Role of cellular metabolism in palate morphogenesis

细胞代谢在上颚形态发生中的作用

基本信息

批准号：
10192706
负责人：
Junichi Iwata
金额：
$ 37.05万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10192706
关键词：
7-dehydrocholesterol 7-dehydrocholesterol reductase Affect Cell Proliferation Cell membrane Cells Cholesterol Cholesterol Homeostasis Cilia Cleft Palate Clinical Complex Congenital Abnormality Craniofacial Abnormalities DNA Sequence Alteration Deformity Development Diagnosis Diet Embryo Environmental Risk Factor Erinaceidae Etiology Exhibits Foundations Gel Gene Expression Gene Expression Regulation Genes Genetic Human Impairment In Vitro Infant Knowledge Label Lead Limb structure Link Live Birth Lovastatin Mediating Membrane Mesenchymal Metabolic Metabolism MicroRNAs Micrognathism Morphogenesis Movement Mus Mutant Strains Mice Mutation Nose Palate Pathogenesis Patients Penetrance Pharmacology Phenocopy Phenotype Physiological Plant Roots Play Post-Transcriptional Regulation Post-Translational Protein Processing Pregnancy Prevalence Prevention Process Production Proteins Proteomics Ptosis Quantitative Reverse Transcriptase PCR Rab8 protein Regulation Reverse Transcriptase Polymerase Chain Reaction Risk Factors Role S-Phase Fraction SHH gene SMO gene Severities Smith-Lemli-Opitz Syndrome Sterols Structure Syndrome Testing Tooth structure Untranslated RNA Up-Regulation Vesicle Work base cholesterol biosynthesis craniofacial craniofacial development desaturase enhanced green fluorescent protein experimental study in vivo inhibitor/antagonist innovation lathosterol malformation mutant novel strategies overexpression palatogenesis protein expression public health relevance rab11 protein receptor smoothened signaling pathway sonic hedgehog receptor transcriptome sequencing

项目摘要

Project Abstract Cleft palate is one of the most common congenital birth defects, with a prevalence of 1/700 live births worldwide. Human linkage studies have shown that either genetic mutations related to cholesterol metabolism or abnormal maternal cholesterol diets lead to craniofacial deformities such as cleft palate. However, it is largely unknown how disturbances in cholesterol production result in cleft palate. In our preliminary studies, we found that mice with loss of sterol-C5-desaturase (Sc5d) displayed cleft palate with complete penetrance through decreased cell proliferation during palate formation. Sonic hedgehog (SHH) signaling, which is crucial for normal palate formation, was compromised in Sc5d mutant mice. The primary cilium, an antenna-like structure receiving hedgehog signals on the plasma membrane, was deformed in palatal mesenchymal cells of Sc5d mutant mice. We also found that posttranscriptional protein modification and expression of non-coding RNAs was altered in the palate of Sc5d mutant mice. Interestingly, while cholesterol synthesis is inhibited similarly in mice deficient for the 7-dehydrocholesterol reductase (Dhcr7) gene, which is crucial for cholesterol synthesis right after SC5D, these mice display cleft palate with a penetrance lower than 10%. Based on this foundation, in this project we will test the hypothesis that lathosterol, a cholesterol precursor that is elevated in Sc5d mutant mice, plays crucial roles in the pathogenesis of cleft palate. We have three specific aims; (1) To determine how a specific cholesterol intermediate interferes with SHH signaling by testing how lathosterol, a cholesterol intermediate accumulated in Sc5d mutant mice, interferes with hedgehog receptor movement and primary cilium formation; (2) To identify altered proteins and modifications in the developing palate of Sc5d mutant mice by conducting proteomic analyses using the palate of Sc5d mutant, Dhcr7 mutant, and control mice; (3) To identify non-coding RNAs and their regulated genes influenced by impaired cholesterol metabolism through analysis of the regulatory mechanism(s) of microRNAs (short non-coding RNAs) and their regulation of genes associated with cleft palate that are specifically altered in Sc5d mutant mice. Building on our strong preliminary work, we expect this study will systematically investigate the roles of cholesterol metabolism (at the cellular, metabolic, proteomic, and post- transcriptional regulation levels) in cleft palate in mice, and the results will lead to innovations in the prevention, diagnosis, and treatment of cholesterol-related craniofacial birth defects.

项目摘要腭裂是最常见的先天性出生缺陷之一，全世界活产婴儿的患病率为 1/700。人类连锁研究表明，要么是与胆固醇代谢相关的基因突变，要么是异常母亲的胆固醇饮食会导致颅面畸形，例如腭裂。然而，很大程度上不为人所知胆固醇生成紊乱如何导致腭裂。在我们的初步研究中，我们发现小鼠甾醇-C5-去饱和酶 (Sc5d) 缺失，显示腭裂，细胞完全外显上颚形成过程中的增殖。声波刺猬 (SHH) 信号传导，对正常味觉至关重要 Sc5d 突变小鼠的形成受到损害。初级纤毛，一种类似天线的结构，接收 Sc5d 突变小鼠的腭间充质细胞中，质膜上的刺猬信号发生了变形。我们还发现转录后蛋白质修饰和非编码RNA的表达发生了改变 Sc5d突变小鼠的上颚。有趣的是，虽然在缺乏胆固醇的小鼠中，胆固醇合成也受到类似的抑制。对于 7-脱氢胆固醇还原酶 (Dhcr7) 基因，该基因对于 SC5D 后胆固醇的合成至关重要，这些小鼠表现出腭裂，外显率低于 10%。基于这个基础，在这个项目中我们将检验以下假设：Lathosterol（一种在 Sc5d 突变小鼠中升高的胆固醇前体）发挥着至关重要的作用在腭裂发病机制中的作用。我们有三个具体目标； (1) 确定特定胆固醇的含量中间体通过测试拉索甾醇（一种胆固醇中间体）如何在体内积累来干扰 SHH 信号传导 Sc5d突变小鼠，干扰hedgehog受体运动和初级纤毛形成； (2) 识别通过进行蛋白质组学改变了 Sc5d 突变小鼠发育中的蛋白质和修饰使用 Sc5d 突变体、Dhcr7 突变体和对照小鼠的上颚进行分析； (3) 鉴定非编码RNA和通过分析调节基因，了解其受胆固醇代谢受损的影响 microRNA（短非编码 RNA）的机制及其对腭裂相关基因的调节 Sc5d 突变小鼠中发生了特别改变。基于我们强有力的前期工作，我们期望这项研究将系统地研究胆固醇代谢的作用（在细胞、代谢、蛋白质组和后代谢方面）转录调控水平）在小鼠腭裂中的应用，其结果将带来预防方面的创新，胆固醇相关颅面出生缺陷的诊断和治疗。