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 left裂是最常见的先天性先天缺陷之一，在全球范围内，有1/700的活产率。人类链接研究表明，与胆固醇代谢有关的遗传突变或异常孕产妇胆固醇饮食会导致颅面畸形，例如唇裂。但是，这在很大程度上未知胆固醇产生的障碍如何导致口感裂。在我们的初步研究中，我们发现小鼠随着sterol-c5-尿素酶（SC5D）的损失显示裂纹口感，完全渗透通过降低的细胞味形成过程中的增殖。 Sonic刺猬（SHH）信号传导，这对于正常口感至关重要在SC5D突变小鼠中造成了形成。原代纤毛，一种天线状结构，接收质膜上的刺猬信号在SC5D突变小鼠的palatal间充质细胞中变形。我们还发现，未编码RNA的转录后蛋白质修饰和表达在 SC5D突变小鼠的口感。有趣的是，尽管胆固醇合成在不足的小鼠中被类似地抑制对于7-脱氢胆固醇还原酶（DHCR7）基因，这对于SC5D之后的胆固醇至关重要，这些小鼠的裂纹裂纹低于10％。基于这个基础，在这个项目中我们将检验以下假设：lathosterol是一种升高SC5D突变体小鼠的胆固醇前体在left裂的发病机理中的作用。我们有三个特定的目标；（1）确定特定胆固醇如何中间体通过测试胆固醇中间体积聚的脂肪酚如何通过测试脂肪酚来干扰SHH信号传导。 SC5D突变小鼠，会干扰刺猬受体运动和原发性纤毛形成。（2）识别通过传导蛋白质组学，SC5D突变小鼠发育中的pa的蛋白质改变了蛋白质和修饰使用SC5D突变体，DHCR7突变体和对照小鼠的口感进行分析；（3）识别非编码RNA和他们受调节的胆固醇代谢受损影响受损的受调节基因 microRNA（短非编码RNA）的机理及其对与left裂相关的基因的调节在SC5D突变小鼠中特异性改变。在我们的强大初步工作的基础上，我们希望这项研究将系统地研究胆固醇代谢的作用（在细胞，代谢，蛋白质组学和后小鼠c裂的转录调节水平），结果将导致预防的创新，与胆固醇相关的颅面先天缺陷的诊断和治疗。