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: 10814125
• 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/10814125
• 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; Disparity; Drug usage; Elderly; Enrollment; Epithelial Cells; Epithelium; 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; Prostate Cancer therapy; Proteins; Proteolysis; Publishing; Receptor Inhibition; Receptor Up-Regulation; 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; 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.

COVID-19 构成巨大的健康威胁，尤其是对美国比例过高的个人 退伍军人社区。男性的 COVID-19 死亡率高于女性；而这个差距至少是 部分由于吸烟率较高等因素，荷尔蒙关联很可能存在，并且可以快速测试 通过重新利用现有药物进行治疗。 COVID-19、SARS-CoV-2 (CoV-2) 的病毒病原体、 通过病毒刺突 (S) 蛋白附着在人气道上皮上，该蛋白与血管紧张素转换蛋白结合 宿主细胞表面的酶 2 (ACE2)。病毒进入需要丝氨酸蛋白酶切割 S 蛋白 TMPRSS2，是雄激素受体 (AR) 的已知转录靶标。肺上皮细胞（目标 CoV-2 感染）表达转录活性 AR。我们假设 AR 上调 TMPRSS2 肺上皮细胞，从而促进病毒进入和感染。我们建议 FDA 批准 AR 拮抗剂将减少 CoV-2 的进入和传播，并可迅速重新用于治疗 COVID-19。白细胞介素6 是中度和重度 COVID-19 病例中释放的主要细胞因子，已发表和我们的 初步数据表明IL-6增强AR转录活性。因此，我们还将审查 白细胞介素 6 (IL-6) 对 TMPRRS2 的 AR 调节的贡献。为了以强有力的方式促进这些研究， 我们建议通过使用荧光素酶表达来分离 SARS-CoV-2 进入机制 含有 SARS-CoV-2 S 蛋白的假病毒颗粒。这样的报告系统具有很高的重现性， 多功能性和动态范围，允许对大范围的病毒进行快速、准确和具体的评估 BSL2+条件下进入原代人肺上皮细胞和肺腺癌细胞系的调节因子 状况。我们将测试 AR 抑制是否会减少 TMPRSS2 和必需的 S 蛋白加工 从而减少 CoV-2 进入宿主肺上皮细胞。随后，我们将确认结果 在经批准的 BSL3 设施中使用活 SARS-Cov-2 开发有前途的化合物的子集。安全有效的增强现实 拮抗剂已被 FDA 批准用于前列腺癌，这项研究将为重新调整这些拮抗剂的用途提供依据 用于 COVID-19 临床试验的药物。 糖皮质激素受体 (GR) 与以下物质共享一个共同的 DNA 反应元件共有序列： AR。此外，在晚期前列腺癌中，GR 上调了 TMPRSS2。因此，在 同时，我们将检查 TMPRSS2 是否受糖皮质激素（皮质醇）调节并被 GR 阻断 人肺上皮模型中的拮抗剂。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）的死亡风险最高 新冠肺炎。世界卫生组织已提供临时指南，以避免在新冠肺炎患者中使用糖皮质激素 19名严重急性呼吸窘迫综合征患者。因此，了解 GR 调节 TMPRSS2 对于重新利用 FDA 批准的 TMPRSS2 抑制剂治疗 COVID-19 也至关重要。我们的 目的是： (1) 评估类固醇激素受体（AR 和 GR）对人类原发性中 TMPRSS2 的调节 气道和肺上皮细胞和肺腺癌细胞系模型和（2）检查能力 FDA 批准的 AR 和 GR 拮抗剂可阻止 CoV-2 进入人类初级气道和肺部并降低其感染性 上皮细胞。 美国退伍军人代表了受新冠肺炎 (COVID-19) 严重影响的几个群体。年长的美国退伍军人是 由于吸烟、糖尿病、心脏病等较高的合并症而特别容易受到伤害 高血压。鉴于贫困人口的倾向，退伍军人社区的负担也成比例地更高 与女性感染 COVID-19 后的男性结果相比。由于抗雄激素疗法 经过测试，FDA 批准用于前列腺癌治疗，并且也已安全用于患有乳腺癌的女性 癌症，我们的研究有可能对所有退伍军人产生直接影响。