Defining the molecular tolerance-to-damage landscape of SMARCA4 helicase genetic alterations.

Evaluating the impact of genomic variation is essential for identifying underlying mechanistic causes of human diseases. The spectrum of neurodevelopmental disorders is driven by diverse genetic alterations with genes like SMARCA4 being prototypical examples. There have been significant hurdles to implementing the protein-specific and mechanism-informed variation effect predictors that are anticipated to have the highest yield of mechanistic information. Yet, there is a pressing need, for example, within SMARCA4 where 98% of the 2780 reported variants lack a disposition and remain of uncertain significance (VUS). Further, the field has yet to identify each variant's specific molecular mechanism, which will inform targeted therapeutic development strategies. In this study we developed a mechanistic structure-informed helicase-specific variant effect predictor by leveraging diverse information with state-specific calculations. Our approach has 100% recall of pathogenic variants while classifying 87.23% of VUS into damaging (55.74%, n=262) versus tolerated effects (31.49%, n=148), including those with conflicting interpretations. This analysis reveals significant enrichment of integrated functional metrics, such as conservation and solvent exposure, that parallel allele frequences in health populations, and emphasizes the robustness of the method. Thus, we have demonstrated a novel approach for the development of mechanism-informed protein-specific interpretation of human genetic information.