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Principal Investigator: Amaia Lujambio
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2021
Award: $620,173
Funding agency: National Cancer Institute
Chronic liver disease and its common sequela cirrhosis, are growing public health concerns, and major risk
factors for the development of hepatocellular cancer (HCC). HCC incidence and mortality is growing in the
USA. Optimal medical therapies for HCC are lacking. FDA approved agents are modestly effective. Immune
checkpoint inhibitors (ICIs) including PD-1 inhibitors Nivolumab and Pembrozilumab, have been both
approved and show good response rates but only in a subset of HCC cases. Biomarkers of response remain
unknown. Recent years have seen a revolution in HCC GWAS studies to identify molecular drivers. Mutations
in CTNNB1 activate b-catenin in the Wnt pathway & seen in up to 37% of HCCs. However, b-catenin activation
alone does not lead to HCC. Analysis of 2 large HCC cohorts (TCGA & French) revealed CTNNB1 mutations to
co-occur with alterations in MET, MYC, TERT, NFE2L2, MLL2, ARID2 & APOB. Overexpression/activation of
Met along with CTNNB1 mutations is seen in ~11% of HCCs. Coexpression of these genes in a subset of
hepatocytes using sleeping beauty transposon/transposase (SB) & hydrodynamic tail vein injection (HTVI) led
to HCC by 6 weeks in mice (hMet-b-catenin model). Gene expression analysis confirmed 70% similarity
between hMet-b-catenin model and HCC patient subset with Met activation & CTNNB1 mutations. In aim 1,
we will generate and characterize mouse models using SB/Crispr and HTVI to co-express mutant CTNNB1 and
other genes frequently co-altered in subsets of human HCC including MYC, TERT, NFE2L2, MLL2, ARID2, &
APOB. We already show HCC development in Met-b-catenin, MYC-b-catenin, TERT-b-catenin & NFE2L2-b-
catenin, while others are ongoing. Comparison of gene expression between mouse models and human HCC
subsets will validate the relevance of these models justifying a more comprehensive cellular & molecular
characterization for innovative therapies. We will also test dependence of all mutant CTNNB1-mouse models to
b-catenin by using of lipid nanoparticles (LNP) containing CTNNB1-siRNA (CTNNB1-LNP) to suppress b-
catenin and assess response as we have shown for Kras-b-catenin (akin to hMet-b-catenin) model. In aim 2, we
will focus on b-catenin-glutamine synthetase (GS)-glutamine-mTORC1 axis in mutant-CTNNB1 HCC, recently
discovered and reported by us (Publication in Cell Metabolism). All mutant-CTNNB1 driven HCC models with
all co-occurrences will be tested for response to mTORC1 inhibitors like Everolimus and RM-006 (novel
exclusive mTORC1 inhibitor) and to upstream GS via genetic deletion of GS in established HCCs or via use of
irreversible GS inhibitor L-methionine sulfoximine (MSO) and glutaminase inhibitor CB-839 that hampers
production of glutamate, a substrate for GS to generate glutamine. In aim 3, we will investigate how to make b-
catenin-active HCCs shown by us to be resistant to ICIs (publication in Cancer Discovery), sensitive to ICIs
through use of combination therapy. Using an immunogenic Myc-lucOS-mutant-b-catenin HCC model, that is
resistant to PD-1 inhibitor, we will test role of concomitant b-catenin suppression via CTNNB1-LNP, mTORC1
inhibition via Everolimus or GS inhibition via MSO, as sensitizing agents to PD-1 inhibitor. Thus overall, our
proposal will develop clinically relevant and validated preclinical models utilizing mutant b-catenin as one
cooperating oncogene and demonstrate role of mutant b-catenin in regulating tumor metabolism and immune
microenvironment. Completion of ours study will thus provide justification for targeting b-catenin in a notable
subset of HCC patients through innovative therapies and eventually pave the way for personalized medicine.
