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.

项目总结 目前，在体外生产可移植的造血干细胞(HSCs)可以使成千上万的患者受益 无法接受血液疾病的治疗。长期目标是能够在临床上产生 从人多能干细胞(PSCs)体外培养出可存活的HSCs。这一成就将彻底改变 许多血液疾病可以被治疗，理论上可以减轻患者不得不等待发现 骨髓移植“匹配”，因为已经存在使用造血干细胞的良好临床路线图 在一种治疗环境中。我们的中心假设是基于我们对 体内HSCs(有功能)和体外来源的‘Pre-HSCs’(无功能)在基因上是相同的。通过消除 遗传背景作为可变性的来源，以确定在功能上的关键分子差异 在体外产生的造血干细胞中存在缺陷，我们能够发现在调控方面的新差异 受miRNAs调控的通路。在强劲的初步数据指导下，本申请的总体目标是 在体外分化过程中调节关键的miRNAs，从而提供一种获得功能性 来自人类PSCs的HSCs。这一目标将在两个具体目标下实现：(1)确定miRNA的变化 在人PSC介导的造血分化过程中。在这个目标中，关键的miRNA表达水平将是 在PSC-造血分化试验中进行分析；以及(2)检测miRNAs对 在体外和体内对造血潜能的影响。在这一目标中，将修改关键的miRNA表达水平 在PSC-造血祖细胞分化试验中，分析其成功生成完全 体外培养有功能的造血干细胞。我们期望我们的战略方针将产生积极的翻译影响 通过研究一种在体外产生造血干细胞的新途径，从而导致长期的造血移植 能力，同时还为造血领域提供了亟需的全面数据集资源 为在体外成功获得临床可行的造血干细胞奠定了基础。