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Zhu M, Fang Y, Sun Y, Li S, Yu J, Xiong B, Wen C, Zhou B, Huang B, Yin H, Xu H. Sonogenetics in the Treatment of Chronic Diseases: A New Method for Cell Regulation. ADVANCED SCIENCE (WEINHEIM, BADEN-WURTTEMBERG, GERMANY) 2024; 11:e2407373. [PMID: 39488795 PMCID: PMC11672274 DOI: 10.1002/advs.202407373] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/28/2024] [Revised: 10/21/2024] [Indexed: 11/04/2024]
Abstract
Sonogenetics is an innovative technology that integrates ultrasound with genetic editing to precisely modulate cellular activities in a non-invasive manner. This method entails introducing and activating mechanosensitive channels on the cell membrane of specific cells using gene delivery vectors. When exposed to ultrasound, these channels can be manipulated to open or close, thereby impacting cellular functions. Sonogenetics is currently being used extensively in the treatment of various chronic diseases, including Parkinson's disease, vision restoration, and cancer therapy. This paper provides a comprehensive review of key components of sonogenetics and focuses on evaluating its prospects and potential challenges in the treatment of chronic disease.
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Affiliation(s)
- Mingrui Zhu
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Yan Fang
- Department of Ultrasound, Huashan HospitalFudan UniversityShanghai200040P. R. China
| | - Yikang Sun
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Shaoyue Li
- Department of Medical Ultrasound, Center of Minimally Invasive Treatment for TumorShanghai Tenth People's HospitalUltrasound Research and Education InstituteClinical Research Center for Interventional MedicineSchool of MedicineTongji UniversityShanghai200072P. R. China
| | - Jifeng Yu
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Bing Xiong
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Congjian Wen
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Boyang Zhou
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Bin Huang
- Zhejiang HospitalHangzhou310013P. R. China
| | - Haohao Yin
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
| | - Huixiong Xu
- Department of UltrasoundInstitute of Ultrasound in Medicine and EngineeringZhongshan HospitalFudan UniversityShanghai200032P. R. China
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Methods for CRISPR-Cas as Ribonucleoprotein Complex Delivery In Vivo. Mol Biotechnol 2023; 65:181-195. [PMID: 35322386 DOI: 10.1007/s12033-022-00479-z] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/04/2021] [Accepted: 03/14/2022] [Indexed: 01/18/2023]
Abstract
The efficient delivery of CRISPR-Cas components is still a key and unsolved problem. CRISPR-Cas delivery in the form of a Cas protein+sgRNA (ribonucleoprotein complex, RNP complex), has proven to be extremely effective, since it allows to increase on-target activity, while reducing nonspecific activity. The key point for in vivo genome editing is the direct delivery of artificial nucleases and donor DNA molecules into the somatic cells of an adult organism. At the same time, control of the dose of artificial nucleases is impossible, which affects the efficiency of genome editing in the affected cells. Poor delivery efficiency and low editing efficacy reduce the overall potency of the in vivo genome editing process. Here we review how this problem is currently being solved in scientific works and what types of in vivo delivery methods of Cas9/sgRNA RNPs have been developed.
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3
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O'Sullivan G, Philips JG, Rasko JEJ. Clinical gene technology in Australia: building on solid foundations. Med J Aust 2022; 217:65-70. [PMID: 35842935 PMCID: PMC9540640 DOI: 10.5694/mja2.51629] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/13/2021] [Revised: 02/28/2022] [Accepted: 03/08/2022] [Indexed: 11/17/2022]
Affiliation(s)
| | | | - John EJ Rasko
- Royal Prince Alfred Hospital Sydney NSW
- University of Sydney Sydney NSW
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Mendonça RH, Zanoteli E. Gene therapy in neuromuscular disorders. ARQUIVOS DE NEURO-PSIQUIATRIA 2022; 80:249-256. [PMID: 35976325 PMCID: PMC9491441 DOI: 10.1590/0004-282x-anp-2022-s135] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 04/04/2022] [Accepted: 04/29/2022] [Indexed: 06/15/2023]
Abstract
Monogenic neuromuscular disorders are potentially treatable through gene therapy. Using viral vectors, a therapeutic transgene aims to restore normal levels of a protein not produced by the defective gene, or to silence a gene whose expression leads to toxic effects. Spinal Muscular Atrophy (SMA) is a good example of a monogenic disease that currently has an AAV9-based vector gene therapy as a therapeutic option. In this review, we intend to discuss the viral vectors and their mechanisms of action, in addition to reviewing the clinical trials that supported the approval of gene therapy (AVXS-101) for SMA as well as neuromuscular diseases that are potentially treatable with gene replacement therapy.
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Affiliation(s)
- Rodrigo Holanda Mendonça
- Universidade de São Paulo, Faculdade de Medicina, Hospital das Clínicas, Departamento de Neurologia, São Paulo, SP, Brazil
| | - Edmar Zanoteli
- Universidade de São Paulo, Faculdade de Medicina, Hospital das Clínicas, Departamento de Neurologia, São Paulo, SP, Brazil
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Shinkuma S. Advances in gene therapy and their application to skin diseases: A review. J Dermatol Sci 2021; 103:2-9. [PMID: 34049771 DOI: 10.1016/j.jdermsci.2021.05.004] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/22/2020] [Revised: 05/18/2021] [Accepted: 05/18/2021] [Indexed: 10/21/2022]
Abstract
With recent advances in genetic engineering technology, gene therapy is now being considered as a treatment not only for congenital diseases but also acquired diseases, such as cancer. Gene therapeutic agents for hereditary immune disorders, haemophilia, retinal diseases, neurodegenerative diseases, and lymphoma have been approved in the United States and Europe. In the field of dermatology, clinical trials of gene therapy have been conducted, because the skin is an easily accessible organ that represents an attractive tissue for gene therapy. In recent years, gene therapy has been attempted for a variety of skin diseases, such as genodermatoses (including epidermolysis bullosa and Netherton syndrome), cutaneous lymphoma, and malignant melanoma. As a result, it is difficult to grasp the current status of gene therapy in dermatology. This review focuses on each of the gene-transfer techniques currently in use and describes the current status of gene therapy for skin diseases using each technology.
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Affiliation(s)
- Satoru Shinkuma
- Department of Dermatology, Nara Medical University School of Medicine, Kashihara, Japan.
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Bulcha JT, Wang Y, Ma H, Tai PWL, Gao G. Viral vector platforms within the gene therapy landscape. Signal Transduct Target Ther 2021; 6:53. [PMID: 33558455 PMCID: PMC7868676 DOI: 10.1038/s41392-021-00487-6] [Citation(s) in RCA: 711] [Impact Index Per Article: 177.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/03/2020] [Revised: 10/05/2020] [Accepted: 10/23/2020] [Indexed: 01/30/2023] Open
Abstract
Throughout its 40-year history, the field of gene therapy has been marked by many transitions. It has seen great strides in combating human disease, has given hope to patients and families with limited treatment options, but has also been subject to many setbacks. Treatment of patients with this class of investigational drugs has resulted in severe adverse effects and, even in rare cases, death. At the heart of this dichotomous field are the viral-based vectors, the delivery vehicles that have allowed researchers and clinicians to develop powerful drug platforms, and have radically changed the face of medicine. Within the past 5 years, the gene therapy field has seen a wave of drugs based on viral vectors that have gained regulatory approval that come in a variety of designs and purposes. These modalities range from vector-based cancer therapies, to treating monogenic diseases with life-altering outcomes. At present, the three key vector strategies are based on adenoviruses, adeno-associated viruses, and lentiviruses. They have led the way in preclinical and clinical successes in the past two decades. However, despite these successes, many challenges still limit these approaches from attaining their full potential. To review the viral vector-based gene therapy landscape, we focus on these three highly regarded vector platforms and describe mechanisms of action and their roles in treating human disease.
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Affiliation(s)
- Jote T Bulcha
- Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA
- Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, USA
| | - Yi Wang
- Department of Pathophysiology, West China College of Basic medical sciences & Forensic Medicine, Sichuan University, Chengdu, China
| | - Hong Ma
- Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA
| | - Phillip W L Tai
- Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA.
- Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, USA.
- VIDE Program, University of Massachusetts Medical School, Worcester, MA, USA.
| | - Guangping Gao
- Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA, USA.
- Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, USA.
- Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA, USA.
