hui Wei
- Department of General Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
| | - Yong Yang
- Department of General Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
| | - Xiao-Wan Wang
- Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China
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Stamati K, Redondo PA, Nyga A, Neves JB, Tran MGB, Emberton M, Cheema U, Loizidou M. The anti-angiogenic tyrosine kinase inhibitor Pazopanib kills cancer cells and disrupts endothelial networks in biomimetic three-dimensional renal tumouroids. J Tissue Eng 2020; 11:2041731420920597. [PMID: 32489578 PMCID: PMC7238304 DOI: 10.1177/2041731420920597] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2020] [Accepted: 03/30/2020] [Indexed: 12/31/2022] Open
Abstract
Pazopanib is a tyrosine kinase inhibitor used to treat renal cell carcinoma. Few in vitro studies investigate its effects towards cancer cells or endothelial cells in the presence of cancer. We tested the effect of Pazopanib on renal cell carcinoma cells (CAKI-2,786-O) in two-dimensional and three-dimensional tumouroids made of dense extracellular matrix, treated in normoxia and hypoxia. Finally, we engineered complex tumouroids with a stromal compartment containing fibroblasts and endothelial cells. Simple CAKI-2 tumouroids were more resistant to Pazopanib than 786-O tumouroids. Under hypoxia, while the more 'resistant' CAKI-2 tumouroids showed no decrease in viability, 786-O tumouroids required higher Pazopanib concentrations to induce cell death. In complex tumouroids, Pazopanib exposure led to a reduction in the overall cell viability (p < 0.0001), disruption of endothelial networks and direct killing of renal cell carcinoma cells. We report a biomimetic multicellular tumouroid for drug testing, suitable for agents whose primary target is not confined to cancer cells.
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Affiliation(s)
- Katerina Stamati
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
| | - Patricia A Redondo
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
| | - Agata Nyga
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
| | - Joana B Neves
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
- Specialist Centre for Kidney Cancer,
Royal Free London NHS Foundation Trust, London, UK
| | - Maxine GB Tran
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
- Specialist Centre for Kidney Cancer,
Royal Free London NHS Foundation Trust, London, UK
| | - Mark Emberton
- Research Department of Targeted
Intervention, Division of Surgery & Interventional Science, University College
London, London, UK
- Department of Urology, University
College London Hospitals NHS Foundation Trust, London, UK
| | - Umber Cheema
- Research Department of Targeted
Intervention, Division of Surgery & Interventional Science, University College
London, London, UK
| | - Marilena Loizidou
- Research Department of Surgical
Biotechnology, Division of Surgery & Interventional Science, University College
London, London, UK
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Husman D, Welzel PB, Vogler S, Bray LJ, Träber N, Friedrichs J, Körber V, Tsurkan MV, Freudenberg U, Thiele J, Werner C. Multiphasic microgel-in-gel materials to recapitulate cellular mesoenvironments in vitro. Biomater Sci 2020; 8:101-108. [DOI: 10.1039/c9bm01009b] [Citation(s) in RCA: 16] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
Cell-instructive biohybrid microgel-in-gel materials can guide the faithful in vitro reconstitution of tissues.
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26
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González Díaz EC, Sinha S, Avedian RS, Yang F. Tissue-engineered 3D models for elucidating primary and metastatic bone cancer progression. Acta Biomater 2019; 99:18-32. [PMID: 31419564 DOI: 10.1016/j.actbio.2019.08.020] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/07/2019] [Revised: 07/12/2019] [Accepted: 08/09/2019] [Indexed: 12/14/2022]
Abstract
Malignant bone tumors are aggressive neoplasms which arise from bone tissue or as a result of metastasis. The most prevalent types of cancer, such as breast, prostate, and lung cancer, all preferentially metastasize to bone, yet the role of the bone niche in promoting cancer progression remains poorly understood. Tissue engineering has the potential to bridge this knowledge gap by providing 3D in vitro systems that can be specifically designed to mimic key properties of the bone niche in a more physiologically relevant context than standard 2D culture. Elucidating the crucial components of the bone niche that recruit metastatic cells, support tumor growth, and promote cancer-induced destruction of bone tissue would support efforts for preventing and treating these devastating malignancies. In this review, we summarize recent efforts focused on developing in vitro 3D models of primary bone cancer and bone metastasis using tissue engineering approaches. Such 3D in vitro models can enable the identification of effective therapeutic targets and facilitate high-throughput drug screening to effectively treat bone cancers. STATEMENT OF SIGNIFICANCE: Biomaterials-based 3D culture have been traditionally used for tissue regeneration. Recent research harnessed biomaterials to create 3D in vitro cancer models, with demonstrated advantages over conventional 2D culture in recapitulating tumor progression and drug response in vivo. However, previous work has been largely limited to modeling soft tissue cancer, such as breast cancer and brain cancer. Unlike soft tissues, bone is characterized with high stiffness and mineral content. Primary bone cancer affects mostly children with poor treatment outcomes, and bone is the most common site of cancer metastasis. Here we summarize emerging efforts on engineering 3D bone cancer models using tissue engineering approaches, and future directions needed to further advance this relatively new research area.
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27
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Pape J, Magdeldin T, Ali M, Walsh C, Lythgoe M, Emberton M, Cheema U. Cancer invasion regulates vascular complexity in a three-dimensional biomimetic model. Eur J Cancer 2019; 119:179-193. [PMID: 31470251 DOI: 10.1016/j.ejca.2019.07.005] [Citation(s) in RCA: 27] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/03/2019] [Revised: 07/03/2019] [Accepted: 07/05/2019] [Indexed: 11/20/2022]
Abstract
INTRODUCTION There is a growing appreciation for including a complex, vascularised stroma in three-dimensional (3D) tumour models to recapitulate the native tumour microenvironment in situ. METHODS Using a compartmentalised, biomimetic, 3D cancer model, comprising a central cancer mass surrounded by a vascularised stroma, we have tested the invasive capability of colorectal cancer cells. RESULTS We show histological analysis of dense collagen I/laminin scaffolds, forming necrotic cores with cellular debris. Furthermore, cancer cells within this 3D matrix form spheroids, which is corroborated with high EpCAM expression. We validate the invasive growth of cancer cells into the stroma through quantitative image analysis and upregulation of known invasive gene markers, including metastasis associated in colon cancer 1, matrix metalloproteinase 7 and heparinase. Tumouroids containing highly invasive HCT116 cancer masses form less complex and less branched vascular networks, recapitulating 'leaky' vasculature associated with highly metastatic cancers. Angiogenic factors regulating this were vascular endothelial growth factor A and hepatocyte growth factor active protein. Where vascular networks were formed with less invasive cancer masses (HT29), higher expression of vascular endothelial cadherin active protein resulted in more complex and branched networks. To eliminate the cell-cell interaction between the cancer mass and stroma, we developed a three-compartment model containing an acellular ring to test the chemoattractant pull from the cancer mass. This resulted in migration of endothelial networks through the acellular ring accompanied by alignment of vascular networks at the cancer/stroma boundary. DISCUSSION This work interrogates to the gene and protein level how cancer cells influence the development of a complex stroma, which shows to be directly influenced by the invasive capability of the cancer.
