Combining BET protein inhibitors with radiation in HPV oropharyngeal cancer

BET 蛋白抑制剂与放射治疗 HPV 口咽癌的联合治疗

• 批准号: 10578185
• 负责人: Gopal Iyer
• 金额: $ 21.19万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-08 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10578185
• 关键词: Address; Apoptosis; Blood; Bromodomain; Bromodomains and extra-terminal domain inhibitor; Cancer Etiology; Cell Cycle; Cell Line; Cells; Cessation of life; Characteristics; Chemicals; Cisplatin; Clinical Trials; Complete Blood Count; Country; DNA; DNA Crosslinking; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Disorder; DNA Repair Gene; Data; Deglutition Disorders; Dependence; Diagnosis; Dose; Double Strand Break Repair; Drug Targeting; Enteral Feeding; Family; Gene Expression; Generations; Genes; Genetic Transcription; Genomic Instability; Genomics; Goals; Growth; HPV oropharyngeal cancer; Human Papillomavirus; Immunohistochemistry; In Vitro; Incidence; Individual; Kinetics; Lead; Length; Low Dose Radiation; Malignant Epithelial Cell; Malignant neoplasm of cervix uteri; Measurement; Measures; Modification; Molecular; Oropharyngeal; Oropharyngeal Neoplasms; Oropharyngeal Squamous Cell Carcinoma; Output; Pathway interactions; Patient Selection; Patients; Pattern; Population; Proliferating; Proteins; Quality of life; RNA; Radiation; Radiation Dose Unit; Radiation therapy; Research; Series; Specificity; Subgroup; Surrogate Markers; Survival Analysis; Survival Rate; Testing; Therapeutic; Tissues; Toxic effect; Transcriptional Regulation; Treatment Protocols; Treatment-related toxicity; Tumor Cell Line; Tumor Volume; USP6 gene; United States; Viral; Viral Genes; Viral Genome; Viral Markers; Virus Integration; Woman; biomarker selection; cancer cell; comorbidity; design; efficacy testing; established cell line; experimental study; gene repression; human papilloma virus oropharyngeal squamous cell carcinoma; improved; in vivo; inhibitor; knock-down; malignant oropharynx neoplasm; member; men; mouse model; neoplastic cell; new therapeutic target; patient derived xenograft model; pleiotropism; preservation; programs; recruit; response; secondary endpoint; standard of care; targeted treatment; transcriptome sequencing; treatment strategy; tumor; tumor eradication; tumor growth; tumor progression; tumor xenograft; viral DNA

PROJECT SUMMARY/ABSTRACT: During the past decade, oropharyngeal cancer surpassed cervical cancer, becoming the most common cancer caused by human papillomavirus (HPV). Nearly 20,000 oropharyngeal cancer cases are diagnosed among men and women in the United States annually. Despite favorable long-term survival, current non-targeted cisplatin/radiation treatment protocols lead to significant treatment toxicities. De-escalating radiation doses with alternative targeted therapies would be critical for limiting treatment-related toxicities and improving patient quality of life. To address this, we propose to reduce the transcriptional output of viral genes and DNA damage response (DDR) by reducing its dependence on a transcriptional co-regulator bromodomain protein, BRD4, expressed in the oropharyngeal tissue. We propose that DDR deficiencies can be induced by second-generation bromodomain-specific chemical inhibitors in HPV oropharyngeal tumors. This chemically induced DDR deficient state delays the kinetics of DNA repair, thereby causing genomic instability. With tumors in this fragile state, administering optimal radiation doses will result in genomic catastrophe leading to eradicating tumors. We established that a first-generation inhibitor that targets both bromodomains BD1 and BD2 of BRD4 preferentially reduced viral and DDR gene expression of a subset of HPV tumors, which harbored disrupted viral genomes over non-disrupted viral genomes. However, pan-BD domain inhibition could give rise to pleiotropic effects. We used second-generation domain-specific inhibitors introduced in 2020 on HPV tumor cell lines to refine the pan- BD inhibition approach. We observed preferential domain-specific transcriptional regulation. While BD2 domain inhibition downregulated DDR response in disrupted viral tumors, BD1 upregulated anti-viral gene expression in non-disrupted viral tumors. These results guide us to postulate that patients selected upfront for disrupted viral genomes can be matched for BD domain-specific inhibitor and de-escalated radiation treatments. We will use in vitro and in vivo approaches to address a) efficacy of BD domain-specific inhibitors towards creating a DDR- deficient state by measuring viral gene expression and quantifying DDR response kinetics, and test its on-target efficacy in BRD4 knockdown and BD1 and BD2 domain-specific deleted tumor cells, b) Optimize the dose of radiation sufficiently to preserve the efficacy of BD domain-specific inhibition in patient-derived xenograft mice models with secondary endpoint analyses of survival, tumor volume growth, apoptosis, complete blood count panel for toxicity and immunohistochemistry for proliferation and DNA repair proteins.

项目总结/摘要： 在过去的十年中，口咽癌超过宫颈癌，成为最常见的癌症 人乳头瘤病毒（HPV）近20，000例口咽癌病例在男性中被诊断出来 和美国妇女每年。尽管长期生存率良好，但目前的非靶向 顺铂/放射治疗方案导致显著的治疗毒性。降低辐射剂量， 替代性靶向治疗对于限制治疗相关毒性和改善患者预后至关重要。 生活质量为了解决这个问题，我们建议减少病毒基因的转录输出和DNA损伤 通过减少其对转录共调节因子溴结构域蛋白BRD 4的依赖， 在口咽组织中表达。我们认为，DDR缺陷可以由第二代 溴结构域特异性化学抑制剂在HPV口咽肿瘤中的应用这种化学诱导的DDR缺陷 状态延迟DNA修复的动力学，从而导致基因组不稳定。对于这种脆弱状态的肿瘤， 给予最佳的放射剂量将导致基因组灾难，从而导致根除肿瘤。我们 确定了优先靶向BRD 4的溴结构域BD 1和BD 2的第一代抑制剂 降低了一部分HPV肿瘤的病毒和DDR基因表达，这些肿瘤含有破坏的病毒基因组 而不是未被破坏的病毒基因组。然而，pan-BD结构域抑制可引起多效性效应。我们 使用2020年在HPV肿瘤细胞系上引入的第二代结构域特异性抑制剂，以完善泛 BD抑制方法。我们观察到优先域特异性转录调控。而BD 2结构域 抑制在破坏的病毒肿瘤中下调DDR反应，BD 1在破坏的病毒肿瘤中上调抗病毒基因表达。 未破坏的病毒肿瘤。这些结果指导我们假设，预先选择破坏病毒的患者 基因组可以与BD结构域特异性抑制剂和逐步降低的放射治疗相匹配。我们将在 a）BD结构域特异性抑制剂对产生DDR-1的功效， 通过测量病毒基因表达和量化DDR反应动力学来检测缺陷状态，并测试其靶向 在BRD 4敲低和BD 1和BD 2结构域特异性缺失的肿瘤细胞中的功效，B）优化 足以保持BD结构域特异性抑制在患者来源的异种移植小鼠中的功效的辐射 模型，次要终点分析包括生存期、肿瘤体积生长、细胞凋亡、全血细胞计数 毒性和增殖及DNA修复蛋白免疫组化检测。