Preclinical development and clinical monitoring of adoptive immune therapy

过继免疫疗法的临床前开发和临床监测

• 批准号: 8938452
• 负责人: Frances Hakim
• 金额: $ 17.2万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8938452
• 关键词: Adoptive Transfer; Allogenic; Antigen-Presenting Cells; Autologous; Biological Assay; Biological Preservation; Blood; Blood Circulation; Bone Marrow; CD19 gene; CD8B1 gene; Cell Transplants; Cell physiology; Cells; Cessation of life; Clinic; Clinical; Clinical Trials; Core Facility; Custom; Development; Documentation; Donor Lymphocyte Infusion; Dose; Engraftment; Equipment; Experimental Designs; Generations; Genes; Goals; Helper-Inducer T-Lymphocyte; Immune; Immunologics; Immunotherapy; Inflammatory; Infusion procedures; Interferons; Interleukin-4; Laboratories; Lymphocyte; Lymphokines; Lymphoma; Malignant Neoplasms; Medicine; Methods; Molecular; Monitor; Multiple Myeloma; Outcome; Patient Monitoring; Patients; Plasma; Population; Procedures; Process; Production; Protocols documentation; Quality Control; RNA; Reagent; Relapse; Reporting; Residual state; Schedule; Services; Sirolimus; Site; Structure; Supporting Cell; T cell therapy; T-Cell Receptor; T-Lymphocyte; Tissues; Training; Transfection; Transfusion; Translations; Transplant Recipients; Transplantation; Transplantation Immunology; Tumor Tissue; Up-Regulation; Validation; Work; anergy; arm; bis(3-bis(4-chlorophenyl)methyl-4-dimethylaminophenyl)amine; cell injury; chimeric antigen receptor; cytokine; design; experimental analysis; graft vs host disease; immune activation; irradiation; laboratory facility; member; monocyte; nano-string; neoplastic cell; novel; pre-clinical; receptor; reconstitution; response; tumor

The Preclinical Development and Clinical Monitoring Facility (PDCMF) of the Experimental Transplantation and Immunology Branch supports the development and implementation of new protocols involving adoptive T cell therapies through preclinical development, translational implementation of clinical products and preservation and analysis of patient blood and tissues during clinical trials. Five novel protocols involving adoptive transfer of T cells have been implemented in recent years as a result of this process. (1) In 04-C-0055 Arm 4A, Daniel Fowler initially utilized expanded donor-derived CD4 helper cells grown for 12 days in IL-4 and rapamycin (T.Rapa.12) to enhance donor engraftment and reduce GVHD. The Preclinical Service supported implementation of clinical trial Arms 4B, utilizing a shorter 6 day expansion period to generate cells with increased anti-tumor potency (T.Rapa.6) and Arm 4C to implement an altered schedule of post transplant rapamycin. Following completion of each trial arm, we have characterized the early immune reconstitution following infusion of the T.Rapa product and have assessed the T cell receptor repertoire diversity of the infused T.Rapa cells and the transplant recipients at day 60 through spectratyping. We have assessed lymphokine production capacity post transplant to evaluate the extent and durability of the cytokine shift resulting from the infusion of Th2-rapa cells These studies have been reported in 2013 (Fowler et al, Blood, 2013). (2) The second novel product involves expansion of autologous T cells (both CD4 and CD8) in the presence of IFNalpha and rapamycin (T1.rapa), to generate a cell product that potentially has activity against residual myeloma plasma cells. In protocol 11-C-0016 (P.I. Claude Sportes/Daniel Fowler), patients with undergo an autologous transplant followed by infusion of T1.Rapa cells; through expanding the scale of product manufacturing, serial infusions of T1.Rapa cells are currently being implemented. Clinical production of these cells was developed through our staff in Cell Processing. Following initiation of this trial, we have monitored serial changes in lymphocyte populations in blood and in bone marrow and have provided plasma assays of inflammatory cytokines to monitor patient responses at each dose escalation. (3) We have supported the implementation of the first trial of the use of donor-derived anti-CD19 Chimeric Antigen Receptor (CAR) T cells in patients with relapsed or persistent lymphoma following allogeneic transplant (Protocol 10-C-0052, P.I. James Kochenderfer). We have tracked the presence of CAR T cells following adoptive transfer and have characterized expression of markers of activation and anergy. These studies have demonstrated the expansion of anti-CD19 CAR T cells in the blood concurrent with the onset of anti-tumor activity approximately one week after adoptive transfer (Kochenderfer et al, Blood 2013). (4) Furthermore, we have completed the translational implementation of a new CAR construct with activity against the BCMA receptor expressed on myeloma cells (Carpenter et al, Clin Cancer Res, 2013). We worked closely with the P.I. to establish the procedures for transfection of the CAR construct and expansion of cells, support functional assays of CAR T cell interferon production and validated the final GMP grade construct for use in a newly initiated trial. (5)Finally we are using cellular and molecular assays to assess a novel therapy (09-C-0224, P.I.: Nancy Hardy) utilizing irradiation of selected tumor sites followed by donor lymphocyte infusions (a standard method of immune therapy). Monitoring focuses on whether the irradiation and subsequent localized deaths of tumor cells has produced activation of T effectors and antigen presenting cells and increased anti-tumor immune activation. We have developed a multiplex RNA quantitation assay (Nanostring) using a custom panel of probes we designed to assess upregulation of genes induced by interferon (IFN) and genes responding to cell damage. Although developed to monitor changes in circulating monocytes in graft versus host disease (GVHD) (described in Project ZIC BC 010934), we have determined that these genes are upregulated following donor lymphocyte infusions and infusions of expanded donor T cells. We are implementing these assays on blood and tumor tissue after adoptive immune therapy.