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8
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Negahdaripour M. Fifty Years of Witnessing Biomedical Science Developments. IRANIAN JOURNAL OF MEDICAL SCIENCES 2020; 45:403-404. [PMID: 33281257 PMCID: PMC7707628 DOI: 10.30476/ijms.2020.47043] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Download PDF] [Subscribe] [Scholar Register] [Indexed: 11/24/2022]
Affiliation(s)
- Manica Negahdaripour
- Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.,Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
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9
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Bajan S, Hutvagner G. RNA-Based Therapeutics: From Antisense Oligonucleotides to miRNAs. Cells 2020; 9:E137. [PMID: 31936122 PMCID: PMC7016530 DOI: 10.3390/cells9010137] [Citation(s) in RCA: 260] [Impact Index Per Article: 52.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2019] [Revised: 12/23/2019] [Accepted: 12/30/2019] [Indexed: 02/07/2023] Open
Abstract
The first therapeutic nucleic acid, a DNA oligonucleotide, was approved for clinical use in 1998. Twenty years later, in 2018, the first therapeutic RNA-based oligonucleotide was United States Food and Drug Administration (FDA) approved. This promises to be a rapidly expanding market, as many emerging biopharmaceutical companies are developing RNA interference (RNAi)-based, and RNA-based antisense oligonucleotide therapies. However, miRNA therapeutics are noticeably absent. miRNAs are regulatory RNAs that regulate gene expression. In disease states, the expression of many miRNAs is measurably altered. The potential of miRNAs as therapies and therapeutic targets has long been discussed and in the context of a wide variety of infections and diseases. Despite the great number of studies identifying miRNAs as potential therapeutic targets, only a handful of miRNA-targeting drugs (mimics or inhibitors) have entered clinical trials. In this review, we will discuss whether the investment in finding potential miRNA therapeutic targets has yielded feasible and practicable results, the benefits and obstacles of miRNAs as therapeutic targets, and the potential future of the field.
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Affiliation(s)
- Sarah Bajan
- Faculty of Science, University of Technology Sydney, Sydney, NSW 2000, Australia
- Health and Sport Science, University of Sunshine Coast, Sunshine Coast, QLD 4556, Australia
| | - Gyorgy Hutvagner
- School of Biomedical Engineering Faculty of Engineering and Information Technology, University of Technology Sydney, Sydney, NSW 2000, Australia
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Attitudes of clinical geneticists and certified genetic counselors to genome editing and its clinical applications: A nation-wide questionnaire survey in Japan. J Hum Genet 2019; 64:945-954. [PMID: 31273322 DOI: 10.1038/s10038-019-0635-z] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2019] [Revised: 04/24/2019] [Accepted: 06/10/2019] [Indexed: 11/08/2022]
Abstract
Genome editing of the human embryo using CRISPR/Cas9 has the potential to prevent hereditary diseases from being transmitted to the next generation. However, attitudes to this technology have not been examined sufficiently among the genetic professionals who will use it in the near future. We conducted a questionnaire survey of Japanese clinical geneticists and certified genetic counselors. Differences were observed between them in their recognition of this technology and impressions on its difficulty and cost. Both groups worried about misuse of it, with insufficient information and rules. As key elements for such rules, they considered ethics, safety, and purpose. Most disapproved of modifying physical traits as an enhancement, though they hoped for the treatment of severe diseases. At current clinical sites, they tended to adopt a prudent attitude by mentioning only the possibility of genome editing in the future. Academic policies and legislation are required, especially for application in human embryos, through a consensus of professionals and general citizens. Furthermore, professionals should maintain awareness of new developments and regularly reexamine attitudes for the ongoing development of more suitable rules, education systems, and clinical protocols. As preparation for changes, opportunities to address ethical issues and initiate discussions are also required.
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Kotcherlakota R, Vydiam K, Jeyalakshmi Srinivasan D, Mukherjee S, Roy A, Kuncha M, Rao TN, Sistla R, Gopal V, Patra CR. Restoration of p53 Function in Ovarian Cancer Mediated by Gold Nanoparticle-Based EGFR Targeted Gene Delivery System. ACS Biomater Sci Eng 2019; 5:3631-3644. [PMID: 33405744 DOI: 10.1021/acsbiomaterials.9b00006] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
Targeted gene delivery of wild type tumor suppressor gene p53 is a promising approach to inhibit the progression of ovarian cancer. Although several gene delivery vehicles have been reported earlier, there is paucity for targeted delivery of wild type p53 to ovarian cancer using gold nanoparticles. As it is well-known that EGFR (epidermal growth factor receptor) is overexpressed in ovarian cancer, in this study we hypothesized that the FDA approved monoclonal antibody C225 (cetuximab) that targets EGFR could be used for targeted delivery of wild type p53 gene. With this impetus, we devised an approach wherein cationic gold nanoparticles (AuNPs) were employed to generate gold nanoparticle-based drug delivery system (DDS, Au-C225-p53DNA where p53DNA is pCMVp53 plasmid) that was formulated and characterized by biochemical and biophysical methods. The nanoconjugate complexed with DNA (Au-C225-p53DNA) is serum-stable and protects the bound DNA from digestion by DNase-I. Additionally, in vitro reporter gene expression assays demonstrated efficient and specific gene transfection in EGFR overexpressing SK-OV-3 cells. Further, the intraperitoneal administration of Au-C225-p53DNA in SK-OV-3 xenograft mouse model displayed significant tumor targeting and tumor regression. Altogether, these studies indicated a promising nanoparticle-based approach for targeting ovarian cancers caused by mutated p53.
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Affiliation(s)
- Rajesh Kotcherlakota
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India.,Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India
| | - Kalyan Vydiam
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India
| | - Durga Jeyalakshmi Srinivasan
- CSIR-Centre for Cellular and Molecular Biology (Council of Scientific and Industrial Research), Uppal Road, Hyderabad 500007, Telangana India
| | - Sudip Mukherjee
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India.,Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India
| | - Arpita Roy
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India
| | - Madhusudana Kuncha
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India
| | - T Nageswara Rao
- Mass and Analytical Division, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana, India
| | - Ramakrishna Sistla
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India.,Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India
| | - Vijaya Gopal
- CSIR-Centre for Cellular and Molecular Biology (Council of Scientific and Industrial Research), Uppal Road, Hyderabad 500007, Telangana India.,Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India
| | - Chitta Ranjan Patra
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Uppal Road, Tarnaka, Hyderabad 500007, Telangana India.,Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India
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12
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Yuan Y, Gu Z, Yao C, Luo D, Yang D. Nucleic Acid-Based Functional Nanomaterials as Advanced Cancer Therapeutics. SMALL (WEINHEIM AN DER BERGSTRASSE, GERMANY) 2019; 15:e1900172. [PMID: 30972963 DOI: 10.1002/smll.201900172] [Citation(s) in RCA: 69] [Impact Index Per Article: 11.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/10/2019] [Revised: 03/04/2019] [Indexed: 06/09/2023]
Abstract
Nucleic acid-based functional nanomaterials (NAFN) have been widely used as emerging drug delivery nanocarriers for cancer therapeutics. Considerable works have demonstrated that NAFN can effectively load and protect therapeutic agents, and particularly enable targeting delivery to the tumor site and stimuli-responsive release. These outstanding performances are due to NAFN's unique properties including inherent biological functions and sequence programmability as well as biocompatibility and biodegradability. In this Review, the recent progress on NAFN as advanced cancer therapeutics is highlighted. Three main cancer therapy approaches are categorized including chemo-, immuno-, and gene-therapy. Examples are presented to show how NAFN are rationally and exquisitely designed to address problems in cancer therapy. The challenges and future development of NAFN are also discussed toward future more practical biomedical applications.
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Affiliation(s)
- Ye Yuan
- Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, P. R. China
| | - Zi Gu
- School of Chemical Engineering and Australian Centre for NanoMedicine, University of New South Wales, Sydney, NSW, 2052, Australia
| | - Chi Yao
- Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, P. R. China
| | - Dan Luo
- Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY, 14853, USA
- CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, 215123, P. R. China
| | - Dayong Yang
- Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, P. R. China
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13
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Loading of PNA and Other Molecular Payloads on Inorganic Nanostructures for Theranostics. Methods Mol Biol 2019; 1811:65-77. [PMID: 29926446 DOI: 10.1007/978-1-4939-8582-1_5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/28/2022]
Abstract
Peptide Nucleic Acids (PNAs) are oligonucleotide mimics that can be used as drugs as they can interact with DNA and RNA targets in organisms. Loading PNAs into inorganic nanocarriers can improve their cellular uptake and co-delivering them together with drugs can improve the therapy efficacy by synergic effects. Furthermore, the functionalization of the carriers with labels allows theranostics, and the possibility to monitor the efficacy of the therapy in real time. The present protocol describes the synthesis of Zeolites-L nanocrystals and mesoporous silica nanoparticles and their loading with cationic PNAs and other smaller molecular weight payloads towards theranostics applications.
