T-cell Therapy for B-lineage Acute Lymphoblastic Leukemia

B 系急性淋巴细胞白血病的 T 细胞疗法

• 批准号: 8112556
• 负责人: Laurence J.N. Cooper
• 金额: $ 31.8万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8112556
• 关键词: Ablation; Acute Lymphocytic Leukemia; Address; Adoptive Immunotherapy; Adoptive Transfer; Allogenic; Anatomic Sites; Antigen Receptors; Apoptotic; Applications Grants; Arabinofuranosyluracil; Asses; Autologous; B-Cell Neoplasm; B-Lymphocytes; Back; Binding; Biodistribution; Biological Response Modifier Therapy; Biopsy; Blast Cell; Bone Marrow; Bone marrow biopsy; CD19 Antigens; CD19 gene; CD28 gene; CD3 Antigens; CD80 gene; Cancer Relapse; Cell Lineage; Cell Therapy; Cell surface; Cells; Clinical Trials; Correlative Study; Cytolysis; DNA; Data; Development; Disease remission; Disease-Free Survival; Dose; Electroporation; Engineering; Engraftment; Event; Evolution; Exhibits; Firefly Luciferases; Funding; Ganciclovir; Gene Transfer; Generations; Genes; Grant; Hematopoietic Stem Cell Transplantation; Herpesvirus 1; Human; Image; Immune response; Immunosuppression; Incidence; Infusion procedures; Interleukin-2; Luciferases; Lymphocyte; Major Histocompatibility Complex; Malignant Neoplasms; Mediating; Membrane; Mus; Non-Hodgkin' s Lymphoma; Patients; Phase I Clinical Trials; Plasmids; Positron-Emission Tomography; Probability; Production; Recurrence; Relapse; Residual Neoplasm; Residual Tumors; Resistance; Safety; Sampling; Scheme; Signal Transduction; Site; Sleeping Beauty; Specificity; Specimen; System; T-Cell Activation; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TK Gene; Thymidine Kinase; Time; Toxic effect; Transgenes; Transplantation; Transposase; Treatment Failure; base; chemotherapy; cohort; conventional therapy; cost; cytokine; design; functional status; gene therapy; graft vs host disease; high risk; immunogenicity; improved; in vivo; innovation; killings; leukemia; mouse model; neoplastic cell; new technology; next generation; plasmid DNA; trafficking; transgene expression; tumor; vector

DESCRIPTION (provided by applicant): This revised R01 grant addresses the problem of relapse of B-lineage acute lymphoblastic leukemia (B-ALL) after allogeneic hematopoietic stem-cell transplantation (HSCT). We hypothesize that the incidence of cancer relapse following allogeneic HSCT can be reduced by targeting post-transplant B-ALL minimal residual disease (MRD) with adoptively transferred donor-derived T cells genetically modified to be specific for CD19. To consolidate HSCT, we have designed a next-generation chimeric antigen receptor (CAR), designated CD19RCD28, to redirect specificity of T cells to the B-cell lineage-restricted cell-surface molecule CD19 independent of major histocompatibility complex (MHC). Genetically modified CD19RCD28+ T cells activated through chimeric CD28 and CD3-6 lyse B-ALL, upregulate production of IL-2 and anti-apoptotic genes, in a CAR-regulated manner. The Sleeping Beauty (SB) system has been combined with electroporation to introduce the CAR as well as co-express HSV-1 thymidine kinase (TK) for imaging by positron emission tomography (PET). The studies in Aim #1 will now evaluate whether an all-human CD19-specific CAR can be developed (hCD19RCD28) that provides a fully-competent activation signal as determined by CD19-dependent killing, cytokine production, and sustained proliferation in T cells that have been genetically modified by SB transposition. A xenogeneic mouse model of disseminated B-lineage tumor will be used to ascertain the feasibility and safety of adoptive therapy using non-invasive bioluminescent imaging (BLI) and <PET to longitudinally asses the persistence of the infused CAR+TK+ cells and the anti-tumor effect. Aim #2 will evaluate the safety, feasibility and persistence, of infusing escalating doses of donor-derived hCD19RCD28+ T cells with/without TK expression, after allogeneic HSCT for high-risk CD19+ B-ALL. T cells expressing TK will be imaged by PET. If necessary, ganciclovir (GCV) will be given for conditional ablation of TK+ cells in the event of serious toxicity. Correlative studies in Aim #3 will delineate the magnitude and persistence of transferred T cells at the prescribed T-cell Dose Levels using vector-specific Q-PCR and TCR spectratyping analyses on serially acquired specimens. Other correlative studies will evaluate the trafficking to sampled bone marrow (BM) of adoptively transferred T cells and the functional status of transferred T cells in this anatomic site of MRD. Human PET imaging using 2'-Deoxy-20-[18F]fluoro-5-ethyl-1-2-D-arabinofuranosyluracil ([18F]-FEAU) metabolized/trapped by TK co-expressed in infused CAR+ T cells, will be used to evaluate the distribution of adoptively transferred T cells. In aggregate, the results of the studies will facilitate the evolution of targeting post-HSCT MRD with donor-derived CD19-specific T cells for enhanced disease-free survival of patients with B-ALL. LAY SUMMARY: We will infuse CD19-specific T cells after transplantation to improve survival for patients with acute lymphoblastic leukemia.

