Tumor-Infiltrating Donor Derived Lymphocyte Therapy After Allogeneic Transplant

同种异体移植后肿瘤浸润供体来源的淋巴细胞治疗

• 批准号: 7970006
• 负责人: Frances Hakim
• 金额: $ 19.14万
• 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/7970006
• 关键词: Allogenic; Amendment; Antigens; B lymphoid malignancy; B-Cell Lymphomas; B-Lymphocytes; Biological Assay; Bone Marrow; CD8B1 gene; Cell Count; Cell Culture Techniques; Cell Therapy; Cells; Chimerism; Clinic; Clinical; Clinical Protocols; Clinical Trials; Development; Documentation; Donor Lymphocyte Infusion; Effector Cell; Endotoxins; Excision; Feasibility Studies; Flow Cytometry; Frequencies; Generations; Goals; Hematopoietic Stem Cell Transplantation; Hodgkin Disease; Homing; Homologous Transplantation; Immune; Infusion procedures; Interferon Type II; Investigational Drugs; Link; Lymphocyte; Lymphoma; Malignant Neoplasms; Malignant lymphoid neoplasm; Marrow; Mediating; Methods; Multiple Myeloma; Nodule; Patients; Population; Preclinical Testing; Procedures; Process; Production; Progressive Disease; Protocols documentation; Reagent; Refractory Disease; Relapse; Research; Residual state; Site; Specificity; T-Cell Lymphoma; T-Lymphocyte; TNFSF5 gene; Testing; Therapeutic; Tissues; Toxic effect; Translations; Transplantation; Withdrawal; base; cytokine; efficacy testing; graft vs host disease; immune function; in vivo; lymphocyte product; manufacturing process; meetings; microbial; neoplastic cell; novel; outcome forecast; patient population; perforin; pre-clinical; tumor

We supported the preclinical development of the TDL clinical protocol (P.I. Michael Bishop; 07-C-0064) by establishing the feasibility of expanding TDL in cultures through the use of anti CD3/anti CD8 beads, resulting in a product with high viability, donor origin and T cell content, very low residual B cell numbers, free of endotoxin or contamination; In this process, we developed methods for viably dissociating cells and culturing them under good manufacturing process (GMP) standards. Furthermore we supported the clinical approval process by developing the necessary documentation of standard operating procedures and certificates of analysis for product release, assembling GMP reagents lists, and providing links to established investigational new drug (IND) protocols for manufacture of an clinical product suitable for infusion into patients. Following the approval of this protocol, we have continued to assess T cell expansion and clearance of B cell lymphoma and Hodgkins disease populations in the TDL expansion cultures in the first six patients. We have characterized the TDL product from both preclinical test cultures and from the first clinical trial cultures, using multiparameter flow cytometry and cytokine production assays. We have demonstrated that the TDL expansion cultures result in the disappearance of lymphoma cells and a marked decline in the frequency of regulatory T cells found in the original tumor population. The final product contains more than 90% T cells, primarily T-Bet+ Th1/Tc1 cells, that have elevated expression of effector molecules including CD40L, NKG2D, and perforin, and produce primarily IFN-gamma on stimulation. These assays will form the basis for tests of efficacy and specificity of anti-tumor activity that will be used to optimize the TDL product. Furthermore we have supported a clinical initiative to make this therapy available to a broader patient population by demonstrating the feasibility of using patient bone marrow (rather than surgically excised lymphoma nodules) as a basis for generating TDL in patients with marrow-resident tumor populations. We have successfully expanded replicate cultures from patient marrow collected following allogeneic transplant, demonstrating significant expansion of donor-derived T cells that meet criteria for T cell numbers and viability, donor chimerism, removal of tumor cells and microbial standards. These tests have supported submission of a protocol amendment providing for use of marrow. Finally we have initiated development of alternative culture conditions to optimize not only the numerical expansion of donor-derived T cells from the lymphoma tissue, but also anti-tumor activity and persistence in vivo after re-infusion. We have altered the cytokine milieu of the culture to enhance retention of activated CD8 effectors.

我们通过使用抗CD3/抗CD8珠子来建立扩展TDL在培养物中扩展TDL的可行性，从而支持TDL临床方案的临床前开发（P.I. Michael Bishop; 07-C-0064），从而导致具有高活力，供体的生存力和T细胞含量，非常低的B细胞数量，非常剩余的B细胞数量，自由氧氧蛋白或contantination，Ondiantination，Ondiantination of Endoxin of Endoxin of Endoxin或Contam of form of Endoxin;在此过程中，我们开发了在良好的制造过程（GMP）标准下逐步解离细胞和培养它们的方法。此外，我们通过制定标准操作程序的必要文档和产品释放，组装GMP试剂列表的必要文档，并提供与已建立的研究新药（IND）制造适合于输注患者的临床产品的链接的必要文档。在获得该方案的批准后，我们​​继续评估前六名患者TDL扩张培养物中B细胞淋巴瘤和Hodgkins病种群的T细胞扩张和清除率。我们使用多参数流式细胞术和细胞因子生产测定法表征了临床前测试培养物和第一个临床试验培养物的TDL产物。我们已经证明，TDL膨胀培养物导致淋巴瘤细胞消失，并且在原始肿瘤种群中发现的调节性T细胞的频率显着下降。最终产物含有90％以上的T细胞，主要是T-Bet+ Th1/Tc1细胞，它们的效应分子表达升高，包括CD40L，NKG2D和Perforin，并且主要在刺激上产生IFN-Gamma。这些测定将构成用于优化TDL产品的抗肿瘤活性功效和抗肿瘤活性特异性的基础。此外，我们还支持一项临床倡议，以证明使用患者骨髓（而不是手术切除的淋巴瘤结节）作为在骨髓抑制肿瘤群体中产生TDL的基础，从而使更广泛的患者人群可以使用这种疗法。我们已经成功地扩展了从同种异体移植后收集的患者骨髓中的重复培养物，表明符合T细胞数量和生存能力标准的供体衍生的T细胞显着扩展，供体嵌合，去除肿瘤细胞和微生物标准。这些测试支持提交规程修正案，以使用骨髓。最后，我们启动了替代培养条件的发展，不仅优化了淋巴瘤组织中供体衍生的T细胞的数值扩张，而且还优化了抗肿瘤活性和抗肿瘤活性和重新输注后体内的持久性。我们改变了培养物的细胞因子环境，以增强活化的CD8效应子的保留率。