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个项目摘要/摘要 2 3同种异体造血干细胞移植（HSCT）是许多血液学疾病的治愈选择 4但是它的功效受到移植抗宿主病（GVHD）的限制，这是发病率和死亡率的主要原因 5 HSCT。由于当前的预防性测量20-70％的HSCT患者发展GVHD，并且容易受到影响 6感染，肿瘤复发和继发性恶性肿瘤。临床前和临床研究表明收养转移 调节性T细胞（TREG）的7个在预防GVHD方面非常有效。临床翻译受到低低的阻碍 8 treg频率在血液中，多克隆Treg的抑制能力可变，需要注入高剂量 9可重复抑制疾病。已经证明抗原特异性的Treg比多克隆更有效 在自身免疫模型和固体器官移植中，有10个Tregs，其中有HLA-A2特异性Treg抑制 11针对HLA-A2不同移植物的反应。虽然这种方法是针对最常见的不匹配之一 12％的非裔美国人和34％的西班牙裔患者存在12种抗原，HLA-A2存在。克服这些 13局限性，我将使用嵌合抗原受体（CAR）将Treg重定向到人CD19（HCD19）B细胞 14使用FDA批准的结构，该结构包含4-1BB共刺激域（CAR19）。出乎意料的是，我的 15带有CAR19 Tregs的初步数据表明了GVHD抑制和杀戮的双重和相反的特性 16个白血病细胞，包括移植物与白血病（GVL）。因此，我的目标是使用针对HCD19的特雷格 17在恶性B细胞上介导GVL和非恶性B细胞以耗尽这些宿主抗原呈递细胞 18在临界GVHD诱导阶段。我将使用接收者小鼠的鼠主要MHC不匹配模型 19在B细胞上表达HCD19。我的初步研究表明，CAR19 Tregs提高了GHVD的存活并导致 20 HCD19 B细胞在HSCT后早期耗竭。我们还发现CAR19 Tregs介导针对HCD19的靶向杀死 21体外淋巴瘤细胞与对照treg不同，并保持体内GVL。了解CAR19 Tregs的使用方式 22抑制GVHD的GVL的细胞解析将有助于扩大我们对Treg生物学和设计的理解 23以最大化它们在Allo-HSCT中的使用。我假设Car19 Tregs的抑制能力是双功能的 24过度和有害的免疫反应（GVHD，CRS），并同时诱导GVL效应。发现 25根据这项研究建议将阐明CAR19 Tregs提供更高保护的机制 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）和 27独特地促进抗肿瘤反应。将汽车Treg转向肿瘤抗原的新型方法打开 28一种新的治疗途径，可防止GVHD并改善Allo-HSCT结果。