Discovery and manipulation of transcription factors to restore long term stem cell repopulation in aged bone-marrow

发现和操纵转录因子以恢复衰老骨髓中的长期干细胞增殖

• 批准号: 10334958
• 负责人: Omar O Abudayyeh
• 金额: $ 60.19万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-04 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10334958
• 关键词: Affect; Age; Aging; Animals; Area; Bar Codes; Biological Assay; Biology; Biology of Aging; Blood; Blood Cells; Bone Marrow; Bone Marrow Cells; Cell Aging; Cell Cycle; Cell Lineage; Cells; Chronology; Complex; Computer Models; Coupling; Data; Data Set; Degenerative Disorder; Deterioration; Development; Disease; Engineering; Engraftment; Equilibrium; Evaluation; Eye; Foundations; Gene Expression Profile; Genes; Genetic Transcription; Health; Hematopoietic Stem Cell heterogeneity; Hematopoietic stem cells; Heterogeneity; Human; In Situ; In Vitro; Individual; Inflammation; Inflammatory; Injury; Intervention; Libraries; Lymphoid; Measurement; Methods; Modeling; Modification; Molecular; Molecular Profiling; Mus; Muscle; Myelogenous; Outcome; Output; Phenotype; Physiological; Physiology; Population; Process; RNA; Recovery; Rejuvenation; Resolution; Skin; Testing; Tissues; Training; Transplantation; Validation; Viral; Whole Organism; age effect; age related; aged; base; bone aging; c-myc Genes; cell regeneration; cell type; cellular engineering; cellular transduction; hematopoietic stem cell aging; hematopoietic stem cell niche; hematopoietic stem cell quiescence; hematopoietic stem cell self-renewal; high throughput screening; immune function; in vivo; injury and repair; insight; irradiation; novel; novel therapeutic intervention; novel therapeutics; overexpression; post-transplant; predictive modeling; progenitor; regeneration potential; repaired; response to injury; restoration; screening; self-renewal; senescence; sex; single cell sequencing; single-cell RNA sequencing; small molecule; stem cell aging; stem cell niche; stem cell population; stem cells; therapeutic candidate; tool; tool development; transcription factor; transcriptome; transcriptomics

Project Summary Aging has a complex underlying biology characterized by a progressive loss of cellular and physiological function and this deterioration is strongly correlated with degenerative disease. In the bone marrow, aging markedly reduces the capacity of hematopoietic stem cells (HSCs) to self-renew and differentiate into lymphoid lineages, resulting in hindered immune function and systemic effects on multiple tissues, such as muscle repair after injury. Bone marrow from young recipients has been shown to rejuvenate aged bone marrow as well as systemically in other tissues. However, the exact HSC and progenitor cell states as well as other factors that drive the rejuvenating effects are not well understood. Profiling of HSCs and other bone marrow cell types during aging and an understanding of the transcription factors (TFs)-that control HSC self-renewal and their changes in activity during aging could provide new therapeutic approaches with the potential to reverse both blood-specific and whole-animal effects of aging. TF-based interventions, such as partial reprogramming, have shown promise to promote stem-cell cycling and regeneration. However, current approaches are limited to a small set of predetermined TFs, commonly the Yamanaka factors Oct3/4, Sox2, Klf4 and c-Myc, and have only been demonstrated in a select set of tissues. Furthermore, the diversity of the HSC population, containing both senescent cells and long-term renewing state (LT-HSC) among other subtypes, makes discovery of master regulators challenging, and existing approaches do not address this heterogeneity. We hypothesize that high- resolution single cell RNA sequencing (scRNA-seq) of HSCs from mice of different ages will reveal putative TF regulators of the aging process, and that these candidates can reprogram aged HSCs towards LT-HSC and quiescent states capable of niche restoration to reverse age-associated phenotypes. We will profile molecular signatures of aging in HSCs at single cell resolution and use these data to both develop metrics for aging and nominate TFs to promote LT-HSC restoration and rejuvenation. We will synthesize selected TFs for pooled screening allowing for rapid evaluation of their reprogramming effects in vitro and in vivo. Coupling these pooled perturbations in vivo with scRNA-seq readouts will allow for evaluation of HSC rejuvenation via our aging signatures and measurement of lymphoid/myeloid skew. Candidate TFs that demonstrate the strongest potential for rejuvenation of LT-HSCs will be tested individually and in combination for modification of whole-organism phenotypes, including increased repopulation potential, reduction of inflammatory factors, and improvement of muscle repair in response to injury. The repurposing of novel TFs regulating HSC rejuvenation as new therapeutics for aging-associated disease provides a new framework for cellular engineering. This proposal, coupling transcriptomic readouts and screening for discovery of new regulators of cell states, serves as the foundation for TF-based interventions for disease, both in aging and in broader human health.

项目摘要 衰老具有复杂的基础生物学，其特征是细胞和生理功能的逐渐丧失 这种恶化与退化性疾病密切相关。在骨髓中，衰老明显 降低造血干细胞（HSC）自我更新并分化为淋巴谱系的能力， 导致对多个组织的免疫功能和全身作用的阻碍，例如受伤后的肌肉修复。 已证明来自年轻接受者的骨髓会使老化的骨髓恢复活力 其他组织。但是，确切的HSC和祖细胞状态以及其他驱动的因素 恢复活力的效果尚不清楚。衰老期间HSC和其他骨髓细胞类型的分析 并了解转录因子（TFS） - 控制HSC自我更新及其活动变化 在衰老期间可以提供新的治疗方法，并有可能扭转血液特异性和 衰老的全动物作用。基于TF的干预措施（例如部分重编程）已显示出有望 促进干细胞循环和再生。但是，当前的方法仅限于一小部分 预定的TF，通常是Yamanaka因子Oct3/4，SOX2，KLF4和C-MYC，仅是 在精选的组织中证明。此外，HSC人口的多样性都包含 衰老细胞和长期更新状态（LT-HSC）以及其他亚型，使人发现 监管机构具有挑战性，现有方法并不能解决这种异质性。我们假设高 来自不同年龄的小鼠的HSC的分辨率单细胞RNA测序（SCRNA-SEQ）将揭示假定的TF 老化过程的调节剂，这些候选人可以将老化的HSC对LT-HSC和 能够恢复生态位的静态状态以逆转与年龄相关的表型。我们将介绍分子 在单细胞分辨率下HSC中衰老的签名，并使用这些数据来开发衰老和 提名TFS以促进LT-HSC恢复和恢复活力。我们将合成选定的TFs以进行合并 筛选可以快速评估其在体外和体内的重编程效应。耦合这些汇总 带有SCRNA-SEQ读数的体内扰动将允许通过我们的衰老评估HSC的恢复活力 淋巴/髓样偏斜的特征和测量。表现出最强潜力的候选TFS 为了复兴LT-HSC，将单独测试并结合使用全生物的修改 表型，包括增加重生潜力，减少炎症因素以及改善 响应损伤的肌肉修复。新颖的TFS调节HSC恢复活力的新型TFS的重新使用 衰老相关疾病的治疗剂为细胞工程提供了新的框架。这个建议， 耦合转录组读数和筛选发现细胞状态的新调节剂，作为 基于TF的疾病干预基金会，包括衰老和更广泛的人类健康。