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Qi J, Tatla T, Nissanka-Jayasuriya E, Yuan AY, Stoyanov D, Elson DS. Surgical polarimetric endoscopy for the detection of laryngeal cancer. Nat Biomed Eng 2023; 7:971-985. [PMID: 37012312 PMCID: PMC10427430 DOI: 10.1038/s41551-023-01018-0] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2020] [Accepted: 02/23/2023] [Indexed: 04/05/2023]
Abstract
The standard-of-care for the detection of laryngeal pathologies involves distinguishing suspicious lesions from surrounding healthy tissue via contrasts in colour and texture captured by white-light endoscopy. However, the technique is insufficiently sensitive and thus leads to unsatisfactory rates of false negatives. Here we show that laryngeal lesions can be better detected in real time by taking advantage of differences in the light-polarization properties of cancer and healthy tissues. By measuring differences in polarized-light retardance and depolarization, the technique, which we named 'surgical polarimetric endoscopy' (SPE), generates about one-order-of-magnitude greater contrast than white-light endoscopy, and hence allows for the better discrimination of cancerous lesions, as we show with patients diagnosed with squamous cell carcinoma. Polarimetric imaging of excised and stained slices of laryngeal tissue indicated that changes in the retardance of polarized light can be largely attributed to architectural features of the tissue. We also assessed SPE to aid routine transoral laser surgery for the removal of a cancerous lesion, indicating that SPE can complement white-light endoscopy for the detection of laryngeal cancer.
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Affiliation(s)
- Ji Qi
- Research Center for Humanoid Sensing, Zhejiang Lab, Hangzhou, China.
- Wellcome/EPSRC Centre for Interventional and Surgical Sciences, University College London, London, UK.
- Department of Computer Science, University College London, London, UK.
- Centre For Medical Image Computing, University College London, London, UK.
- Hamlyn Centre for Robotic Surgery, Imperial College London, London, UK.
- Department of Surgery and Cancer, Imperial College London, London, UK.
| | - Taranjit Tatla
- Hamlyn Centre for Robotic Surgery, Imperial College London, London, UK
- Northwick Park Hospital, London North West University Healthcare NHS Trust, London, UK
| | | | - Alan Yilun Yuan
- Department of Electrical and Electronic Engineering, Imperial College London, London, UK
| | - Danail Stoyanov
- Wellcome/EPSRC Centre for Interventional and Surgical Sciences, University College London, London, UK.
- Department of Computer Science, University College London, London, UK.
- Centre For Medical Image Computing, University College London, London, UK.
| | - Daniel S Elson
- Hamlyn Centre for Robotic Surgery, Imperial College London, London, UK.
- Department of Surgery and Cancer, Imperial College London, London, UK.
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2
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Preaudet A, Fung KY, Putoczki TL. Confocal Endomicroscopy Monitoring of Tumor Formation. Methods Mol Biol 2023; 2691:257-262. [PMID: 37355552 DOI: 10.1007/978-1-0716-3331-1_20] [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] [Indexed: 06/26/2023]
Abstract
The utilization of preclinical murine models of colorectal cancer (CRC) has been essential to our understanding of the onset and progression of disease. As the genetic complexity of these models evolves to better recapitulate emerging CRC subtypes, our ability to utilize these models to discover and validate novel therapeutic targets will also improve. This will be aided, in part, by the development of live animal imaging techniques, including confocal endomicroscopy for mice. Here in this chapter, we describe the combined use of standard white light endoscopy and confocal endomicroscopy thereby providing a method to rapidly image and assess changes in the colon of an individual live mouse in real time. These methods permit the generation of high-resolution cross-sectional images of the tumor microenvironment for immediate visualization of cells of interest, avoiding the need for euthanasia and tissue collection across multiple cohorts of mice.
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Affiliation(s)
- Adele Preaudet
- Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia
| | - Ka Yee Fung
- Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia
- Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia
| | - Tracy L Putoczki
- Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
- Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia.
- Department of Surgery, The University of Melbourne, Parkville, VIC, Australia.
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3
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Kennedy GT, Azari FS, Bernstein E, Nadeem B, Chang A, Segil A, Carlin S, Sullivan NT, Encarnado E, Desphande C, Kularatne S, Gagare P, Thomas M, Kucharczuk JC, Christien G, Lacombe F, Leonard K, Low PS, Criton A, Singhal S. Targeted detection of cancer at the cellular level during biopsy by near-infrared confocal laser endomicroscopy. Nat Commun 2022; 13:2711. [PMID: 35581212 PMCID: PMC9114105 DOI: 10.1038/s41467-022-30265-z] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2021] [Accepted: 04/23/2022] [Indexed: 12/21/2022] Open
Abstract
Suspicious nodules detected by radiography are often investigated by biopsy, but the diagnostic yield of biopsies of small nodules is poor. Here we report a method-NIR-nCLE-to detect cancer at the cellular level in real-time during biopsy. This technology integrates a cancer-targeted near-infrared (NIR) tracer with a needle-based confocal laser endomicroscopy (nCLE) system modified to detect NIR signal. We develop and test NIR-nCLE in preclinical models of pulmonary nodule biopsy including human specimens. We find that the technology has the resolution to identify a single cancer cell among normal fibroblast cells when co-cultured at a ratio of 1:1000, and can detect cancer cells in human tumors less than 2 cm in diameter. The NIR-nCLE technology rapidly delivers images that permit accurate discrimination between tumor and normal tissue by non-experts. This proof-of-concept study analyzes pulmonary nodules as a test case, but the results may be generalizable to other malignancies.
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Affiliation(s)
- Gregory T Kennedy
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Feredun S Azari
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Elizabeth Bernstein
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Bilal Nadeem
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Ashley Chang
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Alix Segil
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Sean Carlin
- Department of Radiology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Neil T Sullivan
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Emmanuel Encarnado
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | - Charuhas Desphande
- Department of Pathology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | | | | | - Mini Thomas
- On Target Laboratories, West Lafayette, IN, USA
| | - John C Kucharczuk
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
| | | | | | | | - Philip S Low
- Department of Chemistry, Purdue University, West Lafayette, IN, USA
| | | | - Sunil Singhal
- Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
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4
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Wei X, Li J, Yang X, Dong B, Geng B, Li Z, Hu X, Ding B, Zhang J, Yan M. An enzyme-activated two-photon ratiometric fluorescent probe with lysosome targetability for imaging β-glucuronidase in colon cancer cells and tissue. Anal Chim Acta 2021; 1192:339354. [DOI: 10.1016/j.aca.2021.339354] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2021] [Revised: 11/30/2021] [Accepted: 12/01/2021] [Indexed: 01/22/2023]
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5
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Marra G. An "expressionistic" look at serrated precancerous colorectal lesions. Diagn Pathol 2021; 16:4. [PMID: 33423702 PMCID: PMC7797135 DOI: 10.1186/s13000-020-01064-1] [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: 08/04/2020] [Accepted: 12/27/2020] [Indexed: 01/10/2023] Open
Abstract
Background Approximately 60% of colorectal cancer (CRC) precursor lesions are the genuinely-dysplastic conventional adenomas (cADNs). The others include hyperplastic polyps (HPs), sessile serrated lesions (SSL), and traditional serrated adenomas (TSAs), subtypes of a class of lesions collectively referred to as “serrated.” Endoscopic and histologic differentiation between cADNs and serrated lesions, and between serrated lesion subtypes can be difficult. Methods We used in situ hybridization to verify the expression patterns in CRC precursors of 21 RNA molecules that appear to be promising differentiation markers on the basis of previous RNA sequencing studies. Results SSLs could be clearly differentiated from cADNs by the expression patterns of 9 of the 12 RNAs tested for this purpose (VSIG1, ANXA10, ACHE, SEMG1, AQP5, LINC00520, ZIC5/2, FOXD1, NKD1). Expression patterns of all 9 in HPs were similar to those in SSLs. Nine putatively HP-specific RNAs were also investigated, but none could be confirmed as such: most (e.g., HOXD13 and HOXB13), proved instead to be markers of the normal mucosa in the distal colon and rectum, where most HPs arise. TSAs displayed mixed staining patterns reflecting the presence of serrated and dysplastic glands in the same lesion. Conclusions Using a robust in situ hybridization protocol, we identified promising tissue-staining markers that, if validated in larger series of lesions, could facilitate more precise histologic classification of CRC precursors and, consequently, more tailored clinical follow-up of their carriers. Our findings should also fuel functional studies on the pathogenic significance of specific gene expression alterations in the initiation and evolution of CRC precursor subtypes. Supplementary Information The online version contains supplementary material available at 10.1186/s13000-020-01064-1.
