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Genetic and Chemical Biological Studies of a Novel Wnt Inhibitor Tiki2

新型 Wnt 抑制剂 Tiki2 的遗传和化学生物学研究

基本信息

批准号：
8239039
负责人：
Xi He
金额：
$ 57.18万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-16 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8239039
关键词：
Adult Age Aging-Related Process Alleles Binding Biochemical Biological Biology Biomechanics Bone Development Bone Diseases Bone Growth Bone Regeneration Bone Resorption Bone Tissue Cell Communication Cell Line Cells Chemicals Communication Complex Data Defect Disease Drug Delivery Systems Embryonic Development Enzymes Exhibits Family Financial compensation Fracture Gene Expression Generations Genes Genetic Genetic Transcription Homeostasis Human Human Activities Human Genetics In Vitro Integral Membrane Protein LDL-Receptor Related Proteins Ligands Link Lipids Lipoproteins Mechanics Mediating Methods Modification Mus Mutant Strains Mice Mutation Organ Osteoblasts Osteocytes Osteogenesis Osteoporosis Palmitates Pathway interactions Phenotype Physiological Prevention Property Proteins Regulation Regulatory Pathway Risk Role Screening procedure Signal Transduction Site Staging Stimulus Tamoxifen Therapeutic Therapeutic Intervention Time Tissues Wnt proteins Woman adduct aging population bone bone cell bone mass cell type chemical genetics gain of function mutation genetic manipulation global health in vivo inhibitor/antagonist interest loss of function men mouse model new therapeutic target novel novel therapeutics programs receptor response small molecule therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Osteoporosis and bone fractures are global health problems. Identification of new therapeutic targets and strategies are of paramount importance. Signaling by the Wnt family of secreted proteins has emerged as a key pathway for human bone mass regulation, as loss-of-function and 'gain-of-function' mutations in Wnt coreceptor LRP5 are associated with familial osteoporosis and high bone mass diseases, respectively. Wnt/LRP5 signaling primarily regulates the osteoblast lineage and bone generation, providing a potential avenue for therapeutics that stimulates bone growth. Remarkably secreted Wnt antagonists are expressed in bone and modulate Wnt/LRP5 signaling locally, and thus offer treatment opportunities targeting bone specifically. Indeed Sclerostin, an osteocyte-specific secreted factor that binds to and inhibits LRP5, has become a key drug target for osteoporosis. We have identified a new family of Wnt antagonists, referred to as Tiki proteins, which likely are novel enzymes that post-translationally modify and inactivate Wnt ligands. We have generated Tiki2-/- mutant mice, which surprisingly are viable but exhibit high bone mass, suggesting that Tiki2, like Sclerostin, is an important negative regulator of bone homeostasis. Because of its enzymatic activity human TIKI2 may be an ideal target for small molecule inhibitors for potential therapeutic intervention for osteoporosis. We propose four aims to investigate the role of Tiki2 in bone biology and potential therapeutic implications. In Aim 1, we will characterize the high bone mass phenotype of Tiki2-/- mutant mice, employing histological, biochemical and biomechanical methods. We will also examine in details the expression of Tiki2 in bone cell types, its regulation in bone by anabolic and mechanical stimuli. In Aim 2, we will generate mutant mice with conditional Tiki2 deletion in the bone and in the adulthood via cell type- specific and a tamoxifen-inducible Cre lines. These studies will define the site and stage of Tiki2 action in bone and in the aging process. In Aim 3, we will investigate human TIKI2 (and TIKI1) bone-related functions and biochemical properties in vitro. We will investigate TIKI2 and TIKI1 expression in bone, and study whether TIKI2 and TIKI1 regulates bone formation in human osteoblast-like cell lines, and characterize the specific Wnt proteins that are modified/inactivated by TIKI proteins in bone mass regulation. In Aim 4, we will identify small molecule inhibitors of TIKI2 via chemical compound screening and to evaluate their potential in bone growth stimulation in vitro and in vivo. As a new class of Wnt antagonists with an enzymatic activity, TIKI proteins are ideal targets for small molecule inhibitors for use in experimental manipulation and therapeutic intervention. We will perform a high throughput chemical compound screen to identify Tiki2 inhibitors for their potential applications in stimulating bone growth. These studies together represent a comprehensive analysis of TIKI function in bone biology via genetic, biochemical and chemical biological methods, and may discover potential novel therapeutics for osteoporosis and bone regeneration. PUBLIC HEALTH RELEVANCE: Cell-to-cell communication is vital for human bone mass regulation. Defects in these communication networks underlie bone diseases including osteoporosis. We have identified a new gene, called TIKI2, which may regulate cell communication in bone growth. This proposal aims to understand how TIKI2 regulates bone development and growth in the mouse model and human cells, and to search for chemical inhibitors for TIKI2 for potential use in bone growth stimulation. These studies will likely provide better understanding of human bone biology, and may identify new therapeutic strategies and molecules for potential treatment of osteoporosis and bone fractures.

