Development and validation of a novel 3D human in vitro model of nonalcoholic steatohepatitis to be used for novel therapeutic screening and research on disease mechanism.

ABSTRACT Nonalcoholic fatty liver diseaseNAFLDis a chronic condition that originates as lipid accumulation within hepatocytessteatosisand progresses into nonalcoholic steatohepatitisNASHcharacterized by lipid accumulationinflammationoxidative stressand fibrosisAn estimatedmillion adults across the U Sare thought to have NAFLDwith up tomillion more estimated to have NASHLeft uncheckedthe disease can advance to cirrhosis and hepatocellular carcinomaCurrentlyNASH contributes to one third of hepatocellular carcinomas in the US and is the second leading cause of liver transplants in the USNASH is often referred to as asilentdisease as most patients are asymptomatic until problems arise from cirrhosis and liver failure and diagnosis only possible via invasive liver biopsiesDespite well defined morphological features and decades of intense research worldwidethe mechanisms of NAFLD progression as well as therapeutic approaches and non invasive diagnostics are still resoundingly absentRecentlyOrganovo developed ExViveHuman Liver Tissuean in vitroD bioprinted liver model comprising primary human hepatocyteshepatic stellates and endothelial cells that better mimics native human liver biology over extended time in cultureThe technological advancement ofD bioprinting enables controlledreproducible fabrication ofD liver tissues ex vivo from human cellsyielding a durable liver model with complex composition and architecture that retains metabolic competence and liver specific functions for at leastweeks in cultureThe current Fast Track SBIR application seeks to utilize ExViveHuman Liver Tissue to model the onset and progression of human NAFLD via nutrient overload in the first Phasefollowed by induction of inflammationoxidative stress and fibrosis to mimic human NASH in the second PhaseImportantlyin order to drive adoptionthe steatosis and NASH models will be validated via comparisons of genomic data with clinically confirmed NAFLD patient data in collaboration with key opinion leader DrDavid Brennerand verification of altered drug metabolism and disposition via CYP AandCactivitiesLastlyprevention of NAFLD progression and regression of NASH will be assessed via therapeutic modulation of disease phenotypesTogetherthese in vitro models have the potential to significantly impact liver disease research by enabling the discovery of novel therapeuticsbiomarkers to non invasively delineate NASH from steatosisand allow the safety assessment of drugs in a disease relevant background PROJECT NARRATIVE Nonalcoholic fatty liver diseaseNAFLDis a chronic condition that originates as lipid accumulation within hepatocytessteatosisand progresses into nonalcoholic steatohepatitisNASHcharacterized by lipid accumulationinflammationoxidative stressand fibrosisDespite decades of intense research worldwidethe mechanisms of NAFLD progression as well as therapeutic approaches and non invasive diagnostics for NASH are still lackingThe current Fast Track SBIR application seeks to utilize Organovo s ExViveHuman Liver Tissue to model steatosisPhase Iand its progression to NASHPhase IIin vitrowith verification of the clinical relevance of the models against patient data and modulation of disease phenotype by therapeutic interventionIf successfulthis model will provide an unprecedented tool for the advancement of NAFLD basic researchbiomarker identification and drug developmentsignificantly impacting patient lives