Development of an automated Umbilical Cord Blood Hematopoietic Stem Cell expansion by endothelial cells for transplantation

开发用于移植的内皮细胞自动扩增脐带血造血干细胞的方法

• 批准号: 8980709
• 负责人: Daniel Joseph Nolan
• 金额: $ 18.55万
• 依托单位: ANGIOCRINE BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8980709
• 关键词: Acute Myelocytic Leukemia; Adoption; Adult; Allogenic; Biology; Bioreactors; Blood Cells; Blood Vessels; CD34 gene; Cell Count; Cell Transplantation; Cells; Child; Childhood; Clinical; Clinical Data; Clinical Trials; Coculture Techniques; Consumption; Cyclic GMP; Development; Devices; Disease; Disease-Free Survival; Dose; Economics; Endothelial Cells; Engraftment; Equipment and supply inventories; Evaluation; Exposure to; Failure; Fiber; Foundations; Growth Factor; Harvest; Healthcare; Hematologic Neoplasms; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Human Resources; In Vitro; Left; Liquid substance; Maintenance; Metabolic Diseases; Mind; Modeling; Movement; Mus; Pathway interactions; Patients; Phenotype; Play; Proteins; Proto-Oncogene Proteins c-akt; Qualifying; Reagent; Recovery; Research; Resources; Retroviridae; Role; Safety; Serum; Signal Transduction; Source; Stem cell transplant; Stem cells; Structure; System; Therapeutic; Time; Training; Transplantation; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; Work; base; cell type; flasks; genome integrity; graft vs host disease; improved; in vivo; mortality; outcome forecast; pre-clinical; public health relevance; quantum; stem; stem cell niche; stem cell population; stemness; success

 DESCRIPTION (provided by applicant): Initially, umbilical cord blood (UCB) transplantation was limited to children, given the low cell dose infused. Both related and unrelated cord blood transplants have been performed with high rates of success for a variety of hematologic disorders and metabolic diseases in the pediatric setting. In more recent years the use of UCB transplants has expanded and results for adult umbilical cord blood transplantation have improved, but not without a number of challenges. As little as 10% of the world-wide inventory of UCB may be of insufficient total nucleated cell count (TNC) for both adult and pediatric transplants. The minimal TNC count results in a profound delay in recovery of the patients leaving them susceptible to numerous lethal complications. In order to realize the full potential and impact UCB can have for patients treated for hematological malignancies, ex vivo expansion of the UCB material is necessitated. Current mechanisms for the expansion of Hematopoietic Stem and Progenitor Cells (HSPCs) lack the proper microenvironment, possibly explaining the clinical trial failures. Evidence continues to mount from numerous groups detailing the indispensable role endothelial cells (ECs) have in promoting the HSPC via the secretion of numerous growth factors, termed angiocrine factors. Traditional mechanisms of culturing endothelial cells have not been permissible to recapitulate the in vivo microenvironment generated by ECs. This limitation has been eliminated in ECs by the VeraVec EC platform by the addition of the Ad5 E4ORF1 protein. VeraVec ECs have repeatedly demonstrated their capacity to expand HSPCs with high fidelity and high engraftability in co-culture conditions. Pre-clinical data demonstrates the capacity for the platform to expand UCB 1000-1800 fold over 12 days. Importantly, this expanded material maintains all aspect of the HSPC phenotype including long term engraftment, serial transplant, and multilineage commitment equal to unmanipulated UCB. However, large scale co-cultures of HSPCs and VeraVec ECs are labor intensive, logistically challenging, and consume large quantities of reagents in cGMP compliant facilities. These barriers make the adoption of the VeraVec EC platform for HSPC unlikely. We therefore propose to introduce the co- culture into the TerumoBCT Quantum hollow fiber bioreactor to automate the expansion. The end product will be an automated, large scale, high fidelity expansion without sacrificing any benefits of the VeraVec platform.

 描述（由适用提供）：鉴于液化剂量的低细胞剂量，最初，脐带血（UCB）的移植仅限于儿童。对于各种血液学疾病和小儿环境中的多种血液学疾病和代谢疾病，相关和无关的绳索血液移植都以很高的成功率进行。近年来，UCB移植物的使用已经扩大，成人脐带血移植的结果有所改善，但没有挑战很多。成人和小儿移植的全球UCB全球库存中的10％可能是总核细胞计数（TNC）的不足。最小的TNC计数导致患者恢复的延迟很大，使他们容易受到许多致命并发症的影响。为了实现UCB对血液系统恶性肿瘤治疗的患者的全部潜力和影响，必须在体内扩展UCB材料。当前造血干细胞和祖细胞（HSPC）扩展的机制缺乏适当的微环境，可能会解释临床试验失败。证据继续来自众多小组，详细介绍了内皮细胞（EC）在通过分泌多种生长因子（称为血管分泌因子）的分泌来促进HSPC方面所具有的作用。不允许使用聚类内皮细胞的传统机制概括由EC产生的体内微环境。通过添加AD5 E4ORF1蛋白，Veravec EC平台在EC中消除了这种限制。 Veravec EC反复证明了他们在共同文化条件下具有高保真度和高耐受性的HSPC的能力。临床前数据显示了该平台在12天内扩展UCB 1000-1800倍的能力。重要的是，这种扩展的材料维持了HSPC表型的所有方面，包括长期植入，连续移植和等于未经操纵的UCB的多ineage承诺。但是，HSPC和VERAVEC EC的大规模共同培养是劳动力密集的，逻辑上的挑战，并且在符合CGMP的设施中消耗了大量试剂。这些障碍使得不太可能采用HSPC的Veravec EC平台。因此，我们建议将共培养物引入Terumobct量子空心纤维生物反应器以自动化扩张。最终产品将是自动化的，大规模的高保真扩展，而无需牺牲Veravec平台的任何好处。