描述(申请人提供)：骨质疏松症和骨折是全球性的健康问题。确定新的治疗目标和策略是至关重要的。由于Wnt辅助受体LRP5的功能缺失和功能获得突变分别与家族性骨质疏松症和高骨量疾病相关，Wnt家族分泌蛋白家族的信号转导已成为人类骨量调节的关键途径。WNT/LRP5信号主要调节成骨细胞谱系和骨生成，为刺激骨生长的治疗提供了潜在的途径。显著分泌的Wnt拮抗剂在骨骼中表达，并在局部调节Wnt/LRP5信号，从而提供针对骨骼的治疗机会。事实上，硬化素，一种结合并抑制LRP5的骨细胞特异性分泌因子，已成为治疗骨质疏松症的关键药物靶点。我们已经确定了一个新的Wnt拮抗剂家族，称为Tiki蛋白，这可能是一种新的酶，可以在翻译后修饰和失活Wnt配体。我们已经产生了Tiki2/-突变小鼠，这些小鼠出人意料地存活下来，但表现出高骨量，这表明Tiki2和硬化素一样，是骨稳态的重要负面调节因子。由于其酶活性，人Tiki2可能成为治疗骨质疏松的小分子抑制剂的理想靶点。我们提出了四个目标来研究Tiki2在骨生物学中的作用和潜在的治疗意义。在目标1中，我们将用组织学、生化和生物力学的方法来表征Tiki2/-突变小鼠的高骨量表型。我们还将详细研究Tiki2在骨细胞类型中的表达，以及合成代谢和机械刺激对其在骨中的调节。在目标2中，我们将通过特定细胞类型和三苯氧胺诱导的Cre系，在骨骼和成年期产生有条件Tiki2缺失的突变小鼠。这些研究将确定Tiki2在骨骼和衰老过程中的作用部位和阶段。在目标3中，我们将在体外研究人类Tiki2(和TIKI1)与骨相关的功能和生化特性。我们将研究Tiki2和TIKI1在骨中的表达，研究Tiki2和TIKI1是否调节人成骨样细胞系的骨形成，并研究在骨量调节中被Tiki蛋白修饰/失活的特异性Wnt蛋白。在目标4中，我们将通过化合物筛选来鉴定Tiki2的小分子抑制剂，并评估它们在体外和体内促进骨生长的潜力。Tiki蛋白是一类新型的具有酶活性的Wnt拮抗剂，是小分子抑制剂用于实验操作和治疗干预的理想靶点。我们将进行高通量化合物筛选，以确定Tiki2抑制剂在刺激骨生长方面的潜在应用。这些研究通过遗传学、生化和化学生物学方法对Tiki在骨生物学中的功能进行了全面的分析，并可能发现治疗骨质疏松症和骨再生的潜在新疗法。公共卫生相关性：细胞与细胞之间的通讯对人类骨量调节至关重要。这些通讯网络的缺陷是包括骨质疏松症在内的骨骼疾病的基础。我们已经发现了一种名为Tiki2的新基因，它可能调节骨骼生长过程中的细胞通讯。这项建议旨在了解Tiki2如何在小鼠模型和人类细胞中调节骨发育和生长，并寻找Tiki2的化学抑制剂，以潜在地用于骨生长刺激。这些研究可能会更好地了解人类的骨生物学，并可能为骨质疏松症和骨折的潜在治疗确定新的治疗策略和分子。