Resistance To Targeted Immunotherapies:CART19 as a Paradigm

项目总结/摘要 嵌合抗原受体（CAR）T细胞为癌症的靶向免疫治疗开辟了新时代。的 申请人的导师Carl June博士被公认为基因工程T细胞领域的世界领导者。 血液恶性肿瘤的治疗。尽管抗CD 19 CAR T细胞（CART 19）产生了前所未有的 在B细胞白血病和淋巴瘤患者中，复发是治疗的主要原因 失败这项建议的长期目标是发展一个独立的研究事业，重点是提高 通过研究和中和肿瘤逃避机制进行靶向免疫治疗。的统一目标， 本申请旨在设计用于治疗和预防肿瘤免疫逃逸的新策略 使用CART 19作为模型，研究导致复发的机制。核心假设是， 机制可能导致逃避免疫攻击;因此，针对该问题的组合方法 从多个侧面将防止逃跑。这项研究的基本原理是识别关键的肿瘤逃逸 机制将使我们能够制定具体的补救措施。这一建议的核心假设将 通过追求两个具体目标进行测试：1）描述白血病复发发生的发病机制 通过研究之前存在的次要CD 19阴性耐药克隆， CART 19治疗和通过使用功能性抗逆转录病毒药物鉴定CD 19阳性复发中的关键耐药途径， 2）设计双特异性嵌合抗原受体T细胞，防止肿瘤逃逸。这 研究将是重要的，因为它将有助于深入（理解复发的机制）， （新的潜在治愈性疗法的）广度到针对癌症的免疫武器库。最后， 这些知识有可能垂直推进CAR重定向T细胞免疫治疗领域， 以及其他靶向免疫疗法。这是一个创新的项目，因为它结合了高吞吐量 筛选技术以及体外和体内功能研究，以研究 免疫治疗后的耐药性，从而产生新的治疗方法。拟议的研究活动 是至关重要的发展申请人作为一个独立资助的科学家，重点是细胞 免疫疗法Ruella博士将接受分子生物学、转化医学和 从他的导师那里学习免疫学，并接受下一代测序、癌症生物学和生物信息学方面的培训 来自宾夕法尼亚大学经验丰富的合作者。因此，在培训结束时 在此期间，申请人将获得一套独特的智力和技术技能，使他能够 从多个角度同时解决靶向免疫疗法的耐药性问题。另外这款 该奖项将支持翻译研究的独特培训经验，并将建立一个学术 发现和开发新的CAR T细胞方法的途径。

PROJECT SUMMARY/ABSTRACT Chimeric-antigen receptor (CAR) T cells have opened a new era for targeted immunotherapy of cancer. The applicant's mentor, Dr. Carl June, is recognized as a world leader in the field of genetically-engineered T cell therapy for hematologic malignancy. Although anti-CD19 CAR T cells (CART19) generate unprecedented responses in patients with B-cell leukemia and lymphoma, relapse represents the major cause of treatment failure. The long term goal of this proposal is to develop an independent research career focused on improving targeted immunotherapy by studying and neutralizing tumor evasion mechanisms. The unifying objective of this application is to design novel strategies for the treatment and prevention of tumor immunoescape based on the mechanisms leading to relapse, using CART19 as a model. The central hypothesis is that multiple mechanisms can lead to escape from immune attack; therefore, combined approaches targeting the problem from multiple sides will prevent escape. The rationale for this research is that identification of key tumor escape mechanisms would allow us to develop specific remedies for these. The central hypothesis of this proposal will be tested by pursuing two specific aims: 1) to characterize the pathogenesis of leukemia relapses occurring after CD19-targeted immunotherapies by studying minor CD19-negative resistant clones present before CART19 treatment and by identifying key resistance pathways in CD19-positive relapses using functional genomics; 2) To design dual-specific chimeric antigen receptor T cells that will prevent tumor escape. This research will be significant because it will contribute depth (of understanding the mechanisms of relapse) and breadth (of novel potentially curative therapy) to the immunotherapeutic arsenal against cancer. Ultimately, such knowledge has the potential to vertically advance the field of CAR-redirected T cell immunotherapy as well as other targeted immunotherapies. This is project is innovative because it combines high throughput screening techniques together with in vitro and in vivo functional studies to study the possible mechanisms of resistance after immunotherapy and therefore to generate novel treatments. The proposed research activities are crucial to the development of the applicant as an independently-funded scientist with a focus on cellular immunotherapy. Dr. Ruella will receive further training in molecular biology, translational medicine, and immunology from his mentors and training in next-generation sequencing, cancer biology and bioinformatics from experienced collaborators at the University of Pennsylvania. Therefore at the conclusion of the training period, the applicant will have acquired a unique set of intellectual and technical skills that will allow him to attack the problem of resistance to targeted immunotherapies from several angles at once. In addition, this award will support a unique training experience in translational research and will establish an academic pathway for the discovery and development of new CAR T cell approaches.