Development Of Methods For Ex Vivo Cultured And Immunologically And/or Geneticall

离体培养、免疫学和/或遗传学方法的开发

• 批准号: 7733562
• 负责人: David Stroncek
• 金额: $ 7.87万
• 依托单位: CLINICAL CENTER
• 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/7733562
• 关键词: ADA protocol; Allogenic; Antibiotics; Antibodies; Autologous; Bone Marrow; CD8B1 gene; Cells; Child; Chronic Granulomatous Disease; Clinical Trials; Collaborations; Complex; Development; Engraftment; Enrollment; Fibronectins; Graft-Versus-Tumor Induction; Harvest; Hematopoietic; Hour; Immunologics; Immunomagnetic Separation; Immunotoxins; Infusion procedures; Institutes; Investigation; Length; Link; Lymphocyte; Methods; Modeling; Mus; Numbers; Patients; Peripheral Blood Stem Cell; Phase I Clinical Trials; Photosensitizing Agents; Population Heterogeneity; Preparation; Process; Protocols documentation; Research Personnel; Resolution; Serum; Sirolimus; Stem cell transplant; System; T-Lymphocyte; T-Lymphocyte Subsets; Techniques; Th2 Cells; Transduction Gene; Transplantation; United States Food and Drug Administration; United States National Institutes of Health; cell killing; cytokine; gene therapy clinical trial; genetic manipulation; graft vs host disease; improved; interest; leukemia; lymphocyte product; method development; pre-clinical; programs; research study; scale up; transduction efficiency; tumorigenesis; vector; vector-induced

Preclinical development of complex processing systems for ex vivo culture-expanded lymphohematopoietic cells, with subsequent immunologic and/or genetic manipulation, have been carried out in collaboration with a number of NIH institute investigators, as follows: Preparation of allogeneic donor lymphocytes selectively depleted for alloreactive T-cells using an anti-CD25 immunotoxin (collaboration with NHLBI, Stem Cell Transplant Program): In FY2001, we completed development and scale up of this complex process, and in September 2001, initiated a phase I clinical trial of DLI selectively depleted of donor-specific alloreactivity in the setting of allogeneic hematopoietic transplantation. In FY2002, 3 patients were entered on the clinical trial and received their transplants plus SD cells. This year, we also made plans to evaluate 2 cell manipulation strategies as alternatives to the immunotoxin: one, an immunomagnetic separation method and the other, UV light-activated killing of cells after incubation with a photosensitizing agent. During FY2006, the UV light-activating killing of cells as an alternative to immunotoxin was submitted in an NHLBI sponsored IND clinical trial. Preparation of donor Th2 T cells for clinical trials (collaboration with NCI): Development of this process incorporated CD8/CD20 depletion and CD3/CD28 bead stimulation, which produces a lymphocyte product that is 95% CD4+ and < 1% CD8+. The clinical trial was initiated in March 2001, and 27 patients have been treated to date through FY2002. In FY2004, Th2 cells cultured in the presence of sirolimus, in murine models, promoted engraftment of allogeneic HPC with lower occurrence of GVHD. The clinical trial was initiated using CD8/20 depletion process which was later replaced by CD4+ enrichment by immunomagnetic selection during FY2005. During FY2006, the trials using these cultured products continued enrollment. Fibronectin transduction: A method for improved gene transduction using fibronectin-coated bags was previously developed and incorporated into the clinical trial of gene therapy for chronic granulomatous disease; that study was completed in 2001. At the end of FY2001, this method was adapted to a new clinical trial of gene therapy in ADA deficiency that takes advantage of new vectors. To date, very high transduction efficiencies (>80%) have been observed, and 2 patients have been treated. In FY2002, methods were improved (with new cytokines) and validated for gene transduction of autologous PBSC and bone marrow of patients with X-linked SCID, and this clinical trial will begin pending resolution of scientific and FDA investigation of the child with apparent vector-induced oncogenesis. During FY2006, both protocols for ADA deficiency and X-linked SCID were re-initiated. For the X-linked SCID protocol, a method of vector concentration using an ultra-centrifuge was evaluated. Initial experiments showed no difference in transduction efficiency.

与NIH研究所的一些研究人员合作，对体外培养扩增的淋巴造血细胞的复杂处理系统进行了临床前开发，随后进行了免疫学和/或遗传学操作，如下所示： 使用抗CD 25免疫毒素制备选择性去除同种异体反应性T细胞的同种异体供体淋巴细胞（与NHLBI干细胞移植计划合作）：在2001财政年度，我们完成了这一复杂过程的开发和规模扩大，并于2001年9月启动了在同种异体造血移植背景下选择性去除供体特异性同种异体反应性的DLI的I期临床试验。 在2002财政年度，3名患者进入临床试验，并接受了他们的移植加SD细胞。 今年，我们还计划评估2种细胞操作策略作为免疫毒素的替代品：一种是免疫磁性分离方法，另一种是与光敏剂孵育后的紫外光激活细胞杀伤。在2006财年期间，在NHLBI申办的IND临床试验中提交了UV光激活细胞杀伤作为免疫毒素的替代品。 用于临床试验的供体Th 2 T细胞的制备（与NCI合作）：该过程的开发包括CD 8/CD 20耗竭和CD 3/CD 28珠刺激，其产生95% CD 4+和< 1% CD 8+的淋巴细胞产物。临床试验于2001年3月开始，到2002财政年度为止，已有27名患者接受了治疗。在2004财年，在存在西罗莫司的小鼠模型中培养的Th 2细胞促进了同种异体HPC的植入，GVHD的发生率较低。 临床试验开始时使用了CD 8/20去除过程，后来在2005财年期间被免疫磁性选择的CD 4+富集所取代。 在2006财年，使用这些培养产品的试验继续招募。 纤连蛋白转导：先前开发了一种使用纤连蛋白包被袋改善基因转导的方法，并将其纳入慢性肉芽肿病基因治疗的临床试验中;该研究于2001年完成。 在2001财政年度末，该方法适用于利用新载体的ADA缺乏症基因治疗的新临床试验。迄今为止，已经观察到非常高的转导效率（>80%），并且已经治疗了2名患者。在2002财政年度，改进了方法（使用新的细胞因子），并验证了X连锁SCID患者自体PBSC和骨髓的基因转导，该临床试验将开始，等待科学和FDA对具有明显载体诱导肿瘤发生的儿童的研究结果。在2006财年期间，重新启动了ADA缺乏症和X连锁SCID的方案。对于X-连锁SCID方案，评价了使用超离心机的载体浓缩方法。初始实验显示转导效率没有差异。