Utilization of cord blood-expanded stem cells for megakaryocyte and platelet production

利用脐带血扩增干细胞产生巨核细胞和血小板

• 批准号: 9020648
• 负责人: Camelia IancuRubin
• 金额: $ 22.5万
• 依托单位: ALLCELLS, LLC
• 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 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9020648
• 关键词: Alloimmunization; Animal Model; Biological Assay; Blood Cells; Blood Platelet Disorders; Blood Platelets; Bypass; CD34 gene; Cell Therapy; Cells; Clinical; Collection; Complement; Cryopreservation; Dependency; Development; Disease; Engraftment; Epigenetic Process; Evaluation; Event; Face; Feasibility Studies; Foundations; Goals; Government; Hematopoietic stem cells; Hemorrhage; Histocompatibility Antigens Class I; Human; In Vitro; Infection; Inherited; Laboratories; Laboratory Research; Lead; Life; Megakaryocytes; Megakaryocytopoieses; Monitor; Mus; Nature; Patients; Persons; Phase; Physiological; Platelet Count measurement; Platelet Transfusion; Population; Procedures; Production; Protocols documentation; Reaction; Research; Research Design; Risk; Shipping; Ships; Small Business Technology Transfer Research; Source; Staging; Stem cells; Surface; System; Testing; Thrombocytopenia; Thrombopoiesis; Transfusion; Transplantation; Umbilical Cord Blood; Valproic Acid; base; clinical application; clinically relevant; commercial application; commercialization; cost; cytokine; design; differential expression; effective therapy; flasks; genetic manipulation; human embryonic stem cell; immunodeficient mouse model; in vivo; induced pluripotent stem cell; prevent; progenitor; public health relevance; tissue culture; volunteer

 DESCRIPTION (provided by applicant): Platelet transfusions are the most effective treatments for patients with thrombocytopenia. The growing demand for platelet transfusions is often limited by shortage in the platelets (PTL) supply due to dependency on volunteer donors, short shelf life, risk of infections, and alloimmunization. It is therefore critical to create an efficient, donor-independent source of PTLs that could complement-and possibly replace-the current donor- dependent transfusion system. The search for a different source of platelets for transfusion was accelerated by the recent advances in hematopoietic stem cell expansion which became a promising source for generating platelets ex vivo. However, creating a clinically relevant platelet product ex vivo has been hampered by technological and quantitative limitations. The major barrier in generating PTLs suitable for transfusion is the low number of PTLs that can be produced by cultured megakaryocytes (MK) ex vivo and the difficulties in recapitulating the complexity of physiological human thrombopoiesis in culture. The studies proposed in this application are designed to develop an alternative approach by creating a MK cell product that can generate platelets in vivo after its transfusion into patients. Creating such product will not only circumvent the technological hurdles and costs associated with ex vivo PTL production but also provide the foundation for developing a cell-based therapy for thrombocytopenic patients and possibly preclude alloimmunization. In addition, MKs generated from this study will be advanced to a manufacturing phase to create the first commercially available source of CD41+ MKs for use in research. A commercial supply of MKs would dramatically accelerate the pace of research in human thrombopoiesis. This goal will be accomplished by implementing a new culture system protocol in which a MK cell product comprising MK progenitors/precursors, immature MKs, and mature MKs, will be generated from a validated source of cord blood-derived CD34+ hematopoietic stem cells expanded by means of epigenetic reprograming by treatment with valproic acid. These proposed feasibility studies are designed to (1) characterize the resultant MKs phenotypically and functionally, (2) determine the MKs' potency ex vivo and in vivo in transplantation assays in an immunodeficient mouse model, and (3) test the MKs' ability to remain viable and functional following cryopreservation and storage. These studies will lead to development of a platform for developing a commercial MK cell product for clinical use to treat thrombocytopenia and for laboratory research.

 描述（由申请人提供）：血小板输注是血小板减少症患者最有效的治疗方法。由于依赖志愿者捐赠者、保质期短、感染风险和同种免疫，血小板（PTL）供应短缺，对血小板输注的需求不断增长往往受到限制。因此，创建一个有效的、独立于供体的 PTL 来源至关重要，它可以补充并可能取代当前依赖供体的输血系统。造血干细胞扩增的最新进展加速了对用于输血的不同血小板来源的寻找，造血干细胞扩增成为离体生成血小板的有前途的来源。然而，离体制造临床相关的血小板产品一直受到技术和数量限制的阻碍。产生适合输血的 PTL 的主要障碍是离体培养的巨核细胞 (MK) 可以产生的 PTL 数量较少，并且难以在培养物中重现生理性人类血小板生成的复杂性。本申请中提出的研究旨在开发一种替代方法，即创建一种 MK 细胞产品，该产品在输注给患者后可以在体内产生血小板。创建这样的 该产品不仅将规避与离体 PTL 生产相关的技术障碍和成本，而且还为开发针对血小板减少症患者的细胞疗法奠定了基础，并可能排除同种免疫。此外，本研究产生的 MK 将进入制造阶段，以创建第一个用于研究的商业可用 CD41+ MK 来源。 MK 的商业供应将极大地加快人类血小板生成的研究步伐。这一目标将通过实施新的培养系统方案来实现，其中包含 MK 祖细胞/前体、未成熟 MK 和成熟 MK 的 MK 细胞产品将从经过验证的脐带血来源的 CD34+ 造血干细胞来源产生，并通过丙戊酸处理进行表观遗传重编程来扩增。这些拟议的可行性研究旨在（1）表征所得 MK 的表型和功能，（2）在免疫缺陷小鼠模型中的离体和体内移植测定中确定 MK 的效力，以及（3）测试 MK 在冷冻保存和储存后保持活力和功能的能力。这些研究将导致开发一个平台，用于开发商业 MK 细胞产品，用于临床治疗血小板减少症和实验室研究。

项目成果

Pre-clinical development of a cryopreservable megakaryocytic cell product capable of sustained platelet production in mice.

能够在小鼠体内持续产生血小板的可冷冻巨核细胞产品的临床前开发。

• DOI: 10.1111/trf.15546
• 发表时间: 2019
• 期刊: Transfusion
• 影响因子: 2.9
• 作者: Patel,Ami;Clementelli,CaraMarie;Jarocha,Danuta;Mosoyan,Gohar;Else,Cindy;Kintali,Manisha;Fong,Helen;Tong,Jay;Gordon,Ronald;Gillespie,Virginia;Keyzner,Alla;Poncz,Mortimer;Hoffman,Ronald;Iancu-Rubin,Camelia
• 通讯作者: Iancu-Rubin,Camelia