Department of Immunology, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA, 02115, USA.
ICCB-Longwood Screening Facility, Harvard Medical School, 250 Longwood Avenue, Boston, MA, 02115, USA.
Virology. 2021 Aug;560:96-109. doi: 10.1016/j.virol.2021.05.006. Epub 2021 May 22.
Approximately 5% of cancers are caused by high-risk human papillomaviruses. Although very effective preventive vaccines will reduce this cancer burden significantly over the next several decades, they have no therapeutic effect for those already infected and remaining at risk for malignant progression of hrHPV lesions. HPV-associated cancers are dependent upon the expression of the viral E6 and E7 oncogenes. The oncogenic function of hrHPV E6 relies partially on its ability to induce p53 degradation. Since p53 is generally wildtype in hrHPV-associated cancers, p53 stabilization arrests proliferation, induces apoptosis and/or results in senescence. Here we describe a live cell, image-based high-throughput screen to identify compounds that stabilize p53 and/or affect viability in HPV-positive cancer HeLa cells. We validate the robustness and potential of this screening assay by assessing the activities of approximately 6,500 known bioactive compounds, illustrating its capability to function as a platform to identify novel therapeutics for hrHPV.
大约 5%的癌症是由高危型人乳头瘤病毒（HPV）引起的。尽管非常有效的预防性疫苗将在未来几十年内显著降低这种癌症负担，但对于已经感染并仍然存在 HPV 病变恶性进展风险的人来说，它们没有治疗作用。HPV 相关癌症依赖于病毒 E6 和 E7 癌基因的表达。hrHPV E6 的致癌功能部分依赖于其诱导 p53 降解的能力。由于 p53 在 hrHPV 相关癌症中通常为野生型，因此 p53 稳定会阻止增殖、诱导细胞凋亡和/或导致衰老。在这里，我们描述了一种基于活细胞成像的高通量筛选方法，以鉴定稳定 p53 并影响 HPV 阳性癌症 HeLa 细胞活力的化合物。我们通过评估大约 6500 种已知生物活性化合物的活性来验证该筛选测定的稳健性和潜力，这表明它有能力作为一种平台，为 hrHPV 识别新的治疗方法。
