Immunogenicity and Structural Effects of INS Gene Mutations in Type 1 Diabetes Mellitus

Type 1 Diabetes (T1D) is an autoimmune disease driven by the destruction of insulin-producing beta cells. Genetic susceptibility includes variations in the *INS* gene, which encodes insulin. We hypothesized that T1D-associated *INS* mutations may alter insulin’s structure, affect its immunogenicity and contribute to autoimmunity. To investigate this, we conducted the first comprehensive structural analysis of all T1D-linked *INS* missense variants reported in ClinVar, using a multi-faceted computational approach. We curated all reported INS missense variants linked to T1D from ClinVar and analyzed their impact using Molecular Dynamics (MD) simulations. Leveraging the InterPro database, we mapped PR proinsulin’s domain architecture and assessed evolutionary conservation via multiple sequence alignment. MD simulations evaluated each mutation’s effect on insulin stability, using Root Mean Square Deviation (RMSD) for structural shifts and Root Mean Square Fluctuation (RMSF) for flexibility. K-means clustering grouped variants based on these metrics. Among 41 identified INS mutations, several occurred in highly conserved regions, suggesting functional significance. Variants such as T97S, A24V, P52R, L68M and G32S showed increased flexibility, with L68M displaying the highest RMSD, indicating structural destabilization. Based on MD data, we classified mutations as "Unstable," "Flexible," or "Stable." Our findings suggest that structural alterations caused by INS mutations may generate neoantigens, contributing to T1D autoimmunity. This classification provides insight into variant pathogenicity and highlights the importance of conserved regions for insulin function, with potential implications for diagnostics and therapeutics.