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14
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Abstract
PURPOSE OF REVIEW The current knowledge of pathophysiological and molecular mechanisms responsible for the genesis and development of heart failure (HF) is absolutely vast. Nonetheless, the hiatus between experimental findings and therapeutic options remains too deep, while the available pharmacological treatments are mostly seasoned and display limited efficacy. The necessity to identify new, non-pharmacological strategies to target molecular alterations led investigators, already many years ago, to propose gene therapy for HF. Here, we will review some of the strategies proposed over the past years to target major pathogenic mechanisms/factors responsible for severe cardiac injury developing into HF and will provide arguments in favor of the necessity to keep alive research on this topic. RECENT FINDINGS After decades of preclinical research and phases of enthusiasm and disappointment, clinical trials were finally launched in recent years. The first one to reach phase II and testing gene delivery of sarcoendoplasmic reticulum calcium ATPase did not yield encouraging results; however, other trials are ongoing, more efficient viral vectors are being developed, and promising new potential targets have been identified. For instance, recent research is focused on gene repair, in vivo, to treat heritable forms of HF, while strong experimental evidence indicates that specific microRNAs can be delivered to post-ischemic hearts to induce regeneration, a result that was previously thought possible only by using stem cell therapy. Gene therapy for HF is aging, but exciting perspectives are still very open.
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Affiliation(s)
- Khatia Gabisonia
- Institute of Life Sciences, Fondazione Toscana Gabriele Monasterio, Scuola Superiore Sant'Anna, Piazza Martiri della Liberta` 33, 56127, Pisa, Italy
| | - Fabio A Recchia
- Institute of Life Sciences, Fondazione Toscana Gabriele Monasterio, Scuola Superiore Sant'Anna, Piazza Martiri della Liberta` 33, 56127, Pisa, Italy.
- Cardiovascular Research Center, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, USA.
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Abstract
Gene therapy is becoming a real therapeutic chance in some genetic disorders. The first candidates to this approach are genetic diseases which involve hematopoiesis, because of the facility for the collection and the manipulation of hematopoietic progenitors. Apheresis techniques, which are able to collect a great number of mononuclear cells from peripheral blood, are ideal for obtaining a large number of cells which can be transfected. Future uses of gene therapy techniques could be: the treatment of hematopoietic genetic disorders, procedures of gene marking, and the manipulation of normal hematopoietic cells with the objective of increasing their resistance to myelotoxic drugs.
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Affiliation(s)
- P. Servida
- Service of Hematology, IRCCS “San Raffaele” Hospital, Milano - Italy
| | - C. Bordignon
- Service of Hematology, IRCCS “San Raffaele” Hospital, Milano - Italy
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17
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Santos-Carballal B, Fernández Fernández E, Goycoolea FM. Chitosan in Non-Viral Gene Delivery: Role of Structure, Characterization Methods, and Insights in Cancer and Rare Diseases Therapies. Polymers (Basel) 2018; 10:E444. [PMID: 30966479 PMCID: PMC6415274 DOI: 10.3390/polym10040444] [Citation(s) in RCA: 77] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2018] [Revised: 04/04/2018] [Accepted: 04/11/2018] [Indexed: 12/23/2022] Open
Abstract
Non-viral gene delivery vectors have lagged far behind viral ones in the current pipeline of clinical trials of gene therapy nanomedicines. Even when non-viral nanovectors pose less safety risks than do viruses, their efficacy is much lower. Since the early studies to deliver pDNA, chitosan has been regarded as a highly attractive biopolymer to deliver nucleic acids intracellularly and induce a transgenic response resulting in either upregulation of protein expression (for pDNA, mRNA) or its downregulation (for siRNA or microRNA). This is explained as the consequence of a multi-step process involving condensation of nucleic acids, protection against degradation, stabilization in physiological conditions, cellular internalization, release from the endolysosome ("proton sponge" effect), unpacking and enabling the trafficking of pDNA to the nucleus or the siRNA to the RNA interference silencing complex (RISC). Given the multiple steps and complexity involved in the gene transfection process, there is a dearth of understanding of the role of chitosan's structural features (Mw and degree of acetylation, DA%) on each step that dictates the net transfection efficiency and its kinetics. The use of fully characterized chitosan samples along with the utilization of complementary biophysical and biological techniques is key to bridging this gap of knowledge and identifying the optimal chitosans for delivering a specific gene. Other aspects such as cell type and administration route are also at play. At the same time, the role of chitosan structural features on the morphology, size and surface composition of synthetic virus-like particles has barely been addressed. The ongoing revolution brought about by the recent discovery of CRISPR-Cas9 technology will undoubtedly be a game changer in this field in the short term. In the field of rare diseases, gene therapy is perhaps where the greatest potential lies and we anticipate that chitosans will be key players in the translation of research to the clinic.
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Affiliation(s)
| | - Elena Fernández Fernández
- Lung Biology Group, Department Clinical Microbiology, RCSI, Education and Research Centre, Beaumont Hospital, Dublin 9, Ireland.
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18
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López-Marín LM, Rivera AL, Fernández F, Loske AM. Shock wave-induced permeabilization of mammalian cells. Phys Life Rev 2018; 26-27:1-38. [PMID: 29685859 DOI: 10.1016/j.plrev.2018.03.001] [Citation(s) in RCA: 25] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2017] [Revised: 02/12/2018] [Accepted: 02/26/2018] [Indexed: 12/18/2022]
Abstract
Controlled permeabilization of mammalian cell membranes is fundamental to develop gene and cell therapies based on macromolecular cargo delivery, a process that emerged against an increasing number of health afflictions, including genetic disorders, cancer and infections. Viral vectors have been successfully used for macromolecular delivery; however, they may have unpredictable side effects and have been limited to life-threatening cases. Thus, several chemical and physical methods have been explored to introduce drugs, vaccines, and nucleic acids into cells. One of the most appealing physical methods to deliver genes into cells is shock wave-induced poration. High-speed microjets of fluid, emitted due to the collapse of microbubbles after shock wave passage, represent the most significant mechanism that contributes to cell membrane poration by this technique. Herein, progress in shock wave-induced permeabilization of mammalian cells is presented. After covering the main concepts related to molecular strategies whose applications depend on safer drug delivery methods, the physics behind shock wave phenomena is described. Insights into the use of shock waves for cell membrane permeation are discussed, along with an overview of the two major biomedical applications thereof-i.e., genetic modification and anti-cancer shock wave-assisted chemotherapy. The aim of this review is to summarize 30 years of data showing underwater shock waves as a safe, noninvasive method for macromolecular delivery into mammalian cells, encouraging the development of further research, which is still required before the introduction of this promising tool into clinical practice.
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Affiliation(s)
- Luz M López-Marín
- Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, 76230 Querétaro, Qro., Mexico.
| | - Ana Leonor Rivera
- Instituto de Ciencias Nucleares & Centro de Ciencias de la Complejidad, Universidad Nacional Autónoma de México, Ciudad Universitaria, 04510 Ciudad de México, Mexico.
| | - Francisco Fernández
- Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, 76230 Querétaro, Qro., Mexico.
| | - Achim M Loske
- Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, 76230 Querétaro, Qro., Mexico.
