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（由申请人提供）：最初，由于输注的细胞剂量较低，脐带血（UCB）移植仅限于儿童。在儿科环境中，针对各种血液系统疾病和代谢性疾病，相关和不相关的脐带血移植均取得了很高的成功率。近年来，UCB 移植的使用不断扩大，成人脐带血移植的结果也有所改善，但也面临着许多挑战。全球 UCB 库存中仅有 10% 的有核细胞总数 (TNC) 可能不足以用于成人和儿童移植。极少的 TNC 计数会导致患者的康复严重延迟，使他们容易出现多种致命的并发症。为了充分发挥 UCB 对血液恶性肿瘤治疗患者的潜力和影响，需要对 UCB 材料进行离体扩增。目前造血干细胞和祖细胞（HSPC）的扩增机制缺乏适当的微环境，这可能解释了临床试验的失败。来自众多研究小组的证据不断增多，详细说明内皮细胞 (EC) 通过分泌多种生长因子（称为血管分泌因子）在促进 HSPC 中发挥着不可或缺的作用。传统的内皮细胞培养机制不允许重现内皮细胞产生的体内微环境。 VeraVec EC 平台通过添加 Ad5 E4ORF1 蛋白消除了 EC 中的这一限制。 VeraVec EC 已多次证明其在共培养条件下以高保真度和高移植性扩展 HSPC 的能力。临床前数据表明该平台能够在 12 天内将 UCB 扩展 1000-1800 倍。重要的是，这种扩展的材料保持了 HSPC 表型的所有方面，包括长期植入、连续移植和与未操作的 UCB 相同的多谱系承诺。然而，HSPC 和 VeraVec EC 的大规模共培养是劳动密集型、后勤挑战，并且在符合 cGMP 的设施中消耗大量试剂。这些障碍使得 HSPC 不太可能采用 VeraVec EC 平台。因此，我们建议将共培养引入 TerumoBCT Quantum 中空纤维生物反应器中，以实现扩增自动化。最终产品将是自动化、大规模、高保真扩展，而不会牺牲 VeraVec 平台的任何优势。