GENE TRANSFER and SOMATIC CELL ENGINEERING

基因转移和体细胞工程

基本信息

批准号：
7671821
负责人：
MICHEL SADELAIN
金额：
$ 21.67万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-31 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7671821
关键词：
Archives CD34 gene Cancer Patient Cell Line Cell Therapy Cell physiology Cells Clinical Clinical Research Clinical Trials Clone Cells Collaborations Cultured Cells Dendritic Cells Detection Development Diagnostic Ensure Environment Flow Cytometry Gene Expression Gene Transfer Generations Genetic Grant Guanosine Monophosphate HIV Hematopoietic Hematopoietic stem cells Immunization Immunotherapy Individual Laboratories Lentivirus Vector Malignant Neoplasms Measures Memorial Sloan-Kettering Cancer Center Modification Molecular Molecular Medicine Murine leukemia virus Nature Numbers Patients Pharmacogenomics Phase Plasmids Play Polymerase Chain Reaction Population Process Production Protocols documentation Reagent Recombinants Research Research Personnel Retroviral Vector Retroviridae Role Services Site Somatic Cell Southern Blotting Specimen System T-Lymphocyte Technology Testing Therapeutic Time Tissues Titrations Training Translations Vaccines Viral base cell bank cell type cellular engineering cellular transduction cost design genetically modified cells novel therapeutics plasmid DNA pre-clinical preclinical study prevent repository small hairpin RNA transgene expression vector

项目摘要

The Gene Transfer and Somatic Cell Engineering Core (GTS) supports the preclinical translation and clinical implementation of gene transfer studies at MSKCC. The supported projects are highly dependent on achieving efficient gene transfer in primary cells, including hematopoietic progenitor cells, T lymphocytes and dendritic cells. In the upcoming grant cycle, the GTS will mainly focus on the development and optimization of clinical cell engineering processes and on the implementation of clinical trials. The specific aims of the GTS are to carry out and/or coordinate: 1. Expansion and transduction of patient cells in semi-closed systems in collaboration with the investigators for clinical trials utilizing genetically modified cells; 2. Generation and characterization of high-titer producer cell clones, master cell banks (MCB) for clinical studies; 3. Production and titration of 5 to 15 liter batches of clinical viral stocks in semi-closed systems; 4. Production and biosafety testing of clinical grade plasmid DMA vaccine for immunization; 5. Detection of replication-competent retrovirus and other biosafety testing in cultured packaging cell clones (MCB), viral stocks and clinical specimen 6. Detection of oncoretroviral vector integration sites by LM-PCR in patient cells; 7. Cell banking, storage of viral stocks, plasmid DMA vaccine and clinical specimens. In addition, the GTS provides essential advisory and training functions for the generation of research grade reagents at the Center. Investigators are thus advised or trained on 1. How to optimize the transduction of various cell types; 2. How to construct recombinant gamma-retroviral and lentiviral vectors, plasmid DMA vectors, and shRNA encoding retroviral vectors; 3. What packaging cell lines to use; 4. How to transfect vector DMA in packaging cells and select producer cell lines; 5. What tests to perform to analyze gene expression; 6. How to titrate cell-free retroviral stocks by flow cytometry, Southern blot or real time PCR analysis. The GTS is thus a repository for numerous reagents and protocols that are made available to investigators at MSKCC. The centralization of cell transduction, vector production and plasmid DMA manufacturing in the GTS decreases the cost of clinical development, ensures high quality and consistency of molecular and cellular processes, and their availability to all investigators at the Center.

基因转移和体细胞工程核心 (GTS) 支持临床前翻译和 MSKCC 基因转移研究的临床实施。所支持的项目高度依赖在原代细胞（包括造血祖细胞、T淋巴细胞和树突状细胞。在即将到来的资助周期中，GTS将主要关注开发和优化临床细胞工程过程和临床试验的实施。 GTS 的具体目标是执行和/或协调： 1. 患者的扩增和转导与研究人员合作，利用半封闭系统中的细胞进行临床试验修饰细胞； 2. 高滴度生产细胞克隆、主细胞库 (MCB) 的生成和表征用于临床研究； 3. 半封闭式5至15升批次临床病毒储备液的生产和滴定系统； 4、免疫用临床级质粒DMA疫苗的生产及生物安全性检测； 5. 在培养的包装细胞克隆中检测具有复制能力的逆转录病毒和其他生物安全测试 (MCB)、病毒储液和临床标本 6. 通过 LM-PCR 检测肿瘤逆转录病毒载体整合位点患者细胞； 7. 细胞库、病毒库存、质粒 DMA 疫苗和临床标本的储存。在此外，GTS 还为研究级人才的培养提供必要的咨询和培训功能试剂在中心。因此，研究人员在以下方面得到建议或培训： 1. 如何优化转导各种细胞类型； 2. 如何构建重组γ-逆转录病毒和慢病毒载体、质粒DMA 载体和编码逆转录病毒载体的shRNA； 3. 使用什么包装细胞系； 4. 如何转染包装细胞和选择生产细胞系中的载体 DMA； 5. 进行哪些测试来分析基因表达; 6. 如何通过流式细胞术、Southern blot 或实时 PCR 滴定无细胞逆转录病毒原液分析。因此，GTS 是众多试剂和方案的存储库，可用于 MSKCC 的调查员。细胞转导、载体生产和质粒 DMA 制造集中在 GTS 中降低临床开发成本，确保分子和细胞的高质量和一致性流程及其对中心所有调查人员的可用性。