Terms: <Abscission><Antigens><Antioncogene Protein p53><BAY 54-9085><Beta Cadherin-Associated Protein><Beta-1 Catenin><Biological Markers><Biology><Breast Cell Glutaminase><CRE Recombinase><CRISPR><CRISPR/Cas system><CTNNB><CTNNB1><CTNNB1 gene><CUL-2><Cancer Genes><Cancer Model><Cancer Patient><Cancer-Promoting Gene><CancerModel><Cancers><Cell Body><Cells><Cellular Metabolic Process><Cellular Tumor Antigen P53><Characteristics><Checkpoint inhibitor><Cirrhosis><Clinic><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats><Combination immunotherapy><Combined Modality Therapy><Data><Data Bases><Databases><Death Rate><Dependence><Development><EC 3.5.1.2><Enterobacteria phage P1 Cre recombinase><Environment><Excision><Extirpation><FDA approved><FK506 Binding Protein 12-Rapamycin Associated Protein 1><FKBP12 Rapamycin Complex Associated Protein 1><FRAP1><FRAP1 gene><FRAP2><GA Protein><GWA study><GWAS><Gene Expression><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Gln><Glutamate Ammonia Ligase (ADP)><Glutamate-Ammonia Ligase><Glutamates><Glutaminase><Glutamine><Glutamine Synthetase><Hepatic Cancer><Hepatic Cells><Hepatic Parenchymal Cell><Hepatic Transplantation><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatocyte><Hepatoma><Human><Immune><Immune checkpoint inhibitor><Immune response><Immunes><Immunological response><Incidence><Induction Therapy><Infiltration><Injections><Innovative Therapy><KO mice><Knock-out Mice><Knockout Mice><L glutamine amidohydrolase><L-Glutamate><L-Glutamine><Lead><Liver Cells><Liver Cells Carcinoma><Liver Glutaminase><Liver Grafting><Liver Transplant><MLL2><MLL2 gene><Malignant Neoplasms><Malignant Tumor><Malignant neoplasm of liver><Mechanistic Target of Rapamycin><Medical><Methionine Sulfoximine><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Multimodal Therapy><Multimodal Treatment><Murine><Mus><Mutate><Mutation><NEOADJ><Neoadjuvant><Neoadjuvant Therapy><Neoadjuvant Treatment><Nivolumab><Null Mouse><Oncogenes><Oncogenesis><Oncogenic><Oncoprotein p53><Opdivo><Other Genetics><P53><PD 1><PD-1><PD-1 antibody therapy><PD-1 inhibitors><PD-1 therapy><PD1><PD1 antibody therapy><PD1 based treatment><PD1 inhibitors><PRO2286><Pathway interactions><Patients><Pb element><Phenotype><Phosphoprotein P53><Phosphoprotein pp53><Pre-Clinical Model><Precision therapeutics><Preclinical Models><Predisposition><Primary carcinoma of the liver cells><Production><Protein TP53><Public Health><Publications><Publishing><Q Levoglutamide><Q. Levoglutamide><RAFT1><Rapamune><Rapamycin><Regulation><Removal><Reporting><Research Proposals><Resistance><Risk Factors><Role><SDZ RAD><SEER Program><SEER-Surveillance, Epidemiology, and End Results><Sampling><Scientific Publication><Short interfering RNA><Sirolimus><Sleeping Beauty><Small Interfering RNA><Sorafenib><Surgical Removal><Surveillance, Epidemiology and End Results><Surveillance, Epidemiology, and End Results Program><Susceptibility><TCGA><TP53><TP53 gene><TRP53><Tail><Testing><The Cancer Genome Atlas><Therapeutic><Transcript Expression Analyses><Transcript Expression Analysis><Transforming Genes><Transposase><Tumor Cell><Tumor Immunity><Tumor Protein p53><Tumor Protein p53 Gene><Unresectable><Veins><Woman><aPD-1><aPD-1 therapy><aPD-1 treatment><aPD1><aPD1 therapy><aPD1 treatment><anti programmed cell death 1><anti programmed cell death protein 1 inhibitor><anti-PD-1><anti-PD-1 inhibitors><anti-PD-1 therapy><anti-PD-1 treatment><anti-PD1><anti-PD1 inhibitors><anti-PD1 therapy><anti-PD1 treatment><anti-programmed cell death 1 therapy><anti-programmed cell death protein 1><anti-programmed cell death protein 1 therapy><anti-tumor immunity><antiPD-1><antiPD1><antitumor immunity><bacteriophage P1 recombinase Cre><base><beta catenin><bio-markers><biologic marker><biomarker><cancer cell metabolism><cancer immunity><cancer metabolism><cell metabolism><cellular metabaolism><chronic hepatic disease><chronic hepatic disorder><chronic liver disease><chronic liver disorder><cirrhotic><clinical relevance><clinically relevant><cohort><combination therapy><combinatorial><combinatorial immunotherapy><combined modality treatment><combined treatment><comparative efficacy><compare efficacy><data base><design><designing><developmental><dual immunotherapy><effective therapy><effective treatment><everolimus><gain of function mutation><gene expression analysis><gene expression assay><genome mutation><genome wide association><genome wide association scan><genome wide association studies><genome wide association study><genomewide association scan><genomewide association studies><genomewide association study><glutamate synthetase><glutamatergic><glutamine synthase><heavy metal Pb><heavy metal lead><host response><immune check point inhibitor><immune microenvironment><immune system response><immunogen><immunogenic><immunoresponse><immunosuppressive microenvironment><immunosuppressive tumor microenvironment><inhibitor><inhibitor/antagonist><lipid nanoparticle><liver cancer><liver carcinoma><liver transplantation><loss of function mutation><mTOR><mTOR Inhibitor><mTOR inhibition><malignancy><malignant liver tumor><mammalian target of rapamycin><men><men's><mortality><mouse model><multi-modal therapy><multi-modal treatment><murine model><mutant><neoplasm/cancer><neoplastic cell><novel><overexpress><overexpression><p53 Antigen><p53 Genes><p53 Tumor Suppressor><palliation><pathway><patient subgroups><patient subpopulations><patient subsets><patient subtypes><personalization of treatment><personalized medicine><personalized therapy><personalized treatment><precision medicine><precision therapies><precision treatment><precision-based medicine><predicting response><prediction of response><predictive response><predictor of response><programmed cell death 1><programmed cell death protein 1><programmed cell death protein 1 therapy><programmed death 1><promoter><promotor><protein p53><resection><resistant><response><response biomarker><response markers><response prediction><siRNA><sle2><social role><systemic lupus erythematosus susceptibility 2><therapeutic target><transcriptional profiling><tumor><tumor cell metabolism><tumor immune microenvironment><tumor initiation><tumor metabolism><tumor-immune system interactions><tumorigenesis><whole genome association analysis><whole genome association studies><whole genome association study><αPD-1><αPD1><β-catenin>