Customized stem cells for clinical application in blood disorders

定制干细胞用于血液疾病的临床应用

• 批准号: 8771044
• 负责人: JAMES J COLLINS
• 金额: $ 144.95万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-19 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8771044
• 关键词: Address; Autologous; Biological; Biology; Biomechanics; Biomedical Engineering; Cell Differentiation process; Cell model; Cell physiology; Cells; Chemicals; Collaborations; Computational algorithm; Computer Analysis; Data Set; Derivation procedure; Development; Disease; Embryo; Endothelium; Failure; Fishes; Functional RNA; Genetic; Goals; Grant; Hematological Disease; Hematology; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemoglobinopathies; Human; Immune; Immunologics; In Vitro; Infection; Knowledge; Lymphoid; Malignant - descriptor; Marrow; Modeling; Monoclonal Antibody R24; Mus; Pathway interactions; Patients; Pluripotent Stem Cells; Population; Protocols documentation; Regulator Genes; Research; Signal Pathway; Specific qualifier value; Stem cells; Technology; Testing; Therapeutic; Transplantation; Zebrafish; chemical genetics; clinical application; clinically relevant; drug development; embryo stage 2; embryonic stem cell; gene discovery; gene repair; graft vs host disease; induced pluripotent stem cell; insight; leukemia; loss of function; morphogens; mortality; novel; progenitor; public health relevance; stem; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): This proposal merges developmental and chemical biology, computational analysis, and bioengineering to tackle one of the most significant challenges in hematology research-the in vitro derivation of long-term engraftable hematopoietic stem cells from pluripotent stem cells. To date, considerable efforts at directed differentiation o hematopoietic stem cells (HSCs) from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have proven only partially successful in mice and largely unsuccessful in humans. The failure to generate true ESC/iPSC-derived HSCs remains a barrier for exploiting patient-derived iPSCs for modeling blood diseases, and precludes harnessing this potentially transformative technology for therapy. This renewal application will build upon progress made during the first two years of the prior grant period to address the central question: What are the developmental pathways that drive the formation of HSCs in embryos, and how can we exploit this knowledge to direct the differentiation of pluripotent stem cells into clinically relevant hematopoietic lineages? We will apply computational algorithms to enhanced expression datasets generated by single cell RNA-seq of hematopoietic stem and progenitor populations from fish, mouse, and human. We will probe gene regulatory networks that direct hematopoietic and lymphoid development, and test candidate transcription factors, non-coding RNAs, and morphogens hypothesized to govern these networks, alone and in combination, through gain and loss-of-function strategies. In a second thrust, we will perform screens in zebrafish blastomeres and human pluripotent stem cells to discover novel chemical and biological regulators of HSCs, and will incorporate chemicals into cell differentiation protocols to drive HSC commitment and enhance HSC function. Lastly, using bioengineered platforms which mimic the biomechanical forces that act on the embryonic hemogenic endothelium, we will configure "hemogenic endothelium-on-a-chip" to interrogate the effectors and downstream signaling pathways that promote emergence of HSCs and expansion of hematopoietic stem and progenitor cells. Our efforts at defining genetic, chemical and biomechanical mechanisms in hematopoietic development represent complementary as well as synergistic strategies to solve our key challenge of deriving HSCs in vitro.

描述（由申请人提供）：该提案融合了发育生物学和化学生物学、计算分析和生物工程，以解决血液学研究中最重大的挑战之一——从多能干细胞体外衍生出长期可移植的造血干细胞。迄今为止，在从胚胎干细胞（ESC）和诱导多能干细胞（iPSC）定向分化造血干细胞（HSC）方面所做的大量努力已被证明在小鼠中仅部分成功，而在人类中大部分不成功。无法生成真正的 ESC/iPSC 衍生的 HSC 仍然是利用患者衍生的 iPSC 进行血液疾病建模的障碍，并且阻碍了利用这种潜在的变革性技术进行治疗。此次更新申请将建立在上一资助期前两年取得的进展的基础上，以解决核心问题：驱动胚胎中 HSC 形成的发育途径是什么，以及我们如何利用这些知识来指导 HSC 的分化。多能干细胞转化为临床相关的造血谱系？我们将应用计算算法来增强由鱼、小鼠和人类的造血干细胞和祖细胞群的单细胞 RNA-seq 生成的表达数据集。我们将探索指导造血和淋巴发育的基因调控网络，并通过获得和丧失功能策略来测试候选转录因子、非编码RNA和假设可单独或组合控制这些网络的形态发生素。第二个重点是，我们将在斑马鱼卵裂球和人类多能干细胞中进行筛选，以发现 HSC 的新型化学和生物调节剂，并将化学物质纳入细胞分化方案中，以推动 HSC 承诺并增强 HSC 功能。最后，利用模拟作用于胚胎造血内皮细胞的生物力学力的生物工程平台，我们将配置“造血内皮芯片”来探究促进 HSC 出现和造血干细胞扩张的效应器和下游信号通路。祖细胞。我们在定义造血发育中的遗传、化学和生物力学机制方面所做的努力代表了解决体外衍生造血干细胞的关键挑战的互补和协同策略。