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Affiliation(s)
- Giancarlo Marra
- Institute of Molecular Cancer Research, University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
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6
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Heichler C, Scheibe K, Schmied A, Geppert CI, Schmid B, Wirtz S, Thoma OM, Kramer V, Waldner MJ, Büttner C, Farin HF, Pešić M, Knieling F, Merkel S, Grüneboom A, Gunzer M, Grützmann R, Rose-John S, Koralov SB, Kollias G, Vieth M, Hartmann A, Greten FR, Neurath MF, Neufert C. STAT3 activation through IL-6/IL-11 in cancer-associated fibroblasts promotes colorectal tumour development and correlates with poor prognosis. Gut 2020; 69:1269-1282. [PMID: 31685519 DOI: 10.1136/gutjnl-2019-319200] [Citation(s) in RCA: 212] [Impact Index Per Article: 42.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/30/2019] [Revised: 09/22/2019] [Accepted: 10/08/2019] [Indexed: 01/10/2023]
Abstract
OBJECTIVE Cancer-associated fibroblasts (CAFs) influence the tumour microenvironment and tumour growth. However, the role of CAFs in colorectal cancer (CRC) development is incompletely understood. DESIGN We quantified phosphorylation of STAT3 (pSTAT3) expression in CAFs of human colon cancer tissue using a tissue microarray (TMA) of 375 patients, immunofluorescence staining and digital pathology. To investigate the functional role of CAFs in CRC, we took advantage of two murine models of colorectal neoplasia and advanced imaging technologies. In loss-of-function and gain-of-function experiments, using genetically modified mice with collagen type VI (COLVI)-specific signal transducer and activator of transcription 3 (STAT3) targeting, we evaluated STAT3 signalling in fibroblasts during colorectal tumour development. We performed a comparative gene expression profiling by whole genome RNA-sequencing of fibroblast subpopulations (COLVI+ vs COLVI-) on STAT3 activation (IL-6 vs IL-11). RESULTS The analysis of pSTAT3 expression in CAFs of human TMAs revealed a negative correlation of increased stromal pSTAT3 expression with the survival of colon cancer patients. In the loss-of-function and gain-of-function approach, we found a critical role of STAT3 activation in fibroblasts in driving colorectal tumourigenesis in vivo. With different imaging technologies, we detected an expansion of activated fibroblasts in colorectal neoplasias. Comparative gene expression profiling of fibroblast subpopulations on STAT3 activation revealed the regulation of transcriptional patterns associated with angiogenesis. Finally, the blockade of proangiogenic signalling significantly reduced colorectal tumour growth in mice with constitutive STAT3 activation in COLVI+ fibroblasts. CONCLUSION Altogether our work demonstrates a critical role of STAT3 activation in CAFs in CRC development.
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Affiliation(s)
- Christina Heichler
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Kristina Scheibe
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Anabel Schmied
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Carol I Geppert
- Department of Pathology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Benjamin Schmid
- Optical Imaging Center, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Stefan Wirtz
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Oana-Maria Thoma
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany.,Erlangen Graduate School of Advanced Optical Technologies (SAOT), Friedrich Alexander University Erlangen-Nürnberg, Erlangen, Germany
| | - Viktoria Kramer
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Maximilian J Waldner
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Christian Büttner
- Institute of Human Genetics, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Henner F Farin
- German Cancer Consortium (DKTK), Heidelberg, Germany.,Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt am Main, Germany
| | - Marina Pešić
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt am Main, Germany
| | - Ferdinand Knieling
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany.,Department of Pediatrics and Adolescent Medicine, Universitätsklinikum Erlangen Kinder- und Jugendklinik, Erlangen, Germany
| | - Susanne Merkel
- Chirurgische Klinik, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Anika Grüneboom
- Third Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Matthias Gunzer
- Institute of Experimental Immunology and Imaging, University Duisburg-Essen and University Hospital Essen, Essen, Germany
| | - Robert Grützmann
- Chirurgische Klinik, Universitätsklinikum Erlangen, Erlangen, Germany
| | | | - Sergei B Koralov
- Department of Pathology, New York University School of Medicine, New York, New York, USA
| | - George Kollias
- Biomedical Sciences Research Center Alexander Fleming, Vari, Greece
| | - Michael Vieth
- Institute of Pathology, Klinikum Bayreuth, Bayreuth, Germany
| | - Arndt Hartmann
- Department of Pathology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Florian R Greten
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, Frankfurt am Main, Germany
| | - Markus F Neurath
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
| | - Clemens Neufert
- First Department of Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg, Universitätsklinikum Erlangen, Erlangen, Germany
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7
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Development of Dual-Scale Fluorescence Endoscopy for In Vivo Bacteria Imaging in an Orthotopic Mouse Colon Tumor Model. APPLIED SCIENCES-BASEL 2020. [DOI: 10.3390/app10030844] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Colorectal cancer is a representative cancer where early diagnosis and proper treatment monitoring are important. Recently, cancer treatment using bacteria has actively progressed and has been successfully monitored using fluorescence imaging techniques. However, because subcutaneous tumor models are limited in reflecting the actual colorectal cancer situation, new imaging approaches are needed to observe cancers growing in the colon. The fluorescence endoscopic approach is an optimal monitoring modality to evaluate the therapeutic response of bacteria in orthotopic colon cancer. In this study, we developed dual-scaled fluorescence endoscopy (DSFE) by combining wide-field fluorescence endoscopy (WFE) and confocal fluorescence endomicroscopy (CFEM) and demonstrated its usefulness for evaluating bacterial therapy. Firstly, the endoscopic probe of DSFE was developed by integrating the CFEM probe into the guide sheath of WFE. Secondly, colorectal cancer tumor growth and tumors infiltrating the fluorescent bacteria were successfully monitored at the multi-scale using DSFE. Finally, the bacterial distribution of the tumor and organs were imaged and quantitatively analyzed using CFEM. DSFE successfully exhibited fluorescent bacterial signals in an orthotopic mouse colon tumor model. Thus, it can be concluded that the DSFE system is a promising modality to monitor bacterial therapy in vivo.
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8
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Rasti P, Wolf C, Dorez H, Sablong R, Moussata D, Samiei S, Rousseau D. Machine Learning-Based Classification of the Health State of Mice Colon in Cancer Study from Confocal Laser Endomicroscopy. Sci Rep 2019; 9:20010. [PMID: 31882817 PMCID: PMC6934609 DOI: 10.1038/s41598-019-56583-9] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/08/2019] [Accepted: 12/09/2019] [Indexed: 01/26/2023] Open
Abstract
In this article, we address the problem of the classification of the health state of the colon's wall of mice, possibly injured by cancer with machine learning approaches. This problem is essential for translational research on cancer and is a priori challenging since the amount of data is usually limited in all preclinical studies for practical and ethical reasons. Three states considered including cancer, health, and inflammatory on tissues. Fully automated machine learning-based methods are proposed, including deep learning, transfer learning, and shallow learning with SVM. These methods addressed different training strategies corresponding to clinical questions such as the automatic clinical state prediction on unseen data using a pre-trained model, or in an alternative setting, real-time estimation of the clinical state of individual tissue samples during the examination. Experimental results show the best performance of 99.93% correct recognition rate obtained for the second strategy as well as the performance of 98.49% which were achieved for the more difficult first case.
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Affiliation(s)
- Pejman Rasti
- Laboratoire Angevin de Recherche en Ingénierie des Systèmes (LARIS), UMR INRA IRHS, Université d'Angers, Angers, 49000, France
| | - Christian Wolf
- INSA-Lyon, INRIA, LIRIS, CITI, CNRS, Villeurbanne, France
| | - Hugo Dorez
- Univ Lyon, INSA-Lyon, Université Claude Bernard Lyon 1, UJM-Saint Etienne, CNRS, Inserm, CREATIS UMR 5220, U1206, Lyon, 69621, France
| | - Raphael Sablong
- Univ Lyon, INSA-Lyon, Université Claude Bernard Lyon 1, UJM-Saint Etienne, CNRS, Inserm, CREATIS UMR 5220, U1206, Lyon, 69621, France
| | - Driffa Moussata
- Univ Lyon, INSA-Lyon, Université Claude Bernard Lyon 1, UJM-Saint Etienne, CNRS, Inserm, CREATIS UMR 5220, U1206, Lyon, 69621, France
| | - Salma Samiei
- Laboratoire Angevin de Recherche en Ingénierie des Systèmes (LARIS), UMR INRA IRHS, Université d'Angers, Angers, 49000, France
| | - David Rousseau
- Laboratoire Angevin de Recherche en Ingénierie des Systèmes (LARIS), UMR INRA IRHS, Université d'Angers, Angers, 49000, France.
