| NSF Org: |
IOS Division Of Integrative Organismal Systems |
| Recipient: |
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| Initial Amendment Date: | June 30, 2020 |
| Latest Amendment Date: | June 30, 2020 |
| Award Number: | 2031626 |
| Award Instrument: | Standard Grant |
| Program Manager: |
Joanna Shisler
jshisler@nsf.gov (703)292-5368 IOS Division Of Integrative Organismal Systems BIO Directorate for Biological Sciences |
| Start Date: | July 1, 2020 |
| End Date: | June 30, 2021 (Estimated) |
| Total Intended Award Amount: | $138,793.00 |
| Total Awarded Amount to Date: | $138,793.00 |
| Funds Obligated to Date: |
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| History of Investigator: |
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| Recipient Sponsored Research Office: |
1523 UNION RD RM 207 GAINESVILLE FL US 32611-1941 (352)392-3516 |
| Sponsor Congressional District: |
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| Primary Place of Performance: |
1 University of Florida Gainesville FL US 32611-5500 |
| Primary Place of
Performance Congressional District: |
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| Unique Entity Identifier (UEI): |
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| Parent UEI: |
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| NSF Program(s): | COVID-19 Research |
| Primary Program Source: |
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| Program Reference Code(s): |
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| Program Element Code(s): |
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| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.074 |
ABSTRACT
The ongoing COVID-19 pandemic has highlighted the lack of human cell culture models available for studying this virus, and the devastating consequences of this shortcoming as it relates to human health and disease. The proposed project, known as ?CLEARED? for Culture of Living-biopsies for Emerging Airway-pathogens and REspiratory Disease, combines cutting-edge technologies in 3D-printing, soft tissue engineering, artificial-intelligence-enhanced imaging, human lung biology, and virology to understand the spread of COVID-19 in lungs. This will increase the knowledge of SARS-CoV-2 biology and transmission. The researchers have developed the technology to grow portions of lung into living 3D-printed tissue structures that resembles the architecture found in the lung in a liquid-like-solid matrix. Thus, this system more closely resemble the environment in living humans versus standard cell culture. After infecting these samples with SARS-CoV-2 virus, advanced imaging of these ?living biopsies? will be used to study virus spread from cell to cell, and the efficacy of therapeutic treatments. Outcomes of the proposed research include: (i) Validating a standard model system using human lung biopsies and known diagnostics in response to SARS-CoV-OC43 infection; (ii) Determining how the disease develops and spreads in biopsies infected with different human and bat coronavirus strains. It is expected that this system will allow scientists to better understand virus transmission and prevention. This project also supports the training of three graduate students, leading to an increase in future workers to drive the bioeconomy.
The proposing team hypothesizes that controlled perfusion of SARS-CoV-2 in 3D culture models of human respiratory microtissue explants can recapitulate early stages of SARS-CoV-2 infection and COVID-19 disease. To test this hypothesis, PIs will establish a 3D model of viral infection using living microtissue explants of human bronchus and peripheral lung, quantify the early responses to viral infection using a novel 3D tissue culture platform, and determine the spatiotemporal pathogenesis of different human and bat coronaviruses strains. Preliminary data show that SARS-CoV-2 indeed infects the micro-tissues of bronchus and peripheral lung. This is a transdisciplinary team of investigators from Astronomy, Chemistry, Medicine, Engineering, Virology and lung biology. The proposed work is organized by two tasks. Task 1 will validate a standard model system using human lung biopsies and known host-response to SARS-CoV-2 infection. Readouts will include viral titer, cytokine production and spatiotemporal imaging of viral replication in response to coronavirus infection. Task 2 will determine the spatiotemporal pathogenesis of human lung biopsies infected with different human coronavirus strains (HCoV-OC43, HCoV-NL63, SARS-CoV-2) and one bat strain (btCoV-HKU3). The heterogenous nature of biopsies will alter the viral titer and cytokine production of biopsies compared to measurements in cell lines, and will provide superior information about progression and virus spread through tissues than standard cell culture technology. This RAPID award is made by the Physiological and Structural Systems Cluster in the BIO Division of Integrative Organismal Systems, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.
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.
The current biological systems for the study of COVID-19 include cell lines, which differ from the human lung in many respects, and animal hosts to which the virus is not adapted. We developed another alternative for studying pathogenesis and drug susceptibility of SARS-CoV-2 in a cryopreserved bank of human lung tissues.
This project was enabled by the emergent technology of novel cryopreservation medias that can preserve small samples of human lung tissue for on-demand experimental use. By preserving multiple samples from the same donor, we are able to find tissue samples that respond well to a specific drug or treatment, and then use another sample of tissue from the same donor to discover the reason for this response. We have infected tissues from 16 different donors with SARS-CoV-2 and found a lot of variability in the way that the tissue responds to infection and treatment. In some donor tissues treatment with certain drugs, already approved by the FDA, greatly reduce viral titer. However, treatment with the same drug in tissues from other donors has little to no effect. We are still discovering the reasons why there is such a varied response in donor tissues infected with SARS-CoV-2 and have begun to assess how these drugs, which were not previously known to reduce viral titer, can alter the growth and replication of the SARS-CoV-2 virus. As a result of our studies one of the drugs we tested is now in clinical trials as a potential treatment for SARS-CoV-2 (NCT04510194).
A second technology that was critical to this study was the use of a unique culture system that enables the tissue to remain alive and healthy for an extended amount of time. This culture system is primarily composed of small pieces of inert gel that gently support the lung tissue in a 3-dimensional (3D) space. The many spaces that exist between the small pieces of gel facilitate the perfusion of nutrients and virus particles through the tissue, which enable the tissue to remain viable for a longer period of time than traditional culture systems.
The use of improved cryopreservation media and our novel 3D culture system were important tools to advance our understanding of the infection of SARS-CoV-2. These tools have helped us to characterize how SARS-CoV-2 infects human lung tissue and will be useful for future studies related to any respiratory illness. Our next set of studies will focus on the different variants of SARS-CoV-2, including the delta strain and other strains from earlier in the pandemic, and the type of response they induce in human lung tissue.
Last Modified: 07/02/2021
Modified by: Matthew Schaller
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