Molecular Mechanisms of CLCN5 Missense Mutations in Dent Disease Type 1: A Comprehensive Computational Analysis and Clinical Correlations in a Chinese Cohort

Dent's disease, an X‐linked recessive disorder predominantly affecting males, is characterized by nephrocalcinosis, nephrolithiasis, and a high risk of progression to end‐stage renal disease. Dent's disease type 1, accounting for 60% of cases, caused by mutations in the CLCN5 gene encoding the chloride ion channel protein ClC‐5, exhibits significant clinical heterogeneity and variability in disease progression. The lack of hotspot mutations poses challenges for genetic diagnosis and counselling, complicating the prediction of disease outcomes. This study systematically evaluated the functional and structural impacts of 181 CLCN5 missense mutations using computational tools, including PredictSNP, MAGPIE, and molecular dynamics simulations, to propose a robust method for improving genetic counselling and prognosis prediction. Our analysis identified mutations at the dimer interface and chloride selectivity filter as critical disruptors of ClC‐5 function and stability. Notably, molecular dynamics simulations of L200R, P213L, and G512R mutations revealed that L200R significantly destabilized the protein structure. Clinical data from a multicentre cohort of Chinese patients with CLCN5 mutations corroborated our computational predictions, highlighting the essential role of helix O in ClC‐5 function. By integrating bioinformatics analyses with clinical validation, this study provides molecular insights into Dent's disease heterogeneity and proposes a framework for enhancing genetic counselling and prognostic assessment for affected patients.