Investigating the Impact of Telomere Specific Oxidative Base Damage in Cellular Aging

研究端粒特异性氧化碱损伤对细胞衰老的影响

• 批准号: 10292913
• 负责人: Ryan P Barnes
• 金额: $ 4.02万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292913
• 关键词: Acute; Address; Aftercare; Aging; Apoptosis; Base Excision Repairs; Basic Science; Biological Assay; Biology; Cancer cell line; Cell Aging; Cell Line; Cell Proliferation; Cells; Cellular biology; Chromosomes; Chronic; Clinical Research; Clone Cells; DNA; DNA Damage; DNA Double Strand Break; DNA biosynthesis; Detection; Disease; Dyes; Educational workshop; Epithelial Cells; Exhibits; Fellowship; Fibroblasts; Free Radicals; Functional disorder; Future; Genetic; Genetic Structures; Genetic Transcription; Genome; Genome Stability; Genomic Instability; Growth; Guanine; Health Promotion; Human; Immune; Immune signaling; Immune system; International; Interphase Cell; Ionizing radiation; Knowledge; Lentivirus; Lesion; Light; Link; Malignant Neoplasms; Mediating; Mentors; Metabolism; Metaphase; Modeling; Molecular; Morphology; Normal Cell; Nucleoproteins; OGG1 gene; Oncogenes; Outcome; Oxidative Stress; Pathology; Pathway interactions; Peptides; Pharmacology; Phototherapy; Physiological; Physiology; Play; Pollution; Reporting; Research Project Grants; Rest; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Solar Energy; Somatic Cell; Source; Stimulator of Interferon Genes; Stress; Structure; System; TERF1 gene; TP53 gene; Techniques; Telomerase; Telomere Shortening; Telomere-Binding Proteins; Telomeric Repeat Binding Protein 1; Testing; Time; Work; Writing; age related; anticancer research; base; biological adaptation to stress; cancer cell; carcinogenesis; career development; cell growth; cytotoxicity; enzyme pathway; experience; healthspan; human DNA damage; in vivo; innovation; insight; live cell imaging; novel; oxidation; prevent; recruit; repaired; replication stress; response; senescence; single molecule; skills; small molecule inhibitor; symposium; telomere; tool; transcriptome sequencing; tumor

Project Summary/Abstract Telomeres play an essential role at the interface between cellular aging and carcinogenesis because they are highly sensitive to oxidative stress. Oxidative stress, or the excess of free radicals, is a ubiquitous source of DNA damage humans experience from normal metabolism, physical stress, exogenous sources such as pollution and solar radiation, and by-products of immune cell signaling. While a link between telomere attrition and oxidative stress exists, there have been no reports on how direct oxidative base damage to telomeric DNA impacts normal cell biology. Our group has developed the first system to induce the common oxidative lesion 8-oxo-guanine (8oxoG) specifically at telomeres by fusing a fluorogen activated peptide (FAP) to the telomere binding protein TRF1. We have generated clonal cell lines that have homogenous FAP-TRF1 expression at the telomere, and find that after a single induction of telomeric 8oxoG, normal cells exhibit a growth arrest, which we did not observe in two cancer cell lines. The hypothesis of this fellowship is that telomeric 8oxoG is sufficient to produce a DDR in normal cells that induces growth arrest, independent of telomere shortening, that depends on the p53/p21 signaling axis. Moreover, following induction of telomeric 8oxoG we observe an increase in the number of micronuclei as early as one day after treatment. This project will also address how these micronuclei are forming, if they are dependent on DNA replication, and if they are sensed by the cGAS/STING axis. Competition of this project will delineate for the first time the contribution of telomeric oxidative stress to cellular outcomes related to aging and cancer. This work will have general applications to all humans, but will also inform the biology of age-related diseases that are associated with increased levels of oxidative stress, and may promote health-span in either case.This fellowship also allows for the acquisition of new skills related to the aims of the project, and has outlines specific mentors for each. This includes single molecule analyses of telomere replication, live-cell imaging, RNA-sequencing, and detection of signaling molecules related to the immune system. Moreover, career development is also a focus of this fellowship. It includes plans to attend workshops related to experimental techniques and scientific writing, in addition to attending national and international conferences to present the findings of this research project.

项目总结/文摘

项目成果

Targeted Formation of 8-Oxoguanine in Telomeres.

• DOI: 10.1007/978-1-0716-2063-2_9
• 发表时间: 2022-01-01
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Barnes, Ryan P;Thosar, Sanjana A;Opresko, Patricia L
• 通讯作者: Opresko, Patricia L

Premature aging is associated with higher levels of 8-oxoguanine and increased DNA damage in the Polg mutator mouse.

• DOI: 10.1111/acel.13669
• 发表时间: 2022-09
• 期刊: AGING CELL
• 影响因子: 7.8
• 作者: Yu, Tenghui;Slone, Jesse;Liu, Wensheng;Barnes, Ryan;Opresko, Patricia L.;Wark, Landon;Mai, Sabine;Horvath, Steve;Huang, Taosheng
• 通讯作者: Huang, Taosheng