Targeting Leukemia with Memory CD8+ T cells Transduced to Express High-Affinity W

描述（由申请人提供）：候选人的目标是成为一个实验室的首席研究员，该实验室有能力进行转化研究，重点研究调节免疫系统以靶向癌症的新方法。由于她已经获得了实现这一目标的大部分相关培训，K08将使她在获得独立助理教授身份之前完成最后的步伐。具体来说，该候选药物致力于解决高风险白血病患者异体造血细胞移植（HCT）后复发的问题，因为这仍然是治疗失败和死亡的一个重要原因。WT1是一种在白血病细胞中过度表达的蛋白，在白血病干细胞（LSC）中表达水平最高，由hla匹配的供者产生，在HCT后给予WT1特异性CD8+ t细胞克隆，无明显毒性，并观察到优先定位于骨髓。然而，在一些患者中，由于缺乏长期的体内t细胞持久性，对WT1靶点的渴望度低，抗白血病疗效受到限制，并且可能通过同时靶向其他表位而增强。为了解决这些障碍，该项目建议在I/II期试验中，从匹配的供体中产生EBV或cmv特异性记忆T细胞，并在输注高风险AML， CML和MDS患者之前将其转导表达高亲和力的wt1特异性TCR。候选人将评估移植细胞的安全性、持久性、定位和功能，并将这些参数与hct后CR患者预防复发的有效性（临床研究1组）和可检测疾病患者移植细胞的潜在抗白血病活性（临床研究2组）相关联。然后，她建议通过测试来自白血病供体的非基因修饰T细胞产品是否可以用于未来的临床试验来评估安全性，探索以新发现的LSC和癌睾丸抗原CCNA1为靶点的T细胞治疗的可行性。同时，她将筛选ccna1特异性的高亲和度/亲和度CD8+ T细胞克隆，并分离其TCR作为“现成”试剂的潜在用途。如果证明单独靶向任一抗原的安全性，这些试剂可以在未来的临床试验中同时使用。除了基于流式细胞术的方法、生物统计学、分子免疫学和FDA相关的基因修饰临床试验监管行为外，候选人还将接受最近开发的基于质谱的新方法的高级培训，以同时评估输注t细胞的多个参数。研究和培训活动将主要在Fred Hutchinson癌症研究中心进行，该中心为基础、临床和转化研究提供了极好的环境，特别是在HCT领域。候选人将由Phil Greenberg博士指导，他是一位经验丰富且成功的导师，在转化免疫学和免疫治疗领域拥有成熟的职业生涯。拟议的转化研究旨在为AML、MDS和CML患者开发有效的、无毒的治疗方法，确定成功根除白血病的免疫参数，并指导针对LSC蛋白的高亲和力T细胞的设计和安全部署。具体目标是：1。在I/II期试验中评估供体来源的C4-CTL的安全性和潜在疗效

DESCRIPTION (provided by applicant): The candidate's objective is to become the principal investigator of a laboratory equipped to perform translational studies focusing on novel ways to modulate the immune system to target cancer. As she has obtained a large part of the relevant training to achieve this goal, the K08 would allow her to complete the final strides before achieving independent Assistant Professor status. Specifically, the candidate aspires to address relapse after allogeneic hematopoietic cell transplantation (HCT) in patients with high-risk leukemias as this remains a significant cause of treatment failure and death. CD8+ T-cell clones specific for WT1, a protein that is over-expressed in leukemic cells with the highest levels expressed in leukemic stem cells (LSC), generated from HLA-matched donors have been administered after HCT without significant toxicity and observed to preferentially localize to the marrow. However, anti-leukemic efficacy was limited in some patients by the lack of long-term in vivo T-cell persistence, low avidity for the WT1 target, and may have been enhanced by simultaneously targeting additional epitopes. To address these obstacles, this project proposes in a Phase I/II trial, to generate EBV- or CMV-specific memory T cells from matched donors and transduce them to express a high affinity WT1-specific TCR before infusion in patients with high-risk AML, CML and MDS post-HCT. The candidate will assess the safety, persistence, localization and function of the transferred cells and correlate the parameters with the efficacy o preventing relapse in post-HCT patients who are in CR (Clinical Study Arm 1), and the potential antileukemic activity of transferred cells in patients with detectable disease (Clinical Study Arm 2). She then proposes to explore the feasibility of targeting the newly identified LSC and cancer testis antigen CCNA1 with T cell therapy by testing if non-gene modified T-cell products from leukemic donors can be generated for use in future clinical trials to assess safety. In parallel, she will screen for CCNA1-specific high-avidity/affinity CD8+ T cell clones, and isolate their TCR for potential use as 'off the shelf' reagents. If safety of targeting either antigen alone is demonstrated, these reagents could be used concurrently in future clinical trials. The candidate will receive advanced training in recently developed novel mass spectrometry- based methods to assess multiple parameters simultaneously on infused T-cells in addition to flow cytometry- based methods, biostatistics, molecular immunology, and FDA related regulatory conduct of gene-modified clinical trials. The research and training activities will be primarily conducted at the Fred Hutchinson Cancer Research Center, which is a superb environment for basic, clinical and translational research, particularly in the field of HCT. The candidate will be mentored by Dr Phil Greenberg, an experienced and successful mentor who has an established career in the field of translational immunology and immunotherapy. The proposed translational studies are aimed at developing potent, non-toxic treatments for patients with AML, MDS and CML, determining immunologic parameters for successful leukemia eradication, and guiding the design and safe deployment of high-affinity T cells targeting LSC proteins. The Specific Aims are: 1. Evaluate in a phase I/II trial the safety and potential efficacy of donor-derived C4-CTL in preventing relapse in remission patients at high risk of AML/MDS/CML recurrence. 2. Evaluate in a Phase I/II the safety, efficacy and potential limitations to the anti-leukemic activity of donr- derived C4-CTL in patients with MRD or recurrent AML/MDS/CML. 3. Explore the potential for targeting CCNA1 with T cell therapy as an alternate target antigen to WT1.