Multi-functional cellular therapies to overcome tumor heterogeneity and limit toxicity in acute myeloid leukemia

项目摘要/摘要 急性髓系白血病(AML)是一种破坏性疾病，5年总存活率只有30%。作为鲜血 癌症，急性髓细胞白血病似乎准备从工程细胞疗法和双特异性 抗体。这些药物对B细胞恶性肿瘤产生了巨大的影响。然而，扩展这一点 急性髓细胞白血病的成功面临两个重大障碍：1)缺乏高度针对疾病的目标，在 目前的AML治疗靶点导致严重的细胞因子释放综合征和严重的骨髓清除术， 排除了有效的治疗指数；以及2)显著的肿瘤内异质性，其中以单一的 表面抗原很可能导致抗原阴性复发。在实践中，这两个挑战几乎没有留下什么 治疗目标足以在大多数患者中实现治愈。在这里，我的长期目标是开发一种新的 克服这些障碍的治疗方法。这项提议将由我本人领导，并由 Arun Wiita博士是一位血液恶性肿瘤和耐药性方面的专家，他得到了Dr。 高级细胞工程专家Kole Roybal和抗体工程专家Jim Wells博士。 具体地说，我假设使用SNIPR，这是新描述的下一代人性化平台 基于SynNotch细胞工程技术(朱等人，Cell2022)，将使 多功能T细胞治疗可消除诱导化疗后残留的AML，清除复发- 驱动白血病干细胞，提高治愈率，同时避免系统疗法的毒性。我的 主要的设计支柱包括1)开发专门用于白血病干细胞的工程化T细胞 利基；2)局部的，但不是系统的，SNIPR T细胞分泌多种蛋白质疗法来消融 异种白血病干细胞和急性髓系白血病在这个利基内和附近发生。在具体目标1中，我测试了可行性 SNIPR工程T细胞在体外和体内的初步效力 针对3个AML靶点的功能性双特异性抗体。在具体目标2中，我使用PDX模型来测试 SNIPR T细胞可以选择性地定位于骨髓，CD70是最佳的白血病干细胞 “触发器”。成功完成这些目标将为后续工作奠定基础，而不是本报告范围之外的工作 建议使用更多的体外和体内模型来验证CD70传感、多双特异性... 分泌T细胞，比其他疗法更彻底地消除AML，同时将毒性降至最低。 此外，这种方法最终将能够将其他治疗有效载荷输送到白血病干细胞 细胞利基，理想地以前所未有的精确度瞄准这一导致疾病的细胞群体。在这里取得成功 也可能勾勒出一种新的治疗策略，用于跨癌症，以消除 微环境，同时保留正常细胞。完成本提案中的工作，与 相关的培训计划将提供关键的专业知识和经验，以支持我成为 独立内科医生兼科学家专注于转化性免疫疗法研究。

PROJECT SUMMARY/ABSTRACT Acute myeloid leukemia (AML) is a devastating disease with only 30% 5-year overall survival. As a blood cancer, AML appears poised to benefit from the revolution in engineered cellular therapies and bispecific antibodies. These agents are making an enormous impact in B-cell malignancies. However, extending this success to AML faces two significant hurdles: 1) lack of highly disease-specific targets, where engaging current AML therapeutic targets leads to severe cytokine release syndrome and severe myeloablation, precluding an effective therapeutic index; and 2) significant intratumoral heterogeneity, where targeting a single surface antigen is likely to lead to antigen-negative relapse. In practice, these two challenges leave few therapeutic targets sufficient to achieve cures in most patients. Here, my long-term goal is to develop a new therapeutic approach to overcome these hurdles. This proposal will be led by myself under the sponsorship of Dr. Arun Wiita, an expert in hematologic malignancies and drug resistance, with collaborative support from Dr. Kole Roybal, expert in advanced cellular engineering; and Dr. Jim Wells, expert in antibody engineering. Specifically, I hypothesize that employing SNIPR, the newly-described, next-generation, humanized platform based on the SynNotch cellular engineering technology (Zhu et al., Cell 2022), will enable development of a multi-functional T-cell therapy that can eliminate residual AML after induction chemotherapy, clearing relapse- driving leukemic stem cells and increasing cure rates, while avoiding toxicities of systemic therapeutics. My major design pillars include 1) develop engineered T-cells that home specifically to the leukemic stem cell niche; and 2) locally, but not systemically, have SNIPR T-cells secrete multiple protein therapeutics to ablate heterogenous leukemic stem cells and AML blasts in and near this niche. In Specific Aim 1, I test the feasibility and initial in vitro and in vivo potency of SNIPR engineered T-cells designed to simultaneously secrete functional bispecific antibodies against 3 AML targets. In Specific Aim 2, I use PDX models to test whether SNIPR T-cells can selectively home to the bone marrow niche, with CD70 as the optimal leukemic stem cell “trigger”. Successful completion of these Aims will set the stage for subsequent work, outside the scope of this proposal, to validate, using additional in vitro and in vivo models, the ability of CD70-sensing, multi-bispecific- secreting T-cells to eliminate AML more completely than other therapies while minimizing toxicities. Furthermore, this approach will ultimately enable delivery of other therapeutic payloads to the leukemic stem cell niche, ideally targeting this disease-driving cellular population with unprecedented precision. Success here may also outline a new therapeutic strategy for use across cancers, to eliminate heterogeneous tumor in the microenvironment while sparing normal cells. Completion of the work in this proposal in conjunction with the associated training plan will provide critical expertise and experience to support my goal of becoming an independent physician-scientist focused on translational immunotherapy research.