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9
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Rong J, Zhang L, Liao W, Xie Y, Lu N, Shu X. The Value of Confocal Laser Endoscopy in Assessing the Quality of Duodenal Ulcer Healing. Lasers Surg Med 2019; 51:701-708. [PMID: 31074497 DOI: 10.1002/lsm.23098] [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] [Accepted: 04/16/2019] [Indexed: 11/06/2022]
Abstract
BACKGROUND AND OBJECTIVE Confocal laser endomicroscopy (CLE) is a novel endoscopic technique that can image cells and subcellular layers of the gastric mucosa in vivo. We aimed to investigate the value of CLE in assessing the quality of ulcer healing (QOUH) and preliminarily establish evaluation criteria. MATERIALS AND METHODS Patients with duodenal ulcers were enrolled. After duodenal ulcer healing, we compared the value of CLE and white light endoscopy (WLE) in assessing the QOUH by using the histopathological diagnosis as the gold standard. At the same time, immunohistochemistry was performed to examine the expressions of transforming growth factor β1 (TGF-β1) and fibroblast growth factor 2 (FGF-2) in normal and scar tissues. RESULTS In assessing the QOUH classified as poor, good, and excellent by the pathological classification, the sensitivity of WLE was 57.14%, 50%, and 47.06%, the specificity was 87.80%, 52.38%, and 81.58%, and the accuracy was 80.00%, 50.91%, and 70.91%, respectively. Meanwhile, the sensitivity of CLE was 73.33%, 85.19%, and 92.31%, the specificity was 95%, 85.71%, and 92.86%, and the accuracy was 89.09%, 85.45%, and 92.73%, respectively. The κ value for the correlation with pathological diagnosis grade was 0.38 for WLE vs. 0.74 for CLE. The assessment of the QOUH in the CLE image classification showed great improvement compared with that in the WLE image classification. The image classification of CLE was not associated with the immunohistochemical expression of TGF-β1 or FGF-2 according the Spearman rank correlation (P > 0.05). CONCLUSION Compared with WLE, CLE has a higher value in assessing the QOUH. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.
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Affiliation(s)
- Jianfang Rong
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006 Jiangxi, China
| | - Liang Zhang
- Department of Medical, The First Affiliated Hospital of Jiangxi Medical College, Shangrao, 334000 Jiangxi, China
| | - Wangdi Liao
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006 Jiangxi, China
| | - Yong Xie
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006 Jiangxi, China
| | - Nonghua Lu
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006 Jiangxi, China
| | - Xu Shu
- Department of Gastroenterology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006 Jiangxi, China
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10
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Dilipkumar A, Al‐Shemmary A, Kreiß L, Cvecek K, Carlé B, Knieling F, Gonzales Menezes J, Thoma O, Schmidt M, Neurath MF, Waldner M, Friedrich O, Schürmann S. Label-Free Multiphoton Endomicroscopy for Minimally Invasive In Vivo Imaging. ADVANCED SCIENCE (WEINHEIM, BADEN-WURTTEMBERG, GERMANY) 2019; 6:1801735. [PMID: 31016109 PMCID: PMC6468963 DOI: 10.1002/advs.201801735] [Citation(s) in RCA: 45] [Impact Index Per Article: 7.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/04/2018] [Revised: 01/29/2019] [Indexed: 05/24/2023]
Abstract
Multiphoton microscopy of cellular autofluorescence and second harmonic generation from collagen facilitates imaging of living cells and tissues without the need for additional fluorescent labels. Here, a compact multiphoton endomicroscope for label-free in vivo imaging in small animals via side-viewing needle objectives is presented. Minimal invasive imaging at cellular resolution is performed in colonoscopy of mice without surgical measures and without fluorescent dyes as a contrast agent. The colon mucosa is imaged repeatedly in the same animal in a mouse model of acute intestinal inflammation to study the process of inflammation at the tissue level within a time period of ten days, demonstrating the capabilities of label-free endomicroscopy for longitudinal studies for the first time.
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Affiliation(s)
- Ashwathama Dilipkumar
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Alaa Al‐Shemmary
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Lucas Kreiß
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Kristian Cvecek
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Institute of Photonic TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Konrad‐Zuse‐Str. 3–591052ErlangenGermany
| | - Birgitta Carlé
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Ferdinand Knieling
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
- Department of Pediatrics and Adolescent MedicineUniversity Hospital ErlangenLoschgestr. 1591054ErlangenGermany
| | - Jean Gonzales Menezes
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Oana‐Maria Thoma
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Michael Schmidt
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Institute of Photonic TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Konrad‐Zuse‐Str. 3–591052ErlangenGermany
| | - Markus F. Neurath
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Maximilian Waldner
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
- Department of Internal Medicine 1University Hospital ErlangenUlmenweg 1891054ErlangenGermany
| | - Oliver Friedrich
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
| | - Sebastian Schürmann
- Institute of Medical BiotechnologyFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 391052ErlangenGermany
- Erlangen Graduate School in Advanced Optical TechnologiesFriedrich‐Alexander‐Universität Erlangen‐Nürnberg (FAU)Paul‐Gordan‐Str. 791052ErlangenGermany
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11
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Li T, Hui H, Hu C, Ma H, Yang X, Tian J. Multiscale imaging of colitis in mice using confocal laser endomicroscopy, light-sheet fluorescence microscopy, and magnetic resonance imaging. JOURNAL OF BIOMEDICAL OPTICS 2019; 24:1-8. [PMID: 30701723 PMCID: PMC6985686 DOI: 10.1117/1.jbo.24.1.016003] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 09/30/2018] [Accepted: 01/15/2019] [Indexed: 06/09/2023]
Abstract
The objective of our study is to develop a multimodality approach by combining magnetic resonance imaging (MRI) and optical imaging methods to assess acute murine colitis at the macro- and microscopic level. In vivo MRI is used to measure the cross-sectional areas of colons at the macroscopic level. Dual-color confocal laser endomicroscopy (CLE) allows in vivo examination of the fluorescently labeled epithelial cells and microvessels in the mucosa with a spatial resolution of ∼1.4 μm during ongoing endoscopy. To further validate the structural changes of the colons in three-dimensions, ex vivo light-sheet fluorescence microscopy (LSFM) is applied for in-toto imaging of cleared colon sections. MRI, LSFM, and CLE findings are significantly correlated with histological scoring (p < 0.01) and the inflammation-associated activity index (p < 0.01). Our multimodality imaging technique permits visualization of mucosa in colitis at different scales, which can enhance our understanding of the pathogenesis of inflammatory bowel diseases.
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Affiliation(s)
- Tianmeng Li
- Northeastern University, Sino-Dutch Biomedical and Information Engineering School, Shenyang, China
- Chinese Academy of Sciences, Institute of Automation, CAS Key Laboratory of Molecular Imaging, Beijing, China
- Institute of Automation, Beijing Key Laboratory of Molecular Imaging, Beijing, China
| | - Hui Hui
- Chinese Academy of Sciences, Institute of Automation, CAS Key Laboratory of Molecular Imaging, Beijing, China
- Institute of Automation, Beijing Key Laboratory of Molecular Imaging, Beijing, China
- University of Chinese Academy of Sciences, Beijing, China
| | - Chaoen Hu
- Chinese Academy of Sciences, Institute of Automation, CAS Key Laboratory of Molecular Imaging, Beijing, China
- Institute of Automation, Beijing Key Laboratory of Molecular Imaging, Beijing, China
| | - He Ma
- Northeastern University, Sino-Dutch Biomedical and Information Engineering School, Shenyang, China
| | - Xin Yang
- Chinese Academy of Sciences, Institute of Automation, CAS Key Laboratory of Molecular Imaging, Beijing, China
- Institute of Automation, Beijing Key Laboratory of Molecular Imaging, Beijing, China
| | - Jie Tian
- Chinese Academy of Sciences, Institute of Automation, CAS Key Laboratory of Molecular Imaging, Beijing, China
- Institute of Automation, Beijing Key Laboratory of Molecular Imaging, Beijing, China
- University of Chinese Academy of Sciences, Beijing, China
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12
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Assessment of MMP-2/-9 expression by fluorescence endoscopy for evaluation of anastomotic healing in a murine model of anastomotic leakage. PLoS One 2018; 13:e0194249. [PMID: 29566031 PMCID: PMC5863981 DOI: 10.1371/journal.pone.0194249] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2017] [Accepted: 02/27/2018] [Indexed: 12/12/2022] Open
Abstract
BACKGROUND Disturbance of intestinal wound closure leads to insufficient anastomotic healing and is associated with considerable morbidity following colorectal resections. Matrix metalloproteinases (MMPs) play a crucial role in regulation of wound closure. Here fluorescence endoscopy was evaluated for assessment of MMP-2/-9 expression during failed intestinal anastomotic healing. METHODS Distal colonic anastomoses were performed as a model for disturbed healing in 36 Balb/c mice. Healing was evaluated endoscopically, macroscopically, and histologically after 1, 3 and 5 days. For detection of MMP-2/-9 expression fluorescence endoscopy (FE) was used following i.v.-administration of a Cy5.5-labeled MMP-2/-9 specific tracer. FE was complemented by quantification of the fluorescence signal using the MS-FX PRO Optical Imaging System. An overall leakage score was calculated and correlated with the results of FE. RESULTS With increasing incidence of anastomotic leakage from POD1 (17%) to POD5 (83%) the uptake of the MMP tracer gradually increased (signal-to-noise ratio (SNR), POD1: 17.91 ± 1.251 vs. POD3: 30.56 ± 3.03 vs. POD5: 44.8 ± 4.473, P<0.0001). Mice with defective anastomotic healing showed significantly higher uptake compared to non-defective (SNR: 37.37± 3.63 vs. 26.16± 3.635, P = 0.0369). White light endoscopy and FE allowed evaluation of anastomotic healing and visualization of mucosal MMPs in vivo. Using FE based detection of MMPs in the anastomosis, an overall positive predictive value of 71.4% and negative predictive value of 66.6% was calculated for detection of anastomotic leakage. CONCLUSION During disturbed anastomotic healing increased expression of MMP-2/-9 was observed in the anastomotic tissue. Fluorescence endoscopy for detection of MMP-2/-9 during the healing process might be a promising tool for early identification of anastomotic leakage.
