CD166 Regulates Human and Murine Hematopoietic Stem Cell Function and the Hematopoietic Niche

CD166 调节人和小鼠造血干细胞功能和造血生态位

• 批准号: 10339403
• 负责人: Melissa A Kacena
• 金额: $ 35.44万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10339403
• 关键词: ALCAM gene; Activities of Daily Living; Biology; Cell Communication; Cell surface; Cells; Competence; Cytokine Signaling; Data; Defect; ENG gene; Elements; Endosteum; Endothelial Cells; Engraftment; Epigenetic Process; Genes; Genetic Models; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Home; Human; Immobilization; Immunoglobulins; Incubated; Investigation; Knockout Mice; Ligands; Maintenance; Marrow; Mediating; Mesenchymal Stem Cells; Mitochondria; Mus; Osteoblasts; PECAM1 gene; PTPRC gene; Pathway interactions; Phenotype; Platelet-Derived Growth Factor alpha Receptor; Play; Proteins; Publishing; Regulation; Research; Research Personnel; Role; SLAM protein; Signal Pathway; Stem cell pluripotency; Surface; Testing; Time; Translational Research; Trees; Wild Type Mouse; base; blood treatment; cell type; exhaustion; fitness; hematopoietic stem cell niche; hematopoietic stem cell self-renewal; improved; in vitro Assay; in vivo; innovation; knockout animal; member; nestin protein; receptor; single-cell RNA sequencing; stem; stem cell function; stem cells

The phenotypic makeup and functional capacity of murine and human hematopoietic stem cells (HSC) has been exhaustively investigated making HSC one of the most-characterized groups of cells in mammalian biology. Still, not a single functional marker has been previously identified that is common to both murine and human HSC while being also expressed on at least three important cellular components of the hematopoietic niche (HN), namely osteoblasts (OB), endothelia cells (EC), and mesenchymal stem cells (MSC). Our focus on CD166 began when we first established that CD166 identifies OB that mediate a hematopoiesis enhancing activity. We also published that CD166 is expressed on normal murine and human HSC and plays a key role in determining their functional capacity and the competence of the HN. We recently confirmed that CD166 is expressed on a subset of EC and nestin+ MSC. Since CD166 is capable of mediating hetero- and homophilic cell-cell interactions, we propose that CD166 serves as both ligand and receptor of HSC interactions with cellular elements of the niche and is an important regulator of the competence of the HN and of stem cell function. HSC from CD166-/- mice engraft poorly in lethally irradiated hosts. On the other hand CD166-/- mice do not support the engraftment of LT-HSC suggesting that loss of CD166 abrogates the competence of the HN. Marrow-homed LSK cells from wildtype (WT) donors lodged closer to the endosteum of host mice than similar cells from knockout (KO) animals suggesting that engraftment is adversely impacted by the lack of HSC-OB homophilic CD166-interactions7. Single-cell RNAseq analysis of WT and CD166-/- CD150+CD48- LSK cells incubated with immobilized CD166 protein revealed that engagement of CD166 on these cells activated cytokine signaling, epigenetic pathways, stem cell pluripotency genes, and mitochondria-related signaling pathways. Based on these findings, we hypothesize that CD166 is a functional marker required for HSC- niche interactions that support the competence of the hematopoietic niche. We also believe that preferential CD166 homophilic interactions in the niche contribute to HSC regulation and maintenance of the hematopoietic potential of stem cells. We will test these hypotheses by investigating 2 aims: 1) Test the hypothesis that CD166 expression on OB, EC, and MSC in the hematopoietic niche is required for the competence of the hematopoietic niche and 2) Test the hypothesis that hematopoietic stem cells lacking CD166 homophilic interactions with OB, EC, and MSC in the niche fail to develop normally and acquire an engraftment defect. The significance of this proposal is that it will examine the role of a new and unique functional marker of human and murine HSC and will investigate the impact of CD166 modulation on stem cell function. The novelty of this proposal stems from its potential to implicate CD166 in the regulation of the HN and HSC that reside in and interact with CD166-rich cellular elements of this microenvironment. These studies have the potential to define how homophilic CD166 interactions promote HSC self-renewal.

小鼠和人造血干细胞（HSC）的表型组成和功能能力具有 已被彻底研究，使HSC成为哺乳动物中最具特征的细胞群之一， 生物学尽管如此，以前还没有鉴定出一种单一的功能性标记物是鼠和人共同的。 人HSC同时也在造血干细胞的至少三种重要细胞组分上表达， 小生境（HN），即成骨细胞（OB）、内皮细胞（EC）和间充质干细胞（MSC）。我们专注于 当我们首次确定CD 166识别OB介导造血增强时，CD 166开始。 活动我们还发表了CD 166在正常鼠和人HSC上表达，并在HSC的增殖和分化中起关键作用。 确定他们的职能能力和东道国的能力。我们最近证实，CD 166是 在EC和巢蛋白+ MSC的子集上表达。由于CD 166能够介导异嗜性和同嗜性 细胞-细胞相互作用，我们提出，CD 166作为HSC相互作用的配体和受体， 细胞元件的生态位，是一个重要的调节能力的HN和干 细胞功能来自CD 166-/-小鼠的HSC在致死辐射的宿主中移植不良。CD166-/- 小鼠不支持LT-HSC的植入，这表明CD 166的缺失消除了LT-HSC的能力。 的HN。来自野生型（WT）供体的骨髓归巢的LSK细胞比来自野生型（WT）供体的骨髓归巢的LSK细胞更靠近宿主小鼠的骨内膜。 来自敲除（KO）动物的类似细胞表明，植入受到缺乏 HSC-OB嗜同性CD 166-相互作用7. WT和CD 166-/-CD 150 + CD 48- LSK的单细胞RNAseq分析 与固定化的CD 166蛋白孵育的细胞显示，这些细胞上的CD 166的接合激活了 细胞因子信号传导、表观遗传途径、干细胞多能性基因和与细胞因子相关的信号传导 途径。基于这些发现，我们假设CD 166是HSC-1表达所需的功能性标志物。 支持造血生态位能力的生态位相互作用。我们也相信 在小生境中优先的CD 166嗜同性相互作用有助于HSC的调节和维持 干细胞的造血潜能。我们将通过调查2个目标来测试这些假设：1）测试 假设造血微环境中OB、EC和MSC上的CD 166表达是造血干细胞增殖所必需的， 能力的造血生态位和2）测试的假设，造血干细胞缺乏 CD 166与OB、EC和MSC在小生境中的嗜同性相互作用不能正常发育， 植入缺陷。这项建议的意义在于，它将审查一个新的、独特的 人和小鼠HSC的功能标记，并将研究CD 166调节对干细胞的影响 功能该提议的新奇源于其可能涉及CD 166对HN的调节。 和HSC，它们驻留在该微环境中富含CD 166的细胞成分中并与之相互作用。这些研究 有可能确定嗜同性CD 166相互作用如何促进HSC自我更新。