Targeting PI3K/BRD4 and Hedgehog Pathway in Alcohol Associated Liver Disease

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PROJECT SUMMARY This K01 application aims to promote the applicant's development to become a multi-disciplinarily trained and independent academic researcher. This application's collective strengths are defined in these 3 major areas: 1) Credentials: PI's application builds upon his productive track-record to achieve scientific independence in the field of drug delivery and liver fibrosis, including alcohol-associated liver disease (AALD). The PI's goals include enhancing grantsmanship, leadership, and team management skills to become an independent, tenure-track academic researcher. In addition to technical training, PI intends to build skills in communicating with faculty members, industry members, and students. 2) Training Environment: Drs. Mahato (mentor) and Kharbanda, and Cheng (co-mentors) are world-class researchers and fully committed to advancing the PI's career. Particular emphasis will be given to grant writing, collaboration building, and applying for external funding. The mentoring committee has strong credentials in developing the next generation of successful academic scientists. The knowledge and experience gained during this award period will help the candidate generate data to apply for an R21/R01 grant. 3) Innovative Research: PI's central hypothesis is that alcohol-induced activation of inflammatory pathways is mediated by AKT and BRD4 and synergize with the hedgehog (Hh) pathway to promote AALD. Simultaneous inhibition of these pathways using targeted nanomedicine could alleviate the progression of AALD and related morbidities. The PI has generated the following preliminary results; (a) Conditioned media from the EtOH treated primary human hepatocytes significantly stimulated hepatic stellate cells (HSCs) as determined by α-SMA expression levels and upregulated GLI1/2 activity; (b) The protein cMYC is upregulated in patient samples diagnosed with alcoholic hepatitis; (c) Treatment with novel inhibitors MDB5 (Hh) and SF2523 (PI3K/BRD4) showed decreased their target genes in HSC-T6 cells; (d) EtOH diet-fed mice show increased p- AKT, cMYC, and GLI1/2 protein levels in the liver compared to mice on the control diet; (e) Treatment with MDB5 and SF2523 decreased liver injury markers ALT and AST, and lowered GLI1, cMYC, and p-AKT protein levels in ethanol-fed mice; (f) Novel cleavable amphiphilic peptide (CAP) was self-assembled into nanoparticles (NPs) of size range 100± 10 nm loaded with both the drugs (payload 5% w/w); (g) CAP-NPs were sensitive to fibroblast activation protein (FAP-α), and NPs dissembled in presence of recombinant FAP-α. Based on these robust results, the proposal has the following specific aims: Aim 1. Establish the role of AKT/BRD4 dual inhibitor SF2523 and Hh inhibitor MDB5 in AALD. Aim 2. Formulate and characterize HSC targeted CAP-NP loaded with SF2523 and MDB5. Aim 3. Determine therapeutic efficacy of SF2523 and MDB5 loaded pPB-CAP-NPs in ALD mice. The new knowledge and nanomedicine generated in this proposal may open new therapeutic avenues for the treatment of AALD.