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13
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Roper J, Tammela T, Akkad A, Almeqdadi M, Santos SB, Jacks T, Yilmaz ÖH. Colonoscopy-based colorectal cancer modeling in mice with CRISPR-Cas9 genome editing and organoid transplantation. Nat Protoc 2018; 13:217-234. [PMID: 29300388 PMCID: PMC6145089 DOI: 10.1038/nprot.2017.136] [Citation(s) in RCA: 73] [Impact Index Per Article: 10.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Most genetically engineered mouse models (GEMMs) of colorectal cancer are limited by tumor formation in the small intestine, a high tumor burden that limits metastasis, and the need to generate and cross mutant mice. Cell line or organoid transplantation models generally produce tumors in ectopic locations-such as the subcutaneous space, kidney capsule, or cecal wall-that do not reflect the native stromal environment of the colon mucosa. Here, we describe detailed protocols to rapidly and efficiently induce site-directed tumors in the distal colon of mice that are based on colonoscopy-guided mucosal injection. These techniques can be adapted to deliver viral vectors carrying Cre recombinase, CRISPR-Cas9 components, CRISPR-engineered mouse tumor organoids, or human cancer organoids to mice to model the adenoma-carcinoma-metastasis sequence of tumor progression. The colonoscopy injection procedure takes ∼15 min, including preparation. In our experience, anyone with reasonable hand-eye coordination can become proficient with mouse colonoscopy and mucosal injection with a few hours of practice. These approaches are ideal for a wide range of applications, including assessment of gene function in tumorigenesis, examination of tumor-stroma interactions, studies of cancer metastasis, and translational research with patient-derived cancers.
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Affiliation(s)
- Jatin Roper
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
- Division of Gastroenterology, Tufts Medical Center, Boston, Massachusetts, USA
- Molecular Oncology Research Institute, Tufts Medical Center, Boston, Massachusetts, USA
| | - Tuomas Tammela
- Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA
| | - Adam Akkad
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
| | - Mohammad Almeqdadi
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
| | - Sebastian B Santos
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
| | - Tyler Jacks
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
- Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA
| | - Ömer H Yilmaz
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA
- Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA
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14
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Canli Ö, Nicolas AM, Gupta J, Finkelmeier F, Goncharova O, Pesic M, Neumann T, Horst D, Löwer M, Sahin U, Greten FR. Myeloid Cell-Derived Reactive Oxygen Species Induce Epithelial Mutagenesis. Cancer Cell 2017; 32:869-883.e5. [PMID: 29232557 DOI: 10.1016/j.ccell.2017.11.004] [Citation(s) in RCA: 249] [Impact Index Per Article: 31.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/10/2017] [Revised: 08/22/2017] [Accepted: 11/07/2017] [Indexed: 01/15/2023]
Abstract
Increased oxidative stress has been suggested to initiate and promote tumorigenesis by inducing DNA damage and to suppress tumor development by triggering apoptosis and senescence. The contribution of individual cell types in the tumor microenvironment to these contrasting effects remains poorly understood. We provide evidence that during intestinal tumorigenesis, myeloid cell-derived H2O2 triggers genome-wide DNA mutations in intestinal epithelial cells to stimulate invasive growth. Moreover, increased reactive oxygen species (ROS) production in myeloid cells initiates tumor growth in various organs also in the absence of a carcinogen challenge in a paracrine manner. Our data identify an intricate crosstalk between myeloid cell-derived ROS molecules, oxidative DNA damage, and tumor necrosis factor α-mediated signaling to orchestrate a tumor-promoting microenvironment causing invasive cancer.
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Affiliation(s)
- Özge Canli
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - Adele M Nicolas
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - Jalaj Gupta
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - Fabian Finkelmeier
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany; Department of Gastroenterology, Hepatology and Endocrinology, Frankfurt Medical School, 60590 Frankfurt/Main, Germany
| | - Olga Goncharova
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - Marina Pesic
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - Tobias Neumann
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany
| | - David Horst
- Institute of Pathology, University of Munich, 80337 Munich, Germany; German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany
| | - Martin Löwer
- TRON - Translational Oncology at the University Medical Center of Johannes Gutenberg University gGmbH, 55131 Mainz, Germany
| | - Ugur Sahin
- German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; TRON - Translational Oncology at the University Medical Center of Johannes Gutenberg University gGmbH, 55131 Mainz, Germany; University Medical Center of the Johannes Gutenberg University, 55131 Mainz, Germany
| | - Florian R Greten
- Institute for Tumor Biology and Experimental Therapy, Georg-Speyer-Haus, 60596 Frankfurt/Main, Germany; German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
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15
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Simultaneous Detection of EGFR and VEGF in Colorectal Cancer using Fluorescence-Raman Endoscopy. Sci Rep 2017; 7:1035. [PMID: 28432289 PMCID: PMC5430917 DOI: 10.1038/s41598-017-01020-y] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/03/2016] [Accepted: 03/24/2017] [Indexed: 12/14/2022] Open
Abstract
Fluorescence endomicroscopy provides quick access to molecular targets, while Raman spectroscopy allows the detection of multiple molecular targets. Using a simultaneous fluorescence-Raman endoscopic system (FRES), we herein demonstrate its potential in cancer diagnosis in an orthotopically induced colorectal cancer (CRC) xenograft model. In the model, epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) were targeted with antibody-conjugated fluorescence and surface-enhanced Raman scattering (F-SERS) dots. FRES demonstrated fast signal detection and multiplex targeting ability using fluorescence and Raman signals to detect the F-SERS dots. In addition, FRES showed a multiplex targeting ability even on a subcentimeter-sized CRC after spraying with a dose of 50 µg F-SERS dots. In conclusion, molecular characteristics of tumor cells (EGFR in cancer cell membranes) and tumor microenvironments (VEGF in the extracellular matrix) could be simultaneously investigated when performing a colonoscopy.
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16
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Beack S, Cho M, Kim YE, Ahn GO, Hahn SK. Hyaluronate-Peanut Agglutinin Conjugates for Target-Specific Bioimaging of Colon Cancer. Bioconjug Chem 2017; 28:1434-1442. [PMID: 28345902 DOI: 10.1021/acs.bioconjchem.7b00126] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023]
Abstract
Colon cancer is one of the most common death-related cancers in the world. For treating colon cancer, it is crucial to detect and remove malignant lesions early. Here, we developed hyaluronate (HA)-peanut agglutinin (PNA) conjugates for the bioimaging of colon cancer. The HA-PNA conjugates were successfully synthesized by the coupling reaction between aldehyde-modified HA and the N-terminal amine group of PNA. For diagnostic imaging, rhodamine B (RhoB) was chemically conjugated onto PNA in HA-PNA conjugates. After intraluminal injection of HA-PNA-RhoB conjugates into tumor-bearing mice, small-sized colon cancers could be effectively visualized by ex vivo imaging with an in vivo imaging system (IVIS) and a two-photon microscope. With these results taken together, we could confirm the feasibility of HA-PNA-RhoB conjugates as a bioimaging agent for detecting colon cancers.
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Affiliation(s)
- Songeun Beack
- Department of Materials Science and Engineering and ‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH) , 77 Cheongam-ro, Nam-gu, Pohang 37673, Korea
| | - Minsoo Cho
- Department of Materials Science and Engineering and ‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH) , 77 Cheongam-ro, Nam-gu, Pohang 37673, Korea
| | - Young-Eun Kim
- Department of Materials Science and Engineering and ‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH) , 77 Cheongam-ro, Nam-gu, Pohang 37673, Korea
| | - G-One Ahn
- Department of Materials Science and Engineering and ‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH) , 77 Cheongam-ro, Nam-gu, Pohang 37673, Korea
| | - Sei Kwang Hahn
- Department of Materials Science and Engineering and ‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH) , 77 Cheongam-ro, Nam-gu, Pohang 37673, Korea
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17
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Robust graph representation of images with underlying structural networks. Application to the classification of vascular networks of mice’s colon. Pattern Recognit Lett 2017. [DOI: 10.1016/j.patrec.2016.07.022] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/23/2022]
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18
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Sun TY, Haberman AM, Greco V. Preclinical Advances with Multiphoton Microscopy in Live Imaging of Skin Cancers. J Invest Dermatol 2016; 137:282-287. [PMID: 27847119 DOI: 10.1016/j.jid.2016.08.033] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/11/2016] [Revised: 08/09/2016] [Accepted: 08/24/2016] [Indexed: 01/13/2023]
Abstract
Conventional, static analyses have historically been the bedrock and tool of choice for the study of skin cancers. Over the past several years, in vivo imaging of tumors using multiphoton microscopy has emerged as a powerful preclinical tool for revealing detailed cellular behaviors from the earliest moments of tumor development to the final steps of metastasis. Multiphoton microscopy allows for deep tissue penetration with relatively minor phototoxicity, rendering it an effective tool for the long-term observation of tumor evolution. This review highlights some of the recent preclinical insights gained using multiphoton microscopy and suggests future advances that could enhance its power in revealing the mysteries of skin tumor biology.
