Multifunctional Nanoparticle Platform to Prevent Alcohol-Associated HCC Development - ABSTRACT
Hepatocellular carcinoma (HCC), the fastest rising cause of cancer-related deaths worldwide with a 5-year
survival of <20%, affects more than 41,000 individuals in the United States every year. Heavy alcohol
consumption leading to fatty liver, hepatitis and cirrhosis has been identified as a key risk factor in HCC
development. Current therapies against alcohol liver disease (ALD) and associated fibrosis are non-specific and
ineffective. Alcohol abstinence remains the gold standard for ALD treatment to prevent progression to HCC,
however this is often hampered by poor compliance. The goal of this proposal is to develop a novel
multifunctional nanoparticle (NP) platform (lipid-PLGA NPs) for treatment of alcohol-associated fibrosis via
targeted activation of G-protein-coupled bile acid receptor (Gpbar1) and anti-fibrotic drug delivery. The NPs will
also release collagenase to facilitate greater NP penetration into the fibrotic liver tissue. We hypothesize that (i)
NP-mediated targeted activation of Gpbar1 – a membrane protein expressed in Kupffer cells (KCs) and not
hepatocytes, will suppress NF-kβ and STAT3 signaling responsible for HCC development, and (ii) targeted
Gpbar1 activation and concurrent anti-fibrotic drug release will synergistically inhibit profibrotic biomarker
expressions and cytokine signaling, leading to attenuation of fibrosis. Preliminary investigations by our
collaborative research team confirmed that the NPs can selectively accumulate in the KCs in in vivo mouse
models. Our proposed aims are: (1) Characterization and in vitro evaluation of liver tissue penetration properties
of the collagenase-containing lipid-PLGA NPs. Physicochemical characterization will be done to ensure that the
lipid-PLGA NPs will have optimal properties for accumulation in the liver. A 3D multicellular spheroid model of
alcohol-induced liver fibrosis will be used to evaluate cytocompatibility, optimum uptake concentrations, and
tissue penetration by the NPs in vitro. (2) Gpbar1 agonist incorporation and in vivo elucidation of safety,
biodistribution and Gpbar1 targeting capabilities of the NPs. Gpbar1 targeting, safety and anti-fibrotic effects of
the lipid-PLGA NPs will be investigated using a widely studied and reported carbon tetrachloride-plus-alcohol
induced mouse models of liver fibrosis. (3) In vitro and in vivo evaluation of synergistic effects of Gpbar1-targeting
lipid-PLGA NPs given in combination with anti-fibrotic therapies. In this aim, the synergistic effects of Gpbar1
activation and interleukin-17A signaling inhibition on fibrosis attenuation will be determined following
encapsulation of anti-fibrotic therapies within the lipid-PLGA NPs. NP efficacy will be evaluated using histology,
biomarker analysis and collagen assays. As a first step towards assessing the translational potential of the
formulation, we will then investigate the therapeutic effects of the NPs using novel ALD liver fibrosis-on-a-chip
developed using primary murine cells. This innovative project will lead to a paradigm shift in the development
and testing of new therapeutic strategies against chronic liver diseases to prevent their progression to HCC.
