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，同种免疫患者的罕见表型血液通常无法获得。使用 当我们的实验室等实验室通过培养红细胞 (cRBC) 来满足这些临床需求时，我们获得了动力： 具备了大规模生产cRBC的条件。目前，包括我们在内的多个实验室正在 解决将 cRBC 建立为临床产品的众多障碍。 2011 年，Douay 实验室证明，2.5x1010 个自体 cRBC（5 mL 血液）由 动员的CD34pos细胞在体内具有正常的存活率。然而，这项开创性的研究有四个警告：1) 动员不太可能被接受作为生成用于临床或诊断用途的 cRBC 的程序，2) 生成的 cRBC 量（约 5 mL 血液）比输血所需的最小剂量低一个对数 儿童（50 mL 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 功效，4) 生产 cRBC 的高成本限制了其作为产品的临床应用。 该提案将通过提供临床有用剂量的 cRBC 的原理验证来解决这些问题 可以从废弃的干细胞来源中产生并用于同种免疫患者的输血 对于罕见表型的人来说，由于没有其他选择，生产 cRBC 的成本被认为是合理的。 该提案基于基础科学家（Migliaccio 博士和 Bieker 博士、伊坎博士）之间的密切合作 医学院和西奈山）和临床医生（输血医学部 Klein 和 Flegel 博士） 美国国立卫生研究院）。这些研究人员将进行实验，旨在确定白细胞减少是否通过- 目前从定期献血中丢弃的具有罕见表型的产品适合生成 2.5x1011 cRBC（目标 1）使用基于“基因编辑”的新颖培养策略，这将提高产量，同时 降低生产成本（目标 2）。 我们相信，记录在案的 Migliaccio 博士和 Bieker 博士以及两位博士之前的合作 新合作者（Funnel 博士和 Zon），研究与该应用程序相关的科学问题以及长期研究 Klein 博士和 Flegel 博士对镰状细胞性贫血患者的临床管理一直感兴趣，包括那些 具有罕见的表型，确保了该提案的科学和临床方面的协同发展。 这对于最大限度地提高有效生产足够数量的 cRBC 的可能性是必要的。 用于首次人类同种异体输血。