Customized stem cells for clinical application in blood disorders

描述（由申请人提供）：本提案汇集了一个跨学科的合作者团队，以解决一个主要挑战：实现患者特异性诱导多能干细胞（iPSCs）在血液病中的临床应用。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，研究疾病机制，并作为治疗血液疾病患者的平台。为了实现这一目标，本提案旨在发现胚胎中驱动造血干细胞（hsc）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因子和化学物质。我们将采用几种新颖和综合的方法，包括：1)应用预测计算算法对来自高度纯化的胚胎HSC群体的基因表达数据进行分析，以发现指导造血和淋巴细胞发育的基因网络；2)筛选斑马鱼胚胎、小鼠和人类多能干细胞，发现新的造血干细胞的化学和生物调节因子；3)利用生物力学力模拟胚胎微环境，构建造血干细胞和祖细胞群体的生物工程平台；4)从原发性免疫缺陷患者中提取iPSC，将基因型与淋巴样表型相关联，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，并制定临床规模造血群体分化的方案。成功生成可移植的、基因上不受干扰的造血干细胞将是一个重大突破，它将增强iPSC在造血疾病建模中的效用，并为基因修复与造血干细胞移植治疗的结合建立一个转化平台。

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.