Abstract C106: Validated novel Immuno-therapeutic targets for lung cancer revealed by SpliceIOTM- Innovative platform that identifies disease-specific alternative splicing

Abstract Alternative splicing, long associated with the generation of novel disease-specific isoforms, is now increasingly recognized as a key driver of cancer progression. Envisagenics has developed a powerful software platform, SpliceIOTM, that harnesses RNA-seq data to identify tumor specific alternatively spliced isoforms with high therapeutic potential for Immuno oncology (IO) therapies. By accurately predicting tumor-specific prevalence, plasma membrane localization, epitope accessibility, and biological relevance, SpliceIO enables the discovery of novel, prevalent, and safe cell surface expressed targets for immunotherapeutic drug development. Lung cancers, especially non-small cell lung cancer with its high burden of alternative splicing, provides an ideal indication to discover tumor-specific isoforms that will serve as high-impact therapeutic targets. Using SpliceIO, Envisagenics has identified and experimentally validated multiple alternative splicing-derived targets in non-small cell lung cancer. The lead SpliceIO-identified candidates exhibit 25–30% prevalence in lung tumors with negligible expression in vital organs, which is confirmed by quantitative polymerase chain reaction (qPCR) across tumor and healthy tissues. RNA-seq analyses also demonstrate these candidates to have concomitant expression in additional solid tumor indications such as glioblastoma, head and neck and esophageal cancers. Further validation by RNA in situ hybridization (RNA-ISH) supports the specificity and intratumor homogeneity of our top targets. Additionally, localization to the plasma membrane has been confirmed by expressing isoforms as GFP-tagged constructs and analyzing transfected cells with flow cytometry and fluorescence microscopy. Evaluation of internalization propensity of lead isoforms by co-localization with subcellular compartment markers, highlights their suitability for antibody-drug conjugate (ADC) development. Isoform-specific antibodies targeting unique epitopes were generated and used to confirm lead candidate epitopes for specificity and selectivity. In summary, Envisagenics has established a robust discovery pipeline through SpliceIO to identify and validate tumor-specific epitopes, laying a strong foundation for clinical advancement of transformative IO therapeutics. Citation Format: Sakshi Gera, Miguel Manzanares, Kendall Anderson, Hasan Zumrat, Nicole Williams, Taylor Floyd, Sanjana Shah, Shaleigh Stanton, Gauri Ganesh, Devon Trahan, Martin Akerman, Gayatri Arun. Validated novel Immuno-therapeutic targets for lung cancer revealed by SpliceIOTM- Innovative platform that identifies disease-specific alternative splicing [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2025 Oct 22-26; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2025;24(10 Suppl):Abstract nr C106.