# Tracing the origin of regenerative and scarring fibroblasts during wound healing with single-cell technologies

> **NIH NIH R21** · WASHINGTON UNIVERSITY · 2021 · $394,123

## Abstract

PROJECT SUMMARY
Wound fibroblasts play a crucial role during wound healing by producing extracellular matrix (ECM), providing a
physical framework for cell repopulation, replenishing the skin's structural strength. While wound fibroblasts
promote skin healing, in adults this process is associated with changes that lead to scarring and can result in
non-functional repair. Here, we hypothesize that the comparison of wound fibroblast differentiation during
regeneration and scarring conditions will identify novel molecular mechanisms to diminish
scarring. Contrasting the differentiation paths of wound fibroblasts in regenerative and scarring processes will
increase our understanding of the molecular mechanisms that drive these processes and could highlight new
targets for therapeutic approaches. In this proposal, we aim to overcome previous technical
limitations surrounding wound fibroblast heterogeneity and a lack of specific markers by adapting and deploying
novel single-cell genomic technologies in an established wound healing model. This approach will enable the
high-resolution deconstruction of the intrinsic and extrinsic components driving wound fibroblast differentiation
to heterogeneous states during regeneration and scarring. First, in Aim 1, we will adapt a single-cell tracking
tool developed in our laboratory, CellTagging, for deployment in the context of regenerative and scarring in
vivo wound healing. CellTagging is the unique labeling of cells using virus-delivered barcodes that are expressed
as transcripts, enabling capture of lineage information in parallel with cell identity and function. We will adapt our
CellTagging technology to track the wound fibroblast origins and differentiation during wound regenerative and
scarring healing. This strategy will consist of labeling fibroblasts recovered from embryonic (promote
regeneration) and adult (promote scarring) uninjured skin, followed by their transplant into a host and subsequent
wounding. Here, we aim to characterize heterogeneity and origin of wound fibroblasts tracing their lineage back
to fibroblast populations in uninjured skin. We will use our proprietary gene regulatory network reconstruction
algorithm, CellOracle, to identify key regulatory factors driving wound fibroblast differentiation during
regeneration and scarring conditions. In our complementary Aim 2, we propose to investigate wound fibroblast
interactions with their cellular microenvironment in situ. To achieve this, we will perform multiplex error-robust
fluorescence in situ hybridization (MERFISH) on sections from wounds transplanted with embryonic
(regenerative) or adult (scarring) fibroblasts to identify cell populations based on the expression of specific
markers in the wound bed. Using established computational tools, we will assign cell identities and states to
individual cells within the tissue, creating a map of gene expression projected onto wound bed histology. This
will allow us to identify key interactin...

## Key facts

- **NIH application ID:** 10216730
- **Project number:** 1R21AR077825-01A1
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** Samantha Annette Morris
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $394,123
- **Award type:** 1
- **Project period:** 2021-09-24 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10216730

## Citation

> US National Institutes of Health, RePORTER application 10216730, Tracing the origin of regenerative and scarring fibroblasts during wound healing with single-cell technologies (1R21AR077825-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10216730. Licensed CC0.

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