TMPRSS2 as a potential target for treatments of COVID-19 and respiratory infectious viruses in lung

TMPRSS2 作为治疗 COVID-19 和肺部呼吸道感染病毒的潜在靶点

• 批准号: 10454033
• 负责人: Ya-Wen Chen
• 金额: $ 70.55万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10454033
• 关键词: 2019-nCoV; 3-Dimensional; ACE2; Address; Age; Air; Animal Model; Antibodies; Apoptosis; Binding; Binding Proteins; COVID-19; COVID-19 impact; COVID-19 susceptibility; COVID-19 treatment; Case Study; Cell Line; Cell Survival; Cell surface; Cells; Cessation of life; Clinical; Complex; DNA Sequence Alteration; Data; Disease; Distal; Endocytosis; Ensure; Environment; Epithelial Cells; Family; Goals; Hamsters; Human; Human Characteristics; Immunologics; In Vitro; Individual; Infection; Inflammation; Influenza A virus; Lead; Liquid substance; Lung; Mediating; Mesocricetus auratus; Modeling; Molecular; Monoclonal Antibodies; Mus; Mutate; Organoids; Passive Immunization; Pathogenesis; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Population; Process; Production; Proteins; Pulmonary Pathology; RNA Viruses; Regulation; Resistance; Role; SARS coronavirus; SARS-CoV-2 entry inhibitor; SARS-CoV-2 infection; SARS-CoV-2 inhibitor; SARS-CoV-2 spike protein; Scientist; Serine Proteinase Inhibitors; Serpins; Structure; Sum; System; TMPRSS2 gene; Testing; Time; Tissues; Transgenic Organisms; Vaccines; Variant; Viral; Virus; Virus Diseases; Work; airway epithelium; alveolar type II cell; cell type; cytotoxicity; dosage; drug candidate; efficacy evaluation; efficacy testing; human disease; human pluripotent stem cell; in vivo; individual patient; infection rate; inhibiting antibody; inhibitor; insight; lung injury; neutralizing antibody; novel; novel therapeutic intervention; novel therapeutics; pandemic disease; prevent; receptor; respiratory; respiratory virus; response; stem cell differentiation; stem cells; therapeutic candidate; transmission process; trend; unvaccinated; vaccine hesitancy; vaccine-induced immunity; viral entry inhibitor

Project Summary In early 2020, a new virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), generated headlines due to its unprecedented rate of transmission. SARS-CoV-2 caused the first reported cases of coronavirus disease 2019 (COVID-19) in December 2019 and continues to spread worldwide. As a family of RNA viruses, SARS-CoV-2 is prone to mutate at a rate up to a million times faster than its hosts1,2. These rapid genomic alterations have already generated highly transmissible variants, and have raised concerns that the virus will evade vaccine-induced immunity. In addition, a large percentage of the global population remains unvaccinated, due to the challenges of production and mass distribution, vaccine hesitancy, and pending approval status for patients under age 12. Therefore, an effective antiviral has the potential to relieve suffering for millions—not only helping individual patients recover and reducing the number of deaths, but also limiting the number of positive carriers and thereby curbing the spread of the pandemic. This proposal aims to develop an efficient antiviral to impede the virus’ entry into cells, specifically into lung alveolar type II (AT2) cells, the stem cells of the distal lung. Thanks to recent studies, we know which “door” (a receptor called ACE2) and “key” (a protease called TMPRSS2) the virus uses to enter cells. Our goal is to remove the key so the virus cannot open the door and enter host cells. We will use a conventional air-liquid interface (ALI) culture that is representative of the in vivo airway and a recently developed 3-dimensional (3D) in vitro lung organoid model that recapitulates many aspects of lung structure and the cellular environment and that has been used to study respiratory viruses, including SARS-CoV-2. These systems represent tissues better than cell lines, but offers the benefit of being less complex than tissue explants or animal models. In addition, we have generated a panel of highly sensitive and specific mouse monoclonal antibodies (mAbs) directed against TMPRSS2. In preliminary studies, the lead TMPRSS2 mAb, AL20, shows no signs of cytotoxicity with a trend towards inhibition of SARS-CoV-2 pseudovirus entry in cell lines and in lung organoids. Furthermore, we have identified at least two serine protease inhibitors (serpins) that form complexes with TMPRSS2, and the presence of these complexes is inversely correlated with the SARS-CoV-2 infection rate. These findings lead to our hypothesis that targeting TMPRSS2 can inhibit SARS-CoV-2 viral entry and spread. To test our hypothesis, we will first test the efficacy of AL20 for blocking the entry of SARS-CoV-2 into AT2 cells in lung organoids and in airway epithelial cells in ALI cultures, and elucidate the underlying mechanisms. We will then evaluate the effects of serpins on TMPRSS2 activity and SARS-CoV-2 viral entry and spread. Finally, to explore the feasibility of advancing AL20 to human trials, we will test humanized AL20 in a SARS-CoV-2 hamster model. Syrian golden hamsters are naturally susceptible to SARS-CoV-2 infection that recapitulates the clinical, virological, histopathological, and immunological characteristics of human disease, enabling study of its pathogenesis, transmission, and passive immunization effect. Transgenic human ACE2 is not required for SARS-CoV-2 infection, ensuring that the cell types infected are highly relevant. These studies will provide critical insights into the mechanisms whereby TMPRSS2 regulates SARS-CoV-2 entry, and suggest potential therapeutic candidates against COVID-19. The proposed work has the potential to impact the lives of millions of individuals affected by COVID-19 and other respiratory viruses, such as influenza A, that use TMPRSS2 to enter cells.

