Differential Expression of Insulin-Like Growth Factor 1 and Wnt Family Member 4 Correlates With Functional Heterogeneity of Human Dermal Fibroblasts

TY - JOUR

T1 - Differential Expression of Insulin-Like Growth Factor 1 and Wnt Family Member 4 Correlates With Functional Heterogeneity of Human Dermal Fibroblasts

AU - Culley, Oliver J

AU - Louis, Blaise

AU - Philippeos, Christina

AU - Oulès, Bénédicte

AU - Tihy, Matthieu

AU - Segal, Joe M

AU - Hyliands, Della

AU - Jenkins, Gail

AU - Bhogal, Ranjit K

AU - Siow, Richard C

AU - Watt, Fiona M

N1 - Funding Information: This work was supported by the UK Medical Research Council (MR/PO18823/1) and the Wellcome Trust (206439/Z/17/Z). OC gratefully acknowledges funding via a BBSRC Unilever iCASE studentship. We also acknowledge funding from the Department of Health via the National Institute for Health Research comprehensive Biomedical Research Centre award to Guy’s and St. Thomas’ National Health Service Foundation Trust in partnership with King’s College London and King’s College Hospital NHS Foundation Trust. Funding Information: We gratefully acknowledge the research contributions of Simon Broad, Davide Danovi, Arsham Ghahramani, Eamonn Morrison, Victor Augusti Negri, Angela Oliveira Pisco, Mark Soldin, Kalle Sipil?, Tanya Shaw, and Sebastiaan Zijl. Funding. This work was supported by the UK Medical Research Council (MR/PO18823/1) and the Wellcome Trust (206439/Z/17/Z). OC gratefully acknowledges funding via a BBSRC Unilever iCASE studentship. We also acknowledge funding from the Department of Health via the National Institute for Health Research comprehensive Biomedical Research Centre award to Guy?s and St. Thomas? National Health Service Foundation Trust in partnership with King?s College London and King?s College Hospital NHS Foundation Trust. Publisher Copyright: © Copyright © 2021 Culley, Louis, Philippeos, Oulès, Tihy, Segal, Hyliands, Jenkins, Bhogal, Siow and Watt. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/4/6

Y1 - 2021/4/6

N2 - Although human dermis contains distinct fibroblast subpopulations, the functional heterogeneity of fibroblast lines from different donors is under-appreciated. We identified one commercially sourced fibroblast line (c64a) that failed to express α-smooth muscle actin (α-SMA), a marker linked to fibroblast contractility, even when treated with transforming growth factor-β1 (TGF-β1). Gene expression profiling identified insulin-like growth factor 1 (IGF1) as being expressed more highly, and Asporin (ASPN) and Wnt family member 4 (WNT4) expressed at lower levels, in c64a fibroblasts compared to three fibroblast lines that had been generated in-house, independent of TGF-β1 treatment. TGF-β1 increased expression of C-X-C motif chemokine ligand 1 (CXCL1) in c64a cells to a greater extent than in the other lines. The c64a gene expression profile did not correspond to any dermal fibroblast subpopulation identified by single-cell RNAseq of freshly isolated human skin cells. In skin reconstitution assays, c64a fibroblasts did not support epidermal stratification as effectively as other lines tested. In fibroblast lines generated in-house, shRNA-mediated knockdown of IGF1 increased α-SMA expression without affecting epidermal stratification. Conversely, WNT4 knockdown had no consistent effect on α-SMA expression, but increased the ability of fibroblasts to support epidermal stratification. Thus, by comparing the properties of different lines of cultured dermal fibroblasts, we have identified IGF1 and WNT4 as candidate mediators of two distinct dermal functions: myofibroblast formation and epidermal maintenance.

AB - Although human dermis contains distinct fibroblast subpopulations, the functional heterogeneity of fibroblast lines from different donors is under-appreciated. We identified one commercially sourced fibroblast line (c64a) that failed to express α-smooth muscle actin (α-SMA), a marker linked to fibroblast contractility, even when treated with transforming growth factor-β1 (TGF-β1). Gene expression profiling identified insulin-like growth factor 1 (IGF1) as being expressed more highly, and Asporin (ASPN) and Wnt family member 4 (WNT4) expressed at lower levels, in c64a fibroblasts compared to three fibroblast lines that had been generated in-house, independent of TGF-β1 treatment. TGF-β1 increased expression of C-X-C motif chemokine ligand 1 (CXCL1) in c64a cells to a greater extent than in the other lines. The c64