Related Experiment Video
Updated: Aug 12, 2025
Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
Transcriptomic analysis provides insight into the mechanism of IKKβ-mediated suppression of HPV18E6-induced cellular
Quincy P Collins1, Michael J Grunsted1, Dahiana Arcila1,2
1Department of Biology, University of Oklahoma, Norman, OK 73019, USA.
Abstract:
High-risk human papillomaviruses (HPVs) 16 and 18 are responsible for more than 70% of cervical cancers and majority of other HPV-associated cancers world-wide. Current treatments for these cancers have limited efficacy, which in turn has resulted in disease recurrence and poor survival rates in advanced disease stages. Hence, there is a significant need for development of novel molecularly-targeted therapeutics. This can only be achieved through improved understanding of disease mechanism. Recently, we developed a Drosophila model of HPV18E6 plus human E3 ubiquitin ligase (hUBE3A) and demonstrated that the E6-induced cellular abnormalities are conserved between humans and flies. Subsequently, we demonstrated that reduced level and activity of IKKβ, a regulator of NF-κB, suppresses the cellular abnormalities induced by E6 oncoprotein and that the interaction of IKKβ and E6 is conserved in human cells. In this study, we performed transcriptomic analysis to identify differentially expressed genes that play a role in IKKβ-mediated suppression of E6-induced defects. Transcriptome analysis identified 215 genes whose expression was altered due to reduced levels of IKKβ. Of these 215 genes, 151 genes showed annotations. These analyses were followed by functional genetic interaction screen using RNAi, overexpression, and mutant fly strains for identified genes. The screen identified several genes including genes involved in Hippo and Toll pathways as well as junctional complexes whose downregulation or upregulation resulted in alterations of E6-induced defects. Subsequently, RT-PCR analysis was performed for validation of altered gene expression level for a few representative genes. Our results indicate an involvement for Hippo and Toll pathways in IKKβ-mediated suppression of E6 + hUBE3A-induced cellular abnormalities. Therefore, this study enhances our understanding of the mechanisms underlying HPV-induced cancer and can potentially lead to identification of novel drug targets for cancers associated with HPV.
Insights
Researchers identified key genes and pathways, including Hippo and Toll, involved in suppressing human papillomavirus (HPV)-induced cervical cancer defects. This discovery offers potential new drug targets for HPV-associated cancers.
Area of Science:
- Oncology
- Virology
- Molecular Biology
Background:
- High-risk human papillomaviruses (HPVs) 16 and 18 cause over 70% of cervical cancers and other HPV-associated malignancies.
- Current treatments for HPV-associated cancers have limited efficacy, necessitating novel molecularly-targeted therapeutics.
- Understanding HPV disease mechanisms is crucial for developing new treatments.
Purpose of the Study:
- To identify genes and pathways involved in the suppression of HPV18E6 oncoprotein-induced cellular abnormalities.
- To investigate the role of IKKβ in mediating the suppression of E6-induced defects.
- To explore potential novel therapeutic targets for HPV-associated cancers.
Main Methods:
- Utilized a Drosophila model of HPV18E6 and human E3 ubiquitin ligase (hUBE3A).
- Performed transcriptomic analysis to identify differentially expressed genes upon reduced IKKβ levels.
- Conducted functional genetic interaction screens (RNAi, overexpression, mutant strains) and RT-PCR validation.
Main Results:
- Transcriptome analysis identified 215 differentially expressed genes, with 151 annotated.
- Genetic screens revealed involvement of Hippo and Toll pathways, and junctional complexes in modulating E6-induced defects.
- Confirmed the role of Hippo and Toll pathways in IKKβ-mediated suppression of E6 + hUBE3A-induced abnormalities.
Conclusions:
- The study elucidates the molecular mechanisms underlying HPV-induced cancer, highlighting the involvement of Hippo and Toll pathways.
- Identified IKKβ as a key regulator in suppressing HPV-induced cellular abnormalities.
- Provides a foundation for developing novel drug targets for HPV-associated cancers.
