Endoplasmic reticulum–resident α-glucosidase II drives non-small cell lung cancer progression via regulation of secretory glycoproteins
Shike Wang, Na Ding, Angelo Chen, Derrick Cardin, Yuting Xu, Kate Grimley, William K. Russell, Jun Xu, Jonathan M. Kurie, Guan-Yu Xiao, Xiaochao Tan
Shike Wang, Na Ding, Angelo Chen, Derrick Cardin, Yuting Xu, Kate Grimley, William K. Russell, Jun Xu, Jonathan M. Kurie, Guan-Yu Xiao, Xiaochao Tan
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Research Article Cell biology Oncology

Endoplasmic reticulum–resident α-glucosidase II drives non-small cell lung cancer progression via regulation of secretory glycoproteins

Abstract

Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality worldwide, yet its molecular drivers are not fully defined. Emerging evidence highlights the importance of tumor-stroma interactions mediated by secreted glycoproteins. However, the mechanisms by which cancer cells regulate the secretion of these protumorigenic proteins remain largely unknown. Endoplasmic reticulum–resident (ER-resident) N-glycan–processing enzymes regulate proper protein folding, a prerequisite for glycoproteins to exit the ER and undergo secretion. By evaluating their prognostic significance in lung tumors and conducting functional screening in lung cancer cells, we identify α-glucosidase II (α-Glc II) as a key regulator of NSCLC progression. α-Glc II promotes tumor growth and dissemination in a glucosidase activity–dependent manner in orthotopic mouse lung tumor model. Genetic disruption of α-Glc II induced ER stress and reduced cell proliferation and motility. Mechanistically, α-Glc II–mediated N-glycan modification regulated the ER-to-Golgi trafficking and secretion of specific oncogenic glycoproteins, including lysyl hydroxylase 2 (LH2), Tissue Inhibitor of Metalloproteinase 1 (TIMP1), and TGF-β, which are known to be associated with extracellular matrix remodeling. These findings uncover a role for ER glycosylation machinery in shaping the NSCLC secretome and highlight α-Glc II as a potential therapeutic target.

Authors

Shike Wang, Na Ding, Angelo Chen, Derrick Cardin, Yuting Xu, Kate Grimley, William K. Russell, Jun Xu, Jonathan M. Kurie, Guan-Yu Xiao, Xiaochao Tan

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Figure 3

GANAB promotes tumor growth and metastasis in a enzymatic activity-dependent manner.

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GANAB promotes tumor growth and metastasis in a enzymatic activity-depen...
(A) Validation of CRISPR-CAS9 mediated GANAB knockout (KO) in 344SQ cells by WB. P, parental cells. (B) Numbers of orthotopic lung tumors (left) and metastases to mediastinal nodes and contralateral lung (right) in syngeneic, immunocompetent mice (129/SV) (dots represent individual mouse numbers) injected orthotopically with P or GANAB-KO 344SQ cells. P values were determined using 1-way ANOVA. (C) Flank tumor weights (left) and numbers of metastases to mediastinal nodes and contralateral lung (right) in 129/SV mice (dots represent individual mouse numbers) injected s.c. with P or GANAB-KO 344SQ cells. P values were determined using 1-way ANOVA. (D) WB confirmation of GANAB expression in A549 cells stably transfected with shRNAs targeting GANAB (shGANAB) or control shRNA (shCTL). (E) Numbers of orthotopic lung tumors (left) and metastases to mediastinal and contralateral lung (right) in nude mice (dots represent individual mouse numbers) injected orthotopically with shCTL- or shGANAB-A549 cells. P values were determined using Student’s t test. (F) Top: Schematic diagram of GANAB domains highlighting the catalytic inactive site (D564). Bottom: WB confirmation of GANAB expression in P- and GANAB-KO–344SQ cells reconstituted without (Vec) or with WT GANAB or D564N mutant GANAB. (G) Numbers of orthotopic lung tumor metastases to mediastinal nodes and contralateral lung in 129/SV mice (dots represent individual mouse numbers) injected orthotopically with P- or GANAB-KO–344SQ cells reconstituted with WT or mutant (D564N) GANAB. P values were determined using 1-way ANOVA. Data indicate the mean ± SEM from a single experiment incorporating biological replicate samples (n ≥ 3) and are representative of at least 2 independent experiments.