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Affiliation(s)
- Judith Pape
- Institute of Orthopaedics and Musculoskeletal Sciences, Division of Surgery and Interventional Science, University College London, Stanmore Campus, Brockley Hill, HA7 4LP, London, United Kingdom
| | - Tarig Magdeldin
- Institute of Orthopaedics and Musculoskeletal Sciences, Division of Surgery and Interventional Science, University College London, Stanmore Campus, Brockley Hill, HA7 4LP, London, United Kingdom
| | - Morium Ali
- Center for Advanced Biomedical Imaging, Paul O'Gorman Building, 72 Huntley Street, University College London, WC1E 6DD, London, United Kingdom
| | - Claire Walsh
- Center for Advanced Biomedical Imaging, Paul O'Gorman Building, 72 Huntley Street, University College London, WC1E 6DD, London, United Kingdom
| | - Mark Lythgoe
- Center for Advanced Biomedical Imaging, Paul O'Gorman Building, 72 Huntley Street, University College London, WC1E 6DD, London, United Kingdom
| | - Mark Emberton
- Faculty of Medical Sciences, University College London, Bloomsbury Campus Maple House, 149 Tottenham Court Road, W1T 7NF, London, United Kingdom
| | - Umber Cheema
- Institute of Orthopaedics and Musculoskeletal Sciences, Division of Surgery and Interventional Science, University College London, Stanmore Campus, Brockley Hill, HA7 4LP, London, United Kingdom.
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Ringuette-Goulet C, Bolduc S, Pouliot F. Modeling human bladder cancer. World J Urol 2018; 36:1759-1766. [PMID: 29948049 DOI: 10.1007/s00345-018-2369-5] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/01/2018] [Accepted: 05/25/2018] [Indexed: 01/22/2023] Open
Abstract
INTRODUCTION Bladder cancer is a major public health concern and the treatment options available are unable to significantly prevent disease recurrence and progression. The need for experimental tumor models to efficiently reproduce the pathology of human cancers has prompted researchers to attempt various approaches. METHODS A PubMed search combining the MeSH bladder cancer and models was executed in March 2017. RESULTS We review the advantages and limitations of currently available in vitro 2D and 3D bladder cancer models as well as in vivo rodent models. To date, despite the description of a variety of animal models (including transplantable, carcinogen-induced and genetically engineered models), the establishment of reliable, simple, practicable and reproducible animal models remains an ongoing challenge. Recently, sophisticated 3D culture systems have been designed to better recapitulate the phenotypic and cellular heterogeneity as well as microenvironmental aspects of in vivo tumor growth, while being more flexible to conduct repeated experiments. CONCLUSION Selecting the most appropriate model for a specific application will maximize the conversion of potential therapies from the laboratory to clinical practice and requires an understanding of the various models available.
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Affiliation(s)
- Cassandra Ringuette-Goulet
- Centre de recherche en organogénèse expérimentale/LOEX, Regenerative Medicine Division, CHU de Québec Research Center, Quebec, QC, Canada
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, Canada
- Oncology Division, CHU de Québec Research Center, Quebec, QC, Canada
| | - Stéphane Bolduc
- Centre de recherche en organogénèse expérimentale/LOEX, Regenerative Medicine Division, CHU de Québec Research Center, Quebec, QC, Canada
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, Canada
| | - Frédéric Pouliot
- Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, Canada.
- Oncology Division, CHU de Québec Research Center, Quebec, QC, Canada.
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Thakuri PS, Liu C, Luker GD, Tavana H. Biomaterials-Based Approaches to Tumor Spheroid and Organoid Modeling. Adv Healthc Mater 2018; 7:e1700980. [PMID: 29205942 PMCID: PMC5867257 DOI: 10.1002/adhm.201700980] [Citation(s) in RCA: 85] [Impact Index Per Article: 12.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/15/2017] [Revised: 09/21/2017] [Indexed: 12/22/2022]
Abstract
Evolving understanding of structural and biological complexity of tumors has stimulated development of physiologically relevant tumor models for cancer research and drug discovery. A major motivation for developing new tumor models is to recreate the 3D environment of tumors and context-mediated functional regulation of cancer cells. Such models overcome many limitations of standard monolayer cancer cell cultures. Under defined culture conditions, cancer cells self-assemble into 3D constructs known as spheroids. Additionally, cancer cells may recapitulate steps in embryonic development to self-organize into 3D cultures known as organoids. Importantly, spheroids and organoids reproduce morphology and biologic properties of tumors, providing valuable new tools for research, drug discovery, and precision medicine in cancer. This Progress Report discusses uses of both natural and synthetic biomaterials to culture cancer cells as spheroids or organoids, specifically highlighting studies that demonstrate how these models recapitulate key properties of native tumors. The report concludes with the perspectives on the utility of these models and areas of need for future developments to more closely mimic pathologic events in tumors.
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Affiliation(s)
- Pradip Shahi Thakuri
- Department of Biomedical Engineering, The University of Akron, Akron, OH, 44325, USA
| | - Chun Liu
- Departments of Radiology, Biomedical Engineering and Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109, USA
| | - Gary D Luker
- Departments of Radiology, Biomedical Engineering and Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109, USA
| | - Hossein Tavana
- Department of Biomedical Engineering, The University of Akron, Akron, OH, 44325, USA
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30
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Mohammad-Hadi L, MacRobert AJ, Loizidou M, Yaghini E. Photodynamic therapy in 3D cancer models and the utilisation of nanodelivery systems. NANOSCALE 2018; 10:1570-1581. [PMID: 29308480 DOI: 10.1039/c7nr07739d] [Citation(s) in RCA: 53] [Impact Index Per Article: 7.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/25/2023]
Abstract
Photodynamic therapy (PDT) is the subject of considerable research in experimental cancer models mainly for the treatment of solid cancerous tumours. Recent studies on the use of nanoparticles as photosensitiser carriers have demonstrated improved PDT efficacy in experimental cancer therapy. Experiments typically employ conventional monolayer cell culture but there is increasing interest in testing PDT using three dimensional (3D) cancer models. 3D cancer models can better mimic in vivo models than 2D cultures by for example enabling cancer cell interactions with a surrounding extracellular matrix which should enable the treatment to be optimised prior to in vivo studies. The aim of this review is to discuss recent research using PDT in different types of 3D cancer models, from spheroids to nano-fibrous scaffolds, using a range of photosensitisers on their own or incorporated in nanoparticles and nanodelivery systems.