实验移植和免疫学分支机构的临床前开发和临床监测设施（PDCMF）支持临床试验中临床产品的转化以及保存以及在临床试验期间患者血液和组织分析的新方案的开发和实施。由于这一过程，近年来已经实施了涉及T细胞继发性转移的五种新颖方案。 （1）在04-C-0055 ARM 4A中，Daniel Fowler最初利用了供体衍生的CD4辅助细胞在IL-4中生长了12天，而雷帕霉素（T.Rapa.12）则可以增强供体植入植物并减少GVHD。临床前服务支持临床试验臂4B的实施，利用较短的6天扩展期以增加抗肿瘤效力（T.RAPA.6）和ARM 4C产生细胞，以实施移植后雷帕霉素的时间表改变。在每个试验臂完成后，我们表征了在输注T.RAPA产品后的早期免疫重建，并评估了注入的T.RAPA细胞的T细胞受体库库多样性和在第60天通过频谱型的移植受者。我们已经评估了移植后的淋巴因子产能力，以评估2013年报道的Th2-Rapa细胞输注的细胞因子转移的程度和耐用性（Fowler等，Blood，2013）。 （2）第二种新型产物涉及在IFNALPHA和雷帕霉素（T1.RAPA）存在下自体T细胞（CD4和CD8）的扩展，以产生潜在具有对残留骨髓瘤血浆细胞活性的细胞产物。在第11-C-0016号方案（P.I. Claude Sportes/Daniel Fowler）中，患有自体移植的患者随后输注T1.Rapa细胞；通过扩大产品制造规模，目前正在实施T1.RAPA细胞的串行输注。这些细胞的临床生产是通过我们的细胞加工的员工开发的。在该试验开始后，我们监测了血液和骨髓中淋巴细胞种群的串行变化，并提供了炎性细胞因子的血浆测定，以监测每次剂量升级时患者反应。 （3）我们支持在同种异体移植后复发或持续性淋巴瘤的患者中使用供体衍生的抗CD19嵌合抗原受体（CAR）T细胞的第一项试验（方案10-C-0052，P.I. James Kochenderfer）。我们已经跟踪了产物转移后的CAR T细胞的存在，并表征了激活和反感标志物的表达。这些研究表明，抗CD19 CAR T细胞在血液同时发生的血液中与抗肿瘤活性的发作大约一周后（Kochenderfer等人，2013年）。 （4）此外，我们已经完成了一种新的汽车构建体的翻译实施，其活性针对在骨髓瘤细胞上表达的BCMA受体（Carpenter等，Clin Cancer Res，2013年）。我们与P.I.为了建立转染CAR构建体和细胞扩展的程序，支持CAR T细胞干扰素生产的功能测定，并验证了最终的GMP级构建体，以在新启动的试验中使用。 （5）最后，我们使用细胞和分子测定法来评估一种新的疗法（09-C-0224，P.I。：Nancy Hardy），利用对选定肿瘤部位的照射，然后是供体淋巴细胞输注（一种标准的免疫治疗方法）。监测的重点是肿瘤细胞的辐射和随后的局部死亡是否产生了T效应子和抗原呈递细胞的激活，并增加了抗肿瘤免疫激活。我们已经使用我们旨在评估干扰素（IFN）诱导的基因的上调和对细胞损伤响应的基因的基因上调的自定义探针图案开发了多重RNA定量测定法（纳米结构）。尽管开发用于监测移植物与宿主疾病（GVHD）中循环单核细胞的变化（在ZIC BC 010934项目中进行了描述），但我们确定供体淋巴细胞输注和供体T细胞的输注后，这些基因在供体淋巴细胞输注后上调。我们正在继承免疫治疗后对血液和肿瘤组织实施这些测定。