项目摘要 在2020年初，一种新的病毒，严重急性呼吸综合征冠状病毒2(SARS-CoV-2)，由于 其史无前例的传播速度。SARS-CoV-2导致2019年首例报告冠状病毒病病例(新冠肺炎) 2019年12月，并继续在全球范围内传播。SARS-CoV-2作为RNA病毒家族，极易发生变异 比它的宿主快一百万倍1，2。这些快速的基因组改变已经产生了高度可传播性 变种，并引起了人们对该病毒将逃避疫苗诱导免疫的担忧。此外，有很大比例的 全球人口仍未接种疫苗，原因是生产和大规模分发的挑战、疫苗的迟疑、 以及12岁以下患者的等待批准状态。因此，有效的抗病毒药物有可能减轻痛苦 对数百万人来说-不仅帮助个别患者康复和减少死亡人数，而且还限制了 控制阳性携带者，从而遏制大流行的蔓延。 这项建议旨在开发一种有效的抗病毒药物来阻止病毒进入细胞，特别是进入肺泡型 II(AT2)细胞，肺远端的干细胞。多亏了最近的研究，我们知道是哪扇门(一种名为ACE2的受体)和 病毒用来进入细胞的“Key”(一种名为TMPRSS2的蛋白酶)。我们的目标是移除密钥，这样病毒就无法打开 打开门，进入宿主牢房。我们将使用传统的气液界面(ALI)培养法来代表活体内的 气道和最近发展起来的三维(3D)体外肺器官模型，它概括了肺的许多方面 结构和细胞环境，并已被用于研究呼吸道病毒，包括SARS-CoV-2。这些 系统比细胞系更好地代表组织，但提供的好处是比组织外植体或动物更简单 模特们。此外，我们还产生了一组高度敏感和特异的鼠单抗(MAbbs) 针对TMPRSS2。在初步研究中，主要的TMPRSS2单抗AL20没有显示出细胞毒性的迹象，有趋势 旨在抑制SARS-CoV-2假病毒在细胞系和肺组织中的侵入。此外，我们已在 至少两个与TMPRSS2形成络合物的丝氨酸蛋白酶抑制剂(Serpins)，并且这些络合物的存在是 与SARS-CoV-2感染率呈负相关。这些发现导致了我们的假设，靶向TMPRSS2可以 抑制SARS-CoV-2病毒的进入和传播。 为了验证我们的假设，我们将首先测试AL20在阻止SARS-CoV-2进入肺AT2细胞方面的有效性 在ALI培养的呼吸道上皮细胞中，并阐明其潜在的机制。然后我们将评估 蛇毒对TMPRSS2活性及SARS-CoV-2病毒侵袭和传播的影响最后，探讨推进的可行性。 AL20到人体试验，我们将在SARS-CoV-2仓鼠模型上测试人源化AL20。叙利亚金黄仓鼠是天然的 易受SARS-CoV-2感染，这概括了临床、病毒学、组织病理学和免疫学 人类疾病的特点，使研究其发病机制、传播和被动免疫效果成为可能。 SARS-CoV-2感染不需要转基因人ACE2，从而确保感染的细胞类型高度相关。 这些研究将为TMPRSS2调节SARS-CoV-2进入的机制提供关键的见解，并建议 针对新冠肺炎的潜在治疗候选药物。这项拟议的工作有可能影响数百万人的生活 受新冠肺炎和其他呼吸道病毒(如甲型流感)影响的个人，使用TMPRSS2进入细胞。