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Affiliation(s)
- Layla Mohammad-Hadi
- Division of Surgery and Interventional Science, Department of Nanotechnology, University College London, Royal Free Campus, Rowland Hill St, London NW3 2PE, UK.
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Ngo MT, Harley BA. The Influence of Hyaluronic Acid and Glioblastoma Cell Coculture on the Formation of Endothelial Cell Networks in Gelatin Hydrogels. Adv Healthc Mater 2017; 6:10.1002/adhm.201700687. [PMID: 28941173 PMCID: PMC5719875 DOI: 10.1002/adhm.201700687] [Citation(s) in RCA: 50] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2017] [Revised: 08/01/2017] [Indexed: 12/16/2022]
Abstract
Glioblastoma (GBM) is the most common and deadly form of brain cancer. Interactions between GBM cells and vasculature in vivo contribute to poor clinical outcomes, with GBM-induced vessel co-option, regression, and subsequent angiogenesis strongly influencing GBM invasion. Here, elements of the GBM perivascular niche are incorporated into a methacrylamide-functionalized gelatin hydrogel as a means to examine GBM-vessel interactions. The complexity of 3D endothelial cell networks formed from human umbilical vein endothelial cells and normal human lung fibroblasts as a function of hydrogel properties and vascular endothelial growth factor (VEGF) presentation is presented. While overall length and branching of the endothelial cell networks decrease with increasing hydrogel stiffness and incorporation of brain-mimetic hyaluronic acid, it can be separately altered by changing the vascular cell seeding density. It is shown that covalent incorporation of VEGF supports network formation as robustly as continuously available soluble VEGF. The impact of U87-MG GBM cells on the endothelial cell networks is subsequently investigated. GBM cells localize in proximity to the endothelial cell networks and hasten network regression in vitro. Together, this in vitro platform recapitulates the close association between GBM cells and vessel structures as well as elements of vessel co-option and regression preceding angiogenesis in vivo.
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Affiliation(s)
- Mai T Ngo
- 193 Roger Adams Laboratory, 600 S. Mathews Ave, Urbana, IL, 61801, USA
| | - Brendan A Harley
- 110 Roger Adams Laboratory, 600 S. Mathews Ave, Urbana, IL, 61801, USA
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Li Y, Yan X, Liu W, Zhou L, You Z, Du Y. 3D Microtissues for Injectable Regenerative Therapy and High-throughput Drug Screening. J Vis Exp 2017:55982. [PMID: 29053690 PMCID: PMC5752368 DOI: 10.3791/55982] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/16/2023] Open
Abstract
To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous microscale cryogels (microcryogels), which can be loaded with a variety of cell types to form 3D microtissues. Herein, we present the protocol to fabricate versatile 3D microtissues and their applications in regenerative therapy and drug screening. Size and shape-controllable microcryogels can be fabricated on an array chip, which can be harvested off-chip as individual cell-loaded carriers for injectable regenerative therapy or be further assembled on-chip into 3D microtissue arrays for high-throughput drug screening. Due to the high elastic nature of these microscale cryogels, the 3D microtissues exhibit great injectability for minimally invasive cell therapy by protecting cells from mechanical shear force during injection. This ensures enhanced cell survival and therapeutic effect in the mouse limb ischemia model. Meanwhile, assembly of 3D microtissue arrays in a standard 384-multi-well format facilitates the use of common laboratory facilities and equipment, enabling high-throughput drug screening on this versatile 3D cell culture platform.
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Affiliation(s)
- Yaqian Li
- Department of Biomedical Engineering, School of Medicine, Tsinghua University; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases
| | - Xiaojun Yan
- Department of Biomedical Engineering, School of Medicine, Tsinghua University
| | - Wei Liu
- Department of Biomedical Engineering, School of Medicine, Tsinghua University
| | - Lyu Zhou
- Department of Biomedical Engineering, School of Medicine, Tsinghua University; School of Life Sciences, Tsinghua University
| | - Zhifeng You
- Department of Biomedical Engineering, School of Medicine, Tsinghua University
| | - Yanan Du
- Department of Biomedical Engineering, School of Medicine, Tsinghua University; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases;
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Tissue-engineered human 3D model of bladder cancer for invasion study and drug discovery. Biomaterials 2017; 145:233-241. [PMID: 28888113 DOI: 10.1016/j.biomaterials.2017.08.041] [Citation(s) in RCA: 41] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2017] [Revised: 08/25/2017] [Accepted: 08/28/2017] [Indexed: 11/22/2022]
Abstract
The tumour microenvironment is critical to both the initiation and maintenance of tumorigenesis. Reconstitution of the microenvironment is a major challenge for in vitro cancer models. Indeed, conventional 2D culture systems cannot replicate the complexity, diversity and dynamic nature of the tumour microenvironment. In this study, we have developed a 3D endotheliazed vesical equivalent by using tissue engineering from primary human cells in which non-invasive or invasive bladder cancer (BCa) cell lines, cultured as compact spheroids, were incorporated. Invasive BCa cells cross the basement membrane and invade the stromal compartment whereas non-invasive BCa cells are confined to the urothelium. Our 3D BCa model could be used as a reliable model for assessing drug responses, potentially reducing or partially replacing animal experiments, and thus should have applications in the identification of novel targets as well as toxicological evaluation of anti-cancer therapies.