描述(申请人提供)：这项修订后的R01资助针对B系急性淋巴细胞性白血病(B-ALL)在异基因造血干细胞移植(HSCT)后复发的问题。我们假设，通过过继转移供者来源的T细胞以针对CD19进行过继转移，可以降低异基因HSCT后癌症复发的发生率。为了巩固HSCT，我们设计了下一代嵌合抗原受体(CAR)，命名为CD19RCD28，将T细胞的特异性重定向到不依赖主要组织相容性复合体(MHC)的B细胞谱系限制性细胞表面分子CD19。转基因CD19RCD28+T细胞通过嵌合CD28和CD3-6溶解B-ALL激活，以CAR调节的方式上调IL-2和抗凋亡基因的产生。睡美人(SB)系统已与电穿孔相结合，引入CAR并共表达HSV-1胸苷激酶(TK)，用于正电子发射断层扫描(PET)成像。AIM#1中的研究现在将评估是否可以开发出一种全人CD19特异性CAR(HCD19RCD28)，该CAR提供由CD19依赖的杀伤、细胞因子产生和已通过SB转座转基因的T细胞的持续增殖所确定的完全有效的激活信号。我们将使用一种播散性B细胞系肿瘤的异种小鼠模型来确定采用非侵入性生物发光成像(BLI)和~lt；PET进行过继治疗的可行性和安全性，以纵向评估注入的CAR+TK+细胞的持久性和抗肿瘤效果。目的#2将评估高危CD19+B-ALL异基因造血干细胞移植后，输注剂量递增的有/无TK表达的hCD19RCD28+T细胞的安全性、可行性和持久性。表达TK的T细胞将被PET成像。如有必要，在发生严重毒性时，将给予更昔洛韦(GCV)用于有条件地消融TK+细胞。AIM#3中的相关研究将通过对连续采集的标本进行载体特异性Q-PCR和TCR分型分析，来描述在规定的T细胞剂量水平下转移的T细胞的大小和持久性。其他相关研究将评估过继移植的T细胞向样本骨髓(BM)的转运，以及移植的T细胞在MRD解剖部位的功能状态。利用2‘-Deoxy-20-[18F]fluoro-5-ethyl-1-2-D-arabinofuranosyluracil([18F]-FeAU)代谢/捕获TK共表达在输注的CAR+T细胞中的人正电子发射计算机断层扫描将用于评估过继转移的T细胞的分布。总而言之，这些研究的结果将促进以供体来源的CD19特异性T细胞为靶点的HSCT后MRD的发展，以提高B-ALL患者的无病生存率。总结：我们将在移植后输注CD19特异性T细胞，以提高急性淋巴细胞白血病患者的存活率。