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Affiliation(s)
- Thomas Yang Sun
- Department of Genetics, Yale School of Medicine, New Haven, Connecticut, USA.
| | - Ann M Haberman
- Departments of Immunobiology and Laboratory Medicine, Yale School of Medicine, New Haven, Connecticut, USA
| | - Valentina Greco
- Department of Genetics, Yale School of Medicine, New Haven, Connecticut, USA; Departments of Dermatology and Cell Biology, Yale Stem Cell Center, Yale Cancer Center, Yale School of Medicine, New Haven, Connecticut, USA.
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19
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Hodgson A, Wier EM, Fu K, Sun X, Wan F. Ultrasound imaging of splenomegaly as a proxy to monitor colon tumor development in Apc(min716/+) mice. Cancer Med 2016; 5:2469-76. [PMID: 27485505 PMCID: PMC5055147 DOI: 10.1002/cam4.842] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2016] [Revised: 07/02/2016] [Accepted: 07/04/2016] [Indexed: 01/13/2023] Open
Abstract
Animal models of colon cancer are widely used to understand the molecular mechanisms and pathogenesis of the disease. These animal models require a substantial investment of time and traditionally necessitate the killing of the animal to measure the tumor progression. Several in vivo imaging techniques are being used in both human clinics and preclinical studies, albeit at high cost and requiring particular expertise. Here, we report that the progression of splenomegaly coincides with and positively correlates to colon tumor development in Apcmin716/+ mice expressing a mutant gene encoding an adenomatous polyposis coli protein truncated at amino acid 716. Ultrasound image‐based spleen size measurement precisely mirrors splenomegaly development in vivo in the tumor‐laden Apcmin716/+ mice. Moreover, the spleen dimensions extracted from the ultrasound sonograms are positively correlated with normalized spleen weight and the number and area of colon tumors. Hence, we propose measuring the spleen size in vivo by ultrasound imaging as a novel approach to estimate splenomegaly development and to indirectly monitor colon tumor development in Apcmin716/+ mice. The widespread use of ultrasound machines in the laboratory setting, coupled with the fact that it is a noninvasive method, make it a straightforward and useful tool for monitoring the experimental progress of colon cancer in mice and determining end points without killing animals strictly for diagnostics purposes.
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Affiliation(s)
- Andrea Hodgson
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, 21025
| | - Eric M Wier
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, 21025
| | - Kai Fu
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, 21025
| | - Xin Sun
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, 21025
| | - Fengyi Wan
- Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, 21025. .,Department of Oncology and Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, 21287.
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Brückner M, Lenz P, Mücke MM, Gohar F, Willeke P, Domagk D, Bettenworth D. Diagnostic imaging advances in murine models of colitis. World J Gastroenterol 2016; 22:996-1007. [PMID: 26811642 PMCID: PMC4716050 DOI: 10.3748/wjg.v22.i3.996] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/29/2015] [Revised: 09/09/2015] [Accepted: 11/13/2015] [Indexed: 02/06/2023] Open
Abstract
Inflammatory bowel diseases (IBD) such as Crohn’s disease and ulcerative colitis are chronic-remittent inflammatory disorders of the gastrointestinal tract still evoking challenging clinical diagnostic and therapeutic situations. Murine models of experimental colitis are a vital component of research into human IBD concerning questions of its complex pathogenesis or the evaluation of potential new drugs. To monitor the course of colitis, to the present day, classical parameters like histological tissue alterations or analysis of mucosal cytokine/chemokine expression often require euthanasia of animals. Recent advances mean revolutionary non-invasive imaging techniques for in vivo murine colitis diagnostics are increasingly available. These novel and emerging imaging techniques not only allow direct visualization of intestinal inflammation, but also enable molecular imaging and targeting of specific alterations of the inflamed murine mucosa. For the first time, in vivo imaging techniques allow for longitudinal examinations and evaluation of intra-individual therapeutic response. This review discusses the latest developments in the different fields of ultrasound, molecularly targeted contrast agent ultrasound, fluorescence endoscopy, confocal laser endomicroscopy as well as tomographic imaging with magnetic resonance imaging, computed tomography and fluorescence-mediated tomography, discussing their individual limitations and potential future diagnostic applications in the management of human patients with IBD.
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21
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From mice to men: Murine models of colorectal cancer for use in translational research. Crit Rev Oncol Hematol 2015; 98:94-105. [PMID: 26558688 DOI: 10.1016/j.critrevonc.2015.10.009] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2015] [Revised: 08/28/2015] [Accepted: 10/27/2015] [Indexed: 12/18/2022] Open
Abstract
Colorectal cancer (CRC) is the third most common carcinoma worldwide and despite advances in treatment, survival for patients with metastatic disease remains poor. With nearly 50% of patients developing metastases, in vivo investigation is essential to improve outcomes for these patients and numerous murine models of CRC have been developed to allow the study of chemoprevention and chemotherapy, in addition to improving our understanding of the pathogenesis of CRC. Selecting the most appropriate murine 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. This review will provide an overview of the murine models currently used in CRC research, discussing the limitations and merits of each and their most relevant application. It is aimed at the developing researcher, acting as a guide to prompt further reading in planning a specific study.
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Interferon Gamma Counteracts the Angiogenic Switch and Induces Vascular Permeability in Dextran Sulfate Sodium Colitis in Mice. Inflamm Bowel Dis 2015; 21:2360-71. [PMID: 26164664 DOI: 10.1097/mib.0000000000000490] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
BACKGROUND Interferon (IFN)-γ is a central pathogenesis factor in inflammatory bowel disease (IBD) with pleiotropic effects on many different cell types. However, as yet, the immune modulatory functions of IFN-γ in IBD have been predominantly investigated. Based on previous studies showing that IFN-γ acts antiangiogenic in colorectal carcinoma, we investigated the effects of IFN-γ on the vascular system in IBD. METHODS Colon tissues of patients with IBD and dextran sulfate sodium-induced colitis in mice were subjected to immunohistochemistry, quantitative real-time polymerase chain reactions, and in situ hybridization to quantify cell activation, angiogenesis, and immune responses. Vascular structure and permeability in mice were analyzed by ultramicroscopy and in vivo confocal laser endomicroscopy. RESULTS We showed a significantly increased blood vessel density in IBD and dextran sulfate sodium colitis. In mice, this was associated with a disorganized blood vessel structure and profound vascular leakage. As compared with genes associated with angiogenesis, genes associated with inflammatory cell activation including IFN-γ were more strongly upregulated in colitis tissues. IFN-γ exerted direct effects on endothelial cells in IBD tissues in vivo, as indicated by the expression of IFN-γ-induced guanylate binding protein 1 (GBP-1). Neutralization of IFN-γ in the acute dextran sulfate sodium colitis model demonstrated that this cytokine exerts endogenous angiostatic activity in IBD and contributes to increased vascular permeability. CONCLUSIONS The dissection of the pleiotropic activities of IFN-γ in IBD provides new insights to the pathological functions of this cytokine and may be of high relevance for the optimization of combination therapy approaches.
