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：
10894975
负责人：
Nicole Renee Sparks
金额：
$ 8.41万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
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 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10894975
关键词：
ATAC-seq Accounting Adverse effects Affect Bioinformatics Biological Biological Process 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 Environment Epigenetic Process Etiology Exposure to Failure Frequencies Gene Expression Generations 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 Repression Research Risk Role Science Signal Pathway Skeletal Development Skeletal system Skeleton 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 biomarker identification bone career development disease diagnosis early embryonic stage 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.

在美国，每 4.5 分钟就有一名患有先天缺陷的婴儿出生。骨骨骼的骨骼缺陷与子宫内环境化学物质暴露有关。我们的实验室已经证明，正确的成骨细胞（骨形成细胞）的发育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，其中毒物诱导差异基因表达成骨细胞分化受到干扰。 MicroRNA (miRNA) 是一种小的非编码 RNA，在表观遗传学上调节基因表达并且是骨骼发育的参与者。目前尚不清楚 miRNA 是否对环境因素产生不良反应，导致骨骼畸形。在这个提案中，我假设毒物诱导的 miRNA 变化在骨骼出生缺陷的表现中起着至关重要的作用。 miRNA 对分化 hESC 的分析先前发现了 10 种因化学物质而下调的 miRNA 暴露抑制成骨细胞分化。我建议 1）验证候选人的职能角色成骨过程中的 miRNA (K99)； 2) 研究候选miRNA的功能效果是否体内复制（K99）； 3) 确定候选影响的mRNA靶基因和信号通路 miRNA (K99)； 4) 研究 miRNA 失调是否是通过表观遗传机制促进的抑制 miRNA 表达 (R00)。拟议的目标将极大地影响对骨骼发育过程中环境诱导的表观遗传毒性。导师博士。马丁·里科马尼奥（生存手术和基因组工程）和 Nicole zur Nieden（hESC、miRNA 和基因组工程）将帮助我接受培训，以成功完成拟议目标并成为一名独立校长研究者。额外的支持将来自博士。马丁·加西亚-卡斯特罗（分子细胞遗传学技术）和帕特里克·阿拉德（表观遗传毒理学）。职业发展计划包括加强我的研究的培训技能，包括下一代测序、生物信息学、子宫内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。计划中的培训和研究将有助于过渡到独立并作为首席研究员取得成功。