Project 2: Regulatory T Cells Generated by Gene Transfer to Prevent GVHD

项目2：基因转移产生的调节性T细胞预防GVHD

• 批准号: 10700002
• 负责人: Maria Grazia Roncarolo
• 金额: $ 33.05万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10700002
• 关键词: Acute Graft Versus Host Disease; Alloantigen; Allogenic; Antigen-Presenting Cells; Antigens; Autologous; Bone Marrow Transplantation; CD4 Positive T Lymphocytes; Cell Therapy; Cells; Childhood; Clinical; Clinical Trials; Complication; Data; Dendritic Cells; Development; Disease; Disparity; Dose; Effectiveness; Evaluation; FDA approved; FOXP3 gene; Future; Gene Transduction Agent; Gene Transfer; Gene Transfer Techniques; Generations; Goals; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histocompatibility; Human; Immune; Immune Tolerance; Immunological Models; Immunologics; In Vitro; Incidence; Inflammatory; Interleukin-10; Lentivirus Vector; Lymphocyte Subset; Lymphoid; Mediating; Modeling; Morbidity - disease rate; Mus; NGFR Protein; Outcome; Patient-Focused Outcomes; Patients; Phase; Phase I Clinical Trials; Phenotype; Population; Pre-Clinical Model; Prevention; Production; Property; Regulatory T-Lymphocyte; Research; Safety; Sampling; Specificity; T cell anergy; T cell response; T-Lymphocyte; Tissues; Transduction Gene; Transplantation; Transplantation Tolerance; anergy; chronic graft versus host disease; comparative; disorder prevention; effective therapy; effector T cell; experimental study; gene transfer vector; graft versus host disease induction; graft vs host disease; graft vs leukemia effect; humanized mouse; immune reconstitution; improved; improved outcome; in vivo; leukemia; leukemia/lymphoma; mortality; mouse model; phase I trial; phase II trial; post-transplant; pre-clinical; pre-clinical research; preclinical study; preclinical trial; prevent; primary endpoint; safety and feasibility; stem cell engraftment; time use; young adult

Project 2: Project Summary A major complication of allogeneic hematopoietic stem cell transplantation (allo-HCT) is acute graft-versus-host disease (aGvHD). aGvHD is an immunologically mediated disease, that contributes substantially to transplant- related morbidity and mortality. The overall incidence of aGvHD is between 35% and 50% depending upon donor-recipient histocompatibility differences and carries approximately a 50% mortality rate. T regulatory cells (Tregs), a phenotypically diverse subset of lymphocytes that promote and maintain immunological tolerance, have been extensively researched as a cellular therapy for aGvHD. Preclinical trials have demonstrated that the presence of IL-10-producing T regulatory type 1 (Tr1) cells or the forkhead box P3 (FOXP3) T regulatory cells (FOXP3+Tregs) correlates with aGvHD modulation and enhanced transplantation tolerance after allo-HCT. T- allo10 cells are a cell product containing donor-derived Tr1 cells primed with host-derived tolerogenic dendritic cells (DC-10) to induce immune tolerance. We have FDA approval (IND# 17292 ) for a Phase 1 clinical trial of escalating doses of T-allo10 cells in pediatric and young adult patients receiving mismatched related or unrelated HCT for hematologic malignancies, and 2 patients have been successfully treated. The primary endpoints of the Phase 1 trial are 1) the feasibility of producing the T-allo10 product in our GMP facility and 2) the safety of the administration of the T-allo10 product. Aim 1 is the completion of this trial including the immunological evaluation of the T-allo10 product and the longitudinal assessment of the HCT patients for the persistence of the infused Tr1 cells using TCR clonotypic analyses and the immune reconstitution. Tr1 containing T-allo10 cells offer enormous promise, but are a mixed cell product also containing T effector cells, that potentially reduce their effectiveness. Although polyclonal FOXP3+Tregs have shown some efficacy in enhancing tolerance post- transplant, the infused cells have limited in vivo survival, and the isolation of pure FOXP3+Tregs is difficult. Therefore, we have developed “second-generation” Tr1 cells and FOXP3+Tregs by lentiviral-vector (LV) gene transfer into CD4+ T effector cells. LV-mediated gene transfer of FOXP3 into CD4+ T cells creates “FOXP3+Treg-like” (CD4FOXP3) cells. Similarly, “Tr1-like” (CD4IL-10) cells can be obtained via LV-IL-10 expression in human CD4+ T effector cells. In preliminary experiments, we have demonstrated that 3rd party polyclonal CD4IL-10 and CD4FOXP3 T cells can suppress GvHD in a humanized model of immune deficient mice. In Aims 2 and 3 we will compare autologous polyclonal and alloAg-specific CD4IL-10 and CD4FOXP3 for their prevention of aGvHD, their impact on hematopoietic stem cell (HSC) engraftment, immune reconstitution, tolerance and GvL activity in preclinical studies. These studies will provide preclinical data toward developing future therapies, that will improve the outcomes of patients receiving allo-HCT.