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Integrating Biological and Mathematical Models to Explain and Overcome Drug Resistance in Cancer. Part 1: Biological Facts and Studies in Drug Resistance. CURRENT STEM CELL REPORTS 2017. [DOI: 10.1007/s40778-017-0097-1] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023]
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Brancato V, Gioiella F, Profeta M, Imparato G, Guarnieri D, Urciuolo F, Melone P, Netti PA. 3D tumor microtissues as an in vitro testing platform for microenvironmentally-triggered drug delivery systems. Acta Biomater 2017; 57:47-58. [PMID: 28483691 DOI: 10.1016/j.actbio.2017.05.004] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/13/2017] [Revised: 04/26/2017] [Accepted: 05/04/2017] [Indexed: 02/02/2023]
Abstract
Therapeutic approaches based on nanomedicine have garnered great attention in cancer research. In vitro biological models that better mimic in vivo conditions are crucial tools to more accurately predict their therapeutic efficacy in vivo. In this work, a new 3D breast cancer microtissue has been developed to recapitulate the complexity of the tumor microenvironment and to test its efficacy as screening platform for drug delivery systems. The proposed 3D cancer model presents human breast adenocarcinoma cells and cancer-associated fibroblasts embedded in their own ECM, thus showing several features of an in vivo tumor, such as overexpression of metallo-proteinases (MMPs). After demonstrating at molecular and protein level the MMP2 overexpression in such tumor microtissues, we used them to test a recently validated formulation of endogenous MMP2-responsive nanoparticles (NP). The presence of the MMP2-sensitive linker allows doxorubicin release from NP only upon specific enzymatic cleavage of the peptide. The same NP without the MMP-sensitive linker and healthy breast microtissues were also produced to demonstrate NP specificity and selectivity. Cell viability after NP treatment confirmed that controlled drug delivery is achieved only in 3D tumor microtissues suggesting that the validation of therapeutic strategies in such 3D tumor model could predict human response. STATEMENT OF SIGNIFICANCE A major issue of modern cancer research is the development of accurate and predictive experimental models of human tumors consistent with tumor microenvironment and applicable as screening platforms for novel therapeutic strategies. In this work, we developed and validated a new 3D microtissue model of human breast tumor as a testing platform of anti-cancer drug delivery systems. To this aim, biodegradable nanoparticles responsive to physiological changes specifically occurring in tumor microenvironment were used. Our findings clearly demonstrate that the breast tumor microtissue well recapitulates in vivo physiological features of tumor tissue and elicits a specific response to microenvironmentally-responsive nanoparticles compared to healthy tissue. We believe this study is of particular interest for cancer research and paves the way to exploit tumor microtissues for several testing purposes.
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Bray LJ, Werner C. Evaluation of Three-Dimensional in Vitro Models to Study Tumor Angiogenesis. ACS Biomater Sci Eng 2017; 4:337-346. [DOI: 10.1021/acsbiomaterials.7b00139] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Affiliation(s)
- Laura J. Bray
- Institute
of Health and Biomedical Innovation, Queensland University of Technology (QUT), 60 Musk Avenue, Kelvin Grove 4059 Queensland Australia
- Mater
Research Institute - University of Queensland (MRI-UQ), Translational Research Institute, 37 Kent Street, Woolloongabba 4102, QLD Australia
| | - Carsten Werner
- Leibniz
Institute of Polymer Research Dresden e.V., Max Bergmann Center of Biomaterials Dresden, Hohe Straße 6, 01069 Dresden, Saxony, Germany
- Center
for Regenerative Therapies Dresden, Technische Universität Dresden, Fetscherstraße 105, 01307 Dresden, Saxony, Germany
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Engineering a vascularised 3D in vitro model of cancer progression. Sci Rep 2017; 7:44045. [PMID: 28276469 PMCID: PMC5343474 DOI: 10.1038/srep44045] [Citation(s) in RCA: 54] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2016] [Accepted: 02/02/2017] [Indexed: 01/10/2023] Open
Abstract
The hallmark of tumours is the ability of cancerous cells to promote vascular growth, to disseminate and invade to distant organs. The metastatic process is heavily influenced by the extracellular matrix (ECM) density and composition of the surrounding tumour microenvironment. These microenvironmental cues, which include hypoxia, also regulate the angiogenic processes within a tumour, facilitating the spread of cancer cells. We engineered compartmentalized biomimetic colorectal tumouroids with stromal surrounds that comprised a range of ECM densities, composition and stromal cell populations. Recapitulating tissue ECM composition and stromal cell composition enhanced cancer cell invasion. Manipulation of ECM density was associated with an altered migration pattern from glandular buds (cellular aggregates) to epithelial cell sheets. Laminin appeared to be a critical component in regulating endothelial cell morphology and vascular network formation. Interestingly, the disruption of vascular networks by cancer cells was driven by changes in expression of several anti-angiogenic genes. Cancer cells cultured in our biomimetic tumouroids exhibited intratumoural heterogeneity that was associated with increased tumour invasion into the stroma. These findings demonstrate that our 3D in vitro tumour model exhibits biomimetic attributes that may permit their use in studying microenvironment clues of tumour progression and angiogenesis.
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Brancato V, Comunanza V, Imparato G, Corà D, Urciuolo F, Noghero A, Bussolino F, Netti PA. Bioengineered tumoral microtissues recapitulate desmoplastic reaction of pancreatic cancer. Acta Biomater 2017; 49:152-166. [PMID: 27916739 DOI: 10.1016/j.actbio.2016.11.072] [Citation(s) in RCA: 53] [Impact Index Per Article: 6.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2016] [Revised: 11/07/2016] [Accepted: 11/30/2016] [Indexed: 02/07/2023]
Abstract
Many of the existing three-dimensional (3D) cancer models in vitro fail to represent the entire complex tumor microenvironment composed of cells and extra cellular matrix (ECM) and do not allow a reliable study of the tumoral features and progression. In this paper we reported a strategy to produce 3D in vitro microtissues of pancreatic ductal adenocarcinoma (PDAC) for studying the desmoplastic reaction activated by the stroma-cancer crosstalk. Human PDAC microtissues were obtained by co-culturing pancreatic cancer cells (PT45) and normal or cancer-associated fibroblasts within biodegradable microcarriers in a spinner flask bioreactor. Morphological and histological analyses highlighted that the presence of fibroblasts resulted in the deposition of a stromal matrix rich in collagen leading to the formation of tumor microtissues composed of a heterotypic cell population embedded in their own ECM. We analyzed the modulation of expression of ECM genes and proteins and found that when fibroblasts were co-cultured with PT45, they acquired a myofibroblast phenotype and expressed the desmoplastic reaction markers. This PDAC microtissue, closely recapitulating key PDAC microenvironment characteristics, provides a valuable tool to elucidate the complex stroma-cancer interrelationship and could be used in a future perspective as a testing platform for anticancer drugs in tissue-on-chip technology. STATEMENT OF SIGNIFICANCE Tumor microenvironment is extremely complex and its organization is due to the interaction between different kind of cells and the extracellular matrix. Tissue engineering could give the answer to the increasing need of 3D culture model that better recapitulate the tumor features at cellular and extracellular level. We aimed in this work at developing a microtissue tumor model by mean of seeding together cancer cells and fibroblasts on gelatin microsphere in order to monitor the crosstalk between the two cell populations and the endogenous extracellular matrix deposition. Results are of particular interest because of the need of heterotypic cancer model that can replicate the complexity of the tumor microenvironment and could be used as drug screening platform.
