Customized stem cells for clinical application in blood disorders

描述(由申请人提供)：这项提议汇集了一个跨学科的合作者团队来解决一个主要的挑战：实现针对患者的诱导多能干细胞(IPSCs)在血液疾病中的临床应用。我们的最终目标是将定制的IPSCs分化为造血干细胞和祖细胞，以准确地模拟人类血液疾病，用于疾病机制的研究，并作为治疗血液病患者的平台。为了实现这一目标，这项提议旨在发现驱动胚胎中造血干细胞(HSCs)形成的发育途径和生物力学原理，并将在体外筛选促进造血成熟的形态来源和化学物质。我们将采用几种新的和集成的方法，包括：1)应用预测计算算法从高度纯化的胚胎HSC群体中获得基因表达数据，以发现指导造血和淋巴发育的基因网络；2)在斑马鱼胚胎和小鼠和人类多能干细胞中筛选，以发现新的HSC的化学和生物调节因子；3)生产造血干和祖细胞群体的生物工程平台，应用生物力学力量模拟胚胎微环境；4)从原发免疫缺陷患者那里获得IPSC，以关联基因和淋巴表型，并通过“体外临床试验”测试基因修复策略；最终，5)获得无转基因的临床级多能干细胞，并制定临床规模的造血群体分化方案。成功构建可植入的、基因稳定的HSC将是一项重大突破，将增强IPSC在血液病模型中的应用，并建立一个将基因修复与HSC移植治疗相结合的翻译平台。

DESCRIPTION (provided by applicant): This proposal brings together an interdisciplinary team of collaborators to address a major challenge: to achieve clinical utility of patient-specific induced pluripotent stem cells (iPSCs) for blood diseases. Our ultimate goal is to differentiate customized iPSCs into hematopoietic stem and progenitor cells to accurately model human blood diseases for research into disease mechanisms, and as a platform for treating patients with blood diseases. To achieve this, this proposal aims to discover the developmental pathways and biomechanical principles that drive the formation of hematopoietic stem cells (HSCs) in embryos, and will screen for morphogens and chemicals that promote hematopoietic maturation in vitro. We will take several novel and integrated approaches including: 1) application of predictive computational algorithms to gene expression data from highly purified embryonic HSC populations to discover the gene networks that direct hematopoietic and lymphoid development; 2) Screens in zebrafish embryos and murine and human pluripotent stem cells to discover novel chemical and biological regulators of HSCs; 3) bioengineered platforms for production of hematopoietic stem and progenitor populations, applying biomechanical forces to mimic the embryonic microenvironment; 4) derivation of iPSC from patients with Primary Immune Deficiency to correlate genotype with lymphoid phenotypes and to test strategies for gene repair through an "in vitro clinical trial"; and ultimately, 5) to derive transgene-free clinical-grade pluripotent stem cells, and develop protocols for differentiation of hematopoietic populations at clinical scale. Success in generating engraftable, genetically unperturbed HSC would be a significant breakthrough that would enhance the utility of iPSC for modeling hematopoietic disease and establish a translational platform for combining gene repair with HSC transplantation therapy.