Crane, Hannah 
ORCID: 0000-0003-1129-0413, Carr, Ian, Hunter, Keith D ORCID: 0000-0002-7873-0877 and El-Khamisy, Sherif F
(2024)
Senataxin modulates resistance to cisplatin through an R-loop mediated mechanism in HPV-associated Head and Neck Squamous Cell Carcinoma.
[Preprint]
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Abstract
<jats:title>Abstract</jats:title><jats:sec><jats:title>Introduction</jats:title><jats:p>Oropharyngeal Squamous Cell Carcinoma (OPSCC) is a site defined subtype of head and neck cancer with two distinct clinical subtypes: HPV-associated (HPV+) and HPV-independent (HPV-); both of which are commonly treated with chemoradiotherapy involving cisplatin. Cisplatin creates DNA crosslinks, which lead to eventual cell death via apoptosis. Clinical outcomes in HPV-OPSCC are poor and although HPV+ has an improved response to therapy, a subset of patients suffer from distant metastases, with a poor prognosis. Therefore, there is a need to understand the molecular basis underlying treatment resistance. A common mechanism of chemotherapy resistance is upregulation of DNA repair, and a major source of endogenous DNA damage are DNA/RNA hybrids, known as R-loops. R-loops are three stranded DNA/RNA hybrids formed in the genome as a by- product of transcription and are normally transient; however, they can persist and become a source of genomic instability. The contribution of R-loops to the development of cisplatin resistance in OPSCC is unknown.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>HPV+ and HPV- cisplatin resistant cell lines were developed, and RNA-sequencing was used to investigate changes in gene expression. Changes in R-loop dynamics were explored using slot blots and DRIP-qPCR. The effect of depleting known R-loop regulators on cisplatin sensitivity was assessed using siRNA. R-loop burden in a cohort of HPV+ and HPV- OPSCC tumours was explored using S9.6 immunohistochemistry.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Development of cisplatin resistant clones led to changes in gene expression consistent with resistance, alongside alterations in the expression of known R-loop regulators. Both HPV+ and HPV- resistant cells had elevated global R-loop levels and in HPV+ resistant cells there was a corresponding upregulation of the R-loop resolving protein, senataxin, which was not observed in HPV- resistant cells. Depletion of senataxin led to increased sensitivity to cisplatin in both HPV+ and HPV- resistant cells, however, the effect was greater in HPV+ cells. Quantification of R-loop levels by S9.6 immunohistochemistry revealed that HPV+ tumours and tumours with bone metastases had a higher R-loop burden.</jats:p></jats:sec><jats:sec><jats:title>Conclusion</jats:title><jats:p>R-loops are involved in modulating sensitivity to cisplatin and may represent a potential therapeutic target.</jats:p></jats:sec>
| Item Type: | Preprint |
|---|---|
| Uncontrolled Keywords: | Biotechnology, Sexually Transmitted Infections, Rare Diseases, Digestive Diseases, Infectious Diseases, Genetics, Dental/Oral and Craniofacial Disease, Cancer, 5 Development of treatments and therapeutic interventions, 5.1 Pharmaceuticals, 2 Aetiology, 2.1 Biological and endogenous factors, Cancer |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 18 Mar 2024 09:50 |
| Last Modified: | 18 Mar 2024 09:50 |
| DOI: | 10.1101/2024.02.22.581374 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3179621 |
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