DEVELOPMENT & EVALUATION OF PRACTICABLE APPROACHES FOR GENERATION OF CYTOTOXIC &

发展

• 批准号: 7318391
• 负责人: Richard John O'REILLY
• 金额: $ 34.46万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7318391
• 关键词: Acute leukemia; Adoptive Immunotherapy; Adoptive Transfer; Alleles; Allogenic; Antigen Receptors; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autologous; Blast Cell; CD19 gene; Cells; Class; Clinical Trials; Cytomegalovirus; Dendritic Cells; Development; Disease; Disease regression; Donor person; Dysmyelopoietic Syndromes; Engraftment; Epitopes; Evaluation; Generations; Growth; Hematopoietic; Hematopoietic stem cells; Human; Human Herpesvirus 4; In Vitro; Life; Mus; Nephroblastoma; Patients; Peptides; Phenotype; Population; Prevention; Reagent; Relapse; Risk; SCID Mice; Series; Staging; Stem cell transplant; T-Cell Receptor; T-Lymphocyte; Transplantation; Viral; Viral Antigens; Virus; Virus Diseases; WT1 Protein; WT1 gene; Xenograft procedure; base; cellular engineering; comparative; cytotoxic; graft vs host disease; immunogenic; in vivo; leukemia; leukemia/lymphoma; mouse model; novel; oncofetal antigen; prevent; receptor; single molecule; success; tumor

Leukemia Relapse remains a significant obstacle to the success of allogeneic HSCT, particularly for patients with acute leukemias and MDS transplanted in advanced stages of disease. Adoptive transfer of donor-derived antigen-specific T cells has emerged as a promising approach for the tretment and prevention of life-threatening viral infections post transplant, and may also be used to provide expandable populations of tumoricidal effector T cells to tumor-beariing hosts. In this project, we propose to explore and develop practicable broadly applicable strategies for generating donor-derived T cells that selectively react againstdeterminants differentially expressed on leukemia cells for adoptive immunotherapy to treat or, ultimately, prevent leukemia relapse in the post transplant peeriod. In Aim 1, we propose to develop and evaluate new strategies for rapid generation of WT1 peptide specific T cells from normal transplant donors expressing at least one of a series of common HLA class I or II alleles by in vitro sensitization with a selectable panel of immediately accessible and replenishable artificial antigen presented cells engineered to express critical costimulatory molecules and single class I or class II alleles shared by the donor which have been either loaded with specific WT1 epitopes or a pool of synthetic overlapping pentadecapeptides spanning the WT1 sequence or transduced to express the WT1 protein. These T cells will then be compared with T cells sensitized with autologous, WT1 peptide loaded dendritic cells as to yield, phenotype, peptide-speciflc reactivity and leukemocidal activity. In Aim 2, we will develop and evaluate in vitro generated and selected EBV or CMV virus-specific T cells transduced to also express either a T cell receptor specific for an immunogenic WT1 peptide presented by a prevalent class I HLA allele, or CD19-specific ScFv- based chimeric antigen receptor and evaluate them for their activity against WT1+ and/or CD19+ leukemias and lymphomas and their viral antigen targets. We hypothesize that introduction of a leukemia reactive WT1-specific TCR or CE19-specific CAR will abrogate the risk of transducing alloreactive T cells and may also enhance persistence of dual receptor T cells through ongoing stimulation in vivo by cells expressing latent viral antigens. In Aim 3, we propose to comparatively evaluate WT1 specific and CD19 specific T vcells generated in aims 1 and 2 for their capacity to migrate to, accumulate and persist in and induce regressions of leukemia xenografts in NOD/SCID mice, and to also assess the effects of the WT1 peptide sensitized T cells and T cells expressing transduced receptors on the engraftment and in vivo expansion of normal hematopoietic cells and leukemia blasts in the permissive NOD/SCIDyc"'" mouse model. Relevance: These studies may yield rapid, practicable and broadly applicable approaches and replenishable reagents for generating leukemia-reactive T cells for adoptive therapy and should provide comparative estimates of the anti-leukemia effects of such T cells essential to plan and prioritize clinical trials

白血病复发仍然是同种异体HSCT成功的重要障碍，特别是对于患者而言 急性白血病和MD在疾病晚期阶段移植。捐助者衍生的收养转移 抗原特异性的T细胞已成为一种有希望的方法来预防和预防生命 移植后病毒感染，也可以用于提供可扩展的肿瘤效应群体 细胞到肿瘤宿主。在这个项目中，我们建议探索和开发可行的广泛适用 生成供体衍生的T细胞的策略，这些细胞有选择地反应针对确定的差异表达 在白血病细胞上进行收养免疫疗法，以治疗或最终防止白血病复发 移植peeriod。在AIM 1中，我们建议制定和评估快速生成WT1的新策略 来自正常移植供体的肽特异性T细胞，表达至少一系列常见HLA I类之一 或II等位基因通过体外敏化，并具有可选的立即可访问和可补充人工的面板 抗原呈现了用于表达关键共刺激分子和单一I类或II类等位基因的细胞 由供体共享的，这些捐赠者要么装有特定的WT1表位或合成重叠池 跨WT1序列或转导的五二肽表达WT1蛋白。这些T细胞将 然后将其与自体，WT1肽载荷的树突状细胞敏感的T细胞进行比较，以使其产生， 表型，肽特异性反应性和白血病活性。在AIM 2中，我们将在体外开发和评估 转导的EBV或CMV病毒特异性T细胞也表达T细胞受体 专门针对由普遍的I级等位基因提出的免疫原性WT1肽，或CD19特异性的SCFV- 基于嵌合抗原受体，并评估它们对WT1+和/或CD19+白血病的活性以及 淋巴瘤及其病毒抗原靶标。我们假设引入白血病反应性WT1特异性 TCR或CE19特异性汽车将消除传输同种异体T细胞的风险，也可能会增强 双重受体T细胞通过表达潜在病毒抗原的细胞在体内持续刺激。 在AIM 3中，我们建议在AIM 1和 2为了迁移，积累和持续存在并诱导白血病异种移植物的回落能力 点头/SCID小鼠，还评估WT1肽敏化的T细胞和表达的T细胞的作用 正常造血细胞和白血病爆炸的植入和体内膨胀上的受体转导受体 在允许的点头/scidyc“”鼠标模型中。相关性：这些研究可能会产生快速，可行的和 广泛适用的方法和可补充的试剂，用于产生白血病反应性T细胞用于收养 治疗，应提供对计划必不可少的T细胞抗白血病作用的比较估计 并优先考虑临床试验