Growth Factor/Extracellular Matrix Interactions During Branching Morphogenesis

分支形态发生过程中生长因子/细胞外基质的相互作用

基本信息

批准号：
8929682
负责人：
Matthew Hoffman
金额：
$ 101.1万
依托单位：
NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8929682
关键词：
Adult Affect Axon Binding Biomedical Engineering Biopsy Blood Vessels Cells Communication Complex Cues Defect Development Duct (organ) structure Engineering Enzymes Epithelial Epithelium Epitopes Excision Extracellular Matrix FGF10 gene Feedback Fibroblast Growth Factor Ganglia Genetic Gland Goals Growth Factor Growth Factor Receptors Head and Neck Neoplasms Heparitin Sulfate Heterogeneity Human In Vitro Inorganic Sulfates Lacrimal gland structure MicroRNAs Modification Morphogenesis Morphology Natural regeneration Nerve Neural Crest Neurons Oral health Organ Organogenesis Patients Peripheral Nervous System Protein Isoforms Quality of life Radiation Repression Role Salivary Salivary Glands Signal Transduction Sjogren&apos s Syndrome Staging Stem cells Structure Structure of parasympathetic ganglion Syndrome Therapeutic Unspecified or Sulfate Ion Sulfates autocrine cell type digital fetal fibroblast growth factor receptor 2b gland development insight nerve supply precursor cell progenitor repaired research study response tumor

项目摘要

We are investigating how heparan sulfate influences FGFR2b signaling in specific progenitor cell types in the epithelium. The exquisite control of growth factor function by HS is dictated by the tremendous structural heterogeneity of its sulfated modifications. It is not known how specific HS structures control growth factor-dependent progenitor expansion during organogenesis. We isolated KIT+ progenitors from fetal salivary glands during a stage of rapid progenitor expansion and profiled HS biosynthetic enzyme expression. Enzymes generating a specific type of 3-O-sulfated-HS (3-O-HS) are enriched in the KIT+ cells, and FGF10/FGFR2b signaling directly regulates their expression. We used bioengineered 3-O-HS to investigate HS function. 3-O-HS binds FGFR2b and stabilizes FGF10/FGFR2b complexes in a receptor- and growth factor-specific manner. Rapid autocrine feedback increases 3-O-HS, KIT and progenitor expansion. In complementary experiments, the knockdown of multiple Hs3st isoforms limits fetal progenitor expansion, but is rescued with bioengineered 3-O-HS, which also increases adult progenitor expansion. We show that rapidly altering a specific 3-O-sulfated epitope provides a cellular mechanism to modulate the response to FGFR2b signaling and control progenitor expansion. We propose that 3-O-HS may expand KIT+ progenitors isolated from biopsies in vitro to use for salivary gland regeneration. We are also identifying the signals that initiate ganglion formation in the salivary gland. Nerves often follow alongside blood vessels to navigate to their targets using similar sets of guidance cues. However, defining the signals that initiate the association between the peripheral nervous system and the salivary epithelium will be important to regenerate or engineer functioning organs. During SMG organogenesis innervation begins when neural crest-derived Schwan cell precursors differentiate into neurons and condense around the epithelial duct to form the parasympathetic ganglion. Subsequently the ganglion neurons extend axons to innervate the epithelium of the gland. We are studying epithelial-derived factors that initiate the neuronal-epithelial communication resulting in neurons condensing around the duct. These epithelial signals are dependent on the localized repression of FGF signaling within the duct. In contrast, enhanced epithelial FGF signaling antagonizes these factors, resulting in defects in epithelial-neuronal communication leading to disrupted ganglia formation and epithelial morphology. This also results in a subsequent depletion of the epithelial progenitor cell reservoir. Our studies demonstrate that epithelial-derived factors are required for gangliogenesis and association with the epithelial duct. Signals that promote gangliogenesis and neuronal-epithelial interactions may inform organ engineering, regeneration, and repair.

我们正在研究上皮细胞中特定祖细胞类型中硫酸盐如何影响FGFR2B信号传导。 HS对生长因子功能的精致控制取决于其硫酸化修饰的巨大结构异质性。尚不清楚特定的HS结构如何控制器官发生过程中生长因子依赖性祖细胞的扩展。在快速祖细胞扩张的阶段，我们从胎儿唾液腺中分离了试剂盒+祖细胞，并介绍了HS生物合成酶的表达。生成特定类型的3-O硫化-HS（3-O-HS）的酶富含在试剂盒+细胞中，而FGF10/FGFR2B信号直接调节其表达。我们使用生物工程的3-O-HS研究HS功能。 3-O-HS结合FGFR2B并以受体和生长因子特异性方式稳定FGF10/FGFR2B复合物。快速自分泌反馈增加了3-O-HS，套件和祖细胞的扩展。在互补实验中，多个HS3同工型的敲低限制了胎儿祖细胞的扩张，但被生物工程的3-O-HS营救，这也会增加成人祖细胞的扩张。我们表明，快速改变特定的3O硫化表位提供了一种细胞机制，以调节对FGFR2B信号传导和控制祖细胞膨胀的响应。我们建议3-O-HS可以扩展体外从活检中分离出来的试剂盒+祖细胞，以用于唾液腺再生。我们还确定了启动唾液腺中神经节形成的信号。神经经常与血管一起使用，使用类似的指导提示导航到目标。但是，定义启动周围神经系统与唾液上皮之间关联的信号对于再生或工程师功能器官非常重要。在SMG器官发生期间，当神经rest衍生的雪旺细胞前体分化为神经元并围绕上皮管凝结以形成副交感神经节时，才开始。随后，神经节神经元延伸轴突以支配腺体的上皮。我们正在研究启动神经元上皮通信的上皮因素，导致神经元在管道周围凝结。这些上皮信号取决于管道内FGF信号的局部抑制。相反，增强的上皮FGF信号传导拮抗了这些因素，从而导致上皮 - 神经元通信的缺陷导致神经节形成和上皮形态破坏。这也导致上皮祖细胞储层的随后耗竭。我们的研究表明，上皮衍生的因素是神经节生成和与上皮管的关联所必需的。促进神经节发生和神经元相互作用的信号可能会为器官工程，再生和修复提供信息。