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: 9282481
• 负责人: ERIC E BOUHASSIRA
• 金额: $ 50.25万
• 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/9282481
• 关键词: Adsorption; African; Alloimmunization; Anemia; Antibodies; Antibody Specificity; Antigens; Asians; Basic Science; Biological Assay; Blood; Blood Banks; Blood Cells; Blood Group Antigens; Blood Transfusion; Blood donor; Cell Count; Cells; Chronic; Chronic Disease; Clinical; Complex; Cooley' s anemia; Development; Engineering; Erythroblasts; Erythrocyte Transfusion; Erythrocytes; European; Event; Foundations; Future; Generations; Genetic; Genetic Variation; Genotype; Goals; Harvest; Hematopoietic; Hematopoietic stem cells; High Prevalence; Human; Immunize; In Vitro; Incidence; Inherited; Laboratories; Life; Medical; Methods; Monoclonal Antibodies; New York; Patients; Performance; Phenotype; Procedures; Process; Production; Reagent; Research; Safety; Savings; Serum; Services; Sickle Cell Anemia; Standardization; Stem Cell Research; Surface Antigens; Technology; Testing; Time; Transfusion; Translating; United States; blood group; clinically significant; cost; design; experimental study; improved; in vivo; induced pluripotent stem cell; innovation; method development; novel; peripheral blood; public health relevance; volunteer

 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(申请人提供)：慢性贫血输血疗法的一个主要问题是亚洲和非洲背景的人的血型抗原与欧洲白人血型抗原的高度遗传多样性，后者是献血者的大多数。红细胞表面的这些多态抗原导致同种异体免疫的高发生率和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异体抗体，对于提供相容血液至关重要。这一应用的主要目的是利用通过扩增造血干细胞和造血祖细胞在体外生产培养的红细胞(CRBC)方面取得的进展，与罕见血型的遗传特征方面的进展相一致，开发试剂cRCs来识别高发献血者抗原的临床显著抗体的存在。试剂cRBC将促进高度同种异体免疫多次输血患者的抗体鉴定，并简化和标准化检测。我们设计了一个由6名具有稀有血型表型的捐赠者组成的小组，这些血型足以解决拥有自身抗体和异体抗体的患者中出现的大多数抗体鉴定问题，并可用于为目前因缺乏适当稀有血液而无法输血的患者输血。在目标1中，我们提出了一种新的培养方法，通过对HSC、HPC和红细胞室的顺序扩增来扩增从外周血中采集的造血细胞或从IPSCs分化而来的造血细胞。我们还建议从这6个捐赠者中培养出诱导多能干细胞，并设计出一种O组、Rh组缺失的IPSCs，这些细胞可用作试剂红细胞，并可用作“通用”输血细胞。从外周血或IPSCs产生的红细胞表面抗原的表达还没有广泛的特征。在目标2中，我们建议将红细胞与来自同一捐赠者的天然红细胞进行比较，以确定它们是否适合用作试剂细胞和输血。我们将重点介绍传统血库检测中红细胞的抗原性、基因分型、储存稳定性和性能。重要的是，我们将生产的试剂红细胞是供不应求的细胞，用于异基因免疫患者的输血。因此，我们计划进行的方法开发和质量测试将为生产足够数量的稀有红细胞用于挽救生命的输血疗法铺平道路，并确定第一个特定应用，生产试剂红细胞，以及第一个人类适应症，在没有其他选择的情况下为SCD患者提供输血支持。