Targeting Cancer-Associated Fibroblasts Prevents Resistance to Chimeric Antigen Receptor T Cell Therapy

项目总结/摘要 近十年来癌症免疫治疗的一个重大突破是嵌合抗原的开发 受体（CAR）T细胞疗法。虽然抗B细胞成熟抗原（BCMA）指导CART细胞治疗， 在复发性/难治性B细胞恶性肿瘤患者中表现出前所未有的初始缓解，或 多发性骨髓瘤（MM），持久的反应是有限的。CART后复发的机制 细胞疗法尚未完全公开。我的提案的长期目标是揭示 对CART细胞疗法的抵抗，并发展一个独立的研究生涯，专注于靶向 肿瘤微环境同时与CART细胞。本申请的统一目标是 根据我的临床经验设计治疗和预防肿瘤免疫逃逸的新策略 相关的初步数据，使用BCMA CART细胞作为模型。中心假设是癌症相关的 成纤维细胞（CAFs）通过复杂机制和肿瘤细胞双重靶向诱导CART功能障碍 并且CAF是安全的并且增强CART细胞疗法的功效。为了验证这个假设，我设计了 三个具体目标：目标#1）确定MM中CAF诱导的BCMA CART细胞功能障碍的机制 2）研究双靶向CART细胞在同基因人源化MM肿瘤中的潜在毒性 微环境（TME）模型;目标#3）利用来自用BCMA CART治疗的患者的样品 研究CAF和CART细胞之间的相互作用。提出这一建议的理由是， 在我建立的MM-TME模型中，CART细胞向肿瘤部位的运输被显著抑制。我的初步 数据还显示CAFs通过分泌TGF-β和抑制性细胞因子抑制CART细胞功能， 以及改变PD-1/PD-L1轴。双重靶向肿瘤细胞和CAFs显著提高了CART细胞 功能协调发展的这项研究将是有意义的，因为它将有助于深入了解（CART细胞 耐药性）和广度（新的潜在治愈性疗法）的免疫策略， 不仅是多发性骨髓瘤，也包括实体瘤。最终，这些发现有可能垂直推进生物医学领域。 CART细胞免疫疗法以及其他靶向免疫疗法。这个项目是创新的，因为它 结合高通量筛选技术来研究耐药后的可能机制， 免疫疗法，并因此产生新的治疗方法。拟议的研究活动对以下方面至关重要： 将申请人发展为独立资助的科学家，专注于细胞免疫疗法。 我将接受导师在分子生物学、转化医学和免疫学方面的进一步培训 以及来自马约诊所经验丰富的合作者的癌症生物学和生物信息学培训。因此，我们认为， 在培训期结束时，我将获得一套独特的智力和技术技能， 这将使我成为一名独立的翻译研究者。

PROJECT SUMMARY/ABSTRACT A major breakthrough in cancer immunotherapy in the last decade is the development of chimeric antigen receptor (CAR) T cell therapy. Although anti-B cell maturation antigen (BCMA) directed CART cell therapy demonstrated unprecedented initial responses in patients with relapse/refractory B-cell malignancies or multiple myeloma (MM), respectively, durable responses are limited. The mechanisms of relapse after CART cell therapy are not fully unveiled yet. The long-term goal of my proposal is to uncover mechanisms of resistance to CART cell therapy and to develop an independent research career focused on targeting the tumor microenvironment simultaneously with CART cells. The unifying objective of this application is to design novel strategies for the treatment and prevention of tumor immunoescape based on my clinically relevant preliminary data, using BCMA CART cell as a model. The central hypothesis is cancer-associated fibroblast (CAFs) induce CART dysfunction through complex mechanisms and that dual targeting of tumor cells and CAFs is safe and enhances the efficacy of CART cell therapy. To test this hypothesis, I have designed three specific aims: Aim #1) Determine the mechanisms of CAF-induced BCMA CART cell dysfunction in MM patients; Aim #2) Study the potential toxicity of dual targeting CART cells in syngeneic humanized MM-tumor microenvironment (TME) models; Aim #3) Utilize samples from patients who were treated with BCMA CART cell therapy to study the interactions between CAFs and CART cells. The rationale for this proposal is that in my established MM-TME model, CART cell trafficking to the tumor site is significantly inhibited. My preliminary data also showed that CAFs suppressed CART cell functions by secreting TGF-β and inhibitory cytokines as well as altering the PD-1/PD-L1 axis. Dual targeting tumor cells and CAFs significantly improved CART cell functions. This research will be significant because it will contribute depth (of understanding the CART cell resistance) and breadth (of novel potentially curative therapy) to the immunotherapeutic strategy against not just MM but also solid tumors. Ultimately, such discoveries have the potential to vertically advance the field of CART cell immunotherapy as well as other targeted immunotherapies. This project is innovative because it combines high throughput screening techniques 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. I will receive further training in molecular biology, translational medicine, and immunology from my mentors and training in cancer biology and bioinformatics from experienced collaborators at the Mayo Clinic. Therefore, at the conclusion of the training period, I will have acquired a unique set of intellectual and technical skills that will allow me to promote myself to become an independent translational researcher.