# Cell-directed gene therapy for pain recovery after surgery and inflammation

> **NIH NIH R01** · WAKE FOREST UNIVERSITY HEALTH SCIENCES · 2024 · $437,659

## Abstract

Summary
We have recently discovered macrophage ED2/CD163 gene overexpression as a novel and safe pain
therapeutic target in the local peripheral immune system in a major surgery rat model. We propose to develop a
cell-directed gene therapy that would correct the underlying local immunological cause of pain resulting from
inflammatory processes, specifically in sub-chronic postoperative pain or inflammatory conditions. Through
unbiased genome-wide transcriptomic analyses in human primary macrophages we identified that ED2/CD163
gene induction modulates tumor necrosis factor alpha (TNFa) and interleukin (IL)-1 beta (IL-1b). CD163
overexpression using a clinically tested nanoparticle designed to target macrophages promoted a more rapid
wound healing in 3D human organotypic skin tissues and prevented sub-chronic postoperative pain behaviors
and reduced local TNFa and IL-1b in rats with skin-muscle incision and retraction (SMIR) surgery. We
hypothesize that ED2/CD163 in macrophages is a safe target for the treatment of inflammatory pain with
opioid sparing effects. We propose a multidimensional research plan including, 1) a sub-chronic surgical pain
model, the SMIR surgery, and a knee inflammatory mode, the Complete Freund Adjuvant (CFA)-induced knee
arthritis; 2) ED2/CD163 gain and loss of function using macrophage-directed nanotechnology; 3) novel, clinically
relevant, and complex operant pain-related behaviors; 4) cellular/molecular, tissue, and transcriptomic outcomes
for mechanistic target engagement; and 5) studies for ED2/CD163’s effects on opioid requirements. We will
implement our plan through these specific aims: 1) Determine that macrophage specific ED2/CD163 gene
induction effectively promotes resolution of inflammatory pain. A mannosilated polyethyleneimine
nanoparticles (Man-PEI) designed to deliver nucleic acids specifically to macrophages will be used to conduct
ED2/CD163 gain (overexpression) or loss (knock down) of function. We will assess classic behaviors (von Frey
and weight bearing), and novel complex and clinically relevant functional activity and attention-related behaviors
developed and validated by our team. 2) Define ED2/CD163 target engagement, i.e. ED2/CD163 as a
signaling driver that dictates the dynamics of macrophage phenotype change and cellular reprogramming
in inflammatory pain. TNFa and IL-1b will be measured as downstream target engagement. Also, we will use
single-cell RNAseq (scRNAseq), cluster, and phenotype trajectory analysis to define how ED2/CD163 impacts
gene expression programs in macrophages infiltrating the inflamed tissue. 3) Establish that macrophage
ED2/CD163 gene induction results in opioid-sparing effects in surgical and inflammatory pain. We will
construct dose responses of morphin in rats with SMIR or arthritis and ED2/CD163 overexpression to measure
opioid-sparing effects. Our project will establish ED2/CD163 as a cell-directed gene therapy for postsurgical pain
that will reduce opioid requiremen...

## Key facts

- **NIH application ID:** 10813720
- **Project number:** 5R01NS122153-03
- **Recipient organization:** WAKE FOREST UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** THOMAS JEFFREY MARTIN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $437,659
- **Award type:** 5
- **Project period:** 2022-01-15 → 2026-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10813720

## Citation

> US National Institutes of Health, RePORTER application 10813720, Cell-directed gene therapy for pain recovery after surgery and inflammation (5R01NS122153-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10813720. Licensed CC0.

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