Engineering iPSC-RBCs for Transfusion

工程 iPSC-RBC 用于输血

• 批准号: 9385217
• 负责人: John D Roback
• 金额: $ 60.57万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9385217
• 关键词: Address; Alloimmunization; Animals; Antibodies; Antigens; Area; Authorization documentation; Autologous; Back; Behavior; Blood Banks; Blood Cells; Blood Group Antigens; Blood Transfusion; CRISPR/Cas technology; Cell Differentiation process; Cell Survival; Cell Therapy; Cellular biology; Characteristics; Clinical; Cyclic GMP; Data; Development; Dissection; Effectiveness; Engineering; Erythrocyte Transfusion; Erythrocytes; Funding; Future; Genes; Genetic Engineering; Grant; Hematopoietic stem cells; Human; Human Biology; Immunology; In Vitro; Infusion procedures; Investigation; Life; Methodology; Methods; Modeling; Morphology; Mouse Strains; Mus; Mutation; National Heart, Lung, and Blood Institute; Patients; Phenotype; Population; Prevention; Procedures; Production; Protocols documentation; Research Personnel; Safety; Savings; Sickle Cell Anemia; Signal Transduction; Stem cells; Techniques; Technology; Testing; Thalassemia; Therapeutic; Therapeutic Uses; Therapeutic procedure; Time; TimeLine; Transfusion; Work; blood product; clinical application; cost; design; genome editing; immunogenicity; in vivo; induced pluripotent stem cell; novel; peripheral blood; permissiveness; pluripotency; response; sterility testing; tool

ABSTRACT: Blood transfusion is the most common therapeutic procedure performed for hospitalized patients. Despite the efficacy of transfusion using standard donor-derived RBCs, we believe the full therapeutic potential of blood transfusion remains untapped. Recent work from several groups has demonstrated that human induced pluripotent stem cells (iPSCs) can be derived from small volumes of peripheral blood, can undergo genome editing to produce precise genetic changes, and can be differentiated into terminally mature RBCs (iPSC-RBCs), all under cGMP-compliant conditions. While iPSC-RBCs may not replace donor-derived RBCs for routine transfusions in the foreseeable future, we believe that iPSC-RBCs that have been genetically engineered to express novel functionalities (eg, prevention of alloimmunization in sickle cell disease patients) could find near term clinical applications in areas where current transfusion therapies are inadequate. In the proper therapeutic niche, engineered iPSC-RBCs would be high-value high-impact products that could sustain high costs due to their unique characteristics. The investigative team proposes to leverage recent advances in iPSC and genomic editing technologies, as well their ongoing NHLBI-funded studies in RBC biology and immunology, to pursue 3 Specific Aims: (1) To optimize cGMP-grade protocols for establishment, propagation, and differentiation of human peripheral blood-derived iPSCs into mature RBCs (iPSC-RBCs); (2A) To engineer iPSCs such that resulting RBCs are negative for multiple blood group antigens, and (2B) To characterize the in vivo survival, morphological maturation, and immunogenicity of iPSC-RBCs when transfused into specialized murine transfusion models; and (3) To obtain FDA IND and institutional IRB approvals, and perform human autologous transfusions of iPSC- RBCs to investigate RBC survival, maturation, and immunogenicity in recipients. The proposed studies have been carefully designed to integrate recently developed techniques (cGMP iPSC methods, gene editing) with specialized capabilities at Emory (EPIC cGMP clean room in the blood bank, in vivo biotinylated RBC tracking, first-in-human cell therapy expertise) to investigate the biology of human iPSC- derived RBCs after transfusion into human recipients. Successful completion of the proposed investigations will lead to novel blood products to meet important unmet clinical needs in transfusion-dependent patients such as those with sickle cell disease.

摘要：输血是住院患者最常见的治疗方法。 尽管使用标准供体来源的红细胞进行输血具有疗效，但我们相信其全部治疗潜力 输血仍有待开发。几个小组最近的工作表明，人类 诱导多能干细胞（iPSC）可以从少量外周血中提取，可以进行 基因组编辑可产生精确的遗传变化，并可分化为最终成熟的红细胞 (iPSC-RBC)，全部在符合 cGMP 的条件下进行。虽然 iPSC-RBC 可能无法替代供体来源的 RBC 在可预见的将来，常规输血中，我们相信经过基因改造的 iPSC-RBC 设计用于表达新功能（例如，预防镰状细胞病患者的同种免疫） 可以在当前输血疗法不足的领域找到近期的临床应用。在 适当的治疗利基，工程 iPSC-RBC 将是高价值、高影响力的产品，可以持续 由于其独特的特性，成本较高。调查小组建议利用最新进展 iPSC 和基因组编辑技术，以及他们正在进行的由 NHLBI 资助的红细胞生物学和 免疫学，追求 3 个具体目标： (1) 优化人类外周血建立、增殖和分化的 cGMP 级方案 将血液来源的 iPSC 转化为成熟红细胞 (iPSC-RBC)； (2A) 改造 iPSC，使所得红细胞对多种血型抗原呈阴性，以及 (2B) 表征输注时 iPSC-RBC 的体内存活、形态成熟和免疫原性 进入专门的小鼠输血模型；和 (3) 获得FDA IND和机构IRB批准，并进行iPSC的人体自体输血- 红细胞用于研究受者红细胞的存活、成熟和免疫原性。 拟议的研究经过精心设计，整合了最近开发的技术（cGMP iPSC 方法、基因编辑）在埃默里大学（血库的 EPIC cGMP 洁净室、体内 生物素化红细胞追踪（人类首创的细胞治疗专业知识）来研究人类 iPSC 的生物学 输注给人类接受者后衍生的红细胞。成功完成拟议的调查将 开发新型血液制品，以满足依赖输血的患者未满足的重要临床需求，例如 患有镰状细胞病的人。