Non-viral genome, epigenome, and transcriptome engineering for clinical CAR-T cell manufacturing

项目总结/摘要 这个应用程序的目标是训练布赖恩博士害羞，在加州圣大学的内科医生科学家 弗朗西斯科，与必要的技能，成为一个独立资助的调查研究，发展， 制造工程细胞疗法。这项研究计划将评估基于CRISPR的非病毒 用于在原代人T细胞中靶向基因组、表观基因组和转录组工程化的方法。这将是 应用于理解DNA修复途径的选择和DNA毒性反应，以提高位点特异性 使用大型治疗性构建体如嵌合抗原受体（CAR）的"敲入"策略， 和有效的方法进行多重T细胞修饰，并建立良好的生产规范（GMP） 用于临床实施该多样化工具集的兼容过程。这项研究是伴随着一个 职业发展和培训计划，将建立在博士的输血医学临床培训， 细胞治疗，以及他在临床前开发和制造实验性 细胞疗法培训将侧重于1）新的细胞工程工具的实施，2）下一代 测序（NGS）技术和大型数据集的分析，3）GMP生产和临床前 开发，4）临床试验设计和标准护理细胞疗法，以及5）实验室管理， 授予写作，演讲和研究策略技能。该职业发展和培训计划将包括 结合正式课程，指导实践培训;会议，会议和研讨会出席; 和指导的共同导师，顾问和合作者的特殊团队。总的来说，这个奖项将 帮助Shy博士为独立的研究生涯做好准备，专注于开发改进的细胞疗法， 治疗癌症和其他人类疾病。

PROJECT SUMMARY / ABSTRACT The goal of this application is to train Dr. Brian Shy, a physician scientist at the University of California San Francisco, with the skills necessary to become an independently-funded investigator studying, developing, and manufacturing engineered cellular therapies. This research proposal will evaluate CRISPR-based non-viral methods for targeted genome, epigenome, and transcriptome engineering in primary human T cells. This will be applied to understand DNA repair pathway choice and DNA toxicity responses in order to improve site-specific “knock-in” strategies with large therapeutic constructs such as chimeric antigen receptors (CARs), develop safe and efficient methods for multiplexed T cell modifications, and establish Good Manufacturing Practice (GMP) compatible processes for clinical implementation of this diverse toolset. This research is accompanied by a career development and training plan that will build on Dr. Shy's clinical training in Transfusion Medicine and Cellular Therapy, and his prior expertise in the pre-clinical development and manufacturing of experimental cellular therapies. Training will focus on 1) implementation of novel cellular engineering tools, 2) next generation sequencing (NGS) technologies and analysis of large datasets, 3) GMP manufacturing and pre-clinical development, 4) clinical trial design and standard-of-care cellular therapies, and 5) laboratory management, grant writing, presentation, and research strategy skills. This career development and training plan will include a combination of formal coursework, mentored practical training; conference, meeting, and workshop attendance; and guidance from an exceptional team of co-mentors, advisors, and collaborators. Altogether, this award will help prepare Dr. Shy for an independent research career focused on developing improved cellular therapies for treatment of cancer and other human diseases.