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19
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Caroline Diana SM, Rekha M. Efficacy of vinyl imidazole grafted cationized pullulan and dextran as gene delivery vectors: A comparative study. Int J Biol Macromol 2017; 105:947-955. [DOI: 10.1016/j.ijbiomac.2017.07.121] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/27/2016] [Revised: 07/01/2017] [Accepted: 07/18/2017] [Indexed: 01/06/2023]
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20
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Li B, Guo W, Zhang F, Liu M, Wang S, Liu Z, Xiang S, Zeng Y. Synthesis and evaluation of L-arabinose-based cationic glycolipids as effective vectors for pDNA and siRNA in vitro. PLoS One 2017; 12:e0180276. [PMID: 28672000 PMCID: PMC5495346 DOI: 10.1371/journal.pone.0180276] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2016] [Accepted: 06/13/2017] [Indexed: 01/14/2023] Open
Abstract
Glycolipids might become a new type of promising non-viral gene delivery systems because of their low cytotoxicity, structural diversity, controllable aqua- and lipo-solubility, appropriate density and distribution of positive charges, high transfer efficiency and potential targeting function. In this study, four kinds of L-arabinose-based cationic glycolipids (Ara-DiC12MA, Ara-DiC14MA, Ara-DiC16MA and Ara-DiC18MA) containing quaternary ammonium as hydrophilic headgroup and two alkane chains as hydrophobic domain were synthesized and characterized. They were observed to have strong affinities for plasmid DNA (pDNA) and siRNA, the pDNA can be completely condensed at N/P ratio less than 2, and the siRNA can be completely retarded at N/P ratio less than 3. The dynamic light scattering (DLS) experiment and atomic force microscopy (AFM) experiment demonstrated that cationic lipids and their lipoplexes possessed suitable particle sizes with near-spherical shape and proper ζ-potentials for cell transfection. The Ara-DiC16MA liposome was found to have good transfection efficacy in HEK293, PC-3 and Mat cells compared with other three kinds of liposomes, and also maintain low cytotoxicity and better uptake capability in vitro. Furthermore, the gene silencing assay showed that Ara-DiC14MA and Ara-DiC16MA liposomes have demonstrated effective delivery and higher gene knockdown activity (>80%) in the above mentioned cells than Lipofectamine 2000. These results indicated Ara-DiC16MA can be developed for efficient and low toxic gene delivery.
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Affiliation(s)
- Bo Li
- National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Wanrong Guo
- National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Fan Zhang
- National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Meiyan Liu
- National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Shang Wang
- Key Laboratory of Protein Chemistry and Developmental Biology of State Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Zhonghua Liu
- The National &Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, Hunan, P.R. China
| | - Shuanglin Xiang
- Key Laboratory of Protein Chemistry and Developmental Biology of State Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha, Hunan, P. R. China
| | - Youlin Zeng
- National & Local Joint Engineering Laboratory for New Petro-chemical Materials and Fine Utilization of Resources, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, P. R. China
- * E-mail:
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21
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Abstract
The potential for cell transplantation is enormous, not only in replacement therapy in conditions such as diabetes but also in approaches to gene therapy and the induction of tolerance to organ transplants. Immunobiological aspects of cell transplantation include: 1) isolation and purification of cells for transplantation, 2) preservation of cells, 3) technical problems of transplantation, 4) the immune response to cell transplants, 5) prevention of the immune response to cell allografts, 6) delivery and regulation of the product of the cell transplant, 7) xenotransplantation, 8) gene therapy. These aspects are briefly discussed.
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Affiliation(s)
- Peter J. Morris
- Nuffield Department of Surgery, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom
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22
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Lu L, Ge Y, Li ZH, Freie B, Clapp DW, Broxmeyer HE. CD34+++ Stem/Progenitor Cells Purified from Cryopreserved Normal Cord Blood can be Transduced with High Efficiency by a Retroviral Vector and Expanded Ex Vivo with Stable Integration and Expression of Fanconi Anemia Complementation C Gene. Cell Transplant 2017; 4:493-503. [PMID: 8520833 DOI: 10.1177/096368979500400510] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022] Open
Abstract
A future possibility for treatment of genetic diseases may be gene therapy using autologous cord blood (CB) stem/progenitor cells. This might require cryopreservation of CB stem/progenitor cells prior to purification, gene transduction, and ex vivo expansion of cells. To address this possibility, nonadherent low density T-lymphocyte depleted (NALT-) cells from fresh or cryopreserved cord blood were sorted for CD34+++ phenotype, transduced with a recombinant retroviral vector encoding Fanconi anemia complementation C (FACC) gene, and cells expanded ex vivo in suspension culture for 7 days with growth factors. The results demonstrate: 1) high recovery of viable cells after thawing; 2) high efficiency purification of CD34+++ cells from NALT- cells prior to and after cryopreservation; 3) high degree of expansion of nucleated cells and immature progenitors from CD34+++ cells before and after cryopreservation; 4) efficient transduction with stable integration and expression of newly introduced genes in cryopreserved and then sorted stem/progenitor cells, as detected prior to and after ex vivo expansion; and 5) high efficiency transduction of single isolated CD34+++ cells obtained from cryopreserved NALT- CB. This information should be of value for future studies evaluating the use of cryopreserved cord blood for gene transfer/gene therapy.
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Affiliation(s)
- L Lu
- Department of Medicine (Hematology/Oncology), Indiana University School of Medicine, Indianapolis 46202, USA
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23
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Carriglio N, Klapwijk J, Hernandez RJ, Vezzoli M, Chanut F, Lowe R, Draghici E, Nord M, Albertini P, Cristofori P, Richards J, Staton H, Appleby J, Aiuti A, Sauer AV. Good Laboratory Practice Preclinical Safety Studies for GSK2696273 (MLV Vector-Based Ex Vivo Gene Therapy for Adenosine Deaminase Deficiency Severe Combined Immunodeficiency) in NSG Mice. HUM GENE THER CL DEV 2017; 28:17-27. [DOI: 10.1089/humc.2016.191] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Affiliation(s)
- Nicola Carriglio
- Pathogenesis and Therapy of Primary Immunodeficiencies Unit, San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, Milan, Italy
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
| | - Jan Klapwijk
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Raisa Jofra Hernandez
- Pathogenesis and Therapy of Primary Immunodeficiencies Unit, San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, Milan, Italy
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
| | - Michela Vezzoli
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
| | - Franck Chanut
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Rhiannon Lowe
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Elena Draghici
- Pathogenesis and Therapy of Primary Immunodeficiencies Unit, San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, Milan, Italy
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
| | - Melanie Nord
- GlaxoSmithKline, Regulatory Affairs, King of Prussia, Pennsylvania
| | - Paola Albertini
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
| | - Patrizia Cristofori
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Jane Richards
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Hazel Staton
- Covance Laboratories Ltd, Harrogate, United Kingdom
| | - Jonathan Appleby
- GlaxoSmithKline, In Vitro In Vivo Translation, Ware, United Kingdom
| | - Alessandro Aiuti
- Pathogenesis and Therapy of Primary Immunodeficiencies Unit, San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, Milan, Italy
- Vita-Salute San Raffaele University, Milan, Italy
| | - Aisha V. Sauer
- Pathogenesis and Therapy of Primary Immunodeficiencies Unit, San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, Milan, Italy
- GLP SR-TIGET Test Facility, Ospedale San Rafaele, Milan, Italy
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Kong Y, Zhang X, Zhao Y, Xue Y, Zhang Y. Uptake of DNA by cancer cells without a transfection reagent. Biol Res 2017; 50:2. [PMID: 28109303 PMCID: PMC5251232 DOI: 10.1186/s40659-017-0107-x] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/02/2016] [Accepted: 01/14/2017] [Indexed: 01/05/2023] Open
Abstract
BACKGROUND Cancer cells exhibit elevated levels of glucose uptake and may obtain pre-formed, diet-derived fatty acids from the bloodstream to boost their rapid growth; they may also use nucleic acid from their microenvironment. The study of processing nucleic acid by cancer cells will help improve the understanding of the metabolism of cancer. DNA is commonly packaged into a viral or lipid particle to be transferred into cells; this process is called transfection in laboratory. Cancer cells are known for having gene mutations and the evolving ability of endocytosis. Their uptake of DNAs might be different from normal cells; they may take in DNAs directly from the environment. In this report, we studied the uptake of DNAs in cancer cells without a transfection reagent. METHODS A group of DNA fragments were prepared with PCR and labeled with isotope phosphorous-32 to test their uptake by Huh 7 (liver cancer) and THLE3 (normal liver cells) after incubation overnight by counting radioactivity of the cells' genomic DNA. Multiple cell lines including breast cancer and lung cancer were tested with the same method. DNA molecules were also labeled with fluorescence to test the location in the cells using a kit of "label it fluorescence in situ hybridization (FISH)" from Mirus (USA). RESULTS The data demonstrated that hepatocellular carcinoma cells possess the ability to take in large DNA fragments directly without a transfection reagent whereas normal liver cells cannot. Huh7 and MDA-MB231 cells displayed a significantly higher Rhodamine density in the cytoplasmic phagosomes and this suggests that the mechanism of uptake of large DNA by cancer cells is likely endocytosis. The efficacy of uptake is related to the DNA's size. Some cell lines of lung cancer and breast cancer also showed similar uptake of DNA. CONCLUSIONS In the present study, we have revealed the evidence that some cancer cells, but not nontumorigenic cells, can take DNA fragments directly from the environment without the aid of the transfecting reagent.
