Clathrin light chain A facilitates small extracellular vesicle uptake to promote hepatocellular carcinoma progression

Abstract

Background Endocytosis is a fundamental process for internalizing small extracellular vesicles (sEVs). The present study aimed to elucidate the role of clathrin light chain A (CLTA) in sEV uptake in hepatocellular carcinoma (HCC). Materials and methods CLTA expression was analyzed by bioinformatics, quantitative PCR and immunohistochemistry. The clinical relevance of CLTA was analyzed by Fisher’s exact test, Kaplan–Meier analysis, and multivariate cox regression model. The functions of CLTA in sEV uptake and cancerous properties were examined by PKH67-sEV uptake, MTT, colony formation, and transwell assays. Mass spectrometry was used to identify the downstream effectors of CLTA. CLTA inhibitor, Pitstop 2, was tested in a mouse model of patient-derived xenografts (PDXs). Results CLTA expression was higher in tumor tissues than in non-tumorous liver tissues and progressively increased from the early to late tumor stage. CLTA overexpression was associated with larger tumor size and poor prognosis in HCC. Cellular CLTA contributed to the sEV uptake, resulting in enhanced cancerous properties. Mechanistically, CLTA increases capping actin protein gelsolin-like (CAPG) expression to facilitate sEV uptake, thereby promoting the proliferation, motility, and invasiveness of HCC cells. What’s more, the CLTA inhibitor Pitstop 2 alone or in combination with sorafenib attenuated tumor growth in mice implanted with PDXs. Conclusions The study reveals the role of CLTA in sEV uptake to promote HCC progression. Inhibition of CLTA and its mediated pathway illuminate a new therapeutic strategy for HCC patients.

Clathrin light chain A‐enriched small extracellular vesicles remodel microvascular niche to induce hepatocellular carcinoma metastasis

“…sEVs derived from metastatic HCC cells have been shown to facilitate premetastatic niche formation and promote tumorigenesis and metastasis (Mao et al., 2020 ). To answer whether CLTA is a functional component in sEVs of HCC cells, sEVs were obtained from vector control (XPack), CLTA overexpressing (XP‐CLTA), non‐target control (CTL‐KD) and CLTA knockdown (CLTA‐KD1 and CLTA‐KD2) clones established in MHCCLM3 and MHCC97L cells (Xu et al., 2023 ) (Figure S3a–d , Figure 2a ). The colony formation ability of HLE and Huh7 was significantly facilitated by both MHCCLM3 and MHCC97L XPack‐sEV compared to PBS treated cells.…”

Hybrid Adipocyte-Derived Exosome Nano Platform for Potent Chemo-Phototherapy in Targeted Hepatocellular Carcinoma

“…11 By carefully extracting the exosome from adipocytes, 19 we secured membranes with preserved expression of proteins characteristic of high fatty acid treatment (Figure 1), which facilitated targeted drug delivery via CD36-auxiliaried clathrin-mediated endocytosis (Figure 2) —a pathway implicated in macrophage and phagocytic activity and known to enhance the uptake of exosomes by HCC cells, thereby promoting tumor progression. 21, 27…”

Extracellular vesicles and cancer stemness in hepatocellular carcinoma – is there a link?

“…TMPRSS2-enriched exosomes were found to suppress the packaging of pro-tumoral nidogen into exosomes and inhibit proliferation and migration of immortalized hepatic cell ( 12 ). In addition, various molecules, including DEAD-box helicase 55 (DDX55), CPE, miR-3129, LINC00161, clathrin light chain A (CTLA), and miR-25, were found to be enriched in cancer-derived exosomes and induce cell proliferation ( 10 , 19 , 21 – 24 ). Signaling alterations upon sEV treatment primarily involved EMT markers, Wnt/β-catenin signaling, and cell cycle pathways ( 19 , 21 , 23 ).…”