Clinical Grade Cultured Rare Red Blood Cells as Reagents and Future Transfusion Support

临床级培养的稀有红细胞作为试剂和未来的输血支持

• 批准号: 9265547
• 负责人: ERIC E BOUHASSIRA
• 金额: $ 52.44万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265547
• 关键词: Clinical; Grade; Cultured; Rare; Red

 DESCRIPTION (provided by applicant): A major problem for transfusion therapy for chronic anemia is the high degree of genetic diversity in blood group antigens in people of Asian and African backgrounds compared to white Europeans, who are the majority of blood donors. These polymorphic antigens on RBCs contribute to the high incidence of allo-immunization and presence of multiple antibodies in the serum of these patients. Identifying the specificity of thes antibodies and determining if they are auto- or allo- antibodies is critical for providing compatibe blood. The major goal of this application is to take advantage of the progress that has been made in the in vitro production of cultured red blood cells (cRBCs) by expansion of hematopoietic stem cells and hematopoietic progenitor cells, coincident with the progress made in the genetic characterization of rare blood groups, to develop reagent cRBCs to identify the presence of clinically significant antibodies to high prevalence donor antigens. Reagent cRBCs will facilitate antibody identification in highly allo-immunized multiply transfused patients, and streamline and standardize testing. We have designed a panel of 6 donors with rare blood group phenotypes that are sufficient to resolve most antibody identification problems seen in patients who have auto- and allo-antibodies, and that could be used to transfuse patients who currently cannot be transfused because of shortage of appropriate rare blood. In Aim 1, we propose to develop new culture methods to expand hematopoietic cells collected from peripheral blood or differentiated from iPSCs by sequential amplification of the HSC, HPC and erythroblast compartments. We also propose to generate induce-pluripotent stem cells from these 6 donors and to engineer a group O, Rh null line of iPSCs that are useful as reagent red cells and that could be used as "universal" cells for transfusion. Expression of surface antigens on cRBCs produced from peripheral blood or from iPSCs has not been extensively characterized. In Aim 2, we propose to compare cRBCs to native red blood cells from the same donor to determine if they would be suitable as reagent cells and for transfusion. We will focus on the antigenic profiles, the genotypes, the storage stability, and the performance of cRBCs in traditional blood bank assays. Importantly, the reagent cRBCs that we will produce are the same cells that are in short supply for transfusion of allo-immunized patients. The method development and the quality testing that we propose to perform will therefore pave the way to the production of sufficient amounts of rare cRBCs for life saving transfusion therapies and define the first specific application, production of reagent red cells, and the first human indication, transfusion support for SCD patients without other options.

 描述（由申请人提供）：慢性贫血输血治疗的一个主要问题是，与占献血者主体的欧洲白人相比，亚洲和非洲背景的人的血型抗原具有高度的遗传多样性。红细胞上的这些多态性抗原导致同种免疫的高发生率以及这些患者血清中存在多种抗体。识别这些抗体的特异性并确定它们是自身抗体还是同种异体抗体对于提供相容性血液至关重要。该应用的主要目标是利用通过扩增造血干细胞和造血祖细胞体外生产培养红细胞 (cRBC) 所取得的进展，与稀有血型遗传表征方面取得的进展相一致，开发​​ cRBC 试剂，以鉴定是否存在针对高流行率供体抗原的具有临床意义的抗体。 cRBC 试剂将有助于高度同种免疫多次输血患者的抗体鉴定，并简化和标准化测试。 我们设计了一组由 6 名具有稀有血型表型的捐献者组成的小组，足以解决在具有自身和同种抗体的患者中出现的大多数抗体识别问题，并且可以用于输血目前因缺乏适当的稀有血液而无法输血的患者。在目标 1 中，我们建议开发新的培养方法，通过顺序扩增 HSC、HPC 和成红细胞室来扩增从外周血收集的造血细胞或从 iPSC 分化而来的造血细胞。我们还建议从这 6 名供体中产生诱导多能干细胞，并设计 O 族、Rh 零系 iPSC，这些细胞可用作试剂红细胞，并可用作输血的“通用”细胞。 外周血或 iPSC 产生的 cRBC 上表面抗原的表达尚未得到广泛表征。在目标 2 中，我们建议将 cRBC 与来自同一供体的天然红细胞进行比较，以确定它们是否适合作为试剂细胞和输血。我们将重点关注传统血库检测中 cRBC 的抗原谱、基因型、储存稳定性和性能。 重要的是，我们将生产的试剂 cRBC 与同种异体免疫患者输血时供应短缺的细胞相同。因此，我们建议进行的方法开发和质量测试将为生产足够量的稀有 cRBC 用于挽救生命的输血疗法铺平道路，并定义第一个具体应用，即试剂红细胞的生产，以及第一个人类适应症，为 SCD 患者提供输血支持，而无需其他选择。