TY - JOUR
T1 - Macrophage TGF-β signaling is critical for wound healing with heterotopic ossification after trauma
AU - Patel, Nicole K.
AU - Nunez, Johanna H.
AU - Sorkin, Michael
AU - Marini, Simone
AU - Pagani, Chase A.
AU - Strong, Amy L.
AU - Hwang, Charles D.
AU - Li, Shuli
AU - Padmanabhan, Karthik R.
AU - Kumar, Ravi
AU - Bancroft, Alec C.
AU - Greenstein, Joey A.
AU - Nelson, Reagan
AU - Rasheed, Husain A.
AU - Livingston, Nicholas
AU - Vasquez, Kaetlin
AU - Huber, Amanda K.
AU - Levi, Benjamin
N1 - Funding Information:
We would like to thank the University of Michigan Center for Molecular Imaging and the Biomedical Research Core Facilities Microscopy Imaging Core for their assistance. We would like to thank Noelle Viss-er and Nicole J. Edwards for technical assistance. MS was funded by Plastic Surgery Foundation National Endowment Award. CDH was supported by Howard Hughes Medical Institute Medical Research Fellowship. BL was funded by NIH 1R01AR079863 and NIH R01AR079171. AKH was funded by the IFOPA ACT grant and CaBRI Research Grant for Rare Diseases.
Funding Information:
We would like to thank the University of Michigan Center for Molecular Imaging and the Biomedical Research Core Facilities Microscopy Imaging Core for their assistance. We would like to thank Noelle Visser and Nicole J. Edwards for technical assistance. MS was funded by Plastic Surgery Foundation National Endowment Award. CDH was supported by Howard Hughes Medical Institute Medical Research Fellowship. BL was funded by NIH 1R01AR079863 and NIH R01AR079171. AKH was funded by the IFOPA ACT grant and CaBRI Research Grant for Rare Diseases.
Publisher Copyright:
© 2022, Patel et al.
PY - 2022/10/24
Y1 - 2022/10/24
N2 - Transforming growth factor–β1 (TGF-β1) plays a central role in normal and aberrant wound healing, but the precise mechanism in the local environment remains elusive. Here, using a mouse model of aberrant wound healing resulting in heterotopic ossification (HO) after traumatic injury, we find autocrine TGF-β1 signaling in macrophages, and not mesenchymal stem/progenitor cells, is critical in HO formation. In-depth single-cell transcriptomic and epigenomic analyses in combination with immunostaining of cells from the injury site demonstrated increased TGF-β1 signaling in early infiltrating macrophages, with open chromatin regions in TGF-β1–stimulated genes at binding sites specific for transcription factors of activated TGF-β1 (SMAD2/3). Genetic deletion of TGF-β1 receptor type 1 (Tgfbr1; Alk5), in macrophages, resulted in increased HO, with a trend toward decreased tendinous HO. To bypass the effect seen by altering the receptor, we administered a systemic treatment with TGF-β1/3 ligand trap TGF-βRII-Fc, which resulted in decreased HO formation and a delay in macrophage infiltration to the injury site. Overall, our data support the role of the TGF-β1/ ALK5 signaling pathway in HO.
AB - Transforming growth factor–β1 (TGF-β1) plays a central role in normal and aberrant wound healing, but the precise mechanism in the local environment remains elusive. Here, using a mouse model of aberrant wound healing resulting in heterotopic ossification (HO) after traumatic injury, we find autocrine TGF-β1 signaling in macrophages, and not mesenchymal stem/progenitor cells, is critical in HO formation. In-depth single-cell transcriptomic and epigenomic analyses in combination with immunostaining of cells from the injury site demonstrated increased TGF-β1 signaling in early infiltrating macrophages, with open chromatin regions in TGF-β1–stimulated genes at binding sites specific for transcription factors of activated TGF-β1 (SMAD2/3). Genetic deletion of TGF-β1 receptor type 1 (Tgfbr1; Alk5), in macrophages, resulted in increased HO, with a trend toward decreased tendinous HO. To bypass the effect seen by altering the receptor, we administered a systemic treatment with TGF-β1/3 ligand trap TGF-βRII-Fc, which resulted in decreased HO formation and a delay in macrophage infiltration to the injury site. Overall, our data support the role of the TGF-β1/ ALK5 signaling pathway in HO.
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U2 - 10.1172/jci.insight.144925
DO - 10.1172/jci.insight.144925
M3 - Article
C2 - 36099022
AN - SCOPUS:85140417090
SN - 2379-3708
VL - 7
JO - JCI Insight
JF - JCI Insight
IS - 20
M1 - e144925
ER -