Optimizing Cancer Immunotherapy Safety and Efficacy using Genome Editing

过继转移的淋巴细胞工程识别肿瘤细胞已显示出巨大的前景， 复发性或难治性B细胞恶性肿瘤患者。在这种方法中，患者来源的T细胞被编程为 在体外与对癌症具有亲和力的工程化T细胞受体（TCR）或嵌合抗原受体（汽车）- 或谱系特异性抗原。随后自体再输注工程化T细胞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善工程化T细胞 通过敲除介导移植物抗宿主病（用于同种异体转移）的基因， 参与肿瘤介导的T细胞疗效抑制，或被有效的临床相关的 克隆抗体然而，到目前为止，还没有对基因组编辑效率或 特异性已经在T细胞中进行。我的大部分博士后工作都集中在开发方法， 表征和提高CRISPR-Cas核酸酶的效用和全基因组特异性，使我成为独一无二的 适合解决这个悬而未决的问题。因此，这项建议的主要目的是：1） 表征和优化T细胞中基因组编辑的效率和特异性，2）利用基因组编辑， 改善T细胞工程中涉及的过程，以及3）利用CRISPR-Cas筛选来增强整体 工程化T细胞的功效。拟议的研究将提供相当深入的可行性， 在T细胞中实施基因组编辑策略，作为提高肿瘤杀伤效力、持久性或 制造业与癌症免疫治疗、癌症生物学、基因组等领域相关的重大发现 编辑和基因治疗是可以预期的。能够使成功的科学培训领域 完成这一建议是T细胞生物学和免疫学的知识，在体内癌症的指导 小鼠建模，并拥有实施全基因组CRISPR-Cas筛选的经验。指导阶段 该奖项将得到基因组编辑技术开发领域世界领导者基思·琼博士的支持，并 Marcela Maus博士，癌症免疫治疗专家。Jesus博士，Maus博士，以及几乎所有其他高级/关键 该项目的人员位于马萨诸塞州总医院或大波士顿科学中心 社区专业和职业发展活动将包括指导培训、责任培训、 进行研究，资助，财务和利益冲突等主题。国家和 将继续参加国际会议，传播拟议研究的结果。的 马萨诸塞州总医院和哈佛的学术和专业发展环境 医学院，与顶级导师团队和科学咨询委员会相结合，将提供最好的 在我向独立过渡的过程中，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.