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Affiliation(s)
- Yanping Kong
- Department of Endocrinology, Dartmouth-Hitchcock Manchester, 100 Hitchcock Way, Manchester, New Hampshire 03104 USA
| | - Xianbo Zhang
- Department of Surgery/Oncology, First Hospital of Shijiazhuang, 36 Fanxi Road, Shijiazhuang, 050011 Hebei China
| | - Yongliang Zhao
- Laboratory of Disease Genomics and Individualized Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, 1 Beichen West Road, Beijing, 10029 China
| | - Yanfang Xue
- Department of Pharmacology, Hebei Medical University, 361 Zhongshan E Rd, Shijiazhuang, 050056 Hebei China
| | - Ye Zhang
- National Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Dongdan Santiao, Beijing, 100005 China
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25
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Abstract
Embryology mirrors phylogeny. The phenotypic expression of the genome is the result of differential gene transcription, the critically timed turning on and off of specific genes by transcription factors to produce cyto-, histo-, and morpho-differentiation that fleetingly reflects evolutionary stages of development during ontogeny. Hox genes regulate transcription of other structural genes and are responsible for patterning of the facial primordia. Cephalic development involves extremely complex morphogenetic mechanisms built on conserved elements that have undergone enormous evolutionary changes. Transient expression of phylogenetic origins characterize ontogeny and are reflected in defective development that may be due to inappropriate expression of Hox genes or distorted or disrupted epignetic processes. The mechanisms by which genetic information is transformed into morphological patterning by the actions of growth factors, morphogenes, and receptors are currently being identified. Biochemical, immunological, and allometric analyses of embryos and fetuses in experimental and descriptive studies are elucidating details of units of craniofacial morphogenesis--faciogenesis, palatogenesis, gnathogenesis, odontogenesis. Three-dimensional model computer-assisted reconstruction of sectioned embryos and fetuses provides a further technique for understanding the complex configurations of tissue migratory patterns and growth sites that account for normal and abnormal craniofaciogenesis.
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Affiliation(s)
- G H Sperber
- Department of Oral Biology, University of Alberta, Edmonton, Canada
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26
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Alajangi HK, Natarajan P, Vij M, Ganguli M, Santhiya D. Role of Unmodified Low Generation - PAMAM Dendrimers in Efficient Non-Toxic Gene Transfection. ChemistrySelect 2016. [DOI: 10.1002/slct.201600576] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/26/2023]
Affiliation(s)
- Hema Kumari Alajangi
- Department of Applied Chemistry and Polymer Technology; Delhi Technological University
| | | | - Manika Vij
- CSIR-Institute of Genomics and Integrative Biology; Mathura Road Delhi
| | - Munia Ganguli
- CSIR-Institute of Genomics and Integrative Biology; Mathura Road Delhi
| | - Deenan Santhiya
- Department of Applied Chemistry and Polymer Technology; Delhi Technological University
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28
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Rhoads RE. Synthetic mRNA: Production, Introduction into Cells, and Physiological Consequences. Methods Mol Biol 2016; 1428:3-27. [PMID: 27236789 DOI: 10.1007/978-1-4939-3625-0_1] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Recent advances have made it possible to synthesize mRNA in vitro that is relatively stable when introduced into mammalian cells, has a diminished ability to activate the innate immune response against exogenous (virus-like) RNA, and can be efficiently translated into protein. Synthetic methods have also been developed to produce mRNA with unique investigational properties such as photo-cross-linking, fluorescence emission, and attachment of ligands through click chemistry. Synthetic mRNA has been proven effective in numerous applications beneficial for human health such as immunizing patients against cancer and infections diseases, alleviating diseases by restoring deficient proteins, converting somatic cells to pluripotent stem cells to use in regenerative medicine therapies, and engineering the genome by making specific alterations in DNA. This introductory chapter provides background information relevant to the following 20 chapters of this volume that present protocols for these applications of synthetic mRNA.
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Affiliation(s)
- Robert E Rhoads
- Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, LA, 71130-3932, USA.
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29
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Garanina EE, Mukhamedshina YO, Salafutdinov II, Kiyasov AP, Lima LM, Reis HJ, Palotás A, Islamov RR, Rizvanov AA. Construction of recombinant adenovirus containing picorna-viral 2A-peptide sequence for the co-expression of neuro-protective growth factors in human umbilical cord blood cells. Spinal Cord 2015; 54:423-30. [PMID: 26439843 DOI: 10.1038/sc.2015.162] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/02/2015] [Revised: 07/18/2015] [Accepted: 07/31/2015] [Indexed: 11/09/2022]
Abstract
STUDY DESIGN Experimental study. OBJECTIVE Several neuro-degenerative disorders such as Alzheimer's dementia, Parkinson's disease and amyotrophic lateral sclerosis (ALS) are associated with genetic mutations, and replacing or disrupting defective sequences might offer therapeutic benefits. Single gene delivery has so far failed to achieve significant clinical improvements in humans, leading to the advent of co-expression of multiple therapeutic genes. Co-transfection using two or more individual constructs might inadvertently result in disproportionate delivery of the products into the cells. To prevent this, and in order to rule out interference among the many promoters with varying strength, expressing multiple proteins in equimolar amounts can be achieved by linking open reading frames under the control of only one promoter. SETTING Kazan, Russian Federation. METHODS Here we describe a strategy for adeno-viral co-expression of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) interconnected through picorna-viral 2A-amino-acid sequence in transfected human umbilical cord blood mono-nuclear cells (hUCB-MCs). RESULTS Presence of both growth factors, as well as absence of immune response to 2A-antigen, was demonstrated after 28-52 days. Following injection of hUCB-MCs into ALS transgenic mice, co-expression of VEGF and FGF2, as well as viable xeno-transplanted cells, were observed in the spinal cord after 1 month. CONCLUSION These results suggest that recombinant adeno-virus containing 2A-sequences could serve as a promising alternative in regenerative medicine for the delivery of therapeutic molecules to treat neurodegenerative diseases, such as ALS.
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Affiliation(s)
- E E Garanina
- Kazan Federal University, Kazan, Russian Federation
| | | | | | - A P Kiyasov
- Kazan Federal University, Kazan, Russian Federation
| | - L M Lima
- Universidade Federal de Viçosa, Viçosa, Brazil
| | - H J Reis
- Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
| | - A Palotás
- Kazan Federal University, Kazan, Russian Federation.,Asklepios-Med (Private Medical Practice and Research Center), Szeged, Hungary
| | - R R Islamov
- Kazan State Medical University, Kazan, Russian Federation
| | - A A Rizvanov
- Kazan Federal University, Kazan, Russian Federation
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30
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Kalaska B, Kaminski K, Sokolowska E, Czaplicki D, Kujdowicz M, Stalinska K, Bereta J, Szczubialka K, Pawlak D, Nowakowska M, Mogielnicki A. Nonclinical evaluation of novel cationically modified polysaccharide antidotes for unfractionated heparin. PLoS One 2015; 10:e0119486. [PMID: 25781030 PMCID: PMC4362941 DOI: 10.1371/journal.pone.0119486] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/16/2014] [Accepted: 01/13/2015] [Indexed: 01/25/2023] Open
Abstract
Protamine, the only registered antidote of unfractionated heparin (UFH), may produce a number of adverse effects, such as anaphylactic shock or serious hypotension. We aimed to develop an alternative UFH antidote as efficient as protamine, but safer and easier to produce. As a starting material, we have chosen generally non-toxic, biocompatible, widely available, inexpensive, and easy to functionalize polysaccharides. Our approach was to synthesize, purify and characterize cationic derivatives of dextran, hydroxypropylcellulose, pullulan and γ-cyclodextrin, then to screen them for potential heparin-reversal activity using an in vitro assay and finally examine efficacy and safety of the most active polymers in Wistar rat and BALB/c mouse models of experimentally induced arterial and venous thrombosis. Efficacy studies included the measurement of thrombus formation, activated partial thromboplastin time, bleeding time, and anti-factor Xa activity; safety studies included the measurement of hemodynamic, hematologic and immunologic parameters. Linear, high molecular weight dextran substituted with glycidyltrimethylammonium chloride groups at a ratio of 0.65 per glucose unit (Dex40-GTMAC3) is the most potent and the safest UFH inhibitor showing activity comparable to that of protamine while possessing lower immunogenicity. Cationic polysaccharides of various structures neutralize UFH. Dex40-GTMAC3 is a promising and potentially better UFH antidote than protamine.
