Generation of cultured RBCs with rare phenotypes for transfusion from sources usually discarded during regular blood donations

产生具有罕见表型的培养红细胞，用于从定期献血期间通常丢弃的来源进行输血

• 批准号: 9789365
• 负责人: JAMES J BIEKER
• 金额: $ 42.38万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789365
• 关键词: Address; Adult; Age; Agonist; Allogenic; Alloimmunization; Anemia; Autologous; Biology; Blood; Blood Donations; Blood donor; Cells; Centrifugation; Child; Chromium; Chronic; Clinical; Clinical Management; Clinical Trials; Collaborations; Databases; Developed Countries; Developing Countries; Development; Dexamethasone; Diagnostic; Donor Selection; Dose; Erythroblasts; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; Filtration; Gender; Generations; Genes; Genetic; Glucocorticoid Receptor; Goals; Hematopoietic stem cells; Human; Individual; Isoantibodies; Label; Laboratories; Lead; Leukocytes; Ligands; Methods; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Polycythemia Vera; Procedures; Process; Production; Reagent; Regulation; Research Personnel; Role; Safety; Scientist; Sickle Cell Anemia; Signal Pathway; Signal Transduction; Source; Stem Cell Factor; Stem cells; Supplementation; Transfusion; Umbilical Cord Blood; United States National Institutes of Health; Vascular blood supply; base; clinical diagnostics; cost; design; experimental study; first-in-human; genetic makeup; immunogenic; improved; in vivo; insight; interest; large scale production; medical schools; novel; phosphoproteomics; trait; transfusion medicine; volunteer

Abstract In developed countries, blood with rare phenotypes for alloimmunized patients is often unavailable. The use of cultured RBCs (cRBCs) to address these clinical needs gained momentum when our laboratory, among others, developed conditions for large scale production of cRBCs. Presently, several laboratories, including ours, are addressing the numerous barriers to establish cRBCs as clinical product. In 2011, the Douay laboratory demonstrated that 2.5x1010 autologous cRBCs (5 mL of blood) generated from mobilized CD34pos cells have normal survival in vivo. However, this ground breaking study has four caveats: 1) mobilization is unlikely to be accepted as procedure to generate cRBCs for clinical or diagnostic use, 2) the amount of cRBCs generated (~5 mL of blood) is one-log lower than the minimal dose required to transfuse a child (50 mL or 2.5x1011 RBCs), 3) normal volunteers are not ideal recipients for the first clinical trial to assess efficacy, and 4) the high costs to produce the cRBC limit their clinical use as product. This proposal will address these caveats by providing proof-of-principle that clinically useful doses of cRBCs can be generated from discarded stem cell sources and used for transfusion of alloimmunized patients with rare phenotypes for whom, having no alternatives, the costs to produce cRBCs is considered reasonable. This proposal is based on a close collaboration among basic scientists (Drs. Migliaccio and Bieker, Icahn School of Medicine and Mount Sinai) and clinicians (Drs. Klein and Flegel, Transfusion Medicine Department of the NIH). These investigators will perform experiments aimed to establish whether leukoreduction by- products currently discarded from regular blood donations with rare phenotypes are suitable to generate 2.5x1011 cRBCs (Aim 1) using novel culture strategies based on “gene editing” that will increase yields while reducing production costs (Aim 2). We believe that the documented previous colloboration between Drs Migliaccio and Bieker, as well as the two new collaborators (Drs. Funnel and Zon), on scientific questions related to this application and the long standing interest of Drs Klein and Flegel in clinical management of Sickle Cell Anemia patients, including those with rare phenotypes, assures the synergistic development of scientific and clinical aspects of this proposal. This is necessary to maximize the likelihood of efficiently producing adequate numbers of cRBCs well-suited for the first-in-man allogenic transfusion.

抽象的 在发达国家中，对于同种免疫患者的血液通常不可用。使用 培养的RBC（CRBC）以满足这些临床需求时，我们的实验室等人等 开发了CRBC大规模生产的条件。目前，包括我们在内的几个实验室 解决将CRBC作为临床产品建立的众多障碍。 2011年，杜伊实验室表明，由2.5x1010自体CRBC（5 mL血液）产生 动员的CD34POS细胞在体内生存正常。但是，这项突破性研究有四个警告：1） 动员不太可能被接受为生成临床或诊断使用的CRBC的程序，2） 产生的CRBC量（血液的〜5 ml）比传播A的最小剂量低一量。 儿童（50毫升或2.5x1011 RBC），3）正常志愿者不是第一个临床试验评估的理想接受者 效率和4）产生CRBC的高成本限制了其作为产品的临床用途。 该建议将通过提供临床上有用剂量的CRBC的原理证明来解决这些警告 可以由废弃的干细胞来源产生，用于输血 罕见的表型，没有其他选择，生产CRBC的成本被认为是合理的。 该提议基于基础科学家之间的密切合作（Migliaccio博士和Bieker，ICAHN 医学院和西奈山）和临床医生（输血医学系Klein和Flegel博士 NIH）。这些研究者将执行旨在确定白细胞化的实验 目前从常规捐赠稀有表型中丢弃的产品适合生成 2.5x1011 CRBC（AIM 1）使用基于“基因编辑”的新文化策略，这些策略将增加产量，同时增加产量 降低生产成本（AIM 2）。 我们认为，Migliaccio博士和Bieker博士以及两个 新的合作者（Dr.Funnel and Zon Drs），关于该应用程序和长期有关的科学问题 Klein博士和Flegel在镰状细胞贫血患者的临床管理中的兴趣，包括 凭借罕见的表型，假定该提案的科学和临床方面的协同发展。 这是最大程度地生产足够数量的CRBC的可能性，这是必不可少的 对于人类的同类异性输血。