Human coronaviruses activate Notch signaling to potentiate virus replication

Dr Chu, Hin   (Principal Investigator (PI))

Dr Shuai Huiping Vivian   (Co-Investigator)

Dr Chan Jasper Fuk Woo   (Co-Investigator)

Professor Yuen Kwok Yung   (Co-Investigator)

36

2022-01-01

1173062

Human coronaviruses activate Notch signaling to potentiate virus replication

human coronaviruses, MERS-CoV, Notch signaling, SARS-CoV, SARS-CoV-2

Virology,Infection/Parasitology

Biology and Medicine (M)

17118621

General Research Fund (GRF)

2021

On-going

1 Objective 1: To characterize the basic biology of Notching signaling in the context of SARS-CoV-2 infection. Extending from our pilot studies on the identification of Notch signaling as a novel pathway for efficient SARS-CoV-2 replication, we aim to thoroughly dissect the role of Notch signaling on SARS-CoV-2 infection in this proposal. In Objective 1, we first seek to understand the fundamental biology of Notch signaling in the context of SARS-CoV-2 infection. (A) First, we will evaluate the expression of Notch signaling components in SARS-CoV-2 susceptible cells before and after SARS-CoV-2 infection. (B) Second, we will investigate the expression and distribution profile of Notch signaling components in human lung tissues. (C) Third, we will explore the expression of Notch signaling components in other SARS-CoV-2-relevant extrapulmonary tissues. 2 Objective 2: To evaluate the physiological impact of inhibiting Notch signaling in in vitro, ex vivo, and in vivo models. Based on the knowledge obtained from Objective 1, we will have a more systemic grasp on the basic biology of Notch signaling relevant to SARS-CoV-2 infection. In Objective 2, we will thoroughly investigate the modulatory effect of Notch signaling on SARS-CoV-2 infection on multiple dimensions. (A) First, we will investigate the impact of inhibiting Notch signaling on SARS-CoV-2 replication in in vitro and ex vivo models. (B) Second, we will investigate the impact of Notch signaling inhibition on SARS-CoV-2 pathogenesis in golden hamster and hACE2 transgenic mouse models. (C) Third, we will evaluate the impact of Notch signaling inhibition on the pathogenesis of other highly pathogenic human coronaviruses including MERS-CoV and SARS-CoV. 3 Objective 3: To investigate the mechanism of how Notch signaling potentiates SARS-CoV-2 replication. Extending from our findings from Objective 1 and 2, we next plan to investigate the mechanism of how Notch signaling potentiates SARS-CoV-2 replication in Objective 3. (A) First, we will investigate if FURIN is responsible for Notch signaling-mediated SARS-CoV-2 replication. (i) To evaluate this possibility, we will determine the expression level of FURIN in the presence of Notch inhibition. (ii) Next, we will evaluate if Notch inhibition will have an impact on SARS-CoV-2 spike cleavage. (iii) In addition, we will check if Notch inhibition will limit SARS-CoV-2 entry. (B) Second, we will investigate if other Notch effectors contribute to Notch signaling-mediated SARS-CoV-2 replication. (i) We will perform gain-of-function and loss-of-function assays on HES and HEY members on their capacity of modulating SARS-CoV-2. (ii) Upon identifying the specific HES and HEY members that could modulate SARS-CoV-2 replication, the underlying mechanism will be further explored.