Novel Strategies for Understanding and Treating Fibrous Dysplasia

理解和治疗纤维发育不良的新策略

基本信息

批准号：
10658595
负责人：
EDWARD C HSIAO
金额：
$ 69.96万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10658595
关键词：
Acceleration Address Affect Artificial Intelligence Automobile Driving Binding Biological Assay Biological Markers Biology Bone Diseases Bone Growth Bone callus Calvaria Cells Clinical Complex Cre driver Cyclic AMP DNA Sequence Alteration Data Disease Dysplasia Foundations Fracture Functional disorder Future G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GNAS gene GTP-Binding Protein alpha Subunits, Gs Gene Expression Profile Genes Genetic Health Hematopoietic stem cells Homeostasis Human Hybrids Hyperactivity Individual Institution Isoproterenol Kinetics Knowledge Lead Lesion Ligands McCune-Albright Syndrome Mediating Medical Modeling Molecular Molecular Probes Mus Musculoskeletal Diseases Mutation Natural regeneration Osteitis Fibrosa Disseminata Osteoblasts Osteocytes Osteogenesis Osteoporosis PTH gene Pathology Pathway interactions Patients Phenotype Physiology Production Property Proteins Public Health Recombinants Role Sampling Signal Pathway Signal Transduction Signaling Molecule Testing Therapeutic Variant WNT Signaling Pathway WNT4 gene WNT9A gene Wnt proteins Work analytical method bone bone cell bone fracture repair bone loss bone repair bone turnover cell type computational intelligence digital effective therapy experience genetic approach guanine nucleotide binding protein improved in vivo evaluation induced pluripotent stem cell mouse model novel novel strategies novel therapeutic intervention novel therapeutics osteogenic overexpression parathyroid hormone-related protein pharmacologic prevent rare condition receptor repaired single-cell RNA sequencing skeletal skeletal stem cell stem cell function stem cell model therapeutic target tool transcriptome transcriptomics treatment strategy

Acceleration Address Affect Artificial Intelligence Automobile Driving Binding Biological Assay Biological Markers Biology Bone Diseases Bone Growth Bone callus Calvaria Cells Clinical Complex Cre driver Cyclic AMP DNA Sequence Alteration Data Disease Dysplasia Foundations Fracture Functional disorder Future G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GNAS gene GTP-Binding Protein alpha Subunits, Gs Gene Expression Profile Genes Genetic Health Hematopoietic stem cells Homeostasis Human Hybrids Hyperactivity Individual Institution Isoproterenol Kinetics Knowledge Lead Lesion Ligands McCune-Albright Syndrome Mediating Medical Modeling Molecular Molecular Probes Mus Musculoskeletal Diseases Mutation Natural regeneration Osteitis Fibrosa Disseminata Osteoblasts Osteocytes Osteogenesis Osteoporosis PTH gene Pathology Pathway interactions Patients Phenotype Physiology Production Property Proteins Public Health Recombinants Role Sampling Signal Pathway Signal Transduction Signaling Molecule Testing Therapeutic Variant WNT Signaling Pathway WNT4 gene WNT9A gene Wnt proteins Work analytical method bone bone cell bone fracture repair bone loss bone repair bone turnover cell type computational intelligence digital effective therapy experience genetic approach guanine nucleotide binding protein improved in vivo evaluation induced pluripotent stem cell mouse model novel novel strategies novel therapeutic intervention novel therapeutics osteogenic overexpression parathyroid hormone-related protein pharmacologic prevent rare condition receptor repaired single-cell RNA sequencing skeletal skeletal stem cell stem cell function stem cell model therapeutic target tool transcriptome transcriptomics treatment strategy

