Document text
Principal Investigator: FANYIN MENG
Organization: RLR VA MEDICAL CENTER
Fiscal Year: 2024
Funding agency: Veterans Affairs
Alcoholic liver disease (ALD) is one of the most common forms of chronic liver injury in the United States.
Chronic ethanol consumption results in toxic metabolites in the liver and increases endothelial senescence
and dysfunction leading to inflammation and liver damage. Aging is linked with the severity and poor
prognosis of various liver diseases including alcoholic liver diseases and is associated gradual alteration of
structure and function in liver tissues and cells including liver sinusoidal endothelial cells. Endothelial
dysfunction is an early pathophysiological hallmark in the development of liver disease. Senescence, the
cellular equivalent of aging, was proposed to be involved in endothelial dysfunction. Indeed, enhanced
cellular senescence was associated with a poor outcome in alcohol-related fatty liver disease, while
increased hepatic senescence mediators such as p21 and TNF-α that were related to liver/endothelial
dysfunction in ALD cirrhosis. Increasing evidence supports that non-coding RNAs (ncRNAs) play a central
role in various cellular pathways by regulating gene expression. Indeed, there is a strong link between
ethanol metabolism and endotoxin/lipopolysaccharide (LPS) induced cellular senescence and endothelial
dysfunction in ALDs. Hepatic sinusoids, connected directly to the portal circulation, serve as the first barrier
against these inflammatory and noxious stimuli. We have novel preliminary data showing that selective
ncRNA/microRNA genes are aberrantly expressed in liver specimens from ALD animals and are regulated
and involved in ethanol metabolism. In particular, we have found: (a) intra-gastric ethanol feeding
significantly increased the expression of cellular senescence initiators EGR1, PAI-1 and Id1 and silenced
p53 effectors E2F1 and IGFBP3; (b) ethanol enhanced microRNA-34a and p53 expressions and altered
their target genes such as SIRT1, PPARα and HNF4α, which leads to senescence phenotypes in liver
sinusoidal endothelial cells (LSECs); (c) Inhibition of miR-34a/p53/TLR4 by anti-miR-34a/p53 Morpholino
and CRISPR/cas9 approaches has successfully recovered alcoholic liver injury in ethanol treated mice in
vivo. Although the combined evidence supports a link between ncRNAs and alcoholic liver disease, there
is a critical need to determine the underlying mechanism whereby ethanol-dependent miRNAs promote
alcoholic liver injury. Our long-term goal is to determine underlying mechanisms contributing to alcohol-
induced liver disease so that new mechanism-based, clinically effective prevention or treatment strategies
can be developed. The objective for this proposal is to determine how ethanol- dependent ncRNAs mediate
endothelial dysfunction and senescence in the progression of alcoholic liver diseases. Our central
hypothesis is that ethanol-dependent miRNAs contribute to alcoholic liver injury through regulation of
cellular senescence and dysfunction in endothelial cells. This hypothesis was formulated based upon the
existing literature and our own preliminary data described above. The following three specific aims are
proposed: First, we will determine the effects of ncRNA mediated cellular senescence and endothelial
dysfunction in senescence accelerated mice with alcoholic liver injury in vivo (Aim 1). Second, we will
identify the downstream targets of p53-miR-34a axis that are involved in endothelial senescence and
dysfunction in miR-34a and p53 knockout mice in vivo (Aim 2). Further, we will determine the effects of
endothelial progenitor cell related ncRNAs enriched extracellular vesicles on anti-senescence/anti-
endothelial dysfunction and recovery of alcoholic liver injury in vivo (Aim 3). At the completion of the
proposed research, it is our expectation to have an important positive impact because a mechanistic
understanding of the role ncRNAs play in alcohol related injury, endothelial cell and senescence mediated
liver disease is likely to provide a foundation for the development of evidence-based clinically useful
approaches to treat or prevent human alcoholic liver diseases including US veterans.
