Targeting immunosuppressive macrophages by CSRP2-regulated CCL28 signaling sensitizes hepatocellular carcinoma to
Changzhou Chen1,2, Sheng Su1,2, Pengcheng Wang3
1Department of Hepatobiliary Surgery and Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, Shanghai, China.
Background:
Despite comparable survival benefit has been obtained, the drug resistance remarkably reduced lenvatinib clinical efficacy. Here, we aimed to identify the potential mechanism by which cysteine and glycine-rich protein 2 (CSRP2) regulates the development of hepatocellular carcinoma (HCC) and participates in the resistance to lenvatinib.
Methods:
We harnessed RNA sequencing, multiplex immunofluorescence staining, and hydrodynamic tail vein (HTV) injection HCC model to systematically explore the function of CSRP2 in HCC progression. To precisely delineate how immunosuppressive macrophages, influenced by CSRP2-regulated C-C motif chemokine ligand 28 (CCL28) signaling, respond to lenvatinib-induced cytotoxicity, we established an in vitro co-culture system and conducted functional cytotoxicity assays.
Results:
Using RNA sequencing, multiplex immunofluorescence staining and HTV injection HCC mouse model, we identified CSRP2 as one of the most significantly upregulated genes in HCC tissues. CSRP2 overexpression drives anti-lenvatinib resistance by inducing high levels of tumor-associated macrophages (TAMs) infiltration and reshaping an immunosuppressive microenvironment. Then flow cytometry, mass spectrometry and chromatin immunoprecipitation were conducted to clarify the underlying mechanism of CSRP2. We showed CSRP2 promotes phosphorylation of activating transcription factor 2 (ATF2) at Thr69/71, leading to the transcriptional activation of CCL28 expression. HCC-derived CCL28 recruits TAMs to drive immunosuppression and anti-lenvatinib tolerance. BI6901, a potent and selective CCR10 antagonist, blocked TAMs recruitment and enhanced T-cell activation. Combining CCR10 inhibition improved the therapeutic benefit of anti-lenvatinib in HCC.
Conclusions:
These results illustrate that CSRP2 regulates the tumor microenvironment to promote HCC growth and drive lenvatinib tolerance via the CSRP2/ATF2/CCL28 axis. Targeting this pathway could synergize with lenvatinib to treat HCC more effectively.
Insights
Cysteine and glycine-rich protein 2 (CSRP2) drives lenvatinib resistance in hepatocellular carcinoma (HCC) by promoting immunosuppressive tumor microenvironments. Targeting the CSRP2/ATF2/CCL28 pathway can enhance lenvatinib efficacy for HCC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Drug resistance significantly limits lenvatinib's clinical efficacy in hepatocellular carcinoma (HCC).
- Understanding the mechanisms of lenvatinib resistance is crucial for improving HCC treatment outcomes.
Purpose of the Study:
- To investigate the role of cysteine and glycine-rich protein 2 (CSRP2) in HCC development and lenvatinib resistance.
- To elucidate the molecular mechanisms by which CSRP2 influences the tumor microenvironment and drug response.
Main Methods:
- RNA sequencing and multiplex immunofluorescence staining were used to identify CSRP2 expression in HCC.
- Hydrodynamic tail vein (HTV) injection HCC models and in vitro co-culture systems were established to study CSRP2 function.
- Flow cytometry, mass spectrometry, and chromatin immunoprecipitation assays were employed to clarify the underlying molecular pathways.
Main Results:
- CSRP2 was identified as a significantly upregulated gene in HCC tissues, driving resistance to lenvatinib.
- CSRP2 overexpression promotes tumor-associated macrophage (TAM) infiltration and creates an immunosuppressive microenvironment.
- CSRP2 activates the ATF2/CCL28 axis, leading to TAM recruitment and enhanced lenvatinib tolerance; CCR10 inhibition improved therapeutic benefit.
Conclusions:
- CSRP2 promotes HCC growth and lenvatinib tolerance by modulating the tumor microenvironment via the CSRP2/ATF2/CCL28 pathway.
- Targeting the CSRP2/ATF2/CCL28 axis offers a potential strategy to synergize with lenvatinib for more effective HCC treatment.
