Role of cellular metabolism in palate morphogenesis

细胞代谢在上颚形态发生中的作用

• 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
• 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

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，全世界活产儿的患病率为1/700。 人类连锁研究表明，基因突变要么与胆固醇新陈代谢有关，要么异常 母亲的胆固醇饮食会导致颅面畸形，如腭裂。然而，它在很大程度上是未知的。 胆固醇生成障碍是如何导致腭裂的。在我们的初步研究中，我们发现老鼠 随着甾醇-C5-去饱和酶(Sc5d)的丢失，表现为完全外露的腭裂，细胞减少 在腭部形成过程中的增殖。Sonic Hedgehog(SHH)信号，这对正常上颌至关重要 形成，在Sc5d突变小鼠中受到影响。初级纤毛，一种天线状结构，接受 Sc5d突变小鼠腭间充质细胞质膜上的Hedgehog信号发生变形。 我们还发现，非编码RNA的转录后蛋白修饰和表达在 Sc5d突变小鼠的味觉。有趣的是，虽然胆固醇合成在缺乏胆固醇的小鼠中也受到类似的抑制 对于7-脱氢胆固醇还原酶(Dhcr7)基因，它在SC5D之后对胆固醇合成至关重要， 这些小鼠的外显率低于10%，表现为腭裂。在此基础上，在这个项目中，我们 将检验这一假设，即在Sc5d突变小鼠中升高的胆固醇前体Lathosterol起着至关重要的作用 在腭裂发病机制中的作用。我们有三个具体的目标：(1)确定特定的胆固醇如何 中间体通过测试胆固醇中间体lathosterol在体内积累的方式干扰SHH信号转导 Sc5d突变小鼠，干扰Hedgehog受体运动和初级纤毛形成；(2)鉴定 Sc5d突变小鼠发育期腭部蛋白质的变化及蛋白质组学研究 利用Sc5d突变体、Dhcr7突变体和对照小鼠的味觉进行分析；(3)鉴定非编码RNA和 胆固醇代谢受损对其调控基因的影响 MicroRNAs的作用机制(S)及其对腭裂相关基因的调控 在Sc5d突变小鼠中被特异性改变的基因。在我们强大的前期工作的基础上，我们期待着这项研究 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 转录调控水平)，这一结果将导致在预防方面的创新， 胆固醇相关头面部出生缺陷的诊断和治疗。