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Genetic targeting of proviral integration in stem cells

干细胞中原病毒整合的基因靶向

基本信息

批准号：
7625013
负责人：
Peter Kurre
金额：
$ 12.91万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-04 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7625013
关键词：
Advisory Committees Affect Area Awareness Biological Models Biology Biometry Cell Line Cells Classification DNA Engraftment Environment Event Exposure to Future Gene Expression Gene Transfer Genes Genetic Genetic Transcription Genomics Goals Gray unit of radiation dose Growth Factor Health Sciences Hematopoiesis Hematopoietic Hematopoietic stem cells Homing In Vitro Insertional Mutagenesis Lentivirus Vector Location Measures Mentors Mentorship Microarray Analysis Modeling Modification Moloney Leukemia Virus Mus Oregon Pattern Population Probability Property Reporting Research Personnel Research Proposals Risk Safety Shapes Site Stem cells Technology Testing Time Training Training Programs Transgenes Tumor Suppressor Genes Universities Work career cellular transduction cytokine gene replacement therapy gene therapy in vivo in vivo Model insight murine retroviral vector preference programs reconstitution response skills stem stem cell biology tool virology

项目摘要

DESCRIPTION (provided by applicant): The aim of this research proposal is to understand how transcriptional activity and its modification can be used to direct HIV-derived lentiviral vector integration in hematopoietic stem cells. Recent awareness of known lentiviral insertion site preferences for transcriptionally active genomic sites and reports of insertional mutagenesis after Moloney murine leukemia virus vector transduction of stem cells have highlighted the need to more fully investigate the biology of proviral integration. I herein propose to develop in vitro and in vivo model systems to explore strategies that should ultimately minimize insertion related risks of HIV-lentiviral gene replacement therapy. We hypothesize that systematic modifications in the transcriptional profile of hematopoietic target cells may serve to "attract" insertion events to specific genomic locations while minimizing mutagenic potential and transcriptional interference at others, such as oncogenes and tumor suppressor loci. Critically, the proposed in vivo studies will aim to reconcile the competing requirements for transcriptional profiling and efficient lentiviral gene transfer with the retention of stem cell properties, including target cell engraftment and long-term repopulation. These studies will provide mechanistic insight into how transcription, be it a surrogate, or a causally related event, affects proviral insertion, and will inform future studies for genomic targeting. The proposed mentored training program will help me acquire additional skills and gain understanding through the mentorship by Dr. David Kabat and Dr. Markus Grompe, and their scientific expertise in virology and stem cell biology, respectively. The proposal is further augmented by input and guidance from advisors/collaborators in the areas of hematopoiesis, DNA array technology, and biostatistics, as well as course work. A career advisory committee will help direct my progress to independence and career advancement. The comprehensive training environment at Oregon Health & Science University (OHSU) and the substantial institutional commitment to my career will allow me to accomplish the goals of this proposal.

描述（由申请人提供）：本研究提案的目的是了解转录活性及其修饰如何用于指导HIV衍生的慢病毒载体整合到造血干细胞中。最近认识到已知的慢病毒插入位点的转录活性基因组位点的偏好和莫洛尼鼠白血病病毒载体转导干细胞后的插入诱变的报告，突出了需要更充分地研究前病毒整合的生物学。我在此建议开发体外和体内模型系统，以探索策略，最终应尽量减少插入相关的风险艾滋病毒-慢病毒基因替代治疗。我们假设造血靶细胞转录谱的系统性修饰可能有助于将插入事件“吸引”到特定的基因组位置，同时最大限度地减少致突变潜力和对其他基因（如癌基因和肿瘤抑制基因座）的转录干扰。至关重要的是，拟议的体内研究将旨在协调转录谱分析和有效的慢病毒基因转移的竞争要求与干细胞特性的保留，包括靶细胞植入和长期再增殖。这些研究将为转录如何影响前病毒插入提供机制性见解，无论是替代物还是因果相关事件，并将为基因组靶向的未来研究提供信息。拟议的指导培训计划将帮助我获得额外的技能，并通过大卫卡巴特博士和马库斯Grompe博士的指导，以及他们在病毒学和干细胞生物学方面的科学专业知识，分别获得理解。以下领域的顾问/合作者的投入和指导进一步充实了该提案：造血，DNA阵列技术和生物统计学，以及课程工作。职业咨询委员会将帮助指导我走向独立和职业发展。俄勒冈州健康与科学大学（OHSU）的综合培训环境和对我职业生涯的实质性机构承诺将使我能够实现本提案的目标。