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Affiliation(s)
- Bartlomiej Kalaska
- Department of Pharmacodynamics, Medical University of Bialystok, Bialystok, Poland
| | - Kamil Kaminski
- Faculty of Chemistry, Jagiellonian University, Krakow, Poland
| | - Emilia Sokolowska
- Department of Pharmacodynamics, Medical University of Bialystok, Bialystok, Poland
| | - Dominik Czaplicki
- Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland
| | | | - Krystyna Stalinska
- Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland
| | - Joanna Bereta
- Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland
| | | | - Dariusz Pawlak
- Department of Pharmacodynamics, Medical University of Bialystok, Bialystok, Poland
| | | | - Andrzej Mogielnicki
- Department of Pharmacodynamics, Medical University of Bialystok, Bialystok, Poland
- * E-mail:
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Gupta K, Singh S, Garg KN. Gene therapy in dentistry: Tool of genetic engineering. Revisited. Arch Oral Biol 2015; 60:439-46. [DOI: 10.1016/j.archoralbio.2014.11.018] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/02/2014] [Revised: 11/26/2014] [Accepted: 11/27/2014] [Indexed: 01/17/2023]
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Song Y, Zhang T, Song X, Zhang L, Zhang C, Xing J, Liang XJ. Polycations with excellent gene transfection ability based on PVP-g-PDMAEMA with random coil and micelle structures as non-viral gene vectors. J Mater Chem B 2015; 3:911-918. [DOI: 10.1039/c4tb01754d] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
PVP-g-PDMAEMA formed random coils in water and PVP-g-PDMAEMA-b-PMMA self-assembled into spherical core–shell micelles. Both displayed excellent pDNA compacting abilities at an extremely low N/P ratio, with PVP-g-PDMAEMA-b-PMMA also showing excellent gear transfection efficiency.
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Affiliation(s)
- Yuhua Song
- Department of Polymer Science and Engineering
- School of Chemical Engineering and Technology
- Tianjin University
- Tianjin
- China
| | - Tingbin Zhang
- Department of Polymer Science and Engineering
- School of Chemical Engineering and Technology
- Tianjin University
- Tianjin
- China
| | - Xiaoyan Song
- College of Material Science and Engineering
- Tianjin Polytechnic University
- Tianjin
- China
| | - Ling Zhang
- Department of Polymer Science and Engineering
- School of Chemical Engineering and Technology
- Tianjin University
- Tianjin
- China
| | - Chunqiu Zhang
- CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety
- National Center for Nanoscience and Technology of China
- Beijing
- China
| | - Jinfeng Xing
- Department of Polymer Science and Engineering
- School of Chemical Engineering and Technology
- Tianjin University
- Tianjin
- China
| | - Xing-Jie Liang
- CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety
- National Center for Nanoscience and Technology of China
- Beijing
- China
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Kumar S, Garg P, Pandey S, Kumari M, Hoon S, Jang KJ, Kapavarapu R, Choung PH, Sobral AJFN, Hoon Chung J. Enhanced chitosan–DNA interaction by 2-acrylamido-2-methylpropane coupling for an efficient transfection in cancer cells. J Mater Chem B 2015; 3:3465-3475. [DOI: 10.1039/c4tb02070g] [Citation(s) in RCA: 43] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/12/2023]
Abstract
Chitosan modification by AMP increases its interaction with DNA leading to a higher DNA delivery in to the cancer cell.
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Affiliation(s)
- Santosh Kumar
- Department of Chemistry
- Faculty of Science & Technology
- University of Coimbra
- Coimbra 3004-535
- Portugal
| | - Pankaj Garg
- Department of Biosystems and Biomaterials Science and Engineering
- Seoul National University
- Seoul 151-921
- Republic of Korea
| | - Shambhavi Pandey
- Department of Biosystems and Biomaterials Science and Engineering
- Seoul National University
- Seoul 151-921
- Republic of Korea
| | - Mridula Kumari
- University Department of Biotechnology
- Vinoba Bhave University
- Hazaribagh-825-301
- India
| | - Seonwoo Hoon
- Department of Biosystems and Biomaterials Science and Engineering
- Seoul National University
- Seoul 151-921
- Republic of Korea
| | - Kyoung-Je Jang
- Department of Biosystems and Biomaterials Science and Engineering
- Seoul National University
- Seoul 151-921
- Republic of Korea
| | - Ravikumar Kapavarapu
- Centre for Neuroscience and Cell Biology
- Institute for Interdisciplinary Research
- University of Coimbra
- Coimbra
- Portugal
| | - Pill-Hoon Choung
- Department of Oral and Maxillofacial Surgery and Dental Research Institute
- School of Dentistry
- Seoul National University
- Seoul 110-774
- Republic of Korea
| | - Abilio J. F. N. Sobral
- Department of Chemistry
- Faculty of Science & Technology
- University of Coimbra
- Coimbra 3004-535
- Portugal
| | - Jong Hoon Chung
- Department of Biosystems and Biomaterials Science and Engineering
- Seoul National University
- Seoul 151-921
- Republic of Korea
- Research Institute for Agriculture and Life Sciences
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34
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Dey D, Maiti C, Maiti S, Dhara D. Interaction between calf thymus DNA and cationic bottle-brush copolymers: equilibrium and stopped-flow kinetic studies. Phys Chem Chem Phys 2015; 17:2366-77. [DOI: 10.1039/c4cp03309d] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Polyplex formation betweenctDNA and PEGylated cationic bottle-brush copolymers: PEG influences the DNA compaction behavior and the kinetics of polyplex formation.
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Affiliation(s)
- Debabrata Dey
- Department of Chemistry
- Indian Institute of Technology Kharagpur
- India
| | - Chiranjit Maiti
- Department of Chemistry
- Indian Institute of Technology Kharagpur
- India
| | - Souvik Maiti
- Proteomics and Structural Biology Unit
- Institute of Genomics and Integrative Biology
- CSIR
- Delhi 110007
- India
| | - Dibakar Dhara
- Department of Chemistry
- Indian Institute of Technology Kharagpur
- India
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Andrade I, Sousa ABDS, da Silva GG. New therapeutic modalities to modulate orthodontic tooth movement. Dental Press J Orthod 2014; 19:123-33. [PMID: 25628089 PMCID: PMC4347420 DOI: 10.1590/2176-9451.19.6.123-133.sar] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2014] [Accepted: 10/14/2014] [Indexed: 12/27/2022] Open
Abstract
Modulation of orthodontic tooth movement (OTM) is desirable not only to patients because it shortens treatment time, but also to orthodontists, since treatment duration is associated with increased risk of gingival inflammation, decalcification, dental caries, and root resorption. The increased focus on the biological basis of tooth movement has rendered Orthodontics a more comprehensive specialty that incorporates facets of all fields of medicine. Current knowledge raises the possibility of using new therapeutic modalities for modulation of OTM, such as corticotomy, laser therapy, vibration (low-intensity pulsed ultrasound), local injections of biomodulators and gene therapy; with the latter being applicable in the near future. They are intended to enhance or inhibit recruitment, differentiation and/or activation of bone cells, accelerate or reduce OTM, increase stability of orthodontic results, as well as assist with the prevention of root resorption. This article summarizes recent studies on each one of these therapeutic modalities, provides readers with information about how they affect OTM and points out future clinical perspectives.
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Affiliation(s)
- Ildeu Andrade
- School of Dentistry, Catholic University of Minas Gerais
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36
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Jing XX, Liu J, Yang BA, Fu SQ, Wu TN, Wang DL. EGFP gene transfection into the synovial joint tissues of rats with rheumatoid arthritis by ultrasound-mediated microbubble destruction. Exp Ther Med 2014; 7:1396-1402. [PMID: 24940446 PMCID: PMC3991517 DOI: 10.3892/etm.2014.1579] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2013] [Accepted: 02/12/2014] [Indexed: 12/29/2022] Open
Abstract
The aim of the present study was to explore the feasibility of enhancing green fluorescent protein (EGFP) gene transfection into the synovial joint tissues of rats with rheumatoid arthritis (RA) by ultrasound-mediated microbubble destruction. An optimal SonoVue dose was determined using 40 normal rats categorized into five groups according to the various doses of microbubbles used. At 1 week after ultrasound irradiation, the rats were sacrificed. Damage to the joint synovial tissues was observed with hematoxylin and eosin histopathological staining under a microscope. A further 44 normal rats were used to establish a rat model of RA, and were then categorized into four groups: EGFP, ultrasound + EGFP, microbubbles + EGFP and ultrasound + microbubbles + EGFP. The last group was irradiated with ultrasound for 10 min following the injection of 300 μl SonoVue and 10 μg EGFP into the joint cavity. Rats were sacrificed after 3 days and synovial tissue was collected from the knee joints for observation of EGFP with fluorescence microscopy and analysis by quantitative polymerase chain reaction. EGFP expression was observed in the synovial tissues of all groups. However, high EGFP expression levels were observed in the ultrasound + microbubbles + EGFP group. No statistically significant differences (P>0.05) were observed in the EGFP expression levels between the EGFP, ultrasound + EGFP and microbubbles + EGFP groups. However, EGFP expression levels in the EGFP, ultrasound + EGFP and microbubbles + EGFP groups significantly differed (P<0.05) from that in the ultrasound + microbubbles + EGFP group. Therefore, ultrasound-mediated microbubble destruction improved EGFP transfection efficiency into the joint synovial tissues of rats with RA.
