Adipocyte-derived FABP4 promotes metabolism-associated steatotic liver–induced hepatocellular carcinoma by driving ITGB1-mediated β-catenin activation
Carmen Oi Ning Leung, Shilpa Gurung, Katherine Po Sin Chung, Rainbow Wing Hei Leung, Martina Mang Leng Lei, Mandy Sze Man Chan, Gregory Kenneth Muliawan, Shakeel Ahmad Khan, Xue Qian Wu, Jun Yu, Hui Lian Zhu, Yin Ying Lu, Stephanie Ma, Xiaoping Wu, Ruby Lai Chong Hoo, Terence Kin Wah Lee
Carmen Oi Ning Leung, Shilpa Gurung, Katherine Po Sin Chung, Rainbow Wing Hei Leung, Martina Mang Leng Lei, Mandy Sze Man Chan, Gregory Kenneth Muliawan, Shakeel Ahmad Khan, Xue Qian Wu, Jun Yu, Hui Lian Zhu, Yin Ying Lu, Stephanie Ma, Xiaoping Wu, Ruby Lai Chong Hoo, Terence Kin Wah Lee
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Research Article Hepatology Oncology

Adipocyte-derived FABP4 promotes metabolism-associated steatotic liver–induced hepatocellular carcinoma by driving ITGB1-mediated β-catenin activation

Abstract

Metabolic dysfunction–associated steatotic liver disease–induced (MASLD-induced) hepatocellular carcinoma (HCC) is an emerging malignancy linked to excessive accumulation of adipose tissue and hepatic fat. Understanding the role of adipocytes in the development of MASLD-induced HCC is crucial. In an in vitro coculture system, differentiated adipocytes were found to enhance cancer stemness and drug resistance in HCC through paracrine signaling. Fatty acid–binding protein 4 (FABP4) was preferentially secreted by adipocytes, and recombinant FABP4 further augmented the cancer stem cell (CSC) properties of HCC cells. Notably, Fabp4–/– mice exhibited a marked delay in the progression of MASLD-HCC, which correlated with the increased HCC risk observed in MASLD patients with elevated FABP4 expression. Mass spectrometry analysis identified integrin β 1 (ITGB1) as a binding partner of FABP4. These data, together with a substantial downregulation of the Wnt/β-catenin pathway in Fabp4–/– mouse tumors, revealed that FABP4 augmented liver CSC functions by activating PI3K/AKT/β-catenin signaling via ITGB1. We developed an anti-FABP4 neutralizing antibody that successfully inhibited FABP4-driven CSC functions and suppressed MASLD-induced HCC. In conclusion, our findings indicate that adipocyte-derived FABP4 plays a critical role in the development of MASLD-induced HCC and targeting the ITGB1/PI3K/AKT/β-catenin signaling cascade may offer a promising approach to combat this aggressive disease.

Authors

Carmen Oi Ning Leung, Shilpa Gurung, Katherine Po Sin Chung, Rainbow Wing Hei Leung, Martina Mang Leng Lei, Mandy Sze Man Chan, Gregory Kenneth Muliawan, Shakeel Ahmad Khan, Xue Qian Wu, Jun Yu, Hui Lian Zhu, Yin Ying Lu, Stephanie Ma, Xiaoping Wu, Ruby Lai Chong Hoo, Terence Kin Wah Lee

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Figure 6

ITGB1 was identified as a receptor of exogenous FABP4 that mediates CSC functions by driving the PI3K/AKT/β-catenin signaling cascade.

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ITGB1 was identified as a receptor of exogenous FABP4 that mediates CSC ...
(A) Workflow for analysis of potential FABP4-binding proteins on the membrane surface of Huh7 cells using biotinylated rhFABP4 (n = 1). (B) Surface proteins detected with ≥2 unique peptides. (C) Interactions between potential FABP4-binding proteins and main players in Wnt/β-catenin signaling. (D) Among HCC patients with MASLD as risk factor in TCGA-LIHC, those with high ITGB1 had poorer overall survival rate than those with low ITGB1 (log-rank test). (E) According to GSE192959, GSE193080, and GSE193066, FABP4 was positively correlated with ITGB1 in patients with MASLD-related HCC and MASLD patients at high risk of HCC (Pearson’s correlation). (F) Reciprocal coimmunoprecipitation demonstrated the interaction between exogenous FABP4 and ITGB1 (n = 2). (G) Western blot analyses of shITGB1 HCC cells upon rhFABP4 treatment. (H) Transactivating activity of β-catenin was examined in shITGB1 HCC cells after treatment with rhFABP4 for 24 hours (n = 3). (I) Limiting dilution sphere analysis showed the role of ITGB1 in regulation of FABP4-driven self-renewal ability (n = 2). NTC, non-target control. (J) Apoptosis of ITGB1-silenced HCC cells pretreated with PBS (rhFABP4 0 ng/mL), rhFABP4 (100 ng/mL), doxorubicin, or sorafenib for 48 hours. n = 4–5. (K) Effect of rhFABP4 on cell migration and invasion upon silencing of ITGB1 (n = 4–6). (L) ITGB1-knockdown and control HCC cells pretreated with either PBS or rhFAPB4 at 100 ng/mL for 24 hours were subcutaneously inoculated into nude mice. Images of xenograft tumors. Scale bars: 1 cm. (M) Graphs showing the tumor masses (n = 5 mice per group). Data represent mean ± SD. *P < 0.01, **P < 0.01, ***P < 0.001, ****P < 0.0001. D: log-rank test; H, J, K, M: 1-way ANOVA followed by Tukey’s multiple-comparison test; I: extreme limiting dilution analysis with χ2 test.