Abstract 3715: Disrupting the effects of KRAS in colorectal cancer: Investigation of protein pathways linking KRAS and PRMT5

Introduction: Numerous therapeutic methods have been explored to inhibit the oncogenic effects of mutant KRAS. Previous in-vitro and in-silico research in our lab indicated that PRMT5 (Protein arginine methyltransferase), a well studied transcription regulator, is a possible surrogate target through which the negative effects of mutant KRAS can be mediated. Our previous preliminary in-silico data developed potential protein pathways linking KRAS and PRMT5 utilizing digital protein models and molecular dynamics simulations. Our current study expands on the preliminary molecular dynamic simulation results, enhancing the understanding of structural and kinetic components of the proteins’ interactions and the strength of the theorized pathways. Methodology: In-silico: The STRING database was used to determine two complementary pathways connecting KRAS and PRMT5. AlphaFold protein structure files downloaded from UniProt were docked with the ClusPro server. Using PyMOL, proteins were separated by chain identities. The Molecular Dynamics Simulation software GROMACS was used to simulate interactions between proteins in a computational topology regulated by Newton’s Laws. GROMACS, gmx_MMPBSA, and XmGrace softwares were used to analyze kinetic and structural components. In-vitro: 4 cell lines, HCT116 and SW620 (KRAS mutant), and Hke3 and LIM420 (KRAS wild-type), were used in this study. Cells were treated with the PRMT5 inhibitor, GSK, for either 6, 9, 24, or 48 hours. Subsequently, quantitative polymerase chain reaction (qPCR) was performed to determine mRNA expression of (HRAS, SNRPE, CLNS1A, WDR77). The difference in fold change between untreated and treated was calculated at each time point. Results: The proteins HRAS, SNRPE, CLNS1A, and WDR77 were found to be significant interactors between KRAS and PRMT5, forming two possible pathways abbreviated: K-H-S-C-P and K-H-S-W-P. RMSD data confirmed complex stability. RMSF data of each protein per complex displayed regions of significant interaction. qPCR data indicated altered expression of pathway mRNA in mutant cells as compared to in wild-type. Conclusion: Targeting the crosstalk between KRAS and PRMT5 may be a suitable method of inhibiting mutant KRAS’ oncogenic effects. Inhibition of PRMT5 has confirmed distinct retroactive effects on the pathway proteins’ gene expression in mutant vs. wild-type cell lines. Therapeutic advances can be made against KRAS using this surrogate target approach with detailed information of pathway interactions. Citation Format: Isaac Silverman, Aaron Shaykevich, Netanel Louloueian, Danbee Chae, Sanjay Goel, Radhashree Maitra. Disrupting the effects of KRAS in colorectal cancer: Investigation of protein pathways linking KRAS and PRMT5 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3715.