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Abstract

Abstract Title

GALNT2 contributes to sarcomatoid differentiation in clear cell renal cell carcinoma, regulating immune escape and tumor microenvironment

Presentation Type

Non-Moderated Poster Abstract

Manuscript Type

Basic Research

Abstract Category *

Oncology: Kidney (non-UTUC)

Author's Information

Number of Authors (including submitting/presenting author) *

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Co-author 1

Shao-hao Chen shaohao.chen@fjmu.edu.cn The First Affiliated Hospital of Fujian Medical University Fuzhou China *

Co-author 2

Jun-ming Zhu 506522068@qq.com The First Affiliated Hospital of Fujian Medical University Fuzhou China -

Co-author 3

Xue-Yi Xue xuexueyi@fjmu.edu.cn The First Affiliated Hospital of Fujian Medical University Fuzhou China -

Co-author 4

Ning Xu drxun@fjmu.edu.cn The First Affiliated Hospital of Fujian Medical University Fuzhou China -

Co-author 5

Co-author 6

Co-author 7

Co-author 8

Co-author 9

Co-author 10

Co-author 11

Co-author 12

Co-author 13

Co-author 14

Co-author 15

Co-author 16

Co-author 17

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Co-author 20

Abstract Content

Introduction

Sarcomatoid renal cell carcinoma (SRCC) is one of the most aggressive forms of renal cell carcinoma (RCC). The prognosis of SRCC is worse than normal RCC, and it is necessary to further elucidate its molecular mechanism and potential therapeutic approaches.

Materials and Methods

Untagged quantitative proteomics was used to analyze pathological tissues from 10 SRCC patients. The specific mechanism by which GALNT2 induces EMT through upregulation of TUBB3 was verified using LinkedOmics database, fluorescence co-localization analysis, lectin pull-down assays, and Western blotting. TIMER 2.0 was used for pan-cancer analysis to explore the relationship between GALNT2 and tumor microenvironment. A stable knockdown GALNT2 RENCA cell line was constructed to validate the impact of GALNT2 on tumor growth and the tumor microenvironment. Targeted inhibition of GALNT2 combined with anti-PD-1 immunotherapy was used to explore therapeutic efficacy in vivo.

Results

A total of 14 differentially expressed proteins were identified in this study. PPI analysis revealed that GALNT2 was a key node. High GALNT2 expression was found to be closely associated with poor prognosis in ccRCC. GALNT2 knockdown inhibited ccRCC cell function and promoted apoptosis. In the LinkedOmics database, TUBB3 was found to have the strongest correlation with GALNT2 among EMT-related genes. Fluorescence co-localization analysis indicated co-localization between GALNT2 and TUBB3 proteins. Lectin pull-down assays demonstrated that GALNT2 regulates O-GlcNAc glycosylation modification. Rescue experiments showed that TUBB3 could counteract the effects of GALNT2 knockdown or overexpression on EMT formation in ccRCC. High GALNT2 expression was closely related to increased CAF infiltration, reduced lymphocyte infiltration, and increased matrix ratio in ccRCC. In vivo tumorigenesis experiments further confirmed these results. Targeted inhibition of GALNT2 combined with BE0146 therapy can suppress tumor immune escape and enhance the efficacy of immune therapy.

Conclusions

GALNT2 regulates TUBB3 glycosylation, induces EMT formation, and promotes ccRCC cell proliferation, migration, and invasion through upregulation of TUBB3. Targeted inhibition of GALNT2 combined with immune therapy can suppress tumor immune escape and enhance the efficacy of immune therapy, thus maximizing tumor growth inhibition.

Keywords

Renal clear cell carcinoma, sarcomatoid differentiation, proteomics, tumor immune microenvironment

Figure 1

https://storage.unitedwebnetwork.com/files/1237/7cd3d38d4be4f9f35fefde3c5460025e.jpg

Figure 1 Caption

Fig. 1. Screening and identification of core genes. (A) Two-dimensional scatter plot of PCA (principal component analysis). (B) Volcanic map of SRCC vs ccRCC. (C) Thermal map of SRCC vs ccRCC. (D)Wayne diagram of differential proteins. (E) Differenti

Figure 2

https://storage.unitedwebnetwork.com/files/1237/ccbe1eab8b2210cae2d20da3cb85d611.jpg

Figure 2 Caption

Fig. 2. Expression analysis of GALNT2 in cells and tissues. (A) Differential expression of GALNT2 between tumor and normal tissues in TCGA database. (B) The correlation between GALNT2 expression and WHO/ISUP grading. (C) The correlation between GALNT

Figure 3

https://storage.unitedwebnetwork.com/files/1237/3419c8a1f48f958b98ef259b882df1d3.jpg

Figure 3 Caption

Fig. 3. GALNT2 Knockdown Inhibits the Proliferation, Migration, and Invasion of Renal Cancer Cells. (A, B) Plate cloning experiment to detect the effect of knocking down GALNT2 on the proliferation ability of ACHN cells and 786-O cells. (C, D) Transw

Figure 4

https://storage.unitedwebnetwork.com/files/1237/f136d31239a76878e5a3d453c92996fd.jpg

Figure 4 Caption

Fig. 4. OE-GALNT2 promotes the proliferation, migration, and invasion ability of ACHN cells. (A) Immunoblotting was used to detect the expression level of GALNT2 in ACHN cell lines infected with GALNT2 overexpressing lentivirus. (B) RT-qPCR was used

Figure 5

https://storage.unitedwebnetwork.com/files/1237/87873a8285ec5f6989e47f02d4e15844.jpg

Figure 5 Caption

Fig. 5. GALNT2 promotes sarcomatoid differentiation of renal clear cell carcinoma through TUBB3. (A) GSEA enrichment analysis of biological processes associated with GALNT2 expression. (B-C) LinkedOmics database analysis of genes related to EMT and G

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