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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要血小板源性生长因子（PDGF）及其相应受体（PDGFRα和PDGFRβ）调节器官发生和组织稳态与发育有关。综合症和成人疾病，如纤维化、心血管疾病和癌症，因此，PDGF受体。最近的报告已提出激活及其下游信号传导作为潜在的治疗靶点。表明 PDGFRβ 信号传导参与调节骨髓 (BM) 微环境和造血干尽管已知骨骼干细胞 (SSC) 表达 PDGFRβ，但具有调节作用。 PDGFRβ 在 HSC 生态位中的作用尚不清楚，我们的目标是确定 PDGFRβ 在 HSC 生态位中的作用。我们的中心假设是 SSC 中的 PDGFRβ 信号调节 SSC 的大小。凭借我们在 PDGF 信号传导和新型遗传方面的专业知识，我们可以了解 HSC 生态位和 BM 的造血功能。方法，该项目将通过以下两个目标来解决中心假设：1）表征 SSC PDGFRβ 信号对 HSC 池维持和骨髓增生性肿瘤 (MPN) 的影响 2) 定义 PDGFRβ 改变对 SSC/基质谱系的功能影响在目标 1 中，他莫昔芬诱导的 Gli1 Cre 重组酶小鼠系 (Gli1-CreER) 将用于有条件地诱导功能获得性 PDGFRβ 敲入等位基因或使内源性 Pdgfrb 基因失活使用 WT 和突变体的 Gli1+ SSC 实验将侧重于评估 BM 和突变体的功能变化。 SSC 中 PDGFRβ 发生改变的血液我们将把来自 PDGFRβ 突变体的 BM 移植到经过辐射的野生型 (WT) 中。小鼠（或反之亦然）以确定造血细胞获得永久功能的程度最后，我们将确定 PDGFRβ 突变环境是否发生改变。使用 JAK2-V617F 突变 BM 进行 BM 移植，调节基质环境改变 MPN 进展在目标 2 中，为了表征 HSC 生态位支持的 SSC 和功能的细胞变化和功能。 PDGFRβ突变体中的基质谱系，我们将使用体外SSC测定来评估SSC的功能变化最后，从 PDGFRβ 突变体的 BM 中分选 SSC。对照将被移植到 WT 肾胶囊中，以确定 SSC PDGFRβ 信号传导对 BM 形成和 HSC 体内定植的调节这些项目的结果将产生新的结果。了解 PDGFRβ 信号介导的基质与 HSC 生态位串扰并促进 BM 和造血系统疾病的潜在治疗靶点。