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Affiliation(s)
- Virginia Brancato
- Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples Federico II, P.le Tecchio 80, Naples, Italy
| | - Valentina Comunanza
- Department of Oncology, University of Torino, SP 142 km 3.95, 10060 Candiolo, Italy; Candiolo Cancer Institute - IRCCS, SP 142 km 3.95, 10060 Candiolo, Italy
| | - Giorgia Imparato
- Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, 80125 Naples, Italy.
| | - Davide Corà
- Department of Oncology, University of Torino, SP 142 km 3.95, 10060 Candiolo, Italy; Candiolo Cancer Institute - IRCCS, SP 142 km 3.95, 10060 Candiolo, Italy
| | - Francesco Urciuolo
- Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, 80125 Naples, Italy
| | - Alessio Noghero
- Department of Oncology, University of Torino, SP 142 km 3.95, 10060 Candiolo, Italy; Candiolo Cancer Institute - IRCCS, SP 142 km 3.95, 10060 Candiolo, Italy
| | - Federico Bussolino
- Department of Oncology, University of Torino, SP 142 km 3.95, 10060 Candiolo, Italy; Candiolo Cancer Institute - IRCCS, SP 142 km 3.95, 10060 Candiolo, Italy
| | - Paolo A Netti
- Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples Federico II, P.le Tecchio 80, Naples, Italy; Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, 80125 Naples, Italy; Department of Chemical, Materials and Industrial Production (DICMAPI), University of Naples Federico II, P.le Tecchio 80, Naples, Italy
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Romero-López M, Trinh AL, Sobrino A, Hatch MMS, Keating MT, Fimbres C, Lewis DE, Gershon PD, Botvinick EL, Digman M, Lowengrub JS, Hughes CCW. Recapitulating the human tumor microenvironment: Colon tumor-derived extracellular matrix promotes angiogenesis and tumor cell growth. Biomaterials 2016; 116:118-129. [PMID: 27914984 DOI: 10.1016/j.biomaterials.2016.11.034] [Citation(s) in RCA: 90] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2016] [Revised: 11/22/2016] [Accepted: 11/23/2016] [Indexed: 12/14/2022]
Abstract
Extracellular matrix (ECM) is an essential and dynamic component of all tissues and directly affects cellular behavior by providing both mechanical and biochemical signaling cues. Changes in ECM can alter tissue homeostasis, potentially leading to promotion of cellular transformation and the generation of tumors. Therefore, understanding ECM compositional changes during cancer progression is vital to the development of targeted treatments. Previous efforts to reproduce the native 3D cellular microenvironment have utilized protein gels and scaffolds that incompletely recapitulate the complexity of native tissues. Here, we address this problem by extracting and comparing ECM from normal human colon and colon tumor that had metastasized to liver. We found differences in protein composition and stiffness, and observed significant differences in vascular network formation and tumor growth in each of the reconstituted matrices, both in vitro and in vivo. We studied free/bound ratios of NADH in the tumor and endothelial cells using Fluorescence Lifetime Imaging Microscopy as a surrogate for the metabolic state of the cells. We observed that cells seeded in tumor ECM had higher relative levels of free NADH, consistent with a higher glycolytic rate, than those seeded in normal ECM. These results demonstrate that the ECM plays an important role in the growth of cancer cells and their associated vasculature.
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Affiliation(s)
- Mónica Romero-López
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - Andrew L Trinh
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - Agua Sobrino
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - Michaela M S Hatch
- Department of Molecular Biology and Biochemistry, School of Biological Sciences, UC Irvine, USA
| | - Mark T Keating
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - Cristhian Fimbres
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - David E Lewis
- Department of Molecular Biology and Biochemistry, School of Biological Sciences, UC Irvine, USA
| | - Paul D Gershon
- Department of Molecular Biology and Biochemistry, School of Biological Sciences, UC Irvine, USA
| | - Elliot L Botvinick
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA; The Edwards Lifesciences Center for Advanced Cardiovascular Technology, UC Irvine, USA
| | - Michelle Digman
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA
| | - John S Lowengrub
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA; Department of Mathematics, School of Physical Sciences, UC Irvine, USA
| | - Christopher C W Hughes
- Department of Biomedical Engineering, The Henry Samueli School of Engineering, UC Irvine, USA; Department of Molecular Biology and Biochemistry, School of Biological Sciences, UC Irvine, USA; The Edwards Lifesciences Center for Advanced Cardiovascular Technology, UC Irvine, USA.
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Pereira JFS, Awatade NT, Loureiro CA, Matos P, Amaral MD, Jordan P. The third dimension: new developments in cell culture models for colorectal research. Cell Mol Life Sci 2016; 73:3971-89. [PMID: 27147463 PMCID: PMC11108567 DOI: 10.1007/s00018-016-2258-2] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2016] [Revised: 04/20/2016] [Accepted: 04/28/2016] [Indexed: 12/23/2022]
Abstract
Cellular models are important tools in various research areas related to colorectal biology and associated diseases. Herein, we review the most widely used cell lines and the different techniques to grow them, either as cell monolayer, polarized two-dimensional epithelia on membrane filters, or as three-dimensional spheres in scaffold-free or matrix-supported culture conditions. Moreover, recent developments, such as gut-on-chip devices or the ex vivo growth of biopsy-derived organoids, are also discussed. We provide an overview on the potential applications but also on the limitations for each of these techniques, while evaluating their contribution to provide more reliable cellular models for research, diagnostic testing, or pharmacological validation related to colon physiology and pathophysiology.
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Affiliation(s)
- Joana F S Pereira
- Departamento de Genética Humana, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016, Lisbon, Portugal
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal
| | - Nikhil T Awatade
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal
| | - Cláudia A Loureiro
- Departamento de Genética Humana, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016, Lisbon, Portugal
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal
| | - Paulo Matos
- Departamento de Genética Humana, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016, Lisbon, Portugal
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal
| | - Margarida D Amaral
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal
| | - Peter Jordan
- Departamento de Genética Humana, Instituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1649-016, Lisbon, Portugal.
- BioISI-Biosystems and Integrative Sciences Institute, Faculty of Sciences, University of Lisbon, Lisbon, Portugal.
