Optimizing Cancer Immunotherapy Safety and Efficacy using Genome Editing

为识别肿瘤细胞而设计的淋巴细胞的过继转移在 复发或难治性B细胞恶性肿瘤患者。在这种方法中，患者来源的T细胞被编程 在体外使用与癌症有亲和力的工程化T细胞受体(TCR)或嵌合抗原受体(CAR)- 或血统特异性抗原。随后工程T细胞的自体回输使肿瘤靶向成为可能 和根除。最近，已经提出了基因组编辑方法来改进工程化T细胞 通过敲除介导移植物抗宿主病(用于同种异体移植)的基因来发挥作用 参与肿瘤介导的T细胞有效性抑制，或被有效的临床相关识别 单克隆抗体。然而，到目前为止，还没有关于基因组编辑效率或 在T细胞中已经进行了特异性研究。我的大部分博士后工作都集中在开发方法上 表征和改进CRISPR-CAS核酸酶的实用性和全基因组特异性，使我成为独一无二的 适合于解决这一悬而未决的问题。因此，这项建议的主要目的是：1) 表征和优化T细胞基因组编辑的效率和特异性，2)利用基因组编辑来 改进T细胞工程涉及的过程，以及3)利用CRISPR-CAS筛查来增强整体 基因工程T细胞的功效。拟议的研究将提供相当大的洞察力，以了解 在T细胞中实施基因组编辑策略作为提高肿瘤杀伤效率、持久性或 制造业。与癌症免疫治疗、癌症生物学、基因组学领域相关的重大发现 编辑和基因治疗有望实现。其他科学培训领域，将使成功 完成这项建议是T细胞生物学和免疫学的知识，体内癌症的指导 在小鼠中进行建模，并体验实施全基因组CRISPR-CAS筛查。的指导阶段 该奖项将由基因组编辑技术开发的世界领先者Keith Joung博士支持，以及 由癌症免疫治疗专家Marcela Maus博士撰写。荣格博士、莫斯博士和几乎所有其他高级/关键人物 该项目的工作人员位于马萨诸塞州总医院或大波士顿科学医院 社区。专业和职业发展活动将包括指导培训，负责 研究的开展、资助金、财务和利益冲突等主题。国家和 将继续参加国际会议，以传播拟议研究的结果。这个 麻省总医院和哈佛大学的学术和职业发展环境 医学院，结合一支顶级的指导团队和科学咨询委员会，将提供最好的 当我过渡到独立的时候，我有机会接受进一步的培训。

The adoptive transfer of lymphocytes engineered to recognize tumor cells has shown tremendous promise in patients with relapsed or refractory B cell malignancies. In this approach, patient derived T cells are programmed in vitro with engineered T cell receptors (TCRs) or chimeric antigen receptors (CARs) that have affinity for cancer- or lineage-specific antigens. Subsequent autologous reinfusion of the engineered T cells enables tumor targeting and eradication. More recently, genome editing approaches have been proposed to improve engineered T cell performance by knocking out genes that mediate graft-versus-host disease (for allogeneic transfer), that are involved in tumor-mediated suppression of T cell efficacy, or that are recognized by potent clinically relevant monoclonal antibodies. However, to this point no robust characterization of genome editing efficiency or specificity has been performed in T cells. Much of my postdoctoral work has focused on developing methods to characterize and improve the utility and genome-wide specificity of CRISPR-Cas nucleases, making me uniquely suited to address this largely outstanding question. The primary aims of this proposal are therefore: 1) to characterize and optimize the efficiency and specificity of genome editing in T cells, 2) utilize genome editing to improve processes involved in T cell engineering, and 3) leverage CRISPR-Cas screens to enhance the overall efficacy of engineered T cells. The proposed research will provide considerable insight into the feasibility of implementing genome editing strategies in T cells as a means to improve tumor killing efficacy, persistence, or manufacturing. Significant findings relevant to the fields of cancer immunotherapy, cancer biology, genome editing, and gene therapy are expected. Areas of additional scientific training that will enable successful completion of this proposal are knowledge of T cell biology and immunology, mentorship on in vivo cancer modeling in mice, and experience implementing genome-wide CRISPR-Cas screens. The mentored phase of the award will be supported by Dr. Keith Joung, a world-leader in genome editing technology development, and by Dr. Marcela Maus, an expert in cancer immunotherapy. Dr. Joung, Dr. Maus, and nearly all other senior/key personnel of this project are located at the Massachusetts General Hospital or in the greater Boston scientific community. Professional and career development activities will include training in mentorship, responsible conduct of research, grantsmanship, finance, and conflict of interest among other topics. National and international meetings will continue to be attended to disseminate findings from the proposed research. The academic and professional development environment at the Massachusetts General Hospital and Harvard Medical School, combined with a top level mentorship team and scientific advisory committee, will offer the best opportunity for further training as I transition to independence.