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: 10188596
• 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 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188596
• 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; 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; Supplementation; Transfusion; Umbilical Cord Blood; United States National Institutes of Health; Vascular blood supply; base; clinical center; clinical diagnostics; cost; design; experimental study; first-in-human; genetic makeup; immunogenic; improved; in vivo; insight; interest; large scale production; medical schools; novel; phosphoproteomics; stem cells; 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.

摘要 在发达国家，同种免疫患者的罕见表型血液通常不可用。使用 培养红细胞（cRBC），以解决这些临床需求获得了动力，当我们的实验室，除其他外， 开发了大规模生产cRBC的条件。目前，包括我们在内的几个实验室， 解决了将cRBC确立为临床产品的众多障碍。 在2011年，Douay实验室证明，2.5x1010个自体cRBC（5 mL血液）产生自 动员的CD 34阳性细胞在体内存活正常。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为产生用于临床或诊断用途的cRBC的程序，2） 产生的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次临床试验评估的理想受体 功效，以及4）生产cRBC的高成本限制了它们作为产品的临床用途。 该提案将通过提供临床有用剂量的cRBC的原理证明来解决这些警告 可以从废弃的干细胞来源产生，并用于输注同种免疫患者， 罕见的表型，因为没有替代品，生产cRBC的成本被认为是合理的。 这项建议是基于基础科学家之间的密切合作（Migliaccio和Bieker博士，Icahn 医学院和西奈山）和临床医生（Klein和Flegel博士，输血医学科 的NIH）。这些研究人员将进行旨在确定是否通过- 目前从具有罕见表型的常规献血中丢弃的产品适合于产生 2.5x1011 cRBC（目标1）使用基于“基因编辑”的新培养策略， 降低生产成本（目标2）。 我们认为，Migliaccio博士和Bieker博士以及两人之间的先前合作记录 新的合作者（漏斗和Zon博士），对有关这一应用和长期的科学问题， Klein博士和Flegel博士对镰状细胞贫血患者的临床管理一直感兴趣，包括那些 与罕见的表型，保证了该建议的科学和临床方面的协同发展。 这对于最大限度地有效生产足够数量的cRBC的可能性是必要的， 进行首次人体同种异体输血

项目成果

Whirling Platelets Away for Transfusion.

旋转血小板以输血。

• DOI: 10.1016/j.cell.2018.07.018
• 发表时间: 2018-07-26
• 期刊: Cell
• 影响因子: 64.5
• 作者: Iancu-Rubin C;Hoffman R;Migliaccio AR
• 通讯作者: Migliaccio AR

Dexamethasone Predisposes Human Erythroblasts Toward Impaired Lipid Metabolism and Renders Their ex vivo Expansion Highly Dependent on Plasma Lipoproteins.

地塞米松使人类成红细胞的脂质代谢受损，并使其离体增殖高度依赖于血浆脂蛋白。

• DOI: 10.3389/fphys.2019.00281
• 发表时间: 2019
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Zingariello,Maria;Bardelli,Claudio;Sancillo,Laura;Ciaffoni,Fiorella;Genova,MariaLuisa;Girelli,Gabriella;Migliaccio,AnnaRita
• 通讯作者: Migliaccio,AnnaRita