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Affiliation(s)
- Xiang-Xiang Jing
- Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
| | - Jie Liu
- Department of Emergency Orthopedics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
| | - Bing-Ang Yang
- Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
| | - Shao-Qing Fu
- Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
| | - Tang-Na Wu
- Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
| | - Dong-Lin Wang
- Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China
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Dey D, Kumar S, Banerjee R, Maiti S, Dhara D. Polyplex Formation between PEGylated Linear Cationic Block Copolymers and DNA: Equilibrium and Kinetic Studies. J Phys Chem B 2014; 118:7012-25. [DOI: 10.1021/jp501234p] [Citation(s) in RCA: 27] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Affiliation(s)
- Debabrata Dey
- Department
of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302, India
| | - Santosh Kumar
- Proteomics
and Structural Biology Unit, Institute of Genomics and Integrative
Biology, CSIR, Mall Road, Delhi 110007, India
| | - Rakesh Banerjee
- Department
of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302, India
| | - Souvik Maiti
- Proteomics
and Structural Biology Unit, Institute of Genomics and Integrative
Biology, CSIR, Mall Road, Delhi 110007, India
| | - Dibakar Dhara
- Department
of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302, India
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38
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Abstract
Oncolytic virotherapy is an emerging technology that uses engineered viruses to treat malignancies. Viruses can be designed with biological specificity to infect cancerous cells preferentially, and to replicate in these cells exclusively. Malignant cells may be killed directly by overwhelming viral infection and lysis, which releases additional viral particles to infect neighboring cells and distant metastases. Viral infections may also activate the immune system, unmask stealthy tumor antigens, and aid the immune system to recognize and attack neoplasms. Delivery of live virus particles is potentially complex, and may require the expertise of the interventional community.
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39
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Gold nanoparticles for nucleic acid delivery. Mol Ther 2014; 22:1075-1083. [PMID: 24599278 DOI: 10.1038/mt.2014.30] [Citation(s) in RCA: 350] [Impact Index Per Article: 31.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2013] [Accepted: 02/21/2014] [Indexed: 12/11/2022] Open
Abstract
Gold nanoparticles provide an attractive and applicable scaffold for delivery of nucleic acids. In this review, we focus on the use of covalent and noncovalent gold nanoparticle conjugates for applications in gene delivery and RNA-interference technologies. We also discuss challenges in nucleic acid delivery, including endosomal entrapment/escape and active delivery/presentation of nucleic acids in the cell.
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40
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Gene therapy and DNA delivery systems. Int J Pharm 2013; 459:70-83. [PMID: 24286924 DOI: 10.1016/j.ijpharm.2013.11.041] [Citation(s) in RCA: 305] [Impact Index Per Article: 25.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/24/2013] [Revised: 10/31/2013] [Accepted: 11/19/2013] [Indexed: 12/29/2022]
Abstract
Gene therapy is a promising new technique for treating many serious incurable diseases, such as cancer and genetic disorders. The main problem limiting the application of this strategy in vivo is the difficulty of transporting large, fragile and negatively charged molecules like DNA into the nucleus of the cell without degradation. The key to success of gene therapy is to create safe and efficient gene delivery vehicles. Ideally, the vehicle must be able to remain in the bloodstream for a long time and avoid uptake by the mononuclear phagocyte system, in order to ensure its arrival at the desired targets. Moreover, this carrier must also be able to transport the DNA efficiently into the cell cytoplasm, avoiding lysosomal degradation. Viral vehicles are the most commonly used carriers for delivering DNA and have long been used for their high efficiency. However, these vehicles can trigger dangerous immunological responses. Scientists need to find safer and cheaper alternatives. Consequently, the non-viral carriers are being prepared and developed until techniques for encapsulating DNA can be found. This review highlights gene therapy as a new promising technique used to treat many incurable diseases and the different strategies used to transfer DNA, taking into account that introducing DNA into the cell nucleus without degradation is essential for the success of this therapeutic technique.
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41
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Dey D, Kumar S, Maiti S, Dhara D. Stopped-flow kinetic studies of poly(amidoamine) dendrimer-calf thymus DNA to form dendriplexes. J Phys Chem B 2013; 117:13767-74. [PMID: 24087941 DOI: 10.1021/jp406973t] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Poly(amidoamine) (PAMAM) dendrimers are known to be highly efficient nonviral carriers in gene delivery. Dendrimer-mediated transfection is known to be a function of the dendrimer to DNA charge ratio as well as the size of the dendrimer. In the present study, the binding kinetics of four PAMAM dendrimers (G1, G2, G3, and G4) with calf thymus DNA (CT-DNA) has been studied using stopped-flow fluorescence spectroscopy. The effect of dendrimer-to-DNA charge ratio and dendrimer generation on the binding kinetics was investigated. In most cases, the results of dendrimer-CT-DNA binding can be explained by a two-step reaction mechanism: a rapid electrostatic binding between the dendrimer and DNA, followed by a conformational change of the dendrimer-DNA complex that ultimately leads to DNA condensation. It was observed that the charge ratio on the dendrimer and the DNA phosphate groups, as well as the dendrimer generation (size), has a marked effect on the kinetics of binding between the DNA and the dendrimers. The rate constant (k'1) of the first step was much higher compared to that of the second step (k'2), and both were found to increase with an increase in dendrimer concentration. Among the four generations of dendrimers, G4 exhibited significantly faster binding kinetics compared to the three smaller generation dendrimers.
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Affiliation(s)
- Debabrata Dey
- Department of Chemistry, Indian Institute of Technology Kharagpur , West Bengal 721302 India
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42
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Garlick JA, Elias J, Taichman LB. Histochemical Detection of a Gene Transferred by Retrovirus Vector in Cultured Human Keratinocytes. J Histotechnol 2013. [DOI: 10.1179/his.1992.15.4.289] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/31/2022]
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43
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Zhang Y, Aigner A, Agarwal S. Degradable and Biocompatible Poly(N
,N
-dimethylaminoethyl Methacrylate-co
-caprolactone)s as DNA Transfection Agents. Macromol Biosci 2013; 13:1267-75. [DOI: 10.1002/mabi.201300043] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2013] [Revised: 03/26/2013] [Indexed: 11/08/2022]
Affiliation(s)
- Yi Zhang
- Fb. Chemie; Philipps-Universität Marburg; Hans-Meerwein Strasse 35032 Marburg Germany
| | - Achim Aigner
- Rudolf-Boehm-Institute for Pharmacology and Toxicology; Clinical Pharmacology; University of Leipzig; Haertelstrasse 16-18 D-04107 Leipzig Germany
| | - Seema Agarwal
- Macromolecular Chemistry II; Bayreuth Center for Interfaces and Colloids; University of Bayreuth; 95447 Bayreuth Germany
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44
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Zhi D, Zhang S, Cui S, Zhao Y, Wang Y, Zhao D. The Headgroup Evolution of Cationic Lipids for Gene Delivery. Bioconjug Chem 2013; 24:487-519. [DOI: 10.1021/bc300381s] [Citation(s) in RCA: 188] [Impact Index Per Article: 15.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023]
Affiliation(s)
- Defu Zhi
- State Key Laboratory of Fine
Chemicals, Dalian University of Technology, Dalian 116012, China
- State Ethnic Affairs Commission-Ministry
of Education Key Laboratory of Biotechnology and Bio-resources Utilization, Dalian Nationalities University, Dalian 116600, China
| | - Shubiao Zhang
- State Ethnic Affairs Commission-Ministry
of Education Key Laboratory of Biotechnology and Bio-resources Utilization, Dalian Nationalities University, Dalian 116600, China
| | - Shaohui Cui
- State Ethnic Affairs Commission-Ministry
of Education Key Laboratory of Biotechnology and Bio-resources Utilization, Dalian Nationalities University, Dalian 116600, China
| | - Yinan Zhao
- State Ethnic Affairs Commission-Ministry
of Education Key Laboratory of Biotechnology and Bio-resources Utilization, Dalian Nationalities University, Dalian 116600, China
| | | | - Defeng Zhao
- State Key Laboratory of Fine
Chemicals, Dalian University of Technology, Dalian 116012, China
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45
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Sarkar K, Kundu P. Preparation of low molecular weight N-maleated chitosan-graft-PAMAM copolymer for enhanced DNA complexation. Int J Biol Macromol 2012; 51:859-67. [DOI: 10.1016/j.ijbiomac.2012.06.038] [Citation(s) in RCA: 40] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/10/2012] [Revised: 06/22/2012] [Accepted: 06/26/2012] [Indexed: 01/29/2023]
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46
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Feng X, Lv F, Liu L, Yang Q, Wang S, Bazan GC. A highly emissive conjugated polyelectrolyte vector for gene delivery and transfection. ADVANCED MATERIALS (DEERFIELD BEACH, FLA.) 2012; 24:5428-5432. [PMID: 22887832 DOI: 10.1002/adma.201202145] [Citation(s) in RCA: 45] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/29/2012] [Indexed: 06/01/2023]
Abstract
An intrinsically fluorescent cationic polyfluorene (CCP) has been designed, synthesized, characterized, and examined as a plasmid DNA (pDNA) delivery vector. This material facilitates nucleic acid binding, encapsulation and efficient cellular uptake. CCP can effectively protect pDNA against nuclease degradation, which is necessary for gene carriers. Green fluorescent protein (GFP) expression experiments reveal that CCP can achieve efficient delivery and transfection of pDNA encoding GFP gene with 92% efficiency, which surpasses that of commercial transfection agents, lipofectamine 2000 (Lipo) and polyethylenimine (PEI). CCP is also highly fluorescent, with 43% quantum yield in water, and exhibits excellent photostability, which allows for real-time tracking the location of gene delivery and transfection. These features and capabilities represent a major step toward designing and applying conjugated polymers that function in both imaging and therapeutic applications.
