Abstract 6249: Single-cell RNA sequencing investigation of multi-stage NASH-related HCC mouse model

Abstract

Abstract Hepatocellular carcinoma (HCC) is one of the most frequent malignancies and a leading cause of cancer-related death worldwide. Its incidence is also one of the most rapidly increasing in the West. Most (>80%) of HCCs are diagnosed at advanced stage and therefore not operable. Even after surgical resection, the long-term prognosis of HCC remains unsatisfactory due to high recurrence rates. It is also an extremely difficult-to-treat cancer. Most HCC patients have a background of chronic liver disease, including chronic viral infection, excessive alcohol consumption or non-alcoholic fatty liver disease NAFLD)/steatohepatitis (NASH). In June 2023, new nomenclature of NAFLD and NASH have been internationally announced as metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis, respectively. Due to the enforcement of neonatal HBV vaccination program and the availability of effective antiviral treatments on HBV and HCV, the global incidence of viral hepatitis-related HCC has been declining, whereas NAFLD/NASH-related incidence of HCC has been increasing. We established NASH-related HCC mouse model using Western diet (WD) plus carbon tetrachloride (CCl4) method. This model demonstrated steatosis, fibrosis, obesity and HCC with high incidence. It has also been reported to accurately portrait the key pathological features of the patients. Next, we collected liver tissue samples at different timepoints (normal; Week 12: NAFLD/NASH; Week 18: fibrosis/cirrhosis; Week 32: HCC) and analyzed them using single-cell RNA sequencing (scRNA-seq). Single-cell sequencing offers an unprecedented platform to perform systematic and in-depth investigation of the admixed tumor microenvironment (TME) in HCC tumor. We have taken advantage of the unique multi-dimensional capacity of scRNA-seq to delineate multifaceted landscapes and cell-cell communication in multi-stage NASH-related HCC mouse model. After removing the low-quality cells, our multi-stage dataset consisted of >130,000 cells. Using typical cell type markers, we identified major cell types, namely T cells, B cells, myeloid cells, liver sinusoidal endothelial cells, hepatic stellate cells, hepatocytes and HCC tumor cells. We observed a change in the cell type composition during disease progression. Throughout our study, we closely examined the communication level between different cell types to identify any irregularities or disturbances that could lead to NASH-related HCC. Our findings concluded that hepatic stellate cells, despite existing in low proportion at all stages, exhibited the strongest level of interaction with other cell types. Moreover, our data also established the comprehensive multi-stage cellular landscape of NASH-relative HCC. Citation Format: Tina Suoangbaji, Vanilla Xin Zhang, Daniel Wai-Hung Ho, Irene Oi-Lin Ng. Single-cell RNA sequencing investigation of multi-stage NASH-related HCC mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6249.