# Genetically-encoded molecular imaging of genome-edited immune cells

> **NIH NIH R01** · UNIVERSITY OF CINCINNATI · 2024 · $497,870

## Abstract

Project Summary
Adoptive cell transfers constitute a new paradigm in cell-based therapeutics with wide-ranging applications
from neurodegenerative, cardiovascular, autoimmune disorders as well as cancer. Autologous lymphocytes
from the patient are genetically engineered to express receptors to specific antigens on the cell surface of
target cells and reinfused back into the patient for therapeutic action. However, once administered the fate of
cells remains uncertain. In order to answer important questions regarding distribution, turnover and eventual
survival of the cells, techniques to non-invasively monitor these genome-edited cells is necessary. Molecular
imaging, particularly Positron emission tomography is advantageous over traditional diagnostic and imaging
tools because it enables diagnosis and tracking of radiotracers in real-time, is non-invasive and has the highest
sensitivity among clinical imaging modalities. We propose to develop analogues of Green fluorescent
protein/luciferase for nuclear imaging, thus combining the highly desirable elements from both these powerful
modalities towards a chemogenetic nuclear imaging modality. The metallophore-transporter complex found in
bacteria is genetically encoded in the prokaryotic DNA and offers the ideal chemical-biological pair that can be
expressed on mammalian cells to enable nuclear imaging and tracking of these cells. By engineering
mammalian cells to ectopically express the bacterial transporters, we will be able to selectively target and
image the genome -edited cells in vivo using metallophore-radionuclide probes. Bacterial metallophores have
evolved to serve as metal chelators for a wide variety of metals with the majority showing a high binding affinity
for iron. However, several pathogenic bacteria secrete metallophores with the highest binding affinity for
copper(II) (Cu). We, therefore, propose to use 64Cu, a popular radionuclide in Positron Emission Tomography
(PET) imaging to generate metallophore/64Cu complexes as contrast agents. We have identified metallophores
that are able to evade the innate immune system and avoid imminent sequestration. Combined with simple
and one-step processing techniques, they are attractive agents for depth-independent real-time imaging,
tracking and identifying genome-edited cells. Because the native or wild type cells do not express these
transporters, we expect minimal uptake in normal mammalian cells including native bacterial “commensal”
flora, as they remain exclusive to pathogenic bacteria. We hypothesize that through a combination of facile
coordinate complexation chemistry, high selectivity, immune evasiveness and non-endogenous nature,
bacterial metallophore/64Cu will serve as ideal nuclear imaging probes to identify and accurately detect GECs
in vivo. In the first phase we will develop and evaluate the bacteria transporter protein mammalian expression
vector and subsequently will be able to incorporate the vectors into CAR-T-Cells for ...

## Key facts

- **NIH application ID:** 10803318
- **Project number:** 1R01CA279962-01A1
- **Recipient organization:** UNIVERSITY OF CINCINNATI
- **Principal Investigator:** Nalinikanth Kotagiri
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $497,870
- **Award type:** 1
- **Project period:** 2024-09-17 → 2029-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10803318

## Citation

> US National Institutes of Health, RePORTER application 10803318, Genetically-encoded molecular imaging of genome-edited immune cells (1R01CA279962-01A1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10803318. Licensed CC0.

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