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Foersch S, Sperka T, Lindner C, Taut A, Rudolph KL, Breier G, Boxberger F, Rau TT, Hartmann A, Stürzl M, Wittkopf N, Haep L, Wirtz S, Neurath MF, Waldner MJ. VEGFR2 Signaling Prevents Colorectal Cancer Cell Senescence to Promote Tumorigenesis in Mice With Colitis. Gastroenterology 2015; 149:177-189.e10. [PMID: 25797700 DOI: 10.1053/j.gastro.2015.03.016] [Citation(s) in RCA: 40] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/01/2014] [Revised: 03/16/2015] [Accepted: 03/16/2015] [Indexed: 12/14/2022]
Abstract
BACKGROUND & AIMS Senescence prevents cellular transformation. We investigated whether vascular endothelial growth factor (VEGF) signaling via its receptor, VEGFR2, regulates senescence and proliferation of tumor cells in mice with colitis-associated cancer (CAC). METHODS CAC was induced in VEGFR2(ΔIEC) mice, which do not express VEGFR2 in the intestinal epithelium, and VEGFR2(fl/fl) mice (controls) by administration of azoxymethane followed by dextran sodium sulfate. Tumor development and inflammation were determined by endoscopy. Colorectal tissues were collected for immunoblot, immunohistochemical, and quantitative polymerase chain reaction analyses. Findings from mouse tissues were confirmed in human HCT116 colorectal cancer cells. We analyzed colorectal tumor samples from patients before and after treatment with bevacizumab. RESULTS After colitis induction, VEGFR2(ΔIEC) mice developed significantly fewer tumors than control mice. A greater number of intestinal tumor cells from VEGFR2(ΔIEC) mice were in senescence than tumor cells from control mice. We found VEGFR2 to activate phosphatidylinositol-4,5-bisphosphate-3-kinase and AKT, resulting in inactivation of p21 in HCT116 cells. Inhibitors of VEGFR2 and AKT induced senescence in HCT116 cells. Tumor cell senescence promoted an anti-tumor immune response by CD8(+) T cells in mice. Patients whose tumor samples showed an increase in the proportion of senescent cells after treatment with bevacizumab had longer progression-free survival than patients in which the proportion of senescent tumor cells did not change before and after treatment. CONCLUSIONS Inhibition of VEGFR2 signaling leads to senescence of human and mouse colorectal cancer cells. VEGFR2 interacts with phosphatidylinositol-4,5-bisphosphate-3-kinase and AKT to inactivate p21. Colorectal tumor senescence and p21 level correlate with patient survival during treatment with bevacizumab.
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Affiliation(s)
| | - Tobias Sperka
- Fritz Lipmann Institute, Leibniz Institute for Age Research, Jena, Germany
| | | | - Astrid Taut
- Department of Medicine 1, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Karl L Rudolph
- Fritz Lipmann Institute, Leibniz Institute for Age Research, Jena, Germany
| | - Georg Breier
- Department of Pathology, Dresden University of Technology, Dresden, Germany
| | - Frank Boxberger
- Department of Medicine 1, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Tilman T Rau
- Department of Pathology, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Arndt Hartmann
- Department of Pathology, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Michael Stürzl
- Division of Molecular and Experimental Surgery, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Nadine Wittkopf
- Department of Medicine 1, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Lisa Haep
- Division of Molecular and Experimental Surgery, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Stefan Wirtz
- Department of Medicine 1, FAU Erlangen-Nürnberg, Erlangen, Germany
| | - Markus F Neurath
- Department of Medicine 1, FAU Erlangen-Nürnberg, Erlangen, Germany
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Ciocâlteu A, Pirici D, Stefanescu A, Georgescu CV, Şurlin V, Săftoiu A. Endomicroscopy with Fluorescent CD105 Antibodies for "In Vivo" Imaging of Colorectal Cancer Angiogenesis. CURRENT HEALTH SCIENCES JOURNAL 2015; 41:288-292. [PMID: 30538832 PMCID: PMC6246985 DOI: 10.12865/chsj.41.03.17] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 02/27/2015] [Accepted: 03/15/2015] [Indexed: 11/25/2022]
Abstract
The aim of this case report was to evaluate the feasibility of in vivo acquisition of microscopic images using fluorescent CD105 antibodies for molecular imaging in human colorectal cancer. After excluding the presence of tissue autofluorescence, the antibody solution was topically administered through a spray-catheter. The targeted area was analyzed by eCLE and images were recorded. The fractal dimension of tumor vessels and the vessel density were determined using ImageJ software. Immunohistochemistry was used as a gold standard. In vivo CLE analysis of CD105 expression enabled the study of tumor vascular network, revealing a chaotic structure.
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Affiliation(s)
- A Ciocâlteu
- Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, Romania
| | - D Pirici
- Department of Research Methodology, University of Medicine and Pharmacy of Craiova, Romania
| | - A Stefanescu
- Department of Research Methodology, University of Medicine and Pharmacy of Craiova, Romania
| | - C V Georgescu
- Department of Pathology, Emergency County Hospital, Craiova, Romania
| | - V Şurlin
- Department of Surgery, University of Medicine and Pharmacy of Craiova, Romania
| | - A Săftoiu
- Research Center of Gastroenterology and Hepatology, University of Medicine and Pharmacy of Craiova, Romania
- Endoscopy Department, Copenhagen University Hospital Herlev, Copenhagen, Denmark
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25
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Walldorf J, Hermann M, Porzner M, Pohl S, Metz H, Mäder K, Zipprich A, Christ B, Seufferlein T. In-vivo monitoring of acute DSS-Colitis using Colonoscopy, high resolution Ultrasound and bench-top Magnetic Resonance Imaging in Mice. Eur Radiol 2015; 25:2984-91. [PMID: 25981216 DOI: 10.1007/s00330-015-3714-3] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2014] [Revised: 01/07/2015] [Accepted: 03/11/2015] [Indexed: 12/13/2022]
Abstract
OBJECTIVE The aim of this study was to establish and evaluate (colour Doppler-) high-resolution-ultrasound (hrUS) and bench-top magnetic resonance imaging (btMRI) as new methods to monitor experimental colitis. MATERIALS AND METHODS hrUS, btMRI and endoscopy were performed in mice without colitis (n = 15), in mice with acute colitis (n = 14) and in mice with acute colitis and simultaneous treatment with infliximab (n = 19). RESULTS Determination of colon wall thickness using hrUS (32 MHz) and measurement of the cross-sectional colonic areas by btMRI allowed discrimination between the treatment groups (mean a vs. b vs. c - btMRI: 922 vs. 2051 vs. 1472 pixel, hrUS: 0.26 vs. 0.45 vs. 0.31 mm). btMRI, endoscopy, hrUS and colour Doppler-hrUS correlated to histological scoring (p < 0.05), while endoscopy and btMRI correlated to post-mortem colon length (p < 0.05). CONCLUSIONS The innovative in vivo techniques btMRI and hrUS are safe and technically feasible. They differentiate between distinct grades of colitis in an experimental setting, and correlate with established post-mortem parameters. In addition to endoscopic procedures, these techniques provide information regarding colon wall thickness and perfusion. Depending on the availability of these techniques, their application increases the value of in vivo monitoring in experimental acute colitis in small rodents. KEY POINTS • Improved in vivo monitoring might balance interindividual differences in murine colitis. • In monitoring murine colitis, btMRI and hrUS are safe and technically feasible. • Very short examination times underline the usefulness especially of hrUS. • Results of btMRI and hrUS correlate with endoscopic and post-mortem findings.
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Affiliation(s)
- J Walldorf
- Department of Internal Medicine I, Martin Luther University Halle-Wittenberg, Ernst-Grube-Strasse 40, 06120, Halle, Germany,
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Mielke L, Preaudet A, Belz G, Putoczki T. Confocal laser endomicroscopy to monitor the colonic mucosa of mice. J Immunol Methods 2015; 421:81-88. [PMID: 25960174 PMCID: PMC5803490 DOI: 10.1016/j.jim.2015.04.012] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/19/2014] [Revised: 03/27/2015] [Accepted: 04/15/2015] [Indexed: 12/13/2022]
Abstract
The gastrointestinal tract is a unique organ system that provides an epithelial barrier between our underlying immune system and luminal pathogens. Disruption of gastrointestinal homeostasis, as a result of impaired barrier function, is associated with numerous pathologies including inflammatory bowel disease and colorectal cancer. In parallel to the clinical development of endoscopy technologies to monitor and diagnose these pathologies in humans, advanced mouse colonoscopy techniques are being developed. When these technologies are coupled with model systems of human disease, which are essential to our understanding of the pathophysiology of gastrointestinal diseases, the requirement for euthanasia of multiple cohorts of mice is eliminated. Here we highlight the suitability of white light endoscopy to monitor the progression of colitis in mice. We further outline the experimental power of combined standard endoscopy with confocal microendoscopy, which permits visualization of fluorescent markers in a single animal in real-time. Together, these technologies will enhance our understanding of the interplay between components of the gastrointestinal microenvironment and their role in disease.
Monitoring of mucosal damage using white light endoscopy Monitoring of the epithelial barrier using confocal endomicroscopy Monitoring of the vasculature architecture using confocal endomicroscopy
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Affiliation(s)
- Lisa Mielke
- The Walter and Eliza Hall Institute for Medical Research, Melbourne, Australia; The Department of Medical Biology, University of Melbourne, Australia
| | - Adele Preaudet
- The Walter and Eliza Hall Institute for Medical Research, Melbourne, Australia
| | - Gabrielle Belz
- The Walter and Eliza Hall Institute for Medical Research, Melbourne, Australia; The Department of Medical Biology, University of Melbourne, Australia
| | - Tracy Putoczki
- The Walter and Eliza Hall Institute for Medical Research, Melbourne, Australia; The Department of Medical Biology, University of Melbourne, Australia.