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Fong ELS, Harrington DA, Farach-Carson MC, Yu H. Heralding a new paradigm in 3D tumor modeling. Biomaterials 2016; 108:197-213. [PMID: 27639438 DOI: 10.1016/j.biomaterials.2016.08.052] [Citation(s) in RCA: 117] [Impact Index Per Article: 13.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2016] [Revised: 08/26/2016] [Accepted: 08/31/2016] [Indexed: 12/14/2022]
Abstract
Numerous studies to date have contributed to a paradigm shift in modeling cancer, moving from the traditional two-dimensional culture system to three-dimensional (3D) culture systems for cancer cell culture. This led to the inception of tumor engineering, which has undergone rapid advances over the years. In line with the recognition that tumors are not merely masses of proliferating cancer cells but rather, highly complex tissues consisting of a dynamic extracellular matrix together with stromal, immune and endothelial cells, significant efforts have been made to better recapitulate the tumor microenvironment in 3D. These approaches include the development of engineered matrices and co-cultures to replicate the complexity of tumor-stroma interactions in vitro. However, the tumor engineering and cancer biology fields have traditionally relied heavily on the use of cancer cell lines as a cell source in tumor modeling. While cancer cell lines have contributed to a wealth of knowledge in cancer biology, the use of this cell source is increasingly perceived as a major contributing factor to the dismal failure rate of oncology drugs in drug development. Backing this notion is the increasing evidence that tumors possess intrinsic heterogeneity, which predominantly homogeneous cancer cell lines poorly reflect. Tumor heterogeneity contributes to therapeutic resistance in patients. To overcome this limitation, cancer cell lines are beginning to be replaced by primary tumor cell sources, in the form of patient-derived xenografts and organoids cultures. Moving forward, we propose that further advances in tumor engineering would require that tumor heterogeneity (tumor variants) be taken into consideration together with tumor complexity (tumor-stroma interactions). In this review, we provide a comprehensive overview of what has been achieved in recapitulating tumor complexity, and discuss the importance of incorporating tumor heterogeneity into 3D in vitro tumor models. This work carves out the roadmap for 3D tumor engineering and highlights some of the challenges that need to be addressed as we move forward into the next chapter.
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Affiliation(s)
- Eliza L S Fong
- Department of Physiology, National University of Singapore, Singapore; Department of Biomedical Engineering, National University of Singapore, Singapore.
| | | | | | - Hanry Yu
- Department of Physiology, National University of Singapore, Singapore; Mechanobiology Institute, National University of Singapore, Singapore; Institute of Bioengineering and Nanotechnology, Agency for Science, Technology and Research, Singapore; Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, China
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Shologu N, Szegezdi E, Lowery A, Kerin M, Pandit A, Zeugolis DI. Recreating complex pathophysiologies in vitro with extracellular matrix surrogates for anticancer therapeutics screening. Drug Discov Today 2016; 21:1521-1531. [DOI: 10.1016/j.drudis.2016.06.001] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2016] [Revised: 05/17/2016] [Accepted: 06/01/2016] [Indexed: 12/12/2022]
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Wobma H, Vunjak-Novakovic G. Tissue Engineering and Regenerative Medicine 2015: A Year in Review. TISSUE ENGINEERING PART B-REVIEWS 2016; 22:101-13. [PMID: 26714410 DOI: 10.1089/ten.teb.2015.0535] [Citation(s) in RCA: 55] [Impact Index Per Article: 6.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
This may be the most exciting time ever for the field of tissue engineering and regenerative medicine (TERM). After decades of progress, it has matured, integrated, and diversified into entirely new areas, and it is starting to make the pivotal shift toward translation. The most exciting science and applications continue to emerge at the boundaries of disciplines, through increasingly effective interactions between stem cell biologists, bioengineers, clinicians, and the commercial sector. In this "Year in Review," we highlight some of the major advances reported over the last year (Summer 2014-Fall 2015). Using a methodology similar to that established in previous years, we identified four areas that generated major progress in the field: (i) pluripotent stem cells, (ii) microtissue platforms for drug testing and disease modeling, (iii) tissue models of cancer, and (iv) whole organ engineering. For each area, we used some of the most impactful articles to illustrate the important concepts and results that advanced the state of the art of TERM. We conclude with reflections on emerging areas and perspectives for future development in the field.
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Affiliation(s)
- Holly Wobma
- 1 Department of Biomedical Engineering, Columbia University , New York
| | - Gordana Vunjak-Novakovic
- 1 Department of Biomedical Engineering, Columbia University , New York.,2 Department of Medicine, Columbia University , New York
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Blum KM, Novak T, Watkins L, Neu CP, Wallace JM, Bart ZR, Voytik-Harbin SL. Acellular and cellular high-density, collagen-fibril constructs with suprafibrillar organization. Biomater Sci 2016; 4:711-23. [PMID: 26902645 DOI: 10.1039/c5bm00443h] [Citation(s) in RCA: 33] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
Collagen is used extensively for tissue engineering due to its prevalence in connective tissues and its role in defining tissue biophysical and biological signalling properties. However, traditional collagen-based materials fashioned from atelocollagen and telocollagen have lacked collagen densities, multi-scale organization, mechanical integrity, and proteolytic resistance found within tissues in vivo. Here, highly interconnected low-density matrices of D-banded fibrils were created from collagen oligomers, which exhibit fibrillar as well as suprafibrillar assembly. Confined compression then was applied to controllably reduce the interstitial fluid while maintaining fibril integrity. More specifically, low-density (3.5 mg mL(-1)) oligomer matrices were densified to create collagen-fibril constructs with average concentrations of 12.25 mg mL(-1) and 24.5 mg mL(-1). Control and densified constructs exhibited nearly linear increases in ultimate stress, Young's modulus, and compressive modulus over the ranges of 65 to 213 kPa, 400 to 1.26 MPa, and 20 to 150 kPa, respectively. Densification also increased construct resistance to collagenase degradability. Finally, this process was amenable to creating high-density cellularized tissues; all constructs maintained high cell viability (at least 97%) immediately following compression as well as after 1 day and 7 days of culture. This method, which integrates the suprafibrillar assembly capacity of oligomers and controlled fluid reduction by confined compression, supports the rational and scalable design of a broad range of collagen-fibril materials and cell-encapsulated tissue constructs for tissue engineering applications.
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Affiliation(s)
- Kevin M Blum
- Weldon School of Biomedical Engineering, College of Engineering, Purdue University, West Lafayette, IN 47907, USA.
