Gasdermin D regulates the activation of EGFR in colorectal cancer
Ying Li1, Jiayao Chen2, Huijun Liang1
1Wuxi School of Medicine, Jiangnan University, Wuxi, 214122, China.
Background:
Gasdermin D (GSDMD) is a key effector molecule that activates pyroptosis through its N terminal domain (GSDMD-NT). However, the roles of GSDMD in colorectal cancer (CRC) have not been fully explored. The role of the full-length GSDMD (GSDMD-FL) is also not clear. In this study, we observed that GSDMD modulates CRC progression through other mechanisms in addition to activating GSDMD-NT.
Methods:
Clinical CRC samples and human-derived CRC cell lines were used in this study. GSDMD expression was evaluated by RT-qPCR, Western blot and immunohistochemical (IHC) analysis. GSDMD knockdown and overexpression stable cell lines were established by Lentiviral transduction. CCK-8 assay, flow cytometry analysis for cell cycle, Transwell assay, and cell scratch assay were performed in vitro to explore the impact of GSDMD on CRC progression. Mouse subcutaneous transplantation tumor models were constructed to assess the role of GSDMD in vivo. Intestinal epithelial cell (IEC)-specific knockout of Gsdmd mice (GsdmdΔIEC) was used to evaluate the effect of GSDMD on intestinal adenoma formation in AOM-DSS and Apcmin/+ mouse models. RNA sequencing was performed to explore the regulatory pathways associated with the role of GSDMD in CRC cells. Co-Immunoprecipitation (CO-IP), Western blot and immunofluorescence (IF) were conducted to investigate the interactions between GSDMD and EGFR. Exogenous addition of Gefitinib was used to evaluate the effect of GSDMD on autophosphorylation of EGFR at the Tyr1068 site.
Results:
GSDMD was highly expressed in clinical CRC tissues and human-derived CRC cell lines. GSDMD knockdown inhibited the viability, cell cycle changes, invasion ability and migration ability of CRC cell lines in vitro and vivo, whereas GSDMD overexpression had the opposite effects. Intestinal adenoma development was reduced in GsdmdΔIEC mice in both AOM-DSS and Apcmin/+ mouse models. GSDMD-FL interacted with EGFR and promoted CRC progression by inducing autophosphorylation of EGFR at the Tyr1068 site, subsequently activating ERK1/2. Exogenous Gefitinib abrogated the tumorigenic properties of GSDMD.
Conclusions:
GSDMD-FL promotes CRC progression by inducing EGFR autophosphorylation at the Tyr1068 site, subsequently activating the downstream ERK1/2. Inhibition of GSDMD is a potential strategy for the treatment of colorectal cancer.
Insights
Gasdermin D (GSDMD) promotes colorectal cancer (CRC) progression by interacting with EGFR and activating ERK1/2. Inhibiting GSDMD offers a potential therapeutic strategy for treating CRC.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Gasdermin D (GSDMD) is a key pyroptosis effector, but its role in colorectal cancer (CRC) and the function of full-length GSDMD (GSDMD-FL) remain unclear.
- This study investigates GSDMD's multifaceted roles in CRC beyond pyroptosis activation.
Purpose of the Study:
- To elucidate the role of GSDMD in colorectal cancer progression.
- To explore the interaction between GSDMD and Epidermal Growth Factor Receptor (EGFR) in CRC.
- To evaluate GSDMD as a potential therapeutic target for CRC.
Main Methods:
- GSDMD expression analysis in clinical CRC samples and cell lines using RT-qPCR, Western blot, and IHC.
- In vitro and in vivo functional assays including cell viability, migration, invasion, and tumor xenograft models.
- Analysis of intestinal adenoma formation in Gsdmd-deficient mouse models (AOM-DSS and Apcmin/+) and RNA sequencing.
- Investigation of GSDMD-EGFR interaction using CO-IP, Western blot, and immunofluorescence, with Gefitinib treatment to assess EGFR phosphorylation.
Main Results:
- GSDMD is highly expressed in CRC tissues and cell lines, promoting CRC cell viability, invasion, and migration.
- GSDMD-FL interacts with EGFR, promoting its autophosphorylation at Tyr1068 and activating the downstream ERK1/2 pathway, thereby driving CRC progression.
- GSDMD deficiency in mice reduced intestinal adenoma formation, and Gefitinib treatment abrogated GSDMD-driven tumorigenic properties.
Conclusions:
- GSDMD-FL promotes colorectal cancer progression by upregulating EGFR autophosphorylation and activating the ERK1/2 pathway.
- Targeting GSDMD presents a promising therapeutic strategy for colorectal cancer treatment.
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