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27
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Fluorescence-Raman dual modal endoscopic system for multiplexed molecular diagnostics. Sci Rep 2015; 5:9455. [PMID: 25820115 PMCID: PMC4377550 DOI: 10.1038/srep09455] [Citation(s) in RCA: 55] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2014] [Accepted: 03/05/2015] [Indexed: 12/20/2022] Open
Abstract
Optical endoscopic imaging, which was recently equipped with bioluminescence, fluorescence, and Raman scattering, allows minimally invasive real-time detection of pathologies on the surface of hollow organs. To characterize pathologic lesions in a multiplexed way, we developed a dual modal fluorescence-Raman endomicroscopic system (FRES), which used fluorescence and surface-enhanced Raman scattering nanoprobes (F-SERS dots). Real-time, in vivo, and multiple target detection of a specific cancer was successful, based on the fast imaging capability of fluorescence signals and the multiplex capability of simultaneously detected SERS signals using an optical fiber bundle for intraoperative endoscopic system. Human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR) on the breast cancer xenografts in a mouse orthotopic model were successfully detected in a multiplexed way, illustrating the potential of FRES as a molecular diagnostic instrument that enables real-time tumor characterization of receptors during routine endoscopic procedures.
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28
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Near-Infrared Confocal Laser Endomicroscopy Detects Colorectal Cancer via an Integrin αvβ3 Optical Probe. Mol Imaging Biol 2015; 17:450-60. [DOI: 10.1007/s11307-015-0825-9] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
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29
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Du Le VN, Wang Q, Gould T, Ramella-Roman JC, Pfefer TJ. Vascular contrast in narrow-band and white light imaging. APPLIED OPTICS 2014; 53:4061-4071. [PMID: 24979441 DOI: 10.1364/ao.53.004061] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/11/2013] [Accepted: 04/22/2014] [Indexed: 06/03/2023]
Abstract
Narrow-band imaging (NBI) is a spectrally selective reflectance imaging technique that is used clinically for enhancing visualization of superficial vasculature and has shown promise for applications such as early endoscopic detection of gastrointestinal neoplasia. We have studied the effect of vessel geometry and illumination wavelength on vascular contrast using idealized geometries in order to more quantitatively understand NBI and broadband or white light imaging of mucosal tissue. Simulations were performed using a three-dimensional, voxel-based Monte Carlo model incorporating discrete vessels. In all cases, either 415 or 540 nm illumination produced higher contrast than white light, yet white light did not always produce the lowest contrast. White light produced the lowest contrast for small vessels and intermediate contrast for large vessels (diameter≥100 μm) at deep regions (vessel depth≥200 μm). The results show that 415 nm illuminations provided superior contrast for smaller vessels at shallow depths while 540 nm provided superior contrast for larger vessels in deep regions. Besides 540 nm, our studies also indicate the potential of other wavelengths to achieve high contrast of large vessels at deep regions. Simulation results indicate the importance of three key mechanisms in determining spectral variations in contrast: intravascular hemoglobin (Hb) absorption in the vessel of interest, diffuse Hb absorption from collateral vasculature, and bulk tissue scattering. Measurements of NBI contrast in turbid phantoms incorporating 0.1-mm-diameter hemoglobin-filled capillary tubes indicated good agreement with modeling results. These results provide quantitative insights into light-tissue interactions and the effect of device and tissue properties on NBI performance.
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Atreya R, Neumann H, Neufert C, Waldner MJ, Billmeier U, Zopf Y, Willma M, App C, Münster T, Kessler H, Maas S, Gebhardt B, Heimke-Brinck R, Reuter E, Dörje F, Rau TT, Uter W, Wang TD, Kiesslich R, Vieth M, Hannappel E, Neurath MF. In vivo imaging using fluorescent antibodies to tumor necrosis factor predicts therapeutic response in Crohn's disease. Nat Med 2014; 20:313-8. [PMID: 24562382 DOI: 10.1038/nm.3462] [Citation(s) in RCA: 291] [Impact Index Per Article: 26.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2013] [Accepted: 06/03/2013] [Indexed: 02/07/2023]
Abstract
As antibodies to tumor necrosis factor (TNF) suppress immune responses in Crohn's disease by binding to membrane-bound TNF (mTNF), we created a fluorescent antibody for molecular mTNF imaging in this disease. Topical antibody administration in 25 patients with Crohn's disease led to detection of intestinal mTNF(+) immune cells during confocal laser endomicroscopy. Patients with high numbers of mTNF(+) cells showed significantly higher short-term response rates (92%) at week 12 upon subsequent anti-TNF therapy as compared to patients with low amounts of mTNF(+) cells (15%). This clinical response in the former patients was sustained over a follow-up period of 1 year and was associated with mucosal healing observed in follow-up endoscopy. These data indicate that molecular imaging with fluorescent antibodies has the potential to predict therapeutic responses to biological treatment and can be used for personalized medicine in Crohn's disease and autoimmune or inflammatory disorders.
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Affiliation(s)
- Raja Atreya
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Helmut Neumann
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Clemens Neufert
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Maximilian J Waldner
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Ulrike Billmeier
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Yurdagül Zopf
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Marcus Willma
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Christine App
- Department of Biochemistry, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Tino Münster
- Department of Anesthesiology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Hermann Kessler
- Department of Surgery, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Stefanie Maas
- Center for Clinical Studies, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Bernd Gebhardt
- Center for Clinical Studies, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Ralph Heimke-Brinck
- Department of Pharmacy, Erlangen University Hospital, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Eva Reuter
- Department of Pharmacy, Erlangen University Hospital, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Frank Dörje
- Department of Pharmacy, Erlangen University Hospital, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Tilman T Rau
- Department of Pathology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Wolfgang Uter
- Department of Medical Informatics, Biometry and Epidemiology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Thomas D Wang
- Division of Gastroenterology, University of Michigan, Ann Arbor, Michigan, USA
| | | | - Michael Vieth
- Institute of Pathology, Klinikum Bayreuth, Bayreuth, Germany
| | - Ewald Hannappel
- Department of Biochemistry, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
| | - Markus F Neurath
- Medical Clinic 1, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany
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31
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Kyrish M, Dobbs J, Jain S, Wang X, Yu D, Richards-Kortum R, Tkaczyk TS. Needle-based fluorescence endomicroscopy via structured illumination with a plastic, achromatic objective. JOURNAL OF BIOMEDICAL OPTICS 2013; 18:096003. [PMID: 24002190 PMCID: PMC3759804 DOI: 10.1117/1.jbo.18.9.096003] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/30/2013] [Revised: 07/19/2013] [Accepted: 07/31/2013] [Indexed: 05/18/2023]
Abstract
In order to diagnose cancer, a sample must be removed, prepared, and examined under a microscope, which is expensive, invasive, and time consuming. Fiber optic fluorescence endomicroscopy, where an image guide is used to obtain high-resolution images of tissue in vivo, has shown promise as an alternative to conventional biopsies. However, the resolution of standard endomicroscopy is limited by the fiber bundle sampling frequency and out-of-focus light. A system is presented which incorporates a plastic, achromatic objective to increase the sampling and which provides optical sectioning via structured illumination to reject background light. An image is relayed from the sample by a fiber bundle with the custom 2.1-mm outer diameter objective lens integrated to the distal tip. The objective is corrected for the excitation and the emission wavelengths of proflavine (452 and 515 nm). It magnifies the object onto the fiber bundle to improve the system's lateral resolution by increasing the sampling. The plastic lenses were fabricated via single-point diamond turning and assembled using a zero alignment technique. Ex vivo images of normal and neoplastic murine mammary tissues stained with proflavine are captured. The system achieves higher contrast and resolves smaller features than standard fluorescence endomicroscopy.
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Affiliation(s)
- Matthew Kyrish
- Rice University, Department of Bioengineering, 6100 Main Street, Houston, Texas 77005
| | - Jessica Dobbs
- Rice University, Department of Bioengineering, 6100 Main Street, Houston, Texas 77005
| | - Shalini Jain
- MD Anderson Cancer Center, Department of Molecular and Cellular Oncology, 1515 Holcombe Boulevard, Box 108, Houston, Texas 77030
| | - Xiao Wang
- MD Anderson Cancer Center, Department of Molecular and Cellular Oncology, 1515 Holcombe Boulevard, Box 108, Houston, Texas 77030
| | - Dihua Yu
- MD Anderson Cancer Center, Department of Molecular and Cellular Oncology, 1515 Holcombe Boulevard, Box 108, Houston, Texas 77030
| | | | - Tomasz S. Tkaczyk
- Rice University, Department of Bioengineering, 6100 Main Street, Houston, Texas 77005
- Address all correspondence to: Tomasz S. Tkaczyk, Rice University, Department of Bioengineering, 6100 Main Street, Houston, Texas 77005. Tel: 713-348-4362; Fax: 713-348-5877; E-mail:
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32
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Descamps E, Duroure N, Deiss F, Leichlé T, Adam C, Mailley P, Aït-Ikhlef A, Livache T, Nicu L, Sojic N. Functionalization of optical nanotip arrays with an electrochemical microcantilever for multiplexed DNA detection. LAB ON A CHIP 2013; 13:2956-62. [PMID: 23695411 DOI: 10.1039/c3lc50335f] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/24/2023]
Abstract
Optical nanotip arrays fabricated on etched fiber bundles were functionalized with DNA spots. Such unconventional substrates (3D and non-planar) are difficult to pattern with standard microfabrication techniques but, using an electrochemical cantilever, up to 400 spots were electrodeposited on the nanostructured optical surface in 5 min. This approach allows each spot to be addressed individually and multiplexed fluorescence detection is demonstrated. Finally, remote fluorescence detection was performed by imaging through the optical fiber bundle itself after hybridisation with the complementary sequence.
