Enhancing Treg Therapeutic Efficacy in GVHD with CARs

利用 CAR 增强 GVHD 中 Treg 的治疗功效

• 批准号: 10312590
• 负责人: Sara Bolivar Wagers
• 金额: $ 4.02万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-13 至 2024-07-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10312590
• 关键词: Adoptive Transfer; African American; Alleles; Allogenic; Antigen Presentation; Antigen-Presenting Cells; Antigens; Antitumor Response; Autoimmune; B lymphoid malignancy; B-Cell Lymphomas; B-Lymphocytes; Biology; Blood; Bone Marrow; CD19 gene; CD28 gene; Cells; Clinical Research; Cyclophosphamide; Cytolysis; Data; Disease; Dose; FDA approved; Fluorescence; Frequencies; Genome engineering; Goals; Graft Rejection; Granzyme; HLA-A2 Antigen; Hematologic Neoplasms; Hematological Disease; Hematopoietic Stem Cell Transplantation; Hispanics; Human; Image; Immune; Immune response; Immunology; Immunosuppression; Immunotherapy; In Vitro; Infection; Inflammatory; Infusion procedures; Knock-out; Knockout Mice; Laboratories; Left; Leukemic Cell; Lymphoma cell; Malignant - descriptor; Malignant Neoplasms; Measures; Mediating; Modeling; Morbidity - disease rate; Mus; Non-Malignant; Organ; Organ Transplantation; Outcome; Patient-Focused Outcomes; Patients; Pediatric Oncologist; Phase; Physicians; Prevention; Production; Property; Receptor Signaling; Recurrence; Regulatory T-Lymphocyte; Reporter; Research Proposals; Scientist; Second Primary Cancers; Severity of illness; Solid; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Transgenes; Transplant Recipients; Transplantation; Treatment Efficacy; Treg therapy; Tumor Antigens; base; bone marrow failure syndrome; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; curative treatments; cytokine; cytokine release syndrome; design; disorder prevention; graft versus host disease induction; graft vs host disease; graft vs leukemia effect; high risk; improved; in vivo; infection risk; mortality; mouse model; novel; novel strategies; novel therapeutics; perforin; preclinical study; prevent; prophylactic; rat orphan nuclear receptor NR4A1; response; secondary lymphoid organ; side effect; skills; transplant model; tumor

1 Project Summary/Abstract 2 3 Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative option for many hematological disorders 4 but its efficacy is limited by graft-versus-host disease (GVHD), a leading cause of morbidity and mortality after 5 HSCT. With current prophylactic measures 20-70% of HSCT patients develop GVHD and are left susceptible to 6 infection, tumor recurrence, and secondary malignancies. Pre-clinical and clinical studies show adoptive transfer 7 of regulatory T cells (Treg) is highly effective at preventing GVHD. Clinical translation has been hindered by low 8 Treg frequency in the blood, variable suppressive capacity of polyclonal Tregs, and the need to infuse high doses 9 to reproducibly suppress disease. Antigen-specific Tregs have been shown to be more potent than polyclonal 10 Tregs in autoimmune models and in solid organ transplants where HLA-A2 specific Tregs are given to suppress 11 responses against HLA-A2 disparate grafts. While this approach targets one of the most commonly mismatched 12 antigens, HLA-A2 is present in only 20% of African American and 34% of Hispanic patients. To overcome these 13 limitations, I will use chimeric antigen receptors (CAR) to redirect Tregs towards human CD19 (hCD19) B cells 14 using an FDA approved construct containing the 4-1BB co-stimulatory domain (CAR19). Unexpectedly, my 15 preliminary data with CAR19 Tregs demonstrate dual and opposing properties of GVHD suppression and killing 16 of leukemia cells including graft-versus-leukemia (GVL). Thus, my goal is to use Tregs specific for hCD19 present 17 on malignant B cells to mediate GVL and on non-malignant B cells to deplete these host antigen presenting cells 18 during the critical GVHD induction phase. I will use a murine major MHC mismatch model where recipient mice 19 express hCD19 on B cells. My preliminary studies demonstrate CAR19 Tregs improve GHVD survival and cause 20 hCD19 B cell depletion early after HSCT. We also found CAR19 Tregs mediate targeted killing of hCD19 21 lymphoma cells in vitro and maintained GVL in vivo unlike control Tregs. Understanding how CAR19 Tregs utilize 22 cytolysis for GVL while suppressing GVHD will help expand our understanding of CAR Treg biology and design 23 to maximize their use in allo-HSCT. I hypothesize CAR19 Tregs are bifunctional in their ability to suppress 24 excessive and deleterious immune responses (GVHD, CRS) and concurrently induce GVL effects. The findings 25 from this research proposal will elucidate the mechanisms by which CAR19 Tregs confer higher protection than 26 control Tregs or even CAR19 T cells at ameliorating GVHD, preventing cytokine release syndrome (CRS), and 27 uniquely promoting anti-tumor responses. The novel approach of redirecting CAR Tregs to tumor antigens opens 28 a new therapeutic avenue to prevent GVHD and improve allo-HSCT outcomes.

