Skeletal Stem Cell PDGFR-beta Signaling as a New Regulator of the HSC Niche

骨骼干细胞 PDGFR-β 信号传导作为 HSC 生态位的新调节器

• 批准号: 10181025
• 负责人: Hae Ryong Kwon
• 金额: $ 7.05万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10181025
• 关键词: Abnormal Platelet; Address; Adipocytes; Adult; Alleles; Anemia; Biological Assay; Blood; Blood Tests; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Cardiovascular Diseases; Cell Lineage; Cell Maintenance; Cells; Chemoresistance; Chondrocytes; Chondrogenesis; Chromosome abnormality; Colony-Forming Units Assay; Colony-forming units; Coupled; DNA Sequence Alteration; Data; Defect; Development; Disease; Environment; Evaluation; Fibroblasts; Fibrosis; Flow Cytometry; Gene Expression Profiling; Genetic; Goals; Growth Factor; Hematologic Neoplasms; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Specification; Hematopoietic stem cells; Homeostasis; Image; In Vitro; Instruction; JAK2 gene; Kidney; Knock-in; Knowledge; Leukemic Cell; Life; Maintenance; Malignant Neoplasms; Marrow; Mediating; Medicine; Mesenchymal Stem Cells; Multipotent Stem Cells; Mus; Mutant Strains Mice; Mutation; Myelofibrosis; Myeloproliferative disease; Neoplasms; Oncogenic; Organogenesis; Osteoblasts; Osteogenesis; PDGFRB gene; Pathology; Phenotype; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Platelet-Derived Growth Factor alpha Receptor; Platelet-Derived Growth Factor beta Receptor; Process; Production; Receptor Activation; Receptor Signaling; Regenerative capacity; Regulation; Reporting; Research Project Grants; Role; Signal Transduction; Stromal Cells; Supporting Cell; Syndrome; Tamoxifen; Testing; Tissues; Transplantation; Wild Type Mouse; Work; blood formation; bone; capsule; cell transformation; cytokine; driving force; experimental study; gain of function; genetic approach; genetic manipulation; hematopoietic stem cell expansion; hematopoietic stem cell niche; hematopoietic stem cell self-renewal; human disease; improved; in vivo; insight; leukemia; lipid biosynthesis; mouse Cre recombinase; mouse genetics; mouse model; mutant; novel; organ growth; postnatal; receptor; regeneration potential; self-renewal; skeletal stem cell; stem cell engraftment; stem cell function; stemness; targeted treatment; therapeutic target; therapeutically effective

PROJECT SUMMARY/ABSTRACT Platelet-derived growth factor (PDGF) and its corresponding receptors (PDGFRα and PDGFRβ) regulate organogenesis and tissue homeostasis. Abnormal receptor signaling has been implicated in developmental syndromes and adult diseases such as fibrosis, cardiovascular disease, and cancer. Therefore, PDGF receptor activation and its downstream signaling have been proposed as potential therapeutic targets. Recent reports have implicated PDGFRβ signaling in regulating bone marrow (BM) microenvironment and hematopoietic stem cell (HSC) maintenance. Although skeletal stem cells (SSCs) are known to express PDGFRβ, a regulatory role of PDGFRβ in HSC niche is unknown. Our goal is to determine the roles of PDGFRβ in HSC niche maintenance and SSC function. Our central hypothesis is that PDGFRβ signaling in SSCs regulates the size of the HSC niche and blood-forming functions of the BM. With our expertise in PDGF signaling and novel genetic approaches, this project will address the central hypothesis with the two following aims: 1) Characterize the impact of SSC PDGFRβ signaling on the HSC pool maintenance and myeloproliferative neoplasia (MPN) development and 2) Define the functional consequences on SSCs/stromal lineages with altered PDGFRβ signaling. In Aim 1, tamoxifen-inducible Gli1 Cre recombinase mouse line (Gli1-CreER) will be used to conditionally induce a gain-of-function PDGFRβ knock-in allele or to inactivate the endogenous Pdgfrb gene in Gli1+ SSCs. Experiments using WT and mutants will focus on the evaluation of functional changes in BM and blood with altered PDGFRβ in SSCs. We will transplant BM from PDGFRβ mutants to irradiated wild type (WT) mice (or vice versa) to determine the extent at which blood forming cells acquire permanent functional alterations from the PDGFRβ mutant environment. Finally, we will determine whether altered PDGFRβ- regulated stromal environment alters MPN progression using BM transplantation with JAK2-V617F mutant BM cells. In Aim 2, in order to characterize cellular changes and functionality in HSC niche-supportive SSCs and stromal lineages in PDGFRβ mutants, we will use in vitro SSC assays to evaluate functional changes in SSC stemness, differentiation, and cytokine production. Finally, SSCs sorted from BM of PDGFRβ mutants or controls will be transplanted into WT kidney capsules to determine the impact of SSC PDGFRβ signaling on the modulation of BM formation and HSC colonization in vivo. The results of these projects will generate new knowledge of PDGFRβ signaling-mediated stroma-to-HSC niche crosstalk and facilitate the development of potential therapeutic targets on BM and hematopoietic diseases.

项目摘要/摘要 血小板衍生的生长因子（PDGF）及其相应的受体（PDGFRα和PDGFRβ）调节 器官发生和组织稳态。发育中已经实施了异常的接收器信号传导 综合征和成人疾病，例如纤维化，心血管疾病和癌症。因此，PDGF受体 激活及其下游信号已被提出为潜在的治疗靶标。最近的报告 在调节骨髓（BM）微环境和造血茎中实现了PDGFRβ信号传导 单元（HSC）维护。尽管已知骨骼干细胞（SSC）表达PDGFRβ，但一种调节作用 HSC利基市场中的PDGFRβ尚不清楚。我们的目标是确定pDGFRβ在HSC利基市场中的作用 维护和SSC功能。我们的中心假设是SSC中的PDGFRβ信号传导调节 BM的HSC利基和血液形成功能。凭借我们在PDGF信号传导和新遗传方面的专业知识 方法是，该项目将以以下两个目的解决中心假设：1）表征 SSCPDGFRβ信号传导对HSC池维护和骨髓增生性肿瘤（MPN）的影响 开发和2）定义对pDGFRβ改变的SSC/基质谱系的功能后果 信号。在AIM 1中，将使用他莫昔芬诱导的Gli1 CRE重组酶小鼠系（GLI1-CREER）用于 有条件地诱导功能获得的PDGFRβ敲入等位基因或使内源性PDGFRB基因在 GLI1+ SSC。使用WT和突变体的实验将重点介绍BM和BM功能变化的评估 SSC中PDGFRβ改变的血液。我们将BM从PDGFRβ突变体移植到辐照的野生型（WT） 小鼠（反之亦然）确定血液形成细胞获得永久功能的程度 PDGFRβ突变体环境的改变。最后，我们将确定是否改变了PDGFRβ- 通过使用JAK2-V617F突变体BM进行BM移植，调节的基质环境改变MPN的进展 细胞。在AIM 2中，为了表征HSC支持SSC的细胞变化和功能 PDGFRβ突变体中的基质谱系，我们将使用体外SSC分析来评估SSC的功能变化 干性，分化和细胞因子产生。最后，从PDGFRβ突变体BM排序的SSC或 对照将被移植到WT肾脏胶囊中，以确定SSCPDGFRβ信号的影响 BM形成和HSC定植在体内的调节。这些项目的结果将产生新的 了解PDGFRβ信号传导介导的基质到HSC的基裂串扰和支持的发展 BM和造血疾病的潜在治疗靶标。