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

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.

Grant Number: 5R01HL159712-04
NIH Institute/Center: NIH
Principal Investigator: Ya-Wen Chen