DEVELOPMENT & EVALUATION OF PRACTICABLE APPROACHES FOR GENERATION OF CYTOTOXIC &

发展

基本信息

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
项目类别：
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

项目摘要

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 成功的重大障碍，特别是对于患者而言急性白血病和骨髓增生异常综合征（MDS）在疾病晚期进行移植。捐赠者来源的过继转移抗原特异性 T 细胞已成为治疗和预防危及生命的疾病的一种有前途的方法移植后病毒感染，也可用于提供可扩展的杀肿瘤效应 T 群体细胞到携带肿瘤的宿主。在这个项目中，我们建议探索和开发切实可行的广泛适用的生成选择性地对抗差异表达的决定簇的供体来源的 T 细胞的策略对白血病细胞进行过继免疫治疗，以治疗或最终预防白血病术后复发移植期。在目标 1 中，我们建议开发和评估快速生成 WT1 的新策略来自正常移植供体的肽特异性 T 细胞，表达一系列常见 HLA I 类中的至少一种或 II 等位基因，通过体外致敏，使用一组可选择的立即可用和可补充的人工抗原呈递细胞被设计为表达关键共刺激分子和单个 I 类或 II 类等位基因由供体共享，这些供体要么加载了特定的 WT1 表位，要么加载了合成重叠库跨越 WT1 序列或转导以表达 WT1 蛋白的十五肽。这些T细胞将然后与用自体、负载WT1肽的树突状细胞致敏的T细胞进行比较，以得出，表型、肽特异性反应性和杀白血病活性。在目标 2 中，我们将在体外开发和评估生成并选择 EBV 或 CMV 病毒特异性 T 细胞，转染后也表达 T 细胞受体对由流行的 I 类 HLA 等位基因或 CD19 特异性 ScFv 呈递的免疫原性 WT1 肽具有特异性基于嵌合抗原受体并评估它们针对 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 CAR 将消除转导同种异体反应性 T 细胞的风险，还可能增强通过表达潜在病毒抗原的细胞在体内持续刺激，双受体 T 细胞的持久性。在目标 3 中，我们建议比较评估目标 1 和目标中生成的 WT1 特异性和 CD19 特异性 T v 细胞 2 因其具有迁移、积累、持续存在并诱导白血病异种移植物消退的能力 NOD/SCID 小鼠，并评估 WT1 肽致敏 T 细胞和表达 T 细胞的效果转导受体对正常造血细胞和白血病母细胞的植入和体内扩增的影响在许可型 NOD/SCIDyc"'" 小鼠模型中。相关性：这些研究可能会产生快速、实用和广泛适用的方法和可补充的试剂，用于生成白血病反应性 T 细胞用于过继治疗并应提供对计划至关重要的此类 T 细胞的抗白血病作用的比较估计并优先考虑临床试验