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Affiliation(s)
- Emeline Descamps
- Institut des Sciences Moléculaires, UMR 5255 CNRS, Université Bordeaux 1-ENSCBP 16, Avenue Pey-Berland, 33607 Pessac, France
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33
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Cicchi R, Sturiale A, Nesi G, Kapsokalyvas D, Alemanno G, Tonelli F, Pavone FS. Multiphoton morpho-functional imaging of healthy colon mucosa, adenomatous polyp and adenocarcinoma. BIOMEDICAL OPTICS EXPRESS 2013; 4:1204-13. [PMID: 23847743 PMCID: PMC3704099 DOI: 10.1364/boe.4.001204] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/16/2013] [Revised: 05/09/2013] [Accepted: 05/09/2013] [Indexed: 05/11/2023]
Abstract
Two-photon spectral resolved imaging was used to image fresh human biopsies of colon tissue and to characterize healthy colon mucosa, adenomatous polyp and adenocarcinoma by means of a morpho-functional analysis. Morphological examination, performed using endogenous tissue fluorescence, discriminated adenomatous and adenocarcinoma tissues from normal mucosa in terms of cellular asymmetry and nucleus-to-cytoplasm ratio. Good agreement was found between multiphoton images and histological examination performed on the same samples. Further characterization, performed by means of spectral-resolved analysis of NADH and FAD fluorescence, demonstrated an altered metabolic activity in both adenomatous and adenocarcinoma tissues compared to healthy mucosa. This morpho-functional approach may represent a powerful method to be used in combination with endoscopy for in vivo optical diagnosis of colon cancer and may be extended to other tissues.
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Affiliation(s)
- Riccardo Cicchi
- National Institute of Optics, National Research Council (INO-CNR), Largo E. Fermi 6, Florence, I-50125, Italy
- European Laboratory for Non-Linear Spectroscopy (LENS), Via N. Carrara 1, Sesto Fiorentino, I-50019, Italy
| | - Alessandro Sturiale
- Department of Clinical Physiopathology, Surgical Unit, University of Florence, Florence, I-50100, Italy
| | - Gabriella Nesi
- Division of Human Pathology and Oncology, Department of Surgical and Medical Critical Care, University of Florence, Florence, I-50100, Italy
| | - Dimitrios Kapsokalyvas
- European Laboratory for Non-Linear Spectroscopy (LENS), Via N. Carrara 1, Sesto Fiorentino, I-50019, Italy
| | - Giovanni Alemanno
- Department of Clinical Physiopathology, Surgical Unit, University of Florence, Florence, I-50100, Italy
| | - Francesco Tonelli
- Department of Clinical Physiopathology, Surgical Unit, University of Florence, Florence, I-50100, Italy
| | - Francesco S. Pavone
- National Institute of Optics, National Research Council (INO-CNR), Largo E. Fermi 6, Florence, I-50125, Italy
- European Laboratory for Non-Linear Spectroscopy (LENS), Via N. Carrara 1, Sesto Fiorentino, I-50019, Italy
- Department of Physics, University of Florence, Via Giovanni Sansone 1, 50019, Sesto Fiorentino, Italy
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34
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Schwitalla S, Ziegler PK, Horst D, Becker V, Kerle I, Begus-Nahrmann Y, Lechel A, Rudolph KL, Langer R, Slotta-Huspenina J, Bader FG, Prazeres da Costa O, Neurath MF, Meining A, Kirchner T, Greten FR. Loss of p53 in enterocytes generates an inflammatory microenvironment enabling invasion and lymph node metastasis of carcinogen-induced colorectal tumors. Cancer Cell 2013; 23:93-106. [PMID: 23273920 DOI: 10.1016/j.ccr.2012.11.014] [Citation(s) in RCA: 232] [Impact Index Per Article: 19.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/09/2011] [Revised: 09/17/2012] [Accepted: 11/26/2012] [Indexed: 12/14/2022]
Abstract
Loss of p53 is considered to allow progression of colorectal tumors from the adenoma to the carcinoma stage. Using mice with an intestinal epithelial cell (IEC)-specific p53 deletion, we demonstrate that loss of p53 alone is insufficient to initiate intestinal tumorigenesis but markedly enhances carcinogen-induced tumor incidence and leads to invasive cancer and lymph node metastasis. Whereas p53 controls DNA damage and IEC survival during the initiation stage, loss of p53 during tumor progression is associated with increased intestinal permeability, causing formation of an NF-κB-dependent inflammatory microenvironment and the induction of epithelial-mesenchymal transition. Thus, we propose a p53-controlled tumor-suppressive function that is independent of its well-established role in cell-cycle regulation, apoptosis, and senescence.
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Affiliation(s)
- Sarah Schwitalla
- Institute of Molecular Immunology, Klinikum rechts der Isar, Technical University Munich, Munich, Germany
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35
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Shu Q, Adam C, Sojic N, Schmittel M. Electrochemiluminescent polymer films with a suitable redox “turn-off” absorbance window for remote selective sensing of Hg2+. Analyst 2013; 138:4500-4. [DOI: 10.1039/c3an00545c] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
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36
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Fujiya M, Kohgo Y. Image-enhanced endoscopy for the diagnosis of colon neoplasms. Gastrointest Endosc 2013; 77:111-118.e5. [PMID: 23148965 DOI: 10.1016/j.gie.2012.07.031] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/20/2011] [Accepted: 07/18/2012] [Indexed: 02/08/2023]
Affiliation(s)
- Mikihiro Fujiya
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical University, Hokkaido, Japan
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37
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Roper J, Hung KE. Priceless GEMMs: genetically engineered mouse models for colorectal cancer drug development. Trends Pharmacol Sci 2012; 33:449-55. [DOI: 10.1016/j.tips.2012.05.001] [Citation(s) in RCA: 30] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2012] [Revised: 04/11/2012] [Accepted: 05/02/2012] [Indexed: 12/13/2022]
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38
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Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals. Nat Protoc 2012; 7:1456-69. [PMID: 22767088 DOI: 10.1038/nprot.2012.078] [Citation(s) in RCA: 60] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Abstract
Intravital fluorescence microscopy has emerged as a powerful technique to visualize cellular processes in vivo. However, owing to their size, the objective lenses required have limited physical accessibility to various tissue sites in the internal organs of small animals. The use of small-diameter probes using graded-index (GRIN) lenses expands the capabilities of conventional intravital microscopes to minimally invasive imaging of internal organs. In this protocol, we describe the detailed steps for the fabrication of front- and side-view GRIN probes and the integration and operation of the probes in a confocal microscope to enable visualization of fluorescent cells and microvasculature in various mouse organs. Some experience in building an optical setup is required to complete the protocol. We also present longitudinal imaging of immune cells in renal allografts and tumor development in the colon. Fabrication and integration can be completed in 5-7 h, and a typical in vivo imaging session takes 1-2 h.
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39
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Duerr CU, Hornef MW. The mammalian intestinal epithelium as integral player in the establishment and maintenance of host-microbial homeostasis. Semin Immunol 2011; 24:25-35. [PMID: 22138188 DOI: 10.1016/j.smim.2011.11.002] [Citation(s) in RCA: 50] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
Only one single layer of epithelial cells separates the densely colonized and environmentally exposed intestinal lumen from the largely sterile subepithelial tissue. Together with the overlaying mucus and the subepithelial mucosal immune system the epithelium has evolved to maintain homeostasis in the presence of the enteric microbiota. It also contributes to rapid and efficient antimicrobial host defence in the event of infection with pathogenic microorganisms. Both, epithelial antimicrobial host defence and homeostasis rely on signalling pathways induced by innate immune receptors demonstrating the active role of epithelial cells in the host-microbial interplay. The interaction of epithelial cells with professional immune cells illustrates the integrated function within the mucosal tissue. In the present review we focus on structural and functional changes of the intestinal epithelium during the fetal-neonatal transition and infancy and try to delineate its role in the induction and maintenance of host-microbial homeostasis. We also address factors that impair epithelial functions and may lead to disruption of the mucosal barrier, tissue damage and the development of symptomatic disease.
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Affiliation(s)
- Claudia U Duerr
- Institute for Medical Microbiology and Hospital Epidemiology, Hannover Medical School, Carl-Neuberg Str. 1, D-30625 Hannover, Germany
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