1项目摘要/摘要 2. 异基因造血干细胞移植(HSCT)是治疗许多血液疾病的一种选择。 但其疗效受到移植物抗宿主病(GVHD)的限制，移植物抗宿主病是移植物抗宿主病后发病率和死亡率的主要原因 5 HSCT。在目前的预防措施下，20%-70%的HSCT患者会发生移植物抗宿主病，并容易患上 6感染、肿瘤复发、继发性恶性肿瘤。临床前和临床研究显示收养转移 7的调节性T细胞(Treg)对预防移植物抗宿主病(GVHD)非常有效。临床翻译一直受到LOW的阻碍 8Treg在血液中的频率，多克隆Treg的可变抑制能力，以及需要输注高剂量 9.可重复地抑制疾病。抗原特异性树突状细胞已被证明比多克隆更有效。 在自身免疫模型和实体器官移植中给予人类白细胞抗原A2特异性树突状细胞抑制的10个树突状细胞 11对人类白细胞抗原A2异种移植物的反应。虽然这种方法的目标是最常见的不匹配的 12个抗原，只有20%的非裔美国人和34%的西班牙裔患者存在HLA-A2。要克服这些障碍 13限制，我将使用嵌合抗原受体(CAR)将Treg重定向至人类CD19(HCD19)B细胞 14使用FDA批准的含有4-1BB共刺激结构域(CAR19)的构建物。没想到，我的 15个cAR19 Tregs的初步数据显示GVHD抑制和杀伤的双重和相反的特性 包括移植物抗白血病(GVL)在内的16个白血病细胞。因此，我的目标是使用特定于hCD19 Present的Tregs 17在恶性B细胞上介导GVL，在非恶性B细胞上耗尽这些宿主抗原提呈细胞 18在GVHD关键诱导期。我将使用小鼠主要MHC错配模型，在该模型中，受体小鼠 19例B细胞表达hCD19。我的初步研究表明，cAR19Tregs可以改善GHVD的存活率和病因 HSCT后20hCD19 B细胞早期耗竭。我们还发现cAR19Tregs介导了hCD19的靶向杀伤 21淋巴瘤细胞在体外和体内维持GVL不同于对照Tregs。了解cAR19树如何利用 22在抑制GVHD的同时对GVL进行细胞溶解将有助于扩大我们对汽车Treg生物学和设计的理解 23以最大限度地利用它们在allo-HSCT中的应用。我假设cAR19树突状细胞具有双重功能 24过度和有害的免疫反应(GVHD、CRS)并同时诱发GVL效应。调查结果 25这项研究提案将阐明cAR19Tregs提供比 26控制Tregs甚至cAR19 T细胞，以改善GVHD，防止细胞因子释放综合征(CRS)，以及 27独特地促进抗肿瘤反应。将CAR树重定向至肿瘤抗原的新方法打开 28预防GVHD和改善allo-HSCT结局的新治疗途径。