Terms: <(TNF)-α><Absolute ethanol><Acceleration><Aging><Alcohol Chemical Class><Alcoholic><Alcoholic Liver Diseases><Alcoholic steatohepatitis><Alcohols><Animal Diseases><Antioncogene Protein p53><Applications Grants><Body Tissues><Boozer><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cachectin><Cas nuclease technology><Cell Aging><Cell Body><Cell Senescence><Cells><Cellular Aging><Cellular Regulation><Cellular Senescence><Cellular Tumor Antigen P53><Cellular injury><Circulation><Cirrhosis><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Data><Dependent drinker><Development><Dysfunction><E2F Transcription Factor 1><E2F transcription factor 1 protein><E2F transcription factors><E2F-1><E2F-1 protein><E2F1><E2F1 gene><E2F1 protein><ETOH><Endothelial Cells><Endothelium><Endotoxins><Ethanol><Ethanol Metabolism><Ethanol dependence><Ethyl Alcohol><Fibrosis><Foundations><Functional RNA><Functional disorder><Gene Expression><Gene Transcription><Genes><Genetic Transcription><Goals><Grain Alcohol><Grant Proposals><Health><Hepatic><Hepatic Cells><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatocyte><Hepatoma><Homolog of Drosophila TOLL><Human><IBP3><IGFBP3><IGFBP3 gene><Inflammation><Inflammatory><Injury to Liver><Intervention><Intervention Strategies><KO mice><Knock-out Mice><Knockout Mice><Link><Lipopolysaccharides><Literature><Liver><Liver Cells><Liver Cells Carcinoma><Liver Fibrosis><Liver diseases><Macrophage-Derived TNF><Mediating><Mediator><Methylcarbinol><Mice><Mice Mammals><Micro RNA><MicroRNAs><Modern Man><Monocyte-Derived TNF><Morbidity><Morbidity - disease rate><Murine><Mus><Non-Coding><Non-Coding RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Null Mouse><Oncoprotein p53><Outcome><P53><PAI-1><PAI1><PBR3><PLANH1><PPAR alpha><PPAR-α><PPARalpha><PPARα><PRB-Binding Protein E2F-1><Pathway interactions><Patients><Peroxisome Proliferator-Activated Receptor alpha><Peroxisome Proliferator-Activated Receptor α><Phenotype><Phosphoprotein P53><Phosphoprotein pp53><Physiopathology><Plasminogen Activator Inhibitor 1><Play><Preventative strategy><Prevention strategy><Preventive strategy><Primary carcinoma of the liver cells><Prognosis><Protein TP53><RBAP-1><RBBP-3><RBBP3><RBP3><RNA Expression><Recovery><Replicative Senescence><Research><Research Specimen><Retinoblastoma Binding Protein 3><Retinoblastoma-Associated Protein 1><Risk Factors><Role><SIRT1><SIRT1 gene><Serine or Cysteine Proteinase Inhibitor Clade E Member 1><Severities><Sirtuin 1><Specimen><Stimulus><Structure><TLR4><TLR4 gene><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFα><TP53><TP53 gene><TRP53><Therapeutic Effect><Tissues><Toll Homologue><Transcription><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><Tumor Protein p53><Tumor Protein p53 Gene><Type 1 Plasminogen Activator Inhibitor><United States><Untranslated RNA><Vascular Diseases><Vascular Disorder><Veterans><alcohol induced hepatic injury><alcohol induced liver disorder><alcohol induced liver injury><alcohol metabolism><alcohol related liver disease><alcohol-associated liver disease><alcohol-induced hepatic dysfunction><alcohol-induced liver disease><alcohol-induced liver dysfunction><alcohol-mediated liver dysfunction><alcohol-mediated liver injury><alcohol-related injury><alcohol-related liver disease><alcoholic liver injury><blood vessel disorder><cell damage><cell growth regulation><cell injury><cellular damage><chronic EtOH drinking><chronic alcohol consumption><chronic alcohol drinking><chronic alcohol ingestion><chronic alcohol use><chronic ethanol consumption><chronic ethanol drinking><chronic ethanol ingestion><chronic hepatic disease><chronic hepatic disorder><chronic liver disease><chronic liver disorder><chronic liver injury><cirrhotic><damage to cells><developmental><effective therapy><effective treatment><endothelial dysfunction><endothelial progenitor><endothelial progenitor cell><endothelial stem cell><ethanol addiction><ethanol dependency><ethanol dependent><ethanol induced hepatic injury><ethanol induced liver disorder><ethanol induced liver injury><ethanol liver disease><ethanol-induced hepatic dysfunction><ethanol-induced liver disease><ethanol-induced liver dysfunction><ethanol-mediated liver dysfunction><ethanol-mediated liver injury><evidence base><expectation><extracellular vesicles><fatty liver disease><feeding><fibrotic liver><hepatic body system><hepatic damage><hepatic disease><hepatic fibrosis><hepatic injury><hepatic organ system><hepatic sinusoid><hepatopathy><in vivo><injury to cells><interventional strategy><liver carcinoma><liver damage><liver development><liver disorder><liver injury><miRNA><miRNAs><mortality><noncoding><novel><p53 Antigen><p53 Genes><p53 Tumor Suppressor><pathophysiology><pathway><prevent><preventing><problem drinker><protein p53><senescence><senescent><social role><toll-like receptor 4><transcription factor E2F1><treatment strategy><vascular dysfunction><vasculopathy>