A microRNA-mediated approach towards generating functional hematopoietic stem cells from human pluripotent stem cells in vitro

一种 microRNA 介导的方法在体外从人多能干细胞生成功能性造血干细胞

• 批准号: 10810589
• 负责人: Athanasia Panopoulos
• 金额: $ 45.93万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10810589
• 关键词: Achievement; Affect; Biological Assay; Bone Marrow; Bone Marrow Transplantation; Cell physiology; Cell surface; Cells; Clinical; Data; Data Set; Derivation procedure; Detection; Development; Disease; Environment; Fetal Liver; Future; Gene Expression Profiling; Genes; Genetic; Goals; Hematological Disease; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic System; Hematopoietic stem cells; Human; In Vitro; Individual; Knowledge; Lead; Mediating; Methods; MicroRNAs; Mission; Molecular; Mus; Pathway interactions; Patients; Pluripotent Stem Cells; Population; Process; Public Health; Regulatory Pathway; Research; Research Personnel; Resources; Sampling; Scientist; Source; Therapeutic; Tissue-Specific Gene Expression; Tissues; Transplantation; Umbilical Cord Blood; United States National Institutes of Health; Variant; Virus; comparative; differential expression; differentiation protocol; embryonic stem cell; expectation; hematopoietic differentiation; hematopoietic transplantation; hemogenic endothelium; human embryonic stem cell; human pluripotent stem cell; human stem cells; improved; in vivo; induced pluripotent stem cell; insight; novel; novel strategies; prevent; reconstitution; stem cell function; stem cell population; stem cells; transcription factor; transcriptomics; translational impact

PROJECT SUMMARY Producing transplantable hematopoietic stem cells (HSCs) in vitro could benefit thousands of patients currently unable to receive treatment for hematological disorders. The long-term goal is to be able to generate clinically viable HSCs in vitro from human pluripotent stem cells (PSCs). This achievement would revolutionize the way numerous blood diseases could be treated, and could theoretically alleviate patients from having to wait to find a bone marrow transplant ‘match’, since a well-established clinical roadmap already exists for the use of HSCs in a therapeutic setting. Our central hypothesis is based on our strategic transcriptional profiling analysis of genetically identical in vivo HSCs (functional) and in vitro-derived ‘pre-HSCs’ (not functional). By eliminating genetic background as a source of variability, to identify the key molecular differences that underlie the functional deficiencies present in in vitro-generated HSCs, we were able to discover novel differences in regulatory pathways modulated by miRNAs. Guided by strong preliminary data, the overall objective of this application is to modulate key miRNAs during in vitro differentiation to thus provide a novel approach to obtaining functional HSCs from human PSCs. This objective will be pursued under two specific aims: (1) To identify miRNA changes during human PSC-mediated hematopoietic differentiation. In this aim, key miRNA expression levels will be analyzed during PSC-hematopoietic differentiation assays; and (2) To examine the potential of miRNAs to influence hematopoietic potential in vitro and in vivo. In this aim, key miRNA expression levels will be modified during PSC-hematopoietic differentiation assays, and analyzed for their ability to successfully generate fully functional HSCs in vitro. It is our expectation that our strategic approach will have a positive translational impact by examining a novel pathway to generate HSCs in vitro that leads to long-term hematopoietic transplantation capability, while also providing a much-needed comprehensive dataset resource for the hematopoietic field as a whole towards the successful derivation of clinically viable HSCs in vitro.

项目概要 目前，体外生产可移植造血干细胞（HSC）可以使数千名患者受益 无法接受血液疾病治疗的长期目标是能够在临床上产生。 从人类多能干细胞（PSC）中体外培养出可行的 HSC，这一成就将彻底改变这一方式。 许多血液疾病都可以得到治疗，理论上可以减轻患者等待发现的时间 骨髓移植“匹配”，因为 HSC 的使用已经存在完善的临床路线图 在治疗环境中，我们的中心假设基于我们的策略转录谱分析。 通过消除基因相同的体内 HSC（有功能）和体外衍生的“前 HSC”（无功能）。 遗传背景作为变异性的来源，以确定功能性背后的关键分子差异 由于体外生成的 HSC 存在缺陷，我们能够发现监管方面的新差异 在强有力的初步数据的指导下，该应用的总体目标是 在体外分化过程中调节关键 miRNA，从而提供一种获得功能性的新方法 来自人类 PSC 的 HSC 将在两个具体目标下实现：(1) 识别 miRNA 变化。 在人类 PSC 介导的造血分化过程中，关键的 miRNA 表达水平将是 在 PSC 造血分化测定过程中进行分析；以及 (2) 检查 miRNA 的潜力 影响体外和体内的造血潜力为此目的，关键的 miRNA 表达水平将被修改。 在 PSC 造血分化测定期间，并分析其成功生成完全细胞的能力 我们期望我们的战略方法能够产生积极的转化影响。 通过研究体外生成造血干细胞的新途径，从而实现长期造血移植 能力，同时也为造血领域提供了急需的综合数据集资源 整个目标是在体外成功衍生出临床上可行的 HSC。