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López-Dávila V, Magdeldin T, Welch H, Dwek MV, Uchegbu I, Loizidou M. Efficacy of DOPE/DC-cholesterol liposomes and GCPQ micelles as AZD6244 nanocarriers in a 3D colorectal cancer in vitro model. Nanomedicine (Lond) 2016; 11:331-44. [PMID: 26786002 DOI: 10.2217/nnm.15.206] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023] Open
Abstract
AIM In this work, we use cationic organic nanocarriers as chemotherapy delivery platforms and test them in a colorectal cancer 3D in vitro model. MATERIALS & METHODS We used 3beta-(N-[N',N'-dimethylaminoethane]carbamoyl])cholesterol (DC-chol) and dioleoylphosphatidylethanolamine (DOPE) liposomes and N-palmitoyl-N-monomethyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycolchitosan (GCPQ) micelles, to deliver AZD6244, a MEK inhibitor, to HCT116 cells cultured as monolayers and in 3D in vitro cancer models (tumoroids). RESULTS Nanoparticle-mediated drug delivery was superior to the free drug in monolayer experiments and despite their therapeutic effect being hindered by poor diffusion through the cancer mass, GCPQ micelles were also superior in tumoroids. CONCLUSION These results support the role of nanoparticles in improving drug delivery and highlight the need to include 3D cancer models in early phases of drug development.
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Affiliation(s)
- Víctor López-Dávila
- Cancer Nanotechnology Group, University College London, Division of Surgery & Interventional Science, Royal Free Campus, London, NW3 2PF, UK
| | - Tarig Magdeldin
- Cancer Nanotechnology Group, University College London, Division of Surgery & Interventional Science, Royal Free Campus, London, NW3 2PF, UK.,Institute of Orthopaedics & Musculoskeletal Sciences, University College London, Division of Surgery & Interventional Science, Stanmore Campus, HA7 4LP, UK
| | - Hazel Welch
- Cancer Nanotechnology Group, University College London, Division of Surgery & Interventional Science, Royal Free Campus, London, NW3 2PF, UK
| | - Miriam Victoria Dwek
- Department of Biomedical Sciences, Faculty of Science & Technology, University of Westminster, 115 New Cavendish Street, London, W1W 6UW, UK
| | - Ijeoma Uchegbu
- School of Pharmacy, University College London, 29-39 Brunswick Square, London, WC1N 1AX, UK
| | - Marilena Loizidou
- Cancer Nanotechnology Group, University College London, Division of Surgery & Interventional Science, Royal Free Campus, London, NW3 2PF, UK
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Asghar W, El Assal R, Shafiee H, Pitteri S, Paulmurugan R, Demirci U. Engineering cancer microenvironments for in vitro 3-D tumor models. MATERIALS TODAY (KIDLINGTON, ENGLAND) 2015; 18:539-553. [PMID: 28458612 PMCID: PMC5407188 DOI: 10.1016/j.mattod.2015.05.002] [Citation(s) in RCA: 229] [Impact Index Per Article: 22.9] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 05/19/2023]
Abstract
The natural microenvironment of tumors is composed of extracellular matrix (ECM), blood vasculature, and supporting stromal cells. The physical characteristics of ECM as well as the cellular components play a vital role in controlling cancer cell proliferation, apoptosis, metabolism, and differentiation. To mimic the tumor microenvironment outside the human body for drug testing, two-dimensional (2-D) and murine tumor models are routinely used. Although these conventional approaches are employed in preclinical studies, they still present challenges. For example, murine tumor models are expensive and difficult to adopt for routine drug screening. On the other hand, 2-D in vitro models are simple to perform, but they do not recapitulate natural tumor microenvironment, because they do not capture important three-dimensional (3-D) cell-cell, cell-matrix signaling pathways, and multi-cellular heterogeneous components of the tumor microenvironment such as stromal and immune cells. The three-dimensional (3-D) in vitro tumor models aim to closely mimic cancer microenvironments and have emerged as an alternative to routinely used methods for drug screening. Herein, we review recent advances in 3-D tumor model generation and highlight directions for future applications in drug testing.
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Affiliation(s)
- Waseem Asghar
- Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
- Department of Computer Engineering & Electrical Engineering and Computer Science, Florida Atlantic University, Boca Raton, FL 33431, USA
| | - Rami El Assal
- Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
| | - Hadi Shafiee
- Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Division of Biomedical Engineering, Division of Infectious Diseases, Renal Division, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA 02139, USA
| | - Sharon Pitteri
- Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
| | - Ramasamy Paulmurugan
- Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
| | - Utkan Demirci
- Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
- Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Division of Biomedical Engineering, Division of Infectious Diseases, Renal Division, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, MA 02139, USA
- Department of Radiology, Canary Center at Stanford for Cancer Early Detection, Stanford School of Medicine, Stanford University, Palo Alto, CA 94304, USA
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Bandula S, Magdeldin T, Stevens N, Yeung J, Moon JC, Taylor SA, Cheema U, Punwani S. Initial validation of equilibrium contrast imaging for extracellular volume quantification using a three-dimensional engineered tissue model. J Magn Reson Imaging 2015; 43:1224-9. [PMID: 26477540 DOI: 10.1002/jmri.25066] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2015] [Revised: 09/19/2015] [Accepted: 09/22/2015] [Indexed: 01/01/2023] Open
Abstract
BACKGROUND To test the principles underpinning equilibrium contrast imaging estimation of tissue extracellular volume (ECV) fraction, using a three-dimensional (3D) engineered tissue model with known cellular and extracellular volumes. METHODS Six 3D tissue models (tumoroids) consisting of cell cultures within a collagen containing hydrogel were constructed after culture centrifugation and direct measurement of the cell component volume. Measured tumoroid ECV ranged from 0.89 to 1. ECV was calculated after measuring the T1 relaxation time at 3 Tesla using inversion recovery relaxometry (TI 100-1500 ms) within the tumoroids and surrounding medium before and 375 min after spiking the medium with Gadolinium (to achieve a concentration of 1.4 mM/L). Linear regression model prediction of directly measured ECV (ECVm ) by EQ-MRI measured ECV (ECVeq ); and Bland-Altman agreement between measures was assessed. RESULTS The fractional cellular volume measured by EQ-MRI (ECVeq ) within the tumoroids ranged from 0.821 to 0.963. ECVeq was a good predictor of ECVm (R2 = 0.77, P = 0.02). The regression line Y-axis intercept (when X = 0) was 0.045 ± 0.019 with a slope of 1.28 ± 0.35. Bland-Altman comparison demonstrated 95% limits of agreement between -0.002 and 0.114 with a bias (SD) of 0.056 (0.03). CONCLUSION This study supports the principles of ECV estimation using equilibrium contrast MRI, but future development of this model may allow validation over a wider, more physiological ECV range and a greater understanding of the effect of tissue extracellular protein burden on ECV.
