Loss of LRP1B expression drives acquired chemo and radio-resistance in HPV-positive head and neck cancer

Mushfiq H Shaikh; Alice Dawson; Stephenie D Prokopec; John W Barrett; Peter Y F Zeng; Mohammed I Khan; Sarah E B Ryan; Matthew Cecchini; David A Palma; Joe S Mymryk; Paul C Boutros; Anthony C Nichols

doi:10.1016/j.oraloncology.2023.106580

Loss of LRP1B expression drives acquired chemo and radio-resistance in HPV-positive head and neck cancer

Oral Oncol. 2023 Nov:146:106580. doi: 10.1016/j.oraloncology.2023.106580. Epub 2023 Sep 30.

Authors

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada.
² Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada; Department of Pathology and Laboratory Medicine, University of Western Ontario, London, Ontario, Canada.
³ Ontario Institute for Cancer Research, Toronto, Ontario, Canada.
⁴ Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada; Department of Oncology, University of Western Ontario, London, Ontario, Canada.
⁵ Department of Pathology and Laboratory Medicine, University of Western Ontario, London, Ontario, Canada.
⁶ Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada; Department of Oncology, University of Western Ontario, London, Ontario, Canada; Department of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada.
⁷ Department of Human Genetics, University of California, Los Angeles, CA, USA; Department of Urology, University of California, Los Angeles, CA, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA, USA; Institute for Precision Health, University of California, Los Angeles, CA, USA; Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, CA, USA.
⁸ Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada; Department of Oncology, University of Western Ontario, London, Ontario, Canada. Electronic address: anthony.nichols@lhsc.on.ca.

PMID: 37778229
DOI: 10.1016/j.oraloncology.2023.106580

Abstract

Objectives: Although human papillomavirus positive (HPV+) oropharyngeal squamous cell carcinoma (OPSCC) patients typically experience excellent survival, 15-20 % of patients recur after treatment with chemotherapy and radiation. Therefore, there is a need for biomarkers of treatment failure to guide treatment intensity.

Materials and methods: Whole genome sequencing was carried out on HPV+OPSCC patients who were primarily treated with concurrent chemotherapy (cisplatin) and radiation. We then explored whether the loss of LRP1Bwas sufficient to drive an aggressive phenotype, and promote a resistance to cisplatin and radiation therapy both in vitro using HPV+ cell lines (93VU147T, UMSCC47, UWO37 and UWO23) and in vivo.

Results: Through integrative genomic analysis of three HPV+OPSCC tumour datasets, we identified that deletion of LRP1B was enriched in samples that recurred following chemo-radiation. Knockdown using siRNA in four HPV+ cell lines (UWO23, UWO37, UMSCC47 and 93VU147T) resulted in increased proliferation of all cases. CRISPR/Cas9 deletion of LRP1B in the same cell line panel demonstrated increased proliferation, clonogenic growth and migration, as well as resistance to both cisplatin and radiation in LRP1B deleted cells compared to their respective non-targeting control cells. Cell line derived xenograft studies indicated that the LRP1B knockout tumours were more resistant to cisplatin and radiation therapy compared to their controls invivo.

Conclusion: Taken together, our work implicates LRP1B deletion as a potential biomarker for identifying treatment resistant HPV+ OPSCC cases.

Keywords: CRISPR/Cas9; Cell lines; Cisplatin; HPV; Head and neck cancer; OPSCC; Radiation.

MeSH terms

Carcinoma, Squamous Cell* / pathology
Cisplatin / pharmacology
Cisplatin / therapeutic use
Drug Resistance, Neoplasm*
Head and Neck Neoplasms* / drug therapy
Head and Neck Neoplasms* / genetics
Head and Neck Neoplasms* / radiotherapy
Humans
Neoplasm Recurrence, Local
Oropharyngeal Neoplasms*
Papillomavirus Infections* / complications
Papillomavirus Infections* / genetics
Papillomavirus Infections* / pathology
Radiation Tolerance*
Receptors, LDL / therapeutic use
Squamous Cell Carcinoma of Head and Neck / genetics
Squamous Cell Carcinoma of Head and Neck / radiotherapy

Substances

Cisplatin
LRP1B protein, human
Receptors, LDL