PRPF19 Inhibition Attenuates Non-Small Cell Lung Cancer Progression through Impairing Citron Kinase-controlled Cytokinesis and Enhancing Anti-tumor Immune Responses [Prpf19_KP-1_RNAseq]

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related death, primarily due to limited therapeutic targets and the emergence of drug resistance. In this study, we conducted CRISPR knockout screens to identify the pre-mRNA processing factor 19 (PRPF19) as a potential therapeutic target for NSCLC. Our findings revealed that PRPF19 is highly expressed in NSCLC, and its expression is negatively correlated with the clinical outcomes. Through functional validation, we confirmed that genetic ablation of PRPF19 significantly attenuated cancer progression both in vitro and in vivo. Mechanistically, we discovered that PRPF19 plays a crucial role in regulating the precise location of Citron Kinase (CIT) within the contractile ring, thereby sustaining the normal cytokinesis processes. Inhibition of PRPF19 resulted in cytokinesis failure, leading to micronuclei formation and activating the anti-tumor immune response through the TBK1-IFNs pathway. Taken together, our results demonstrate the critical involvement of the PRPF19-CIT axis in NSCLC progression and suggest that targeting PRPF19 represents a promising therapeutic approach for NSCLC patients. To investigate the signaling pathway regulated by PRPF19 in our model, we established KP-1-shPrpf19 stable cell lines utilizing a doxycycline (Dox)-inducible short hairpin RNA (shRNA) system. And then, we performed gene expression profiling analysis for control cell lines (KP-1-shLacZ) and it’s KD cell lines (KP-1-shPrpf19).