Developmental Activation of the Inflammasome Controls Hematopoietic Stem Cell Production

炎症小体的发育激活控制造血干细胞的产生

• 批准号: 10668397
• 负责人: TRISTA E. NORTH
• 金额: $ 52.38万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10668397
• 关键词: Acute; Biological; Biological Assay; Blood; Cell Culture Techniques; Cells; Clinical; Clinical Protocols; Clinical Trials; Complex; Cues; Data; Development; Embryo; Embryonic Development; Endothelium; Engraftment; FDA approved; Foundations; Genes; Genetic; Genetic Epistasis; Goals; Growth; HIF1A gene; Harvest; Hematological Disease; Hematopoietic; Hematopoietic Cell Production; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Immune; Immune system; In Vitro; Inflammasome; Inflammation; Inflammatory; Interleukin-1 beta; Lymphoid Cell; Maintenance; Mediating; Metabolic; Methods; Modeling; Molecular; Organism; Outcome; Physiological; Pluripotent Stem Cells; Population; Process; Production; Productivity; Proliferating; Protocols documentation; Reactive Oxygen Species; Regulation; Reporter; Research; Role; Signal Pathway; Signal Transduction; Specific qualifier value; Sterility; System; Therapeutic; Therapeutic Uses; Transplantation; Validation; Vertebrates; Work; Xenograft procedure; Zebrafish; cell type; extracellular; gain of function; glucose metabolism; hematopoietic stem cell expansion; hematopoietic stem cell fate; hematopoietic stem cell formation; hemogenic endothelium; improved; in vivo; induced pluripotent stem cell; knock-down; loss of function; novel; overexpression; pharmacologic; programs; prospective; self-renewal; spatiotemporal; standard of care; stem cell function; stem cell therapy; stem-like cell; stressor; transcription factor; transcriptome sequencing; vertebrate embryos

SUMMARY Hematopoietic stem cells (HSCs) supply the lifelong foundation of the blood and immune systems. HSCs are therapeutically valuable as HSC transplantation is the standard of care for many hematological diseases. However, treatment availability remains problematic due to immune incompatibility and donor shortage. Likewise, while the number of transplanted HSCs directly impacts outcome, there are currently no established clinical protocols to successfully expand donor-harvested HSCs, nor to differentiate embryonic or induced pluripotent stem cells (iPSCs) into functional HSCs in vitro. Therefore, the identification of novel modifiers of de novo production of HSCs with long-term self-renewal and differentiation capacity is a major unmet clinical need. Despite decades of research, current protocols rely primarily on transcription factor overexpression of force cells into an “HSC-like” program; however, transplantation of these in vitro-derived HSCs yields only limited long-term engraftment and multilineage potential. These observations imply that current in vitro differentiation strategies are missing critical cues which are essential to unlock or maintain full HSC function in vivo. HSCs are first formed in the vertebrate embryo from a unique population of mesodermal precursors termed hemogenic endothelium through a highly conserved process known as endothelial-to-hematopoietic transition. We have previously utilized the zebrafish model to discover novel regulators of HSC formation, including the first therapeutic identified in zebrafish to be used in FDA-approved clinical trials. In addition, we have demonstrated that key extrinsic signals occurring during embryogenesis coordinate the timing and scale of HSC production. More recently, our lab revealed an essential role for inflammatory signaling in developmental HSPC formation. However, the mechanism by which sterile inflammation originates in the embryo has not been determined. Our preliminary data indicate that physiological changes in metabolic state during embryogenesis initiate sterile inflammatory signaling to stimulate HSC commitment and expansion, without loss of multi-potency. Our central hypothesis is that “developmental stressors”, such as onset of metabolic activity, activate the NLRP3-inflammasome to drive sterile IL1β-mediated commitment to HSC production. Our proposed work will demonstrate the essential role of developmental inflammasome activation in establishing HSPC commitment and expansion in vivo in zebrafish embryos and in vitro in human iPSC culture. Defining the molecular signaling pathways that mediate productive HSC formation in vivo will reveal new targets for optimizing the directed expansion and/or de novo production of human HSCs for therapeutic use.

总结 造血干细胞（HSC）提供血液和免疫系统的终身基础。HSCs 是有治疗价值的，因为HSC移植是许多血液病的标准治疗。 然而，由于免疫不相容性和供体短缺，治疗的可用性仍然存在问题。 同样，虽然移植HSC的数量直接影响结果，但目前还没有确定的 成功扩增供体收获的HSC的临床方案，也不分化胚胎或诱导的HSC。 多能干细胞（iPSC）在体外转化为功能性HSC。因此，识别新的修饰de 具有长期自我更新和分化能力的HSC的新生生产是主要的未满足的临床需求。 尽管几十年的研究，目前的协议主要依赖于转录因子过度表达的力细胞 然而，这些体外衍生的HSC的移植仅产生有限的长期存活。 移植和多谱系潜力。这些观察结果表明，目前的体外分化 这些策略缺少关键线索，这些线索对于解锁或维持体内HSC的全部功能至关重要。 HSC首先在脊椎动物胚胎中由独特的中胚层前体群体形成 通过高度保守的内皮-造血过程， 过渡我们之前已经利用斑马鱼模型发现了HSC形成的新调节因子， 包括第一个在斑马鱼中发现的治疗药物，用于FDA批准的临床试验。另外我们 已经证明，在胚胎发生过程中发生的关键外部信号协调的时间和规模， HSC生产。最近，我们的实验室揭示了炎症信号在发育中的重要作用。 HSPC形成。然而，胚胎中无菌性炎症起源的机制尚未被阐明。 测定我们的初步数据表明，在胚胎发生过程中，代谢状态的生理变化 启动无菌炎症信号传导以刺激HSC定型和扩增，而不丧失多能性。 我们的中心假设是，“发育压力源”，如代谢活动的开始，激活了 NLRP 3-炎性体驱动无菌IL 1 β介导的HSC产生。我们建议的工作 将证明发育炎性小体激活在建立HSPC定型中的重要作用 以及在斑马鱼胚胎中的体内扩增和在人iPSC培养物中的体外扩增。分子信号的定义 在体内介导生产性HSC形成的途径将揭示优化定向HSC形成的新靶点。 扩增和/或从头产生人HSC用于治疗用途。

项目成果

Metabolic Regulation of Inflammasome Activity Controls Embryonic Hematopoietic Stem and Progenitor Cell Production.

• DOI: 10.1016/j.devcel.2020.07.015
• 发表时间: 2020-10-26
• 期刊: Developmental cell
• 影响因子: 11.8
• 作者: Frame JM;Kubaczka C;Long TL;Esain V;Soto RA;Hachimi M;Jing R;Shwartz A;Goessling W;Daley GQ;North TE
• 通讯作者: North TE