Molecular and cellular mechanisms underlying Satb2-mediated variation in craniofacial disease

Satb2介导的颅面疾病变异的分子和细胞机制

基本信息

批准号：
10046976
负责人：
Jennifer Leslie Fish
金额：
$ 45.59万
依托单位：
UNIVERSITY OF MASSACHUSETTS LOWELL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-12-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10046976
关键词：
Address Affect Apoptosis Binding Binding Proteins Birth Bone Density Bone Diseases CRISPR/Cas technology Calvaria Cell Cycle Progression Cell Line Cell Separation Cells ChIP-seq Chromatin Cleft Palate Clinical Complex Congenital Abnormality Craniofacial Abnormalities DNA DNA Damage DNA Sequence Alteration DNA biosynthesis Data Defect Dental Development Diagnosis Differentiated Gene Disease Environment Exhibits Funding G1 Phase G2 Phase Gene Expression Genes Genetic Genetic Counseling Genetic Transcription Genotype Human Hydrogen Peroxide Individual Institution Investigation Jaw Lamin Type A Lamins Lead Link Matrix Attachment Regions Mediating Micrognathism Molecular Mus Mutate Mutation Nuclear Osteoblasts Osteogenesis Oxidative Stress Patients Penetrance Phenocopy Phenotype Population Process Publishing Regulation Regulatory Element Research Risk Role S Phase Severities Severity of illness Skeleton Suggestion Syndrome Testing Transcriptional Regulation Variant Work base cell type craniofacial craniofacial bone craniofacial disorder disease phenotype dosage experimental study improved induced pluripotent stem cell insight malformation mutant novel osteoblast proliferation osteogenic outcome forecast progenitor replication stress single-cell RNA sequencing skeletal transcription factor undergraduate student

项目摘要

PROJECT SUMMARY Craniofacial anomalies are among the most common and debilitating human birth defects, affecting 1/500 to 1/2000 births depending on the population. Variation in the severity and penetrance of craniofacial anomalies is a widely observed, but poorly understood, phenomenon. This variation causes complex clinical problems, such as difficulties in the diagnosis, treatment, and genetic counseling of individuals affected by, or susceptible to, craniofacial disorders. In particular, it is widely recognized that similar genetic mutations often express a spectrum of disease phenotypes, but it is still unknown what mechanisms contribute to this variation. For example, human patients with mutations in SATB2 exhibit a range of craniofacial phenotypes, including small lower jaws (micrognathia) and variable penetrance in cleft palate. Similarly, in mice, Satb2 has a dosage-effect on jaw size. Yet, the mechanisms causing such variation are not well understood. Satb2 is a matrix attachment region (MAR)-binding protein that has been shown to regulate osteogenic differentiation through chromatin organization, acting as a high-order transcription factor. Reduction in the expression of osteogenic differentiation genes has been thought to underlie Satb2-mediated craniofacial defects. However, recently published data indicate a more complex role for Satb2 in osteogenesis that includes regulation of pre- osteoblast proliferation and may also involve a role in DNA replication. MARs are implicated in both gene transcription and DNA replication, and Satb2 is highly expressed in S-phase in osteoblasts, suggestive of a novel role in DNA replication. The specific hypothesis to be tested in this proposal is that Satb2 binding at MARs regulates osteogenic proliferation either through regulation of transcription of genes involved in proliferation and/or by regulation of DNA replication. Reductions in Satb2 levels are predicted to decrease the fidelity and increase variation in these processes. This hypothesis will be evaluated through three Specific Aims. Specific Aim 1 will determine how mutations in Satb2 affect gene expression in osteoblast progenitors. Specific Aim 2 will determine how cell cycle progression affects Satb2 localization on MARs. Specific Aim 3 will determine how proliferation in Satb2 mutant pre-osteoblasts is affected by Lamin A/C expression and oxidative stress (factors previously found to be altered in Satb2 mutant cells). Experiments will be performed in both primary and immortalized mouse calvarial cells. Satb2 mutant cells will be compared to wild-type cells from an isogenic background. Data generated from work in this proposal will provide insight into a novel mechanism for Satb2 regulation of osteogenesis. Future research plans include further investigation of genetic, developmental, and environmental contributions to variation in disease phenotypes in patients affected by the SATB2-associated syndrome (SAS) using patient-derived iPSCs. Importantly, this work will also substantially enhance the research environment and provide exciting opportunities for undergraduates to conduct high- impact research, preparing them to enter the biomedical workforce.

项目摘要颅面异常是最常见和令人衰弱的人类出生缺陷，影响1/500至 1/2000分的出生，具体取决于人口。颅面异常的严重程度和外观变化是一个广泛观察的现象，但知之甚少。这种变异会导致复杂的临床问题，例如，受影响或易感的个体的诊断，治疗和遗传咨询困难颅面疾病。特别是，人们广泛认识到，相似的遗传突变通常表达疾病表型的光谱，但尚不清楚哪种机制有助于这种变异。为了例如，SATB2突变的人类患者表现出一系列颅面表型，包括小下颌（微知）和left裂的可变渗透率。同样，在小鼠中，SATB2具有剂量效应下颌大小。然而，引起这种变化的机制尚不清楚。 SATB2是矩阵附件区域（MAR）结合蛋白已被证明通过染色质调节成骨分化组织，充当高级转录因子。降低成骨的表达分化基因被认为是SATB2介导的颅面缺陷的基础。但是，最近已发表的数据表明SATB2在成骨中的作用更为复杂，其中包括调节前的调节成骨细胞增殖，也可能涉及DNA复制中的作用。火星与两个基因有关转录和DNA复制，satb2在成骨细胞中高度表达在DNA复制中的新作用。在此提案中要检验的具体假设是Satb2结合在火星通过调节涉及的基因的转录来调节成骨增殖增殖和/或通过调节DNA复制。预计SATB2水平的降低将降低保真度并增加这些过程的变化。该假设将通过三个特定的评估目标。具体目标1将决定SATB2突变如何影响成骨细胞祖细胞中的基因表达。特定的目标2将决定细胞周期进程如何影响火星上的SATB2定位。具体的目标3将确定SATB2突变体中的增生如何受层粘连蛋白A/C表达和氧化的影响应力（先前发现在SATB2突变细胞中改变的因素）。两者都将进行实验原发性和永生的小鼠钙细胞。 SATB2突变细胞将与来自的野生型细胞进行比较等生背景。该提案中工作产生的数据将为您提供有关一种新型机制的见解 SATB2调节成骨。未来的研究计划包括进一步研究遗传，受到影响的患者的发育和环境贡献使用患者衍生的IPSCS SATB2相关综合征（SAS）。重要的是，这项工作也将大大增强研究环境，并为大学生提供高级的激动人心的机会影响研究，使他们准备进入生物医学劳动力。