NSF Org:	MCB Division of Molecular and Cellular Biosciences
Recipient:	SOUTH DAKOTA STATE UNIVERSITY
Initial Amendment Date:	July 20, 2020
Latest Amendment Date:	July 20, 2020
Award Number:	2030266
Award Instrument:	Standard Grant
Program Manager:	Matt Buechner mbuechne@nsf.gov  (703)292-4675 MCB  Division of Molecular and Cellular Biosciences BIO  Directorate for Biological Sciences
Start Date:	August 1, 2020
End Date:	January 31, 2022 (Estimated)
Total Intended Award Amount:	$199,475.00
Total Awarded Amount to Date:	$199,475.00
Funds Obligated to Date:	FY 2020 = $199,475.00
History of Investigator:	Adam Hoppe (Principal Investigator) adam.hoppe@sdstate.edu
Recipient Sponsored Research Office:	South Dakota State University 940 ADMINISTRATION LN BROOKINGS SD  US  57007-0001 (605)688-6696
Sponsor Congressional District:	00
Primary Place of Performance:	South Dakota State University 1015 Campanile Ave Brookings SD  US  57007-0001
Primary Place of Performance Congressional District:	00
Unique Entity Identifier (UEI):	DNZNC466DGR7
Parent UEI:
NSF Program(s):	Cellular Dynamics and Function
Primary Program Source:	01002021DB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s):	096Z, 7465, 7914, 9150
Program Element Code(s):	111400
Award Agency Code:	4900
Fund Agency Code:	4900
Assistance Listing Number(s):	47.074

ABSTRACT

This project will discover genes that help and/or inhibit the entry of the COVID-19 virus into cells using a new unbiased screening approach. This work will benefit society by creating a more comprehensive view of how COVID-19 infects cells. In addition, this work has the potential to identify host cell antiviral defense genes that can inhibit viral entry, as well as define genes that contribute to antibody protection in immune cells. Here, new CRISPR technology will be applied to screen the entire human genome to find these genes by correlating gene inactivation with increased or decreased viral entry into lung and immune cells. This information will provide the knowledge needed to develop new anti-viral strategies and to understand how environmental, genetic and cellular factors may influence the susceptibility of an individual to infection as well as the severity of the infection. The Broader Impacts of the project include training the next generation of virologists and cell biologists. Graduate students will participate in all phases of the project, including learning state-of-the-art CRISPR technologies. With the help of high school teachers, the work of this project will also be used to produce artistic descriptions that will allow broad dissemination of our findings to students, the scientific community, and the broader public.

This work aims to elucidate host genes that promote and inhibit the entry of the SARS-CoV-2 virus, the causative agent of COVID-19, into epithelial cells and macrophages using new whole-genome CRISPR screening approaches. At present, the ACE2 viral receptor and the host cell protease, TMRPSS2, have been the focus of research on the machinery needed for viral entry. However, other receptors, including CD147 and other proteases can facilitate viral entry indicating that a broader perspective is needed. Conversely, little is known about the role of innate antiviral genes that may restrict viral entry. Rapid adaptation of current CRISPR whole genome screening strategies will enable identification and ranking of all genes contributing to and inhibiting SARS-CoV-2 entry into epithelial cells expressing the ACE2 receptor and macrophages via antibody/Fc receptor or CD147-mediated attachment. Screens conducted in epithelial cells are expected to provide a map of the genes that contribute to and inhibit infection as well as viral spread within the lung. While it is not yet clear the extent to which SARS-CoV-2 can replicate in macrophages, multiple lines of evidence indicate that the virus can infect macrophages in a process aided by the presence of antibodies that may modulate their inflammatory state contributing to COVID-19. The screening strategy of the Project using pseudotyped lentiviral vectors will enable detection of viral entry, absent viral replication, and will provide a description of the genes aiding and inhibiting antibody-dependent viral entry into macrophages. Comprehensive genetic analysis across both cell types will provide an unprecedented view of viral entry and will provide scientific knowledge urgently needed to combat COVID-19.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

 

In this project, we sought to develop new whole genome screening strategies to identify human genes that facilitate or inhibit entry of the virus that causes COVID19.  The goal was to rapidly adapt a whole-genome analysis approach based on CRISPR-mediated gene disruption to in a human cell line.  Moreover, we sought to develop virus-like particles that could be used safely and efficiently.  We were successful in developing the screening method and viral like particles as well as a new and improved strategy for identifying human genes affecting viral entry.  These new tools are being disseminated to the scientific community and should help prepare for rapid responses to new emerging viruses.  Moreover, this study identified human genes that inhibit viral entry unique to COVID19 and as well as genes that inhibit other viral like particles from entering cells.  Further analysis of genes identified in our study is ongoing and are providing new insight into how viruses gain access to cells and helps form a basis for future strategies for inhibiting viral entry. Additionally, the techniques developed in this study are applicable to screening human gene determinates of other enveloped viruses.  This project engaged high-school teachers working on their Master’s in Chemical Education to create new graphics that to communicate this work to high-school students and to the general public.   

 

Last Modified: 05/02/2022
Modified by: Adam D Hoppe

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