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Affiliation(s)
- Steve Bandula
- UCL Centre for Medical Imaging, University College London
| | - Tarig Magdeldin
- Tissue Repair and Engineering Centre, Institute of Orthopaedics and Musculoskeletal Sciences, UCL Division of Surgery and Interventional Science, Stanmore, United Kingdom
| | - Nicola Stevens
- UCL Centre for Medical Imaging, University College London
| | - Jason Yeung
- UCL Centre for Medical Imaging, University College London
| | - James C Moon
- UCL Institute for Cardiovascular Science, University College London
| | | | - Umber Cheema
- Tissue Repair and Engineering Centre, Institute of Orthopaedics and Musculoskeletal Sciences, UCL Division of Surgery and Interventional Science, Stanmore, United Kingdom
| | - Shonit Punwani
- UCL Centre for Medical Imaging, University College London
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Bartlett R, Everett W, Lim S, G N, Loizidou M, Jell G, Tan A, Seifalian AM. Personalized in vitro cancer modeling - fantasy or reality? Transl Oncol 2014; 7:657-64. [PMID: 25500073 PMCID: PMC4311045 DOI: 10.1016/j.tranon.2014.10.006] [Citation(s) in RCA: 30] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2014] [Revised: 10/06/2014] [Accepted: 10/13/2014] [Indexed: 01/06/2023] Open
Abstract
With greater technological advancements and understanding of pathophysiology, “personalized medicine” has become a more realistic goal. In the field of cancer, personalized medicine is the ultimate objective, as each cancer is unique and each tumor is heterogeneous. For many decades, researchers have relied upon studying the histopathology of tumors in the hope that it would provide clues to understanding the pathophysiology of cancer. Current preclinical research relies heavily upon two-dimensional culture models. However, these models have had limited success in recreating the complex interactions between cancer cells and the stroma environment in vivo. Thus, there is increasing impetus to shift to three-dimensional models, which more accurately reflect this phenomenon. With a more accurate in vitro tumor model, drug sensitivity can be tested to determine the best treatment option based on the tumor characteristics. Many methods have been developed to create tumor models or “tumoroids,” each with its advantages and limitations. One significant problem faced is the replication of angiogenesis that is characteristic of tumors in vivo. Nonetheless, if three-dimensional models could be standardized and implemented as a preclinical research tool for therapeutic testing, we would be taking a step towards making personalized cancer medicine a reality.
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Affiliation(s)
- Richard Bartlett
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; UCL Medical School, University College London (UCL), London, UK
| | - William Everett
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; UCL Medical School, University College London (UCL), London, UK
| | - Santi Lim
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; UCL Medical School, University College London (UCL), London, UK
| | - Natasha G
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; UCL Medical School, University College London (UCL), London, UK
| | - Marilena Loizidou
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK
| | - Gavin Jell
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK
| | - Aaron Tan
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; UCL Medical School, University College London (UCL), London, UK; Biomaterials & Advanced Drug Delivery Laboratory (BioADD), Stanford School of Medicine, Stanford University, Stanford, CA, USA
| | - Alexander M Seifalian
- Centre for Nanotechnology & Regenerative Medicine, Research Department of Nanotechnology, UCL Division of Surgery & Interventional Science, University College London (UCL), London, UK; Royal Free London NHS Foundation Trust Hospital, London, UK.
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Tissue-engineered 3D tumor angiogenesis models: potential technologies for anti-cancer drug discovery. Adv Drug Deliv Rev 2014; 79-80:30-9. [PMID: 24819220 DOI: 10.1016/j.addr.2014.05.006] [Citation(s) in RCA: 61] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2013] [Revised: 04/14/2014] [Accepted: 05/02/2014] [Indexed: 01/06/2023]
Abstract
Angiogenesis is indispensable for solid tumor expansion, and thus it has become a major target of cancer research and anti-cancer therapies. Deciphering the arcane actions of various cell populations during tumor angiogenesis requires sophisticated research models, which could capture the dynamics and complexity of the process. There is a continuous need for improvement of existing research models, which engages interdisciplinary approaches of tissue engineering with life sciences. Tireless efforts to develop a new model to study tumor angiogenesis result in innovative solutions, which bring us one step closer to decipher the dubious nature of cancer. This review aims to overview the recent developments, current limitations and future challenges in three-dimensional tissue-engineered models for the study of tumor angiogenesis and for the purpose of elucidating novel targets aimed at anti-cancer drug discovery.
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Ricketts KPM, Cheema U, Nyga A, Castoldi A, Guazzoni C, Magdeldin T, Emberton M, Gibson AP, Royle GJ, Loizidou M. A 3D in vitro cancer model as a platform for nanoparticle uptake and imaging investigations. SMALL (WEINHEIM AN DER BERGSTRASSE, GERMANY) 2014; 10:3954-61. [PMID: 24990320 PMCID: PMC4282585 DOI: 10.1002/smll.201400194] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/22/2014] [Revised: 04/22/2014] [Indexed: 05/24/2023]
Abstract
In order to maximize the potential of nanoparticles (NPs) in cancer imaging and therapy, their mechanisms of interaction with host tissue need to be fully understood. NP uptake is known to be dramatically influenced by the tumor microenvironment, and an imaging platform that could replicate in vivo cellular conditions would make big strides in NP uptake studies. Here, a novel NP uptake platform consisting of a tissue-engineered 3D in vitro cancer model (tumoroid), which mimics the microarchitecture of a solid cancer mass and stroma, is presented. As the tumoroid exhibits fundamental characteristics of solid cancer tissue and its cellular and biochemical parameters are controllable, it provides a real alternative to animal models. Furthermore, an X-ray fluorescence imaging system is developed to demonstrate 3D imaging of GNPs and to determine uptake efficiency within the tumoroid. This platform has implications for optimizing the targeted delivery of NPs to cells to benefit cancer diagnostics and therapy.
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Affiliation(s)
- Kate P M Ricketts
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
| | - Umber Cheema
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
| | - Agata Nyga
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
| | - Andrea Castoldi
- Dipartimento di Elettronica e Informazione, Politecnico di Milano, Edificio 33, via Rimembranze di Lambrate 14, 20133 Milano, Italy and Instituto Nazionale di Fisica NucleareSezione di Milano, Italy
| | - Chiara Guazzoni
- Dipartimento di Elettronica e Informazione, Politecnico di Milano, Edificio 33, via Rimembranze di Lambrate 14, 20133 Milano, Italy and Instituto Nazionale di Fisica NucleareSezione di Milano, Italy
| | - Tarig Magdeldin
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
| | - Mark Emberton
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
| | - Adam P Gibson
- Department of Medical Physics and Bioengineering, University College LondonMalet Place Engineering Building, WC1E 6BT, London, UK
| | - Gary J Royle
- Department of Medical Physics and Bioengineering, University College LondonMalet Place Engineering Building, WC1E 6BT, London, UK
| | - Marilena Loizidou
- Division of Surgery and Interventional Science, University College London9th Floor Royal Free Campus, Rowland Hill Street, NW3 2PF, London, UK
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