Control mechanisms of 5-hydroxymethylcytosine metabolism in human cells

人体细胞5-羟甲基胞嘧啶代谢的控制机制

• 批准号: 10629908
• 负责人: Haeyoung Kim
• 金额: $ 13.8万
• 依托单位: TEXAS A&M UNIVERSITY-KINGSVILLE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629908
• 关键词: Attention; Base Excision Repairs; Biological Assay; Biomedical Research; Blood; Brain; CRISPR/Cas technology; Cell Line; Cell physiology; Cells; CpG dinucleotide; Cytosine; DNA; DNA Damage; DNA Double Strand Break; DNA Methylation; DNA Modification Methylases; DNA Repair; DNA biosynthesis; DNA lesion; Development; Disease; Enzymes; Epigenetic Process; Family; Functional disorder; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Genome; Genome Stability; Glioblastoma; Goals; Human; Induced pluripotent stem cell derived neurons; Knowledge; Light; Link; Lymphoma; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Metabolism; Methylation; Mitotic; Modeling; Neurodegenerative Disorders; Neurons; Nuclear; Nucleotides; Organ; Organism; Outcome; Pathway interactions; Pluripotent Stem Cells; Process; Production; Proteins; Resistance; Role; Site; Survival Rate; System; Techniques; Testing; Thymine DNA Glycosylase; Time; Tissue-Specific Gene Expression; Tissues; age related; anti-cancer therapeutic; cell type; conditional knockout; demethylation; drug discovery; epigenetic marker; epigenetic regulation; high throughput screening; human DNA; human disease; knock-down; mortality; novel; novel marker; novel therapeutics; oxidation; postmitotic; repaired; response; translocase; tumor progression

Project Summary The 5-hydroxymethylcytosine (5hmC) in mammalian DNA is drawing significant attention in epigenetics because of its indispensable roles in gene expression regulation. The loss of 5hmCs in various cancers including glioblastomas and lymphomas has been linked to the poor survival rate as well as the resistance to anti-cancer therapeutics. Although there is amassing evidence of 5hmCs as a novel epigenetic marker, there is very little understanding on how 5hmCs are enriched at specific loci in the genome. In this project, we aim to identify the factors controlling the DNA demethylation pathway, which result in the formation of locus-specific 5hmCs in human neurons. For this purpose, proteins controlling the activity of ten-eleven translocase (TET) family enzymes in the DNA demethylation pathway will be identified. To examine the function of candidate proteins, we will take advantage of human induced-neurons (hiNs) derived from human induced-pluripotent stem cells (hiPSCs). In addition, we will elucidate the role and fate of 5hmCs formed during DNA damage repair process using blue light-inducible CRISPR/Cas9 technique. This robust experimental platform will enable the rapid production of isogenic and homogenous neurons and other cell types. With this cell-based assay system, high-throughput assays can be developed that will advance biomedical researches and drug discoveries. Finally, we expect the successful completion of this project will expand our understanding on DNA demethylation and their roles in epigenetics, development, and various human diseases.

项目总结