Document text
Principal Investigator: DAVID R BRIGSTOCK
Organization: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
Fiscal Year: 2024
Award: $409,839
Funding agency: National Institute on Alcohol Abuse and Alcoholism
ABSTRACT
The broad long term objective is to improve methods of disease treatment in the liver. Hepatic fibrosis is a
major cause of morbidity and mortality that affects millions of people world-wide and is usually a feature of
chronic liver disease such as that caused by excessive alcohol consumption. In the United States, alcoholic
liver disease (ALD) is a leading cause of GI-related deaths, with about half of the 75,000 liver disease deaths
each year being related to alcohol use. A major limitation is the lack of approved therapeutics for treating either
liver fibrosis or ALD. However, a new lead has emerged from our studies of exosomes that are produced by
hepatocytes. Our overall objective is to establish therapeutic uses of exosomes for treating liver disease. Our
Preliminary Data show that hepatocyte exosomes: (i) attenuate expression of genes that regulate fibrogenesis
or activation in cultured primary hepatic stellate cells (HSC); (ii) suppress fibrogenic pathways and reverse
hepatic fibrosis in vivo; (iii) attenuate damage in cultured hepatocytes exposed to ethanol or CCl4, (vi) contain
relatively high levels of miR-532-5p or miR-214 that likely contribute, in part, to their therapeutic actions in
fibrosis or ALD respectively especially as their respective tissue levels in the liver are individually suppressed
during chronic injury; and (v) bind to target hepatocytes or HSC via cell-surface integrin αvβ3 and α5β1. Our
central hypothesis is that miR-532-5p-/miR-214 or specific integrin ligands contribute to, respectively, the
therapeutic actions or cellular binding of hepatocyte exosomes. The Specific Aims to test this hypothesis are:
Aim 1 - Identify miR-532-5p as a therapeutic component of hepatocyte exosomes that modulates
SMAD3 in hepatocytes or HSC by using purified exosomes or cell co-culture assays, employing SMAD3 3’-
UTR reporters, CTGF promoter reporters, expression of SMAD3 downstream targets, siRNA-mediated SMAD3
knockdown, and miR-532-5p over-expression or antagonism to show direct functional regulation of SMAD3 in
each target cell; Aim 2 - Determine the role of hepatocyte exosomes and of exosomal miR-214 or miR-
532 in attenuating ethanol-induced liver injury by demonstrating the therapeutic effect of exosomes in
ethanol diet models in vivo, on ethanol/TNFα- or LPS-mediated pathways in hepatocytes or Kupffer cells
respectively, and by demonstrating that exosomal targeting of CTGF or ICAM-1 by miR-214 or of SMAD 3 by
miR-532 recapitulates exosomal action in vivo and in vitro; and Aim 3 - Identify exosomal binding partners
of cellular integrins by establishing the role of hepatocyte FN, VN, or CTGF in engaging integrin αvβ3 or
α5β1 on HSC or hepatocytes and thereby mediating target cell binding. The rationale is that current methods
of treating liver fibrosis or ALD are inadequate and our approach is a cutting-edge and innovative solution that
harnesses natural disease-suppressing properties of exosomes. The expected outcome will be a novel
exosome-based therapy for treating fibrosis or alcohol-induced cell damage and altered immune function in the
liver. The positive impact will be to improve the health of millions of people globally with chronic liver disease.
