Introduction/Background: Genetic variants beyond FDA-approved drug targets are often identified in non-small cell lung cancer (NSCLC) patients. Although the performances of in silico tools in predicting variant pathogenicity have been analyzed in previous studies, they have not been analyzed for actionable targets of FDA-approved therapies for NSCLC. The aim of this study is to compare the performance of commonly used in silico tools in classifying the pathogenicity of actionable variants in NSCLC. Materials and Methods: We evaluated the performance of the following in silico tools: Polyphen-2 (HumDiv, HumVar), Align-GVGD, MutationTaster2021, CADD, CONDEL, and REVEL. A curated set of targetable NSCLC missense variants (n=236) was used. The overall accuracy, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and Matthews correlation coefficient (MCC) of each in silico tool was determined. Results: The most recently released MutationTaster2021 demonstrated the highest performance in terms of accuracy, specificity, PPV, and MCC, but was outperformed by CADD for both sensitivity and NPV. Although some tools demonstrated high sensitivities, all tools except MutationTaster2021 displayed markedly low overall specificities, as low as 23%. Conclusion: The collective results indicate that the evaluated in silico tools can provide guidance in predicting the pathogenicity of NSCLC missense variants, but are not fully reliable. The tools analyzed in this study could be acceptable to rule out pathogenicity in variants given their higher sensitivities, but are limited when it comes to identifying pathogenicity in variants due to low specificities.

The authors have declared no competing interest.

This study did not receive any funding.

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All data produced in the present study are available upon reasonable request to the authors.