Covid-19: Fast-tracking treatment by exploiting the steroid hormone receptor/TMPRSS2 axis

Covid-19：利用类固醇激素受体/TMPRSS2 轴进行快速治疗

• 批准号: 10341159
• 负责人: Kerry L Burnstein
• 金额: --
• 依托单位: MIAMI VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10341159
• 关键词: 2019-nCoV; ACE2; Acute; Acute Respiratory Distress Syndrome; Address; Adrenal Cortex Hormones; Affect; Agonist; Androgen Antagonists; Androgen Receptor; Antiandrogen Therapy; Binding; COVID-19; COVID-19 mortality; COVID-19 patient; COVID-19 treatment; Cell Line; Cell surface; Clinical Trials; Collaborations; Colorado; Communities; Consensus Sequence; DNA; Data; Diabetes Mellitus; Disease; Drug usage; Elderly; Enrollment; Epithelial; Epithelial Cells; Etiology; FDA approved; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Health; Health Personnel; Heart Diseases; Hormonal; Human; Hydrocortisone; Hypertension; Individual; Interdisciplinary Study; Interleukin-6; Ligands; Link; Luciferases; Lung; Lung Adenocarcinoma; Malignant neoplasm of prostate; Measures; Modeling; Nuclear Receptors; Outcome; Patients; Pattern; Pharmaceutical Preparations; Pharmacological Treatment; Prostate Cancer therapy; Proteins; Proteolysis; Publishing; Receptor Inhibition; Regulation; Reporter; Reproducibility; Research; Resistance; Response Elements; SARS-CoV-2 entry inhibitor; SARS-CoV-2 infection; SARS-CoV-2 spike protein; Serine Protease; Smoking; Stanolone; System; TMPRSS2 gene; Testing; Therapeutic; Universities; Veterans; Viral; Virus; Woman; World Health Organization; advanced prostate cancer; airway epithelium; antagonist; biosafety level 3 facility; castration resistant prostate cancer; chronic inflammatory disease; comorbidity; cytokine; demographics; drug repurposing; glucocorticoid receptor alpha; high risk; human model; loved ones; mRNA Expression; male; malignant breast neoplasm; men; post SARS-CoV-2 infection; protein expression; receptor upregulation; response; severe COVID-19; steroid hormone receptor; therapeutic evaluation; transcriptome; viral entry inhibitor

COVID-19 poses a tremendous health threat, particularly to individuals overrepresented in the US Veteran community. COVID-19 mortality is greater in men than in women; while this disparity is at least partially due to factors such as higher rates of smoking, a hormonal link is likely and can be rapidly tested therapeutically by repurposing existing drugs. The viral etiologic agent of COVID-19, SARS-CoV-2 (CoV-2), attaches to human airway epithelium via the viral spike (S) protein, which binds to angiotensin-converting enzyme 2 (ACE2) on the host cell surface. Viral entry requires S protein cleavage by the serine protease TMPRSS2, which is a known transcriptional target of the androgen receptor (AR). Lung epithelial cells (a target of CoV-2 infection) express transcriptionally active AR. We hypothesize that AR up-regulates TMPRSS2 in lung epithelial cells and thereby promotes viral entry and infectivity. We propose that FDA-approved AR antagonists will decrease CoV-2 entry and spread and can be rapidly repurposed for COVID-19. IL-6 is the major cytokine released in moderate and severe COVID-19 cases and both published and our preliminary data show that IL-6 enhances AR transcriptional activity. We will therefore also examine the contribution of interleukin 6 (IL-6) to AR regulation of TMPRRS2. To facilitate these studies in a robust manner, we propose to isolate the SARS-CoV-2 entry mechanism through the use of luciferase-expressing pseudovirions that harbor the SARS-CoV-2 S protein. Such a reporter system has high reproducibility, versatility and dynamic range, allowing for the rapid, accurate and specific assessment of a large range of viral entry regulators into primary human lung epithelial cells and lung adenocarcinoma cell lines under BSL2+ conditions. We will test whether AR inhibition reduces TMPRSS2 and the requisite S protein processing thereby decreasing CoV-2 entry into host lung epithelial cells. Subsequently, we will confirm results on a subset of promising compounds using live SARS-Cov-2 in an approved BSL3 facility. Safe and effective AR antagonists are FDA approved for prostate cancer and this study will provide rationale to repurpose these drugs for use in clinical trials for COVID-19. The glucocorticoid receptor (GR) shares a common DNA response element consensus sequence with AR. Furthermore, GR upregulation of TMPRSS2 has been shown in advanced prostate cancer. Therefore, in parallel, we will examine whether TMPRSS2 is regulated by glucocorticoids (cortisol) and blocked by a GR antagonist in models of human lung epithelia. Patients taking corticosteroids (including the elderly and individuals with diabetes, hypertension and chronic inflammatory disease) are at the highest risk of death from COVID-19. The World Health Organization has provided interim guidance to avoid glucocorticoids in COVID- 19 patients with severe acute respiratory distress syndrome. Therefore, understanding GR regulation of TMPRSS2 is also essential to repurposing the TMPRSS2-inhibitory FDA-approved agents for COVID-19. Our aims are to: (1) Evaluate steroid hormone receptor (AR and GR) regulation of TMPRSS2 in human primary airway and lung epithelial cells and lung adenocarcinoma cell line models and (2) Examine the capacity of FDA-approved AR and GR antagonists to block CoV-2 entry and infectivity in human primary airway and lung epithelial cells. US Veterans represent several demographics acutely afflicted by COVID-19. Older US Veterans are particularly vulnerable because of higher comorbidities including smoking, diabetes, heart disease and hypertension. Burden on the Veteran community is also proportionally higher given the propensity for poor outcome in men as compared to women following COVID-19 infection. Since the anti-androgen therapies to be tested are FDA approved for prostate cancer treatment and have also been used safely in women with breast cancer, our study has potential for immediate impact to all veterans.

