Role of GI OSTERIX in Gut and Bone Biology

GI OSTERIX 在肠道和骨骼生物学中的作用

基本信息

批准号：
10388883
负责人：
Wentian Yang
金额：
$ 39.41万
依托单位：
RHODE ISLAND HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-28 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10388883
关键词：
Ablation Affect Alleles Binding Sites Biochemical Biological Assay Biology Bone Development Bone Resorption Cell Lineage Cell physiology Cells Cellular Metabolic Process Clinical Medicine Development Enteral Enterocytes Enteroendocrine Cell Epithelial Cells Foundations Foxes Funding Mechanisms Goals Goblet Cells Growth Factor Histology Homeostasis Hormones Human Intake Intestines Investigation Knockout Mice LGR5 gene Maintenance Mediating Metabolic Marker Metabolism Minerals Molecular Mus Musculoskeletal Nutrient OLFM4 gene Organoids Osteoblasts Osteoclasts Osteogenesis Outcome PTPN11 gene Paper Physiology Play Proteins Publications Regulation Reporter Reporting Research Role Signal Pathway Skeletal Development Skeletal bone Skeleton Somatotropin Specific qualifier value Stains Testing Tissues Work Zinc Fingers absorption base bone bone cell bone metabolism cell type crypt cell experimental study feeding gastric inhibitory polypeptide receptor gastrointestinal gastrointestinal epithelium innovation intestinal crypt knockout gene mouse model novel osteoblast differentiation osteogenic osteosarcoma promoter self-renewal skeletal stem cell biomarkers stem cell self renewal stem cells stemness transcription factor

项目摘要

PROJECT SUMMARY: It has been known for decades that the gut profoundly regulates bone mineral homeostasis by influencing the development and function of bone-forming osteoblasts and bone-degrading osteoclasts. At the cellular and molecular levels, how the gut affects the bone and whether skeletal and gastrointestinal (GI) epithelium development is functionally synchronized remains largely unknown. The work outlined in this proposal builds on our novel and exciting discovery that the zinc finger protein OSTERIX, which to this point has been considered a bone-specific master transcription factor, is robustly expressed in the intestinal crypts. OSTERIX expression has not yet been reported in extraskeletal tissues. We discovered OSTERIX in GI epithelium while examining the role of the phosphatase SHP2 in mouse skeletal development using Osterix-Cre-mediated deletion of PTPN11 foxed alleles. Subsequently, we confirmed that the OSTERIX promoter is active and OSTERIX protein is expressed in the GI crypt base stem cells (CBCs). Given OSTERIX’s central role in bone development, we hypothesize that OSTERIX in the GI crypt cells may participate in GI epithelial development and bone mineral homeostasis. Building on these novel findings we will take the R21 funding mechanism and determine how OSTERIX modulates the self-renewal and multiple lineage differentiation of the GI CBCs, and examine whether and how OSTERIX in the GI epithelium modulates osteoblast and osteoclast function and bone mineral homeostasis. In Specific Aim #1 we will test the hypothesis that OSX maintains CBC stemness and regulates multilineage differentiation using genetically modified mice in combination of approaches including marker colocalization, live cell-lineage tracing, organoid culture, histology, and immunohistochemical staining. In Specific Aim #2 we will test the hypothesis that GI OSX regulates OB and osteoclast function and bone mineral homeostasis by modulating the expression of GI epithelial growth factors and hormones. Approaches including new bone formation assays, RNAScope®, histomorphometry, immunostaining, and biochemical assays of bone metabolic markers will be applied in this study.

项目概要：几十年来，人们都知道肠道通过影响骨矿物质稳态来深刻调节骨矿物质稳态。骨形成成骨细胞和骨降解破骨细胞的发育和功能。和分子水平、肠道如何影响骨骼以及骨骼和胃肠 (GI) 上皮是否该提案中概述的工作在功能上是同步的，但在很大程度上仍然未知。关于我们令人兴奋的新发现，即锌指蛋白 OSTERIX，到目前为止，它已被被认为是骨特异性主转录因子，在肠隐窝中强烈表达。我们在胃肠道上皮中发现 OSTERIX 的表达尚未有报道。使用 Osterix-Cre 介导检查磷酸酶 SHP2 在小鼠骨骼发育中的作用随后，我们确认了 PTPN11 狐狸等位基因的缺失，并且 OSTERIX 启动子处于活跃状态。鉴于 OSTERIX 在骨骼中的核心作用，OSTERIX 蛋白在胃肠道隐窝基底干细胞 (CBC) 中表达。发育过程中，我们追寻胃肠道隐窝细胞中的OSTERIX可能参与胃肠道上皮的发育基于这些新发现，我们将采用 R21 资助机制和确定 OSTERIX 如何调节 GI CBC 的自我更新和多谱系分化，以及检查胃肠道上皮中的 OSTERIX 是否以及如何调节成骨细胞和破骨细胞功能，以及在特定目标 #1 中，我们将检验 OSX 维持 CBC 干性的假设。并结合多种方法使用转基因小鼠调节多谱系分化包括标记共定位、活细胞谱系追踪、类器官培养、组织学和在特定目标#2 中，我们将测试 GI OSX 调节 OB 和免疫组织化学染色的假设。通过调节胃肠道上皮生长因子的表达来调节破骨细胞功能和骨矿物质稳态方法包括新骨形成测定、RNAScope®、组织形态测定法、本研究将应用骨代谢标志物的免疫染色和生化测定。