Department of Otolaryngology-Head and Neck Surgery, University of Western Ontario, London, Ontario, Canada.
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.
Oral Oncol. 2023 Nov;146:106580. doi: 10.1016/j.oraloncology.2023.106580. Epub 2023 Sep 30.
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.
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.
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.
Taken together, our work implicates LRP1B deletion as a potential biomarker for identifying treatment resistant HPV+ OPSCC cases.
尽管人乳头瘤病毒阳性（HPV+）口咽鳞状细胞癌（OPSCC）患者的生存预后通常较好，但仍有 15-20%的患者在接受化疗和放疗后会复发。因此，需要寻找治疗失败的生物标志物来指导治疗强度。
对主要接受同期化疗（顺铂）和放疗的 HPV+OPSCC 患者进行全基因组测序。然后，我们探索了 LRP1B 的缺失是否足以驱动侵袭性表型，并在 HPV+细胞系（93VU147T、UMSCC47、UWO37 和 UWO23）和体内中促进对顺铂和放疗的耐药性。
通过对三个 HPV+OPSCC 肿瘤数据集的综合基因组分析，我们发现 LRP1B 的缺失在接受化疗放疗后复发的样本中富集。在四种 HPV+细胞系（UWO23、UWO37、UMSCC47 和 93VU147T）中使用 siRNA 敲低导致所有情况下的增殖增加。在相同的细胞系面板中使用 CRISPR/Cas9 缺失 LRP1B 导致缺失细胞的增殖、集落形成生长和迁移增加，以及对顺铂和放疗的耐药性增强，与各自的非靶向对照细胞相比。细胞系衍生的异种移植研究表明，与对照相比，LRP1B 敲除肿瘤在体内对顺铂和放疗治疗更具抵抗力。
综上所述，我们的工作表明 LRP1B 缺失可能是识别治疗耐药性 HPV+ OPSCC 病例的潜在生物标志物。