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项目摘要

PROJECT SUMMARY Current treatments for many musculoskeletal disorders are often suboptimal. Understanding the signals controlling skeletal homeostasis and repair is of high relevance, since this knowledge is critical for developing effective therapies for common conditions such as osteoporosis and fractures. In this proposal, we study fibrous dysplasia (FD) as a way to understand the regulators of human bone formation, and test if these pathways could then be used to enhance bone repair. FD accounts for 2.5% of all bone lesions and can occur as part of McCune-Albright Syndrome (MAS). FD is caused by genetic mutations in the GNAS locus, leading to a constitutively active Gs-GPCR protein, hence increasing cAMP levels and causing aberrant cellular signaling. Medical treatments for this disfiguring disorder are sorely lacking. This proposal uses new tools, including mouse models, human induced pluripotent stem cells (iPSCs), skeletal stem/progenitor cells, and advanced genetic strategies, to address the critical knowledge gaps and to find novel therapies for these medically significant conditions. We propose three specific aims: Aim 1: Identify novel compounds that directly target Gsα-regulated cAMP and Wnt production. We previously showed that stopping excess Gs-GPCR pathway activity in mice could dramatically reverse FD-like bone lesions. Using a new artificial intelligence computational approach, we found 71 candidate compounds predicted to selectively bind the GsαR201H protein. Preliminary studies demonstrate that some of these show the desired inhibition of GsαR201H-induced basal cAMP production. This Aim takes our top candidates and further characterizes their ability to block cAMP and Wnt activity, as potential molecular tools for manipulating GNAS. We also test if the lead compounds can reverse existing FD lesions in mouse calvarial cultures. Aim 2: Test if Wnt inhibition can prevent FD bone lesions in mice. How the GNAS mutations in FD cause dramatic bone formation is still poorly defined. We recently found that three proteins, Wnt4, Wnt5a, and Wnt9a, are upregulated in both mouse and human FD bone lesions. This Aim tests if blocking these proteins can reverse FD bone lesions in mice, and examines how each Wnt protein impacts FD lesion pathology. Aim 3: Determine the pathways that are dysregulated in human FD bone lesions and assess the roles of WNT signaling in human skeletal stem/progenitor cells during fracture repair. This aim uses advanced genetics on human FD bone samples to identify the malfunctioning cell types that cause FD. We also test how overactivating WNT4, WNT5a, or WNT9a in human skeletal stem/progenitor cells affect bone formation in a fracture healing model. These results will identify new targets for treating FD and for promoting fracture healing. This application address key knowledge gaps about which Wnt signaling molecules drive FD and how they impact human osteogenesis. The proposal comes from an established, strong, collaborative, multi-institutional team with extensive experience in GPCRs, FD, bone biology, and bone analytical methods. 1

项目摘要目前对许多肌肉骨骼疾病的治疗通常是次优的。了解信号控制骨骼稳态和修复是很高的相关性，因为此知识对于发展至关重要对于常见疾病（例如骨质疏松症和裂缝）的有效疗法。在此提案中，我们研究纤维发育不良（FD），是一种理解人骨形成的调节剂的一种方式，并测试是否可以使用这些途径来增强骨修复。 FD占所有骨骼病变的2.5％并且可以作为McCune-Albright综合征（MAS）的一部分发生。 FD是由GNA中的基因突变引起的基因座，导致持续活跃的GS-GPCR蛋白，从而增加营地水平并引起异常细胞信号传导。严重缺乏这种毁容障碍的医疗治疗。该建议使用新的工具，包括小鼠模型，人类诱导的多能干细胞（IPSC），骨骼干/祖细胞，和先进的遗传策略，以解决关键知识差距并找到这些新的疗法具有医学意义的状况。我们提出了三个具体目标：目标1：确定直接靶向GSα调节的CAMP和WNT产生的新型化合物。我们以前表明，在小鼠中停止超过GS-GPCR途径活动可能会急剧逆转FD样骨骼病变。使用新的人工智能计算方法，我们发现了71种候选化合物预测将GSαR201H蛋白结合。初步研究表明，其中一些表明 GSαR201H诱导的基本cAMP生产所需的抑制作用。这个目标将我们的顶级候选人和进一步将其阻止CAMP和WNT活动的能力作为操纵GNA的潜在分子工具。我们还测试了铅化合物是否可以逆转小鼠颅培养物中现有的FD病变。 AIM 2：测试WNT抑制是否可以防止小鼠的FD骨病变。 FD中的GNA突变是如何引起的戏剧性的骨形成仍然很差。我们最近发现三种蛋白质Wnt4，Wnt5a和Wnt9a，在小鼠和人类FD骨骼病变中均上调。该目标测试是否阻止这些蛋白质可以逆转小鼠的FD骨病变，并检查每个Wnt蛋白如何影响FD病变病理。 AIM 3：确定在人FD骨骼病变中失调的途径，并评估断裂修复过程中人类骨骼干/祖细胞中的Wnt信号传导。这个目标使用高级人类FD骨样品的遗传学，以鉴定引起FD的细胞类型。我们还测试了如何在人骨骼干/祖细胞中过度活化的Wnt4，Wnt5a或Wnt9a会影响A中的骨骼形成断裂愈合模型。这些结果将确定用于治疗FD和促进断裂愈合的新目标。此应用程序地址关键知识差距关于哪种Wnt信号分子驱动FD以及它们如何影响人的成骨。该提案来自既定，强大，协作，多机构的在GPCR，FD，骨生物学和骨分析方法方面拥有丰富经验的团队。 1