# In vivo PET imaging of novel engineered AAVs informs capsid design

> **NIH NIH R01** · STANFORD UNIVERSITY · 2020 · $655,422

## Abstract

Abstract of: Proposed COVID-19-related supplement to
 In vivo PET imaging of novel engineered AAVs informs capsid design
There is an urgent need to develop tools to assess viral pathogenesis and the efficacy of potential therapeutics
for novel viruses such as SARS-CoV-2. While organoid and cell-based assays have been broadly used to
assess the candidate receptor of such viruses, these assays cannot answer key questions as to: 1) whether
multiple receptors for the virus exist, 2) the in vivo receptor affinity of the virus and accumulation within the
upper respiratory tract, 3) transport of the virus into the vascular system and ultimately to the heart and
kidneys, and 4) the resulting transfection of these various sites. Our laboratory has previous developed
methods to label adeno-associated viruses and track their transport following systemic injection. We found
that these engineered viruses carry cargo attached to the capsid across the blood brain barrier and the cargo
accumulates deep within the brain. Combined optical and PET studies have suggested that binding of the
virus to its receptor results in transcytosis of the intact capsid. We hypothesize that coronaviruses may
possess similar capabilities to be transported across the lung epithelium. We plan to address key issues by
assessing receptor binding and transduction using PET and optical imaging. Our resulting specific aims are the
following: 1) development and validation of tagging strategies to image pseudotype viral particles at BSL2, 2)
development and validation of reporter gene strategies to image transduction of engineered viruses, and 3)
application and dissemination of these dual strategies to assess viral transport, transduction and susceptibility
to available therapies including a) antibodies, b) protease inhibitors, and c) fusion inhibitors. We propose to
develop and image engineered viruses expressing the spike protein and a reporter gene and track these
viruses within a model of lung fibrosis and a mouse model with a humanized ACE2 receptor. We will leverage
the capabilities to label and track viral capsids and transduction developed within this R01 and key capabilities
of Stanford University. Pseudotype viruses based on vesicular stomatitis virus (VSV) and lentivirus have been
developed with spike proteins corresponding to SARS-CoV or SARS-CoV-2. Further, replicons with intact viral
proteases have been engineered. We propose to collaborate with those developing and testing engineered
viruses and therapeutics at Stanford, including Jan Carette and Catherine Blish in addition to the key personnel
on our parent project. At the conclusion of each phase of this project, we will disseminate strategies for the
incorporation of a PET tag, a reporter gene, and dual PET imaging protocols. We hypothesize that these tools
can be disseminated and rapidly modified to assess both SARS-CoV-2 and future viruses. We will make our
technology available through commercial and scientific partner...

## Key facts

- **NIH application ID:** 10148990
- **Project number:** 3R01EB028646-02S1
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Katherine W Ferrara
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $655,422
- **Award type:** 3
- **Project period:** 2019-08-01 → 2021-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10148990

## Citation

> US National Institutes of Health, RePORTER application 10148990, In vivo PET imaging of novel engineered AAVs informs capsid design (3R01EB028646-02S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10148990. Licensed CC0.

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