权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mitochondrial G-quadruplex structures in health and disease

健康和疾病中的线粒体 G-四链体结构

基本信息

批准号：
10221730
负责人：
Brett A Kaufman
金额：
$ 36.38万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2020-08-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10221730
关键词：
Acute Address Affect Alleles Antibodies Berberine Binding Bioinformatics Biology Biophysics Cell Line Cell Respiration Cells Cellular Structures ChIP-seq Clinical DNA Maintenance DNA Sequence DNA biosynthesis Data Development Disease Enzymes Excision Future G-Quartets Gene Expression Genetic Transcription Goals Guanine Health Heritability Intrabody Knowledge Lead Ligands Location Malignant Neoplasms Metabolic syndrome Mitochondria Mitochondrial DNA Mitochondrial Diseases Mitochondrial Matrix Molecular Nuclear Nucleic Acids Nucleotides Pathogenicity Pathologic Pathology Patient Care Patients Penetrance Physiological Ploidies Property Proteins Publishing Reagent Regulation Resolution Respiration Respiratory physiology Role Severities Specificity Structure Testing Therapeutic Tissue Viability Translations Variant Work chromatin immunoprecipitation early onset fascinate genome integrity helicase heteroplasmy improved innovation mitochondrial genome novel novel strategies respiratory response small molecule tool

项目摘要

PROJECT SUMMARY G-quadruplex structures (G4) arise in guanine-rich sequences and have a high potential for formation in the mitochondrial DNA (mtDNA) due to its strand specific biases in nucleotide content. Preliminary studies and prior published work suggest that G4s impact mitochondrial function, but the evidence remains largely indirect, obscuring the role of these fascinating structures in normal and pathological mitochondrial biology. This proposal will address this gap in knowledge by defining the specific regions of mtDNA that form G4 in the cell and the conditions that promote G4 emergence and stability. The proposal is significant because a fuller understanding of the regulation of mtDNA maintenance and expression may be important in future approaches to diverse pathologies including heritable mitochondrial diseases, metabolic syndromes and sporadic cancers. The proposal is innovative in its development of novel reagents to detect mitochondrial G4s and in its novel approach to the therapy of mitochondrial disorders. The overarching hypothesis is that physiological G4 formation within mtDNA is widespread and regulates mitochondrial transcription and replication. The first specific aim will employ an innovative tool, a mitochondrial-targeted intrabody that binds to G4 sequences, to probe for G4-interacting sequences in the mitochondrial matrix by chromatin immunoprecipitation (ChIP). The sequences of mtG4s will be identified and their relative abundance will be evaluated under a range of conditions. The conditions include basal, elevated and inhibited mitochondrial function, and under G4-activated or G4-inhibited conditions. The role of DNA unwinding enzymes will also be evaluated. The second specific aim will expand upon a recent observation that induced G4 formation, using G4 binding agents, selects against specific pathogenic mtDNA variants that enhance G4 formation. Such variants typically exist in a state known as heteroplasmy, where healthy mtDNA is also present and the ratio between pathogenic and wild type sequence determines penetrance and severity. We will use patient cell lines and patient-derived cybrid cells to evaluate the range of pathogenic variants that may be susceptible to this approach. We will also expand the identification of novel G4 binding compounds that discriminate between pathogenic and wild type alleles. Overall, these studies will contribute mechanistic evidence for specific G4 structure formation in different conditions, connect their formation to the regulation of mtDNA replication and transcription, as well as develop new tools and reagent to positively impact mtDNA content in certain heteroplasmic conditions.

项目总结