Resistance To Targeted Immunotherapies:CART19 as a Paradigm

项目摘要/摘要 嵌合抗原受体(CAR)T细胞开辟了肿瘤靶向免疫治疗的新纪元。这个 申请人的导师卡尔·琼博士被公认为基因工程T细胞领域的世界领先者 恶性血液病的治疗。尽管抗CD19的CAR T细胞(CART19)产生了前所未有的 B细胞白血病和淋巴瘤患者的反应，复发是治疗的主要原因 失败了。这项提议的长期目标是发展一种独立的研究事业，专注于改进 通过研究和中和肿瘤逃逸机制进行靶向免疫治疗。统一的目标是 这一应用是为了设计治疗和预防肿瘤免疫逃逸的新策略 以CART19为模型，探讨导致复发的机制。中心假设是，多个 机制可以导致逃避免疫攻击；因此，针对该问题的组合方法 从多个方面都会阻止逃脱。这项研究基本原理是确定关键的肿瘤逃逸 机制将使我们能够为这些问题制定具体的补救办法。这项提议的中心假设是 通过追求两个特定的目标进行测试：1)描述白血病复发的发病机制 在CD19靶向免疫治疗后，通过研究以前存在的少量CD19阴性抗性克隆 CART19治疗和识别CD19阳性复发的关键耐药途径 基因组学；2)设计双特异性嵌合抗原受体T细胞，以防止肿瘤逃逸。这 研究将具有重要意义，因为它将有助于深入(了解复发的机制)和 针对癌症的免疫治疗武器的广度(新的潜在治疗方法)。最终， 这种知识有可能垂直推进汽车重定向T细胞免疫治疗领域，如 以及其他靶向免疫疗法。这个IS项目具有创新性，因为它结合了高吞吐量 结合体外和体内功能研究的筛选技术，以研究其可能的机制 在免疫治疗后产生抗药性，因此产生新的治疗方法。拟议的研究活动 对申请者的发展至关重要，成为一名独立资助的科学家，专注于细胞 免疫疗法。鲁埃拉博士将接受分子生物学、转化医学和 来自他导师的免疫学和下一代测序、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 从多个角度一下子解决靶向免疫治疗耐药问题。此外，这一点 该奖项将支持在翻译研究方面的独特培训经验，并将建立一个学术 发现和发展新的CAR T细胞途径。

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.