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Affiliation(s)
- Xuli Feng
- Beijing National Laboratory for Molecular Sciences, Key Laboratory of Organic Solids, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, P. R. China
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47
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Chang PL, Hortelano G, Awrey DE, Tse M. Growth of recombinant fibroblasts in alginate microcapsules. Biotechnol Bioeng 2012; 43:925-33. [PMID: 18615439 DOI: 10.1002/bit.260431005] [Citation(s) in RCA: 47] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
To develop a novel strategy of nonautologous somatic gene therapy, we now demonstrate the feasibility of culturing genetically modified fibroblasts within an immunoprotective environment and the optimal conditions required for their continued survival in vitro. When mouse Ltk(-) fibroblasts transfected with the human growth hormone gene were enclosed within permselective microcapsules fabricated from alginate-polylysine-alginate, they continued to secrete human growth hormone at the same rates as the nonencapsulated cells. They also continued to proliferate in vitro for at least 1 month even though their viability gradually declined to about 50%. The viability can be improved by controlling for (a) temperature during encapsulation, (b) duration of treatment with polylysine, (c) duration of liquefying the core alginate with sodium citrate, and (d) cell density at the time of encapsulation. The best conditions leading to improved survival and maximum proliferation of cells within the microcapsules were obtained by encapsulating the cells at 4 to 10 degrees C instead of room temperature, coating the microspheres with polylysine for 6 to 10 min instead of 20 min, liquefying the core alginate by treating with citrate for 20 min instead of 6 to 10 min, and using a concentration of 2 x 10(6) cells/mL of alginate for encapsulation. Under such conditions, normally adherent and genetically engineered mouse fibroblasts survived and proliferated optimally within the microcapsule environment. The encapsulated fibroblasts maintained their level of transgene expression while recombinant gene products such as human growth hormone could diffuse through the microcapsule membrane without impediment. The demonstration that genetically modified fibroblasts can survive and continue to deliver recombinant gene products from within these microcapsules and the optimization for their maximal viability and growth within microcapsules should increase the potential for success in using such microencapsulated recombinant cells for somatic gene therapy. (c) 1994 John Wiley & Sons, Inc.
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Affiliation(s)
- P L Chang
- Department of Pediatrics, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5
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48
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Recent advances in the rational design of silica-based nanoparticles for gene therapy. Ther Deliv 2012. [DOI: 10.4155/tde.12.98] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Gene therapy has attracted much attention in modern society and provides a promising approach for treating genetic disorders, diseases and cancers. Safe and effective vectors are vital tools to deliver genetic molecules to cells. This review summarizes recent advances in the rational design of silica-based nanoparticles and their applications in gene therapy. An overview of different types of genetic agents available for gene therapy is provided. The engineering of various silica nanoparticles is described, which can be used as versatile complexation tools for genetic agents and advanced gene therapy. Several challenges are raised and future research directions in the area of gene therapy using silica-based nanoparticles are proposed.
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49
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MSC and Tumors: Homing, Differentiation, and Secretion Influence Therapeutic Potential. ADVANCES IN BIOCHEMICAL ENGINEERING/BIOTECHNOLOGY 2012; 130:209-66. [PMID: 22990585 DOI: 10.1007/10_2012_150] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
: Mesenchymal stromal/stem cells (MSC) are adult multipotent progenitors with fibroblast-like morphology able to differentiate into adipocytic, osteogenic, chondrogenic, and myogenic lineages. Due to these properties, MSC have been studied and introduced as therapeutics in regenerative medicine. Preliminary studies have also shown a possible involvement of MSC as precursors of cellular elements within tumor microenvironments, in particular tumor-associated fibroblasts (TAF). Among a number of different possible origins, TAF may originate from a pool of circulating progenitors from bone marrow or adipose tissue-derived MSC. There is growing evidence to corroborate that cells immunophenotypically defined as MSC are able to reside as TAF influencing the tumor microenvironment in a potentially bi-phasic and obscure manner: either promoting or inhibiting growth depending on tumor context and MSC sources. Here we focus on relationships between the tumor microenvironment, cancer cells, and MSC, analyzing their diverse ability to influence neoplastic development. Associated activities include MSC homing driven by the secretion of various mediators, differentiation towards TAF phenotypes, and reciprocal interactions with the tumor cells. These are reviewed here with the aim of understanding the biological functions of MSC that can be exploited for innovative cancer therapy.
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50
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Sajomsang W, Gonil P, Ruktanonchai UR, Petchsangsai M, Opanasopit P, Puttipipatkhachorn S. Effects of molecular weight and pyridinium moiety on water-soluble chitosan derivatives for mediated gene delivery. Carbohydr Polym 2012; 91:508-17. [PMID: 23121939 DOI: 10.1016/j.carbpol.2012.08.053] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/14/2012] [Revised: 08/16/2012] [Accepted: 08/16/2012] [Indexed: 11/17/2022]
Abstract
The aim of this study is to investigate the effects of molecular weight, the pyridinium/trimethyl ammonium (Py/Tr) ratio, the nitrogen atoms (N) in the methylated N-(3-pyridylmethyl) chitosan chloride (M3-PyMeChC)/the phosphorus atoms (P) in DNA (N/P) ratio, and the physicochemical properties of nanopolyplexes on transfection efficiency. The water-soluble chitosan derivative, M3-PyMeChC, was used as a non-viral vector to deliver pEGFP-C2 into human hepatoma (Huh7) cell lines. The results revealed that higher molecular weight M3-PyMeChC was able to form complexes completely with DNA at lower N/P ratios than that with lower molecular weights, which led to higher transfection efficiency. Moreover, the M3-PyMeChC with higher Py/Tr ratios showed superior transfection efficiency at lower Py/Tr ratios at all N/P ratios studied. The highest transfection efficiency for the nanopolyplexes occurred for a molecular weight of 82kDa at a N/P ratio of 5. The results indicated that the hydrophobic effect of pyridinium moiety could enhance gene transfection efficiency, which can be attributed to the dissociation of DNA from nanopolyplexes. High Py/Tr ratios in nanopolyplexes tended to decrease cytotoxicity due to delocalization of positive charge into a pyridine ring while high N/P ratios and molecular weight increased cytotoxicity. Our results showed that the vector was able to spread the positive charge by delocalizing it into a heterocyclic ring, suggesting to a promising approach to mediate higher levels of gene transfection.
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Affiliation(s)
- Warayuth Sajomsang
- National Nanotechnology Center, National Science and Technology Development Agency, Pathumthani, Thailand.
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