GDF15 negatively regulates chemosensitivity via TGFBR2-AKT pathway-dependent metabolism in esophageal squamous cell carcinoma

doi:10.1007/s11684-022-0949-7

Front. Med.

2023, Vol. 17

Issue (1) : 119-131 https://doi.org/10.1007/s11684-022-0949-7

RESEARCH ARTICLE

GDF15 negatively regulates chemosensitivity via TGFBR2-AKT pathway-dependent metabolism in esophageal squamous cell carcinoma

Yingxi Du¹, Yarui Ma¹, Qing Zhu², Yong Fu³, Yutong Li¹, Ying Zhang¹, Mo Li¹, Feiyue Feng⁴(

), Peng Yuan⁵(

), Xiaobing Wang¹(

)

¹. State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China
². Department of Clinical Laboratory, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
³. Township Health Center in Tanggou Town, Wuhu 24100, China
⁴. Thoracic Surgery Department, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China
⁵. Department of VIP Medical Services, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China

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Abstract

Treating patients with esophageal squamous cell carcinoma (ESCC) is challenging due to the high chemoresistance. Growth differentiation factor 15 (GDF15) is crucial in the development of various types of tumors and negatively related to the prognosis of ESCC patients according to our previous research. In this study, the link between GDF15 and chemotherapy resistance in ESCC was further explored. The relationship between GDF15 and the chemotherapy response was investigated through in vitro and in vivo studies. ESCC patients with high levels of GDF15 expression showed an inferior chemotherapeutic response. GDF15 improved the tolerance of ESCC cell lines to low-dose cisplatin by regulating AKT phosphorylation via TGFBR2. Through an in vivo study, we further validated that the anti-GDF15 antibody improved the tumor inhibition effect of cisplatin. Metabolomics showed that GDF15 could alter cellular metabolism and enhance the expression of UGT1A. AKT and TGFBR2 inhibition resulted in the reversal of the GDF15-induced expression of UGT1A, indicating that TGFBR2-AKT pathway-dependent metabolic pathways were involved in the resistance of ESCC cells to cisplatin. The present investigation suggests that a high level of GDF15 expression leads to ESCC chemoresistance and that GDF15 can be targeted during chemotherapy, resulting in beneficial therapeutic outcomes.

Keywords GDF15 esophageal squamous cell carcinoma chemoresistance cellular metabolism TGFBR2 AKT

Corresponding Author(s): Feiyue Feng,Peng Yuan,Xiaobing Wang

Just Accepted Date: 04 November 2022 Online First Date: 12 December 2022 Issue Date: 15 March 2023

Cite this article:

Yingxi Du,Yarui Ma,Qing Zhu, et al. GDF15 negatively regulates chemosensitivity via TGFBR2-AKT pathway-dependent metabolism in esophageal squamous cell carcinoma[J]. Front. Med., 2023, 17(1): 119-131.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-022-0949-7
https://academic.hep.com.cn/fmd/EN/Y2023/V17/I1/119

Fig.1 GDF15 was associated with chemoresistance. (A) Expression level of GDF15 from the GEPIA database. (B) GDF15 gene expression between esophageal cancer and normal adjacent tissues from the GEO database. (C) Survival curve of low- and high-GDF15-expression esophageal cancer patients who received chemotherapy. (D) High GDF15 expression was related to a high IC50 (µmol/L) for DDP. (E) Expression of GDF15 in ESCC chemosensitive and chemoresistant patients using IHC.

Fig.2 GDF15 was linked to the chemosensitivity of ESCC to cisplatin in vitro and in vivo. (A) Cell viability was measured in KYSE30 cells with or without GDF15 (5 ng/mL) for two days. Experiments were repeated three times with three replicates. (B) Results of flow cytometry apoptosis analysis of KYSE30 cells exposed to DDP (0.79 ng/mL) and DDP/GDF15 (5 ng/mL) for two days. (C) Cell viability was measured in KYSE150 cells treated with DDP and DDP/anti-GDF15 antibody for two days. (D) Western blot analysis of Bax and Bcl-2 in KYSE150 cells after adding DDP and anti-GDF15 antibody. (E) Representative image of xenograft tumors. (F) Graph showing the change in tumor volume after treatment. The data are presented as the means ± SEM. (G) Weights of dissected xenograft tumors.

Fig.3 GDF15 was involved in drug metabolism. (A, B) Differentially expressed mRNAs in patients with low and high GDF15 expression were categorized using GO and KEGG enrichment analyses. (C) Heatmap showing differentially expressed genes in the GDF15 (5 ng/mL) treatment and control groups. (D) KEGG enrichment analysis of differentially expressed genes between the GDF15 treatment and control groups.

Fig.4 GDF15 changed the metabolome of cancer cells. (A, C) Heatmaps of differential metabolites from the control and experimental groups in the negative (ESI?) ion mode and positive (ESI+) ion mode. (B, D) Enriched pathways between the two groups of differential metabolites in the negative (ESI?) ion mode and positive (ESI+) ion mode. Experimental group cells were treated with GDF15 (5 ng/mL) for two days.

Fig.5 GDF15 regulated the TGFBR2-AKT pathway. (A) Potential signaling pathways associated with GDF15 in the GSCA database. (B) Protein–protein interactions of various proteins predicted using the STRING database. (C, D) AKT expression and phosphorylation levels were detected by Western blot from KYSE30 and KYSE150 cells treated with GDF15 (5 ng/mL) or anti-GDF15 antibody (1 µg/mL). (E, F) Western blot analysis of AKT in KYSE150 cells after the addition of LY21573299 and LY2109761. (G) AKT expression after knockdown of TGFBR2 in KYSE150 cells. (H) AKT expression after overexpression of TGFBR2 in KYSE150 cells. Data are reported as the mean ± SD.

Fig.6 GDF15 regulated the sensitivity of ESCC to cisplatin via UGT1A. (A) qRT-PCR analyses of UGT1A expression in the following treatment groups: vehicle, GDF15 (5 ng/mL), LY294002 (5 μmol/L), LY2109761(5 μmol/L), combination of GDF15 and LY294002, and combination of GDF15 and LY2109761. Data are reported as the mean ± SD. (B, C) Effect of UGT1A silencing on cell proliferation under cisplatin, control, and cisplatin + GDF15 treatment for two days. (D, E) Effect of UGT1A silencing on cell apoptosis under cisplatin, control, and cisplatin + GDF15 treatment for two days.

Fig.7 Schematic depicting the role of GDF15 in resistance to DDP.

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