Terms: <(TNF)-α><Absolute ethanol><Adrenal Cortex Hormones><Affect><Alcohol Chemical Class><Alcohol Drinking><Alcohol associated hepatitis><Alcohol consumption><Alcohol hepatitis><Alcohol induced hepatitis><Alcohol related hepatitis><Alcoholic Hepatitis><Alcoholic Liver Diseases><Alcoholic beverage heavy drinker><Alcohols><Alternative Therapies><Alternative intervention><Animals><Apoptosis><Apoptosis Pathway><Applications Grants><Assay><Attenuated><Behavior><Binding><Bioassay><Biological Assay><Body Tissues><CCN2><CD54 Antigens><CTGF><Cachectin><California><Cause of Death><Cell Body><Cell Survival><Cell Viability><Cell surface><Cells><Cellular Immune Function><Cellular injury><Cessation of life><Chronic><Cicatrix><Cirrhosis><Clinic><Co-culture><Cocultivation><Coculture><Coculture Techniques><Corticoids><Corticosteroids><Data><Death><Deposit><Deposition><Diet><Disease><Disorder><ETOH><EtOH drinking><EtOH use><Ethanol><Ethanol-induced hepatitis><Ethyl Alcohol><FDA approved><Fatty Liver><Fibrillar Collagen><Fibrosis><Gene Expression><Goals><Grain Alcohol><Grant Proposals><Health><Heavy Drinker><Heavy Drinking><Hepatic Cells><Hepatic Cirrhosis><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatic Stellate Cell><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatocyte><Hepatoma><Hypoxia><Hypoxic><ICAM-1><IGF-binding protein-related protein-2><IGFBP-8><IGFBP-rP2><In Vitro><Incidence><Individual><Inflammatory><Injury><Integrin aVBeta3><Integrin alpha-5 beta-1><Integrin alpha-v beta-3><Integrin alpha5beta1><Integrin alphaVbeta3><Integrin α5β1><Integrin αVβ3><Integrins><Integrins Extracellular Matrix><Intercellular adhesion molecule 1><Ito Cell><JV15-2><Knowledge><Kupffer Cells><Lead><Ligands><Liver><Liver Cells><Liver Cells Carcinoma><Liver Cirrhosis><Liver Fibrosis><Liver Steatosis><Liver diseases><MADH3><MADH3 gene><Macrophage-Derived TNF><Mediating><Mediator><Medical><Messenger RNA><Methods><Methylcarbinol><Modeling><Molecular><Molecular Interaction><Monocyte-Derived TNF><Morbidity><Morbidity - disease rate><Mothers Against Decapentaplegic, Drosophila, Homolog of, 3><NIAAA><National Institute on Alcohol Abuse and Alcoholism><Ohio><Outcome><Oxygen Deficiency><Pathology><Pathway interactions><Patients><Pb element><Persons><Phenotype><Platelet Glycoprotein Ic/IIa><Primary carcinoma of the liver cells><Production><Programmed Cell Death><Progressive Disease><Property><Proteins><Receptor Protein><Regulation><Reporter><Risk Factors><Role><SMA- and MAD-Related Protein 3><SMAD3><Scars><Short interfering RNA><Small Interfering RNA><Stellate Sinusoidal Macrophage><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFα><Testing><Therapeutic><Therapeutic Effect><Therapeutic Uses><Tissues><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><UTRs><United States><Universities><Untranslated Regions><VLA-5><VLA-5 Receptors><Vesicle><aVBeta3><alcohol exposed><alcohol exposure><alcohol induced hepatic injury><alcohol induced liver disorder><alcohol induced liver injury><alcohol ingestion><alcohol intake><alcohol product use><alcohol related liver disease><alcohol use><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 liver disease><alcoholic beverage consumption><alcoholic drink intake><alcoholic liver injury><alpha-v beta-3 Integrin Receptors><antagonism><antagonist><attenuate><attenuates><cell damage><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><connective tissue growth factor><cytokine><damage to cells><diets><drink heavily><ethanol consumption><ethanol drinking><ethanol exposed><ethanol exposure><ethanol induced hepatic injury><ethanol induced liver disorder><ethanol induced liver injury><ethanol ingestion><ethanol intake><ethanol liver disease><ethanol product use><ethanol use><ethanol-induced hepatic dysfunction><ethanol-induced liver disease><ethanol-induced liver dysfunction><ethanol-mediated liver dysfunction><ethanol-mediated liver injury><excessive alcohol consumption><excessive alcohol ingestion><excessive alcohol intake><excessive drinking><excessive ethanol ingestion><exosome><exposed to alcohol><exposed to ethanol><exposure to alcohol><exposure to ethanol><extracellular><extreme drinking><fibrogenesis><fibrotic liver><fisp12 protein><healing><heavy alcohol use><heavy metal Pb><heavy metal lead><hepatic body system><hepatic disease><hepatic fibrosis><hepatic organ system><hepatic steatosis><hepatopathy><hepatosteatosis><immune function><improved><in vivo><in vivo Model><injuries><injury to cells><innovate><innovation><innovative><insulin-like growth factor binding protein 8><knock-down><knockdown><liver carcinoma><liver disorder><liver macrophage><mRNA><mortality><nano vesicle><nanovesicle><novel><overexpress><overexpression><pathway><programs><promoter><promotor><receptor><siRNA><side effect><social role><therapeutic candidate>