Customized stem cells for clinical application in blood disorders

描述（由申请人提供）：该提案汇集了一个跨学科的合作者团队，以解决一个重大挑战：实现患者特异性诱导多能干细胞（iPSC）用于血液疾病的临床效用。我们的最终目标是将定制的iPSC分化为造血干细胞和祖细胞，以准确模拟人类血液疾病，用于研究疾病机制，并作为治疗血液疾病患者的平台。为了实现这一目标，该提案旨在发现驱动胚胎中造血干细胞（HSC）形成的发育途径和生物力学原理，并将筛选促进体外造血成熟的形态发生剂和化学物质。我们将采取几种新的综合方法，包括：1）将预测计算算法应用于高度纯化的胚胎HSC群体的基因表达数据，以发现指导造血和淋巴发育的基因网络; 2）在斑马鱼胚胎以及小鼠和人类多能干细胞中筛选，以发现HSC的新化学和生物调节剂; 4）从患有原发性免疫缺陷的患者获得iPSC，以将基因型与淋巴表型相关联，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，并开发用于临床规模的造血群体分化的方案。成功产生可移植的、遗传上未受干扰的HSC将是一个重大突破，这将增强iPSC用于造血疾病建模的效用，并建立将基因修复与HSC移植治疗相结合的翻译平台。 公共卫生相关性：患者来源的诱导多能干细胞（iPSC）代表了研究和治疗的重要新平台，但发现新机制，开发药物筛选和递送细胞将取决于我们将iPSC分化为相关造血干细胞和祖细胞（HSC/HSPC）的能力，这仍然是一个主要障碍。该提案旨在通过发现如何在体外指导HSC/HSPC的分化来实现iPSC的翻译潜力。

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. PUBLIC HEALTH RELEVANCE: Patient-derived induced pluripotent stem cells (iPSCs) represent an important new platform for research and therapy, but discovering novel mechanisms, developing drug screens, and delivering cells will hinge on our capacity to differentiate iPSC into relevant hematopoietic stem and progenitor cells (HSCs/ HSPCs), which remains a major hurdle. This proposal aims to realize the translational potential of iPSCs by discovering how to direct the differentiation of HSCs/ HSPCs in vitro.