A rare germline mutation reverses the suppressive effect of GPC5 thereby promoting lung adenocarcinoma development and tumorigenesis DOI Creative Commons

Zhifa Zheng,

Lina Zhao,

Sen Zhao

et al.

Frontiers in Genetics, Journal Year: 2025, Volume and Issue: 16

Published: April 25, 2025

Background and Objective Glypican-5 (GPC5) has been well-characterized as a tumor suppressor in lung adenocarcinoma (LUAD); however, the functional implications of its germline mutations cancer pathogenesis remain largely unexplored. In this study, we identified characterized pathogenic GPC5 variant (c.776C>T, p.Pro259Leu) within Chinese LUAD pedigree, systematically investigating oncogenic mechanisms through comprehensive molecular cellular analyses. Methods Our investigation employed multifaceted approach beginning with recruitment LUAD-affected family cohort (n=4 patients, 1 healthy control), followed by exome sequencing matched blood FFPE samples. Through rigorous rare analysis, prioritized c.776C>T variant, subsequently validating pathogenicity via integrated computational modeling immunohistochemical profiling. Mechanistic studies A549 H2009 cell lines encompassed: (1) proliferation apoptosis assessment using CCK-8, colony formation, EdU incorporation, flow cytometry; (2) migration invasion evaluation Transwell wound healing assays; (3) EMT/Wnt pathway interrogation Western blot analysis E-cadherin, N-cadherin, Vimentin, β-catenin expression patterns; (4) definitive validation overexpression knockdown experiments. Results Genetic revealed exhibited complete cosegregation phenotype pedigree while being absent control populations (gnomAD frequency: 0.000003989), accompanied significantly reduced tissues. Functional characterization demonstrated that compared to wild-type, mutant conferred aggressive properties: enhanced proliferative capacity, impaired induction, markedly increased migratory potential. Molecular analyses promoted EMT activation nuclear accumulation subsequent upregulation mesenchymal markers. Crucially, siRNA-mediated phenocopied these effects, providing evidence tumor-suppressive function. Discussion findings establish mutation drives progression novel mechanism involving degradation, translocation, consequent activation. These results position critical nodal regulator Wnt/β-catenin signaling suggest may serve valuable biomarkers for hereditary risk assessment. Therapeutically, highlight potential utility Wnt inhibitors managing GPC5-mutant cases, also framework future investigations into glypican members biology.

Language: Английский

A rare germline mutation reverses the suppressive effect of GPC5 thereby promoting lung adenocarcinoma development and tumorigenesis DOI Creative Commons

Zhifa Zheng,

Lina Zhao,

Sen Zhao

et al.

Frontiers in Genetics, Journal Year: 2025, Volume and Issue: 16

Published: April 25, 2025

Background and Objective Glypican-5 (GPC5) has been well-characterized as a tumor suppressor in lung adenocarcinoma (LUAD); however, the functional implications of its germline mutations cancer pathogenesis remain largely unexplored. In this study, we identified characterized pathogenic GPC5 variant (c.776C>T, p.Pro259Leu) within Chinese LUAD pedigree, systematically investigating oncogenic mechanisms through comprehensive molecular cellular analyses. Methods Our investigation employed multifaceted approach beginning with recruitment LUAD-affected family cohort (n=4 patients, 1 healthy control), followed by exome sequencing matched blood FFPE samples. Through rigorous rare analysis, prioritized c.776C>T variant, subsequently validating pathogenicity via integrated computational modeling immunohistochemical profiling. Mechanistic studies A549 H2009 cell lines encompassed: (1) proliferation apoptosis assessment using CCK-8, colony formation, EdU incorporation, flow cytometry; (2) migration invasion evaluation Transwell wound healing assays; (3) EMT/Wnt pathway interrogation Western blot analysis E-cadherin, N-cadherin, Vimentin, β-catenin expression patterns; (4) definitive validation overexpression knockdown experiments. Results Genetic revealed exhibited complete cosegregation phenotype pedigree while being absent control populations (gnomAD frequency: 0.000003989), accompanied significantly reduced tissues. Functional characterization demonstrated that compared to wild-type, mutant conferred aggressive properties: enhanced proliferative capacity, impaired induction, markedly increased migratory potential. Molecular analyses promoted EMT activation nuclear accumulation subsequent upregulation mesenchymal markers. Crucially, siRNA-mediated phenocopied these effects, providing evidence tumor-suppressive function. Discussion findings establish mutation drives progression novel mechanism involving degradation, translocation, consequent activation. These results position critical nodal regulator Wnt/β-catenin signaling suggest may serve valuable biomarkers for hereditary risk assessment. Therapeutically, highlight potential utility Wnt inhibitors managing GPC5-mutant cases, also framework future investigations into glypican members biology.

Language: Английский

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