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.

项目概要颅面异常是最常见、最使人衰弱的人类出生缺陷之一，影响 1/500 至 1/2000 的出生率取决于人口。颅面异常的严重程度和外显率的变化这是一种被广泛观察但了解甚少的现象。这种变异会导致复杂的临床问题，例如对受影响或易感个体的诊断、治疗和遗传咨询方面的困难颅面疾病。特别是，人们广泛认识到，相似的基因突变通常表达疾病表型谱，但仍不清楚是什么机制导致了这种变异。为了例如，携带 SATB2 突变的人类患者表现出一系列颅面部表型，包括小下颌（小颌）和腭裂的外显率变化。同样，在小鼠中，Satb2 也具有剂量效应关于下巴的大小。然而，引起这种变化的机制尚不清楚。 Satb2 是矩阵附件区域（MAR）结合蛋白已被证明可以通过染色质调节成骨分化组织，充当高阶转录因子。成骨细胞表达减少分化基因被认为是 Satb2 介导的颅面缺陷的基础。然而，最近已发表的数据表明 Satb2 在成骨中发挥更复杂的作用，包括调节前成骨细胞增殖也可能参与 DNA 复制。 MAR 与两个基因有关转录和 DNA 复制，Satb2 在成骨细胞的 S 期高表达，提示 DNA复制中的新作用。本提案中要测试的具体假设是 Satb2 结合在 MARs 通过调节相关基因的转录来调节成骨增殖增殖和/或通过DNA复制的调节。 Satb2 水平的降低预计会降低保真度并增加这些过程的变化。该假设将通过三个具体的评估目标。具体目标 1 将确定 Satb2 突变如何影响成骨细胞祖细胞中的基因表达。具体目标 2 将确定细胞周期进展如何影响 Satb2 在 MAR 上的定位。具体目标 3 将确定 Satb2 突变体前成骨细胞的增殖如何受到 Lamin A/C 表达和氧化的影响应激（之前发现 Satb2 突变细胞中发生改变的因素）。实验将在两个方面进行原代和永生化小鼠颅骨细胞。 Satb2 突变细胞将与野生型细胞进行比较同基因背景。本提案中的工作生成的数据将提供对一种新机制的深入了解 Satb2 成骨的调节。未来的研究计划包括进一步研究遗传、发育和环境对受该疾病影响的患者疾病表型变异的贡献使用患者来源的 iPSC 进行 SATB2 相关综合征 (SAS)。重要的是，这项工作也将大大改善研究环境并为本科生提供令人兴奋的机会进行高水平研究影响研究，为他们进入生物医学队伍做好准备。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mutation in Eftud2 causes craniofacial defects in mice via mis-splicing of Mdm2 and increased P53.

Eftud2 突变通过 Mdm2 的错误拼接和 P53 的增加导致小鼠颅面缺陷。

DOI：
10.1093/hmg/ddab051
发表时间：
2021-02-18
期刊：
Human molecular genetics
影响因子：
3.5
作者：
Marie;Anissa Djedid;E. Bareke;Fjodor Merkuri;Rachel Aber;Annie S. Tam;M. Lines;K. Boycott;Peter C. Stirling;J. Fish;J. Majewski;L. Jerome
通讯作者：
L. Jerome

Satb2 regulates proliferation and nuclear integrity of pre-osteoblasts.

Satb2 调节前成骨细胞的增殖和核完整性。