Nonhuman Primate Model for Preclinical Evaluation of Haplotype-Based iPSC Banking for HLA-matched Blood Products

项目摘要/摘要 人诱导多能干细胞向造血细胞分化的研究进展 使IPSC衍生血液产品的临床翻译更接近现实，并突出了 建立新的动物模型用于临床前评估阿司匹林的有效性、安全性和免疫原性 IPSC衍生的血液产品。由于IPSC可以从预期的接收方派生，因此它们提供 有可能产生无限数量的自体血细胞，避免人类白细胞抗原同种异体免疫。然而， 个人化干细胞治疗的高成本和复杂性使其目前在广泛的 申请。创建IPSC库已被提议作为供应人类白细胞抗原匹配血细胞的一种方法。 然而，由于人类白细胞抗原的高度变异性，使用随机捐赠者的IPSC银行仍然不切实际。这个 移植来自人类白细胞抗原纯合子捐献者的IPSCs(基于单倍型的移植策略)已被建议 为细胞提供可扩展的免疫相容细胞供应的有效方法 治疗和最大限度地发挥干细胞库的效用。为此，开发了银行业人类白细胞抗原的实验模型 纯合的ipsc株和评估它们的免疫原性、治疗效果和安全性将是至关重要的。 对未来干细胞库的设计和应用，以生产人类白细胞抗原相容的血液产品。在这里，我们 建立银行主要组织相容性(MHC)纯合子IPSC的非人灵长类动物模型 排队接受输血治疗。拟议的模型将使用毛里求斯食蟹猴，它们是 来自一个很小的创始人群体的后代，具有非常有限的MHC多样性，只有7个 常见单倍型(M1-M7)。这为快速选择MHC纯合子和MHC提供了一个独特的机会 以及血型相同的动物或通过基因筛查明确定义的MHC不匹配的动物。我们会 应用该模型评价MHC纯合子IPSC来源的CD34+CD45+CD38-的实用性和安全性 多能造血祖细胞治疗清髓干细胞后细胞减少症 移植并检验输注来自MHC匹配纯合子的imHPs的假设 IPSCs可减少人类白细胞抗原同种异体免疫。在目标1中，我们将建立一个MHC纯合子NHP模型 MHC相容的IPSC衍生血液产品的临床前评估。在目标2中，我们将评估疗效 在MCM模型中，基于MHC纯合子IPSC的获得性骨髓抑制治疗的安全性。 在目标3中，我们将评估转移后对MHC纯合imHPs的同种异体免疫反应。 MHC屏障。总体而言，拟议的研究将证明使用MHC的实用性和安全性 纯合子iPSC来源的造血细胞用于治疗骨髓抑制和人类白细胞抗原 同种异体免疫减少。

Project Summary/Abstract Recent advances in hematopoietic differentiation from human induced pluripotent stem cells (iPSCs) have brought the clinical translation of iPSC-derived blood products closer to reality and highlight the need for the developing of novel animal models for preclinical evaluation of the efficacy, safety, and immunogenicity of iPSC-derived blood products. Because iPSCs can be derived from the intended recipient, they offer the possibility to generate autologous blood cells in unlimited numbers and avoid HLA alloimmunization. However, the high costs of personalized stem cell therapy and its complexity makes it currently impractical for broad application. Creation of iPSC banks has been proposed as an approach to supply HLA-matched blood cells. Yet, iPSC banking using random donors would remain impractical due to the high variability of HLA. The banking iPSCs from HLA-homozygous donors (haplotype-based banking strategy) has been suggested to be an effective way to provide a scalable off-the-shelf supply of immunologically compatible cells for cellular therapies and maximize the utility of stem cell banking. Thus, developing experimental models for banking HLA homozygous iPSC lines and assessing their immunogenicity, therapeutic efficacy, and safety will be essential to the future design and utility of stem cell banks for manufacturing HLA-compatible blood products. Here, we propose to establish a nonhuman primate model for banking major histocompatibility (MHC) homozygous iPSC lines for transfusion therapies. The proposed model will employ Mauritian cynomolgus macaques, which are descendent from a small founder population and have very limited MHC diversity consisting of only seven common haplotypes (M1-M7). This provides a unique opportunity to rapidly select MHC homozygous and MHC and blood group-identical animals or animals with well-defined MHC mismatches by genetic screening. We will use this model to evaluate the utility and safety of MHC homozygous iPSC-derived CD34+CD45+CD38- multipotent hematopoietic progenitors in the treatment of cytopenia following myeloablative stem cell transplantation and test the hypothesis that transfusion of imHPs derived from MHC-matched homozygous iPSCs reduces HLA alloimmunization. In aim 1, we will establish a MHC homozygous NHP model for preclinical evaluation of MHC compatible iPSC-derived blood products. In aim 2, we will evaluate the efficacy and safety of MHC homozygous iPSC-based therapies for acquired bone marrow suppression in MCM model. In aim 3, we will assess the alloimmune responses toward MHC homozygous imHPs following transfer across MHC barriers. Overall, the proposed research will demonstrate the utility and safety of using MHC homozygous iPSC-derived hematopoietic cells for the treatment of myelosuppression and HLA alloimmunization reduction.