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Exploring the role of hypoxia signaling and its inhibition as a therapy for Facioscapulohumeral muscular dystrophy

探索缺氧信号传导及其抑制作为面肩肱型肌营养不良症治疗的作用

基本信息

批准号：
10463106
负责人：
Justin Cohen
金额：
$ 6.98万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10463106
关键词：
ACTA1 gene Affect Apoptosis Attenuated Autophagocytosis Biological Assay Biological Markers Biopsy CRISPR screen CRISPR-mediated transcriptional activation CRISPR/Cas technology Cell Death Cell Death Inhibition Cell Line Chromosome 4 Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complementary DNA Complex D4Z4 Disease Embryonic Development Energy Metabolism Epigenetic Process FDA approved FRAP1 gene Facioscapulohumeral Muscular Dystrophy Functional disorder Gene Activation Generations Genes Genetic Genetic Diseases Genetic Engineering Genetic Screening Genetic Transcription Genus Hippocampus Goals HIF1A gene Hand Strength Haplotypes Histology Human Hypoxia Hypoxia Pathway Immune Individual Knock-out Knowledge Lifting Link Measurement Measures Mediating Metabolic Metabolic dysfunction Microscopy Modeling Molecular Mus Muscle Muscle Cells Muscle Contraction Muscle function Myopathy Neuromuscular Diseases Oxidative Phosphorylation Oxidative Stress Oxygen Consumption Pathogenicity Pathologic Pathology Pathway interactions Patients Pharmacology Phenotype Physiological Prevalence Proteins Quality of life Reactive Oxygen Species Reporting Repression Reproducibility Research Role SDZ RAD Screening Result Signal Pathway Signal Transduction Skeletal Muscle Techniques Testing Therapeutic Therapeutic Agents Tissues Titrations Toxic effect Transcript Transgenes Transplantation Wait Time Xenograft procedure arm design epigenetic silencing experience extracellular gene therapy genome-wide head impact hypoxia inducible factor 1 in vivo knock-down loss of function mTOR Inhibitor mTOR inhibition metabolic profile mouse model muscle physiology muscular dystrophy mouse model mutation screening new therapeutic target novel overexpression pre-clinical prevent promoter response skeletal muscle wasting small molecule small molecule inhibitor therapeutic evaluation therapeutic target therapy development transcription factor transcriptome sequencing treadmill

项目摘要

Abstract Facioscapulohumeral muscular dystrophy (FSHD) is a neuromuscular disorder for which there is currently no treatment or cure. Affected individuals have difficulty accomplishing everyday tasks such as lifting one’s arms above their head, impacting quality of life. Much research has been done trying to elucidate the mechanism of disease pathology but there has been little congruence apart from apoptosis via DUX4, the toxic protein associated with FSHD. We took advantage of this phenotype to design a genome-wide complimentary CRISPR-Cas9 loss of function and activation screens for genetic modifiers of DUX4 toxicity. These screens identified hypoxia signaling as a novel pathway relating to FSHD pathology with therapeutic potential. Small molecule inhibitors that target modifiers of this pathway, including the FDA-approved compound everolimus, reduced DUX4-mediated cell death and FSHD biomarkers, demonstrating the viability of targeting hypoxia signaling as a potential therapy. In this project we intend to further explore the mechanism of how hypoxia signaling relates to DUX4-mediated pathology (Specific aims 1). The PI3K/Akt/mTOR signaling axis that interacts with hypoxia signaling will be modulated through knock-down and/or pharmacological inhibition for their effect on DUX4-mediated apoptosis. The consequence of autophagy modulation through the mTOR inhibitor everolimus will be explored. The relationship between DUX4-induced hypoxia and metabolic dysfunction will be assessed through RNAseq, Seahorse assays of oxygen consumption and extracellular acidification rates, modulation of oxidative phosphorylation and reactive oxygen species generation. We will additionally test the therapeutic potential of everolimus in a DUX4-inducible mouse model of FSHD (Specific aims 2). We will measure effects on muscle function through treadmill endurance, grip strength and ex vivo contractile force. Using microscopy, we will examine effects on muscle histology and markers of hypoxic signaling while exploring if this pathology is affected by the metabolic profile of the individual muscles. Lastly, effect on FSHD biomarkers will be assessed both in the mouse model and validated using patient biopsy myocytes. If successful, this project will elucidate a new mechanism of FSHD pathology for therapeutic targeting and identify everolimus as a therapeutically promising compound whose FDA status allows for a shorter timeframe before patient availability.

摘要