CD19-Specific CB T-cell Therapy for Patients with B-cell Malignancies

项目3解决(I)机会性病毒感染和(Ii)脐带血移植(UCBT)后B系恶性肿瘤复发的问题。我们推测，过继转移供者来源的多病毒特异性细胞毒性T淋巴细胞(CTL)可以减少异基因UCBT后病毒感染和白血病复发的发生率，这些CTL是针对白血病细胞表达的CD19分子而进行的。为了巩固UCBT，我们设计了一种嵌合抗原受体(CAR)，将T细胞的抗原特异性重定向到B细胞谱系限制的细胞表面分子CD19。来自项目2的病毒特异性CTL将被转导来表达这种CAR，导致MHC非依赖和CAR依赖的激活，通过嵌合CD28和CD3-Zeta来激活Lyse淋巴样细胞，上调IL-2，以及响应CD19的抗凋亡基因。在《特殊目的#1》中提出的研究将评估是否可以使脐带血来源的多病毒特异性CTL对GDI 9具有特异性，以及是否可以在免疫缺陷小鼠中表达HSV-1胸苷激酶转基因以用于正电子发射断层扫描(PET)成像。使用非侵入性生物发光成像(BLI)和微型PET的过继免疫治疗将纵向评估输注细胞的持久性和抗肿瘤效果。特殊目的#2将评估UCBT后输注经修饰以表达CAR CD19的多病毒特异性CTL的可行性、安全性和持久性。相关研究将描述(I)持续的程度和持续时间，(Ii)贩运，以及(Iii)过继转移的CTL的抗病毒和抗白血病作用。在为人类PET T细胞成像做准备的过程中，在特定的目标#3中，我们将开发一个非人类灵长类动物模型，将猴子来源的T细胞与CARS和TK进行基因改造，并进行输注。使用胸苷激酶(TK)代谢的[18FJFEAU]的纵向PET成像将用于评估这些过继转移的猕猴T细胞的分布。总而言之，这些研究的结果将促进以病毒和CD19特异性CTL为靶点的UCBT后MRD的发展，以提高B细胞恶性肿瘤患者的无病生存率。

Project 3 addresses the problems of (i) opportunistic viral infection and (ii) relapse of B-lineage malignancies after umbilical cord blood transplantation (UCBT). We hypothesize that the incidence of viral infection and leukemia relapse following allogeneic UCBT can be reduced by adoptively transferred donor-derived multi virus-specific cytotoxic T lymphocytes (CTL), genetically modified (transduced) to be specific for the CD19 molecule expressed by leukemic cells. To consolidate UCBT we have designed a chimeric antigen receptor (CAR) to redirect antigen specificity of T cells to the B cell lineage-restricted cell-surface molecule CD19 The virus-specific CTLs from Project 2 will be transduced to express this CAR, resulting in MHC-independent and CAR-dependent activation through chimeric CD28 and CD3-zeta to lyse lymphoid cells, upregulate IL-2, and anti-apoptotic genes in response to CD19. The studies proposed in Specific Aim #1 will evaluate whether UCB-derived multivirus-specific CTLs can be rendered specific for GDI 9 and whether HSV-1 thymidine kinase transgene can be expressed for imaging by positron emission tomography (PET) in immunodeficient mice. Adoptive immunotherapy using non-invasive bioluminescent imaging (BLI) and micro PET will longitudinally asses the persistence of the infused cells, and the anti-tumor effect. Specific Aim #2 will evaluate the feasibility, safety, and persistence of infusing multivirus-specific CTLs modified to express CAR CD19-specific after UCBT. The correlative studies will delineate the (i) magnitude and duration of persistence, (ii) trafficking, and (iii) anti-viral and anti-leukemic effects of adoptively transferred CTLs. In preparation for human PET T-cell imaging, in Specific Aim #3, we will develop a nonhuman primate model Monkey-derived T cells will be genetically modified with CARs and TK, and infused. Longitudinal PET imaging using [18FJFEAU, metabolized by thymidine kinase (TK) will be used to evaluate the distribution of these adoptively transferred macaque T cells. In aggregate, the results of the studies will facilitate the evolution of targeting post-UCBT MRD with viral- and CD19-specific CTLs for enhanced disease-free survival of patients with B cell malignancies.