新冠肺炎构成了巨大的健康威胁，特别是对在美国任职人数过多的个人 老兵社区。新冠肺炎的死亡率男性高于女性，而这一差距至少是 部分原因是吸烟率较高等因素，荷尔蒙的联系很可能存在，而且可以快速检测出来。 通过改变现有药物的用途来治疗。新冠肺炎的病毒病原体是SARS-CoV-2(冠状病毒2)， 通过病毒尖峰蛋白(S)附着于人的呼吸道上皮，该蛋白与血管紧张素转换相结合 宿主细胞表面的酶2(ACE2)。病毒的进入需要丝氨酸蛋白酶对S蛋白的切割 TMPRSS2，它是雄激素受体(AR)的已知转录靶点。肺上皮细胞(靶标 CoV-2感染)表达转录活性AR。我们假设AR上调TMPRSS2在 肺上皮细胞，从而促进病毒进入和传染性。我们建议FDA批准 AR拮抗剂将减少CoV-2的进入和传播，并可迅速被重新用于新冠肺炎。IL-6 在中重度新冠肺炎病例中释放的主要细胞因子是已发表和我们的 初步数据显示，IL-6增强AR转录活性。因此，我们还将研究 IL-6在TMPRRS2 AR调节中的作用为了以强有力的方式促进这些研究， 我们建议通过表达荧光素酶来分离SARS-CoV-2的入侵机制。 含有SARS-CoV-2 S蛋白的假病毒粒子。这样的报告系统具有很高的重现性， 多功能性和动态范围，允许快速、准确和具体地评估大范围的病毒 BSL2+对原代人肺上皮细胞和肺腺癌细胞株的进入调节作用 条件。我们将测试AR抑制是否会减少TMPRSS2和必要的S蛋白质加工 从而减少CoV-2进入宿主肺上皮细胞。随后，我们将确认一项 在批准的BSL3设施中使用活的SARS-CoV-2的有希望的化合物的子集。安全有效的AR FDA批准了拮抗剂用于前列腺癌，这项研究将为这些药物的再利用提供理论依据 用于新冠肺炎临床试验的药物。 糖皮质激素受体(GR)与GR具有共同的DNA反应元件序列 阿。此外，在晚期前列腺癌中，TMPRSS2的表达也有上调。因此，在 同时，我们将研究TMPRSS2是否受糖皮质激素(皮质醇)调节并被GR阻断 人肺上皮细胞模型中的拮抗剂。服用皮质类固醇的患者(包括老年人和 患有糖尿病、高血压和慢性炎症性疾病的人)死于 新冠肺炎。世界卫生组织提供了临时指南，以避免COVID中的糖皮质激素。 严重急性呼吸窘迫综合征19例。因此，理解GR对 TMPRSS2对于改变FDA批准的用于新冠肺炎的TMPRSS2抑制药的用途也是必不可少的。我们的 目的：(1)评价TMPRSS2对人原代细胞类固醇激素受体(AR和GR)的调节作用 建立呼吸道和肺上皮细胞及肺腺癌细胞系模型；(2)检测 FDA批准的AR和GR拮抗剂用于阻断CoV-2进入人体主呼吸道和肺的传染性 上皮细胞。 美国退伍军人代表了几个深受新冠肺炎影响的人群。年长的美国退伍军人 特别脆弱，因为吸烟、糖尿病、心脏病和 高血压。考虑到穷人的倾向，退伍军人社区的负担也成比例地更高 新冠肺炎感染后男性与女性的预后比较。因为抗雄激素疗法是 FDA批准了用于前列腺癌治疗的测试，并已安全地用于乳房女性 癌症，我们的研究有可能对所有退伍军人产生立竿见影的影响。