Abstract 5114: Deciphering the implications of THOC7 in medulloblastoma: Unraveling its role in tumor growth and progression

Abstract

Abstract Background: Medulloblastoma (MB) is the most common malignant brain cancer in children. Despite the progress made in treating MB, the 5-year survival rate for high-risk tumors remains poor and risk of recurrence within 2 years of treatment is still high. Unfortunately, patients who do survive have reduced quality of life because of the highly toxic side effects of radiation and chemotherapy. Those facts underline the importance of identifying new drivers of MB and understanding the mechanism by which those drivers may promote MB. Using CRISPR-based dependency analysis, we discovered THO Complex Subunit 7 Homolog (THOC7) as one of the highly differentially expressed genes that may uniquely provide survival advantage to MB cells. THOC7 is a component of the THO subcomplex of the TREX complex, which is known to couple mRNA transcription, processing and nuclear export associated with spliced mRNA. Methods: To understand the role of THOC7 in MB, we silenced the expression of THOC7 using siRNAs in multiple MB cell lines. The knockdown efficiency was confirmed by western blot and qPCR. The effect of THOC7 silencing on the viability of MB cells was assessed by short-term viability and long-term colony formation assays. The effect of THOC7 silencing on MB cell proliferation was measured using an IncuCyte. To assess whether THOC7 may affect MB progression, we performed Transwell migration assay. To understand the mechanism by which THOC7 may play a role in medulloblastomogenesis, we measured the effect of THOC7 silencing on the cell cycle progression and apoptosis. As presence of cancer stem cells is linked to relapse and makes MB refractory to radiation therapy, we asked whether THOC7 may support MB stem cell proliferation using neurosphere assay. Results: Our analysis revealed that MB is highly addicted to THOC7. Importantly, higher expression of THOC7 is strongly correlated with lower overall survival of MB patients. We showed that depletion of THOC7 inhibited the short- and long-term viability as well as proliferation of MB cells. In addition, silencing of THOC7 inhibited migration/invasion of MB cells. THOC7 depletion led to cell cycle arrest and increased apoptosis of MB cells. Furthermore, our neurosphere assay demonstrated that silencing of THOC7 blocked the MB stem cell proliferation. Conclusion: Our results suggest that THOC7 may be a novel driver of MB growth and progression. Inhibition of MB stem cell proliferation in THOC7 depleted cells indicates that THOC7 inhibition may sensitize refractory MB to radiation therapy. Experiments are currently underway to understand the mechanism by which THOC7 may support MB growth and progression. In conclusion, our results showing no effect of THOC7 silencing on normal cell viability suggest that approaches aimed at silencing THOC7 may be a safe and viable approach for treating MB. Citation Format: Shahad M. Abdulsahib, Santosh Timilsina, Saif Nirzhor, Daisy Medina, Prabhakar V. Pitta, Panneerdoss Subbarayalu, Manjeet K. Rao. Deciphering the implications of THOC7 in medulloblastoma: Unraveling its role in tumor growth and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5114.