Analysis of environmentally-sensitive epigenetic machinery during osteogenic differentiation

成骨分化过程中环境敏感的表观遗传机制分析

基本信息

批准号：
10318541
负责人：
Nicole Renee Sparks
金额：
$ 9.82万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
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：
2020-12-15 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318541
关键词：
ATAC-seq Accounting Adverse effects Affect Bioinformatics Biological Biological Markers Biological Process Birth Bone Development Cells Chemical Exposure Chemicals Chromatin Communication Complex Congenital Abnormality Cytogenetic Analysis DNA DNA Methylation DNA methyltransferase inhibition Data Defect Development Development Plans Disease Electroporation Embryo Environment Epigenetic Process Etiology Exposure to Failure Frequencies Gene Expression Genes Genetic Genetic Transcription Genome engineering Genomics Global Change Goals Health Hypermethylation In Vitro Investigation Knock-out Knockout Mice Knowledge Mediating Mentors Messenger RNA Methylation MicroRNAs Mission Modification Molecular Mus Ontology Operative Surgical Procedures Osteoblasts Osteogenesis Outcome Pathway interactions Patients Physiologic calcification Play Prevention Principal Investigator Process Proteins Public Health Regulation Research Risk Role Science Signal Pathway Skeletal Development Skeletal system Skeleton Small RNA System Systems Development Testing Therapeutic Tissue-Specific Gene Expression Toxic Environmental Substances Toxic effect Toxicant exposure Toxicology Training United States United States National Institutes of Health Untranslated RNA Validation Writing advanced disease adverse outcome base bone career development disease diagnosis environmental agent environmental chemical exposure epigenetic regulation experimental study human embryonic stem cell in utero in vivo independency infant death interest knock-down mRNA Expression malformation next generation sequencing non-genetic novel osteoblast differentiation osteogenic overexpression pregnant prenatal exposure promoter public health relevance skeletal skeletal abnormality skeletal disorder skills stem success toxicant transcriptome sequencing

项目摘要

Every 4.5 minutes, a baby is born with a birth defect in the United States. Skeletal defects of the bony skeleton have been associated with environmental chemical exposure in utero. Our lab has shown that proper development of osteoblasts, the bone forming cells, depends on tight regulation of bone-specific genes. We developed a human embryonic stem cell (hESC) system where toxicant-induced differential gene expression perturbed osteoblast differentiation. MicroRNAs (miRNAs) are small non-coding RNAs that epigenetically regulate gene expression and are actors of skeletal development. There is no knowledge whether miRNAs adversely respond to environmental agents to result in skeletal malformations. In this proposal, I hypothesize that toxicant-induced miRNA changes play a critical role in the manifestation of skeletal birth defects. MiRNA profiling on differentiating hESCs previously identified 10 miRNAs downregulated stemming from chemical exposure that repressed osteoblast differentiation. I propose to 1) validate the functional role of candidate miRNAs during osteogenesis (K99); 2) investigate whether the functional effects of the candidate miRNAs are replicated in vivo (K99); 3) determine the mRNA target genes and signaling pathways affected by the candidate miRNAs (K99); and 4) investigate whether miRNA dysregulation is facilitated through epigenetic machinery that represses miRNA expression (R00). The proposed aims will significantly impact the understanding of environmentally induced epigenetic toxicity during skeletal development. Mentors Drs. Martin Riccomagno (survival surgeries and genome engineering) and Nicole zur Nieden (hESCs, miRNAs and genome engineering) will aid in my training for successful completion of the proposed aims and becoming an independent principal investigator. Additional support will come from Drs. Martin Garcia-Castro (molecular cytogenetic techniques) and Patrick Allard (epigenetic toxicology). The career development plan includes training to enhance my research skills, including Next Generation Sequencing, bioinformatics, in utero electroporation, and in vivo skeletal analysis, as well as scholarly skills and professional development, including but not limited to science communication, writing, mentoring, and management skills. The planned training and research will facilitate the transition to independency and success as a principal investigator.

每4.5分钟，一个婴儿天生就有在美国出生缺陷。骨骨骼的骨骼缺陷与子宫内的环境化学暴露有关。我们的实验室表明这是正确的成骨细胞（骨形成细胞）的发展取决于骨特异性基因的严格调节。我们开发了人类胚胎干细胞（HESC）系统，其中有毒物质诱导的差异基因表达干扰成骨细胞的分化。 microRNA（miRNA）是表观遗传的小非编码RNA 调节基因表达，是骨骼发育的参与者。不知道mirnas是否对环境药物的反应不利，导致骨骼畸形。在这个建议中，我假设毒物引起的miRNA变化在骨骼先天缺陷的表现中起着关键作用。 mirna 在区分先前鉴定的10个miRNA下调的hESC上进行了分析。抑制成骨细胞分化的暴露。我建议1）验证候选人的功能作用成骨过程中的miRNA（K99）； 2）研究候选miRNA的功能效应是否是在体内复制（K99）; 3）确定受候选者影响的mRNA靶基因和信号通路 mirnas（K99）; 4）研究miRNA失调是否通过表观遗传机制促进压抑miRNA表达（R00）。拟议的目标将极大地影响在骨骼发育过程中，环境诱导的表观遗传毒性。导师博士。马丁·里卡格诺（Martin Riccomagno）（生存手术和基因组工程）和妮可·祖尔·尼登（Nicole Zur Nieden）（hESC，miRNA和基因组工程）将帮助我培训成功完成拟议的目标并成为独立校长研究者。额外的支持将来自Drs。 Martin Garcia-Castro（分子细胞遗传学技术）和帕特里克·阿拉德（Patrick Allard）（表观遗传学毒理学）。职业发展计划包括培训以增强我的研究技能，包括下一代测序，生物信息学，子宫电穿孔和体内骨骼分析以及学术技能和专业发展，包括但不限于科学沟通，写作，指导和管理技能。计划的培训和研究将促进过渡到独立性和作为首席研究员的成功。