Unbiased mapping of skeletal stem cell function at single cell resolution in homeostasis and injury.

在稳态和损伤中以单细胞分辨率对骨骼干细胞功能进行无偏映射。

• 批准号: 10661359
• 负责人: Philipp Leucht
• 金额: $ 22.37万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661359
• 关键词: Age; Aging; Automobile Driving; Bar Codes; Bioinformatics; Bone Injury; Bone Marrow; Bone Regeneration; Cell Separation; Cell physiology; Cells; Characteristics; Complex; Data; Degenerative polyarthritis; Development; Directories; Disease; Environment; Epiphysial cartilage; Exhibits; Exposure to; Femoral Fractures; Fracture; Future; Gene Expression; Gene Expression Profile; Generations; Genetic; Genetic Induction; Genetic Transcription; Harvest; Homeostasis; In Situ; In Vitro; Individual; Injury; Internet; Knowledge; Label; Location; Maintenance; Maps; Messenger RNA; Multipotent Stem Cells; Mus; Musculoskeletal Diseases; Natural regeneration; Organ; Osteoporosis; Pathway interactions; Periosteum; Phenotype; Play; Population; Population Dynamics; Probability; Regional Anatomy; Resolution; Role; Signal Pathway; Signal Transduction; Site; Skeleton; Structure; Technology; Testing; Therapeutic; Time; Tissues; adult stem cell; bone; bone fracture repair; bone healing; bone health; bone repair; cell behavior; cell injury; cell type; experimental study; improved; in vivo; injury and repair; novel; osteoprogenitor cell; population health; progenitor; programs; prospective; protein expression; regenerative approach; repaired; response; response to injury; self renewing cell; self-renewal; single cell technology; single-cell RNA sequencing; skeletal; skeletal injury; skeletal regeneration; skeletal stem cell; stem cell function; stem cell population; stem cell therapy; stem cells; transcriptome; transcriptomic profiling

Project Summary Tissue integrity is maintained by specialized adult stem cells that replenish old and damaged cells. During aging and in disease states, stem cell function is compromised leading to progressive degeneration of the structure and function of vital organs. This is particularly apparent in the skeleton, which becomes increasingly fragile and prone to fractures as we age. To uncover and counteract the mechanisms of skeletal degeneration and enhance bone repair, we must first define skeletal stem and progenitor cell (SSPC) behavior in vivo and determine the essential regulators of cellular function. This is a technically challenging feat as, in contrast to other organs where stem cells reside within a defined physical location and are readily identified by gene and protein expression, cells with progenitor activity can be found throughout bone and there are no specific markers to identify these in situ. As a result, we still know remarkably little about SSPCs, including whether a multipotent stem cell exists in vivo or if bone is maintained by a pool of lineage-restricted progenitors. Additionally, it is unclear whether progenitors from different locations are functionally equivalent. Recently, we identified several distinct skeletal populations with stem cell characteristics that demonstrate distinct dynamic responses to injury. We hypothesize that these correspond to SSPC subtypes from different anatomical regions of bone and that they play unique roles in skeletal regeneration in homeostasis and injury repair. We will leverage cutting-edge technologies to investigate this hypothesis. In the first part of this proposal, we will perform parallel in vivo clonal functional and transcriptional analysis to identify cells with progenitor activity in an unbiased manner, interrogate the differentiation potential of SSPCs in defined bone regions and distinguish gene expression signatures associated with precise cellular behaviors. These experiments will, for the first time, establish the phenotype and function of individual SSPCs in their native environment and across distinct local niches, providing a holistic overview of the SSPC landscape to enable the identification of novel regulators of stem cell activity and markers to enable the isolation of therapeutically useful cell populations. In the second part of this proposal, we will chart the dynamic response and contribution of individual progenitors to the repair of various types of bone injury. These data will reveal the key cell populations driving discrete stages of bone healing and their transcriptional response to injury, including the signaling pathway signatures enriched in progenitors. Together this study will dissect the complex web of skeletal stem cell states to facilitate the development of targeted strategies to improve bone health and fracture healing.

项目摘要 组织的完整性是由专门的成体干细胞来维持的，这些干细胞补充了衰老和受损的细胞。在老化期间 在疾病状态下，干细胞功能受损，导致结构进行性退化， 重要器官的功能。这在骨骼中尤其明显，骨骼变得越来越脆弱， 随着年龄的增长容易骨折为了揭示和对抗骨骼退化的机制， 骨修复，我们必须首先确定骨骼干细胞和祖细胞（SSPC）在体内的行为，并确定 细胞功能的重要调节剂。这是一个技术上具有挑战性的壮举，与其他器官相比， 干细胞存在于确定的物理位置内并且容易通过基因和蛋白质表达来鉴定， 具有祖细胞活性的细胞可以在整个骨中找到， 原地。因此，我们对SSPCs的了解仍然非常少，包括SSPCs中是否存在多能干细胞。 体内或如果骨由谱系限制的祖细胞库维持。此外，尚不清楚是否 来自不同位置的祖细胞在功能上是等同的。最近，我们发现了几具 具有干细胞特征的群体，表现出对损伤的不同动态反应。我们假设 这些对应于来自骨骼不同解剖区域的SSPC亚型， 在体内平衡和损伤修复中的骨骼再生中的作用。我们将利用尖端技术， 调查这个假设。在本提案的第一部分，我们将进行平行的体内克隆功能和 转录分析，以无偏的方式鉴定具有祖细胞活性的细胞， SSPC在确定的骨区域中的分化潜力并区分基因表达特征 与精确的细胞行为相关。这些实验将首次建立 和功能的个别SSPC在其原生环境和不同的地方利基，提供了一个整体的 SSPC景观概述，以识别干细胞活性和标记物的新型调节剂 从而能够分离治疗上有用的细胞群。在本提案的第二部分，我们将绘制 单个祖细胞对各种类型骨修复的动态响应和贡献 损伤这些数据将揭示驱动骨愈合的离散阶段的关键细胞群及其生物学特性。 转录响应损伤，包括信号转导途径的签名丰富的祖细胞。一起 这项研究将剖析骨骼干细胞状态的复杂网络，以促进靶向干细胞的发展。 改善骨骼健康和骨折愈合的策略。