项目2：项目摘要 同种异体造血干细胞移植（Allo-HCT）的主要并发症是急性移植物 - 抗宿主 疾病（AGVHD）。 AGVHD是一种免疫学介导的疾病，对移植 - 相关的发病率和死亡率。 AGVHD的整体事件在35％至50％之间 供体 - 接触性组织相容性差异和大约50％的死亡率。 T调节细胞 （tregs），一种表型多样的淋巴细胞的子集，促进和维持免疫耐受性， 已被广泛研究为AGVHD的细胞疗法。临床前试验表明 存在IL-10产生的T型Type 1（TR1）细胞或Forkhead Box P3（FOXP3）T调节细胞的存在 （FOXP3+Tregs）与Allo-HCT后的AGVHD调制和增强的移植耐受性相关。 t- Allo10细胞是含有供体衍生的TR1细胞的细胞产物 细胞（DC-10）诱导免疫耐受性。我们有FDA批准（IND＃17292），用于1阶段的临床试验 在接受不匹配或无关的儿科和年轻患者中，小儿和年轻患者的T-Allo10细胞的剂量不断升级 血液学恶性肿瘤和2例患者的HCT已成功治疗。主要终点 第1阶段试验是1）在我们的GMP设施中生产T-Allo10产品的可行性； 2） T-Allo10产品的给药。 AIM 1是该试验的完成，包括免疫评估 T-Allo10产品和HCT患者的纵向评估 TR1细胞使用TCR clonotypic分析和免疫缔合。包含T-Allo10单元的TR1提供 巨大的希望，但是包含T效应细胞的混合细胞产物，可能会减少其 有效性。尽管多克隆Foxp3+Tregs在增强耐受性后表现出一定的有效性。 移植，感染的细胞在体内存活率有限，纯Foxp3+Treg的分离很困难。 因此，我们开发了慢病毒载体（LV）基因的“第二代” TR1细胞和Foxp3+Tregs 转移到CD4+ T效应细胞中。 LV介导的FOXP3转移到CD4+ T细胞中会产生 “ Foxp3+Treg样”（CD4FOXP3）细胞。同样，可以通过LV-IL-10表达获得“ TR1样”（CD4IL-10）细胞 在人CD4+ T效应细胞中。在初步实验中，我们证明了第三方多克隆 CD4IL-10和CD4FOXP3 T细胞可以在人性化的免疫缺陷小鼠模型中抑制GVHD。在目标2中 3我们将比较自体多克隆和AlloAG特异性CD4IL-10和CD4FOXP3的预防 AGVHD，它们对造血干细胞（HSC）植入，免疫重构，耐受性和GVL的影响 临床前研究的活性。这些研究将为开发未来疗法提供临床前数据， 将改善接受Allo-HCT的患者的结局。