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）：该提案合并了发育和化学生物学，计算分析和生物工程，以解决血液学研究中最重要的挑战之一-从多能干细胞中体外衍生出长期可植入的造血干细胞。迄今为止，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）的相当大的努力已经证明在小鼠中仅部分成功，而在人类中大部分不成功。未能产生真正的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，并排除了利用这种潜在的变革性技术进行治疗。这项更新申请将建立在前两年的基础上，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将多能干细胞分化为临床相关的造血谱系？我们将应用计算算法来增强由来自鱼、小鼠和人类的造血干细胞和祖细胞群体的单细胞RNA-seq生成的表达数据集。我们将探测基因调控网络，直接造血和淋巴的发展，并测试候选转录因子，非编码RNA，和形态发生假设，以管理这些网络，单独和组合，通过增益和功能丧失的策略。在第二个推力中，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，以推动HSC的承诺并增强HSC的功能。最后，使用模拟作用于胚胎造血内皮的生物力学力的生物工程平台，我们将配置“芯片上的造血内皮”来询问促进HSC出现和造血干细胞和祖细胞扩增的效应子和下游信号通路。我们在造血发育中定义遗传、化学和生物力学机制的努力代表了互补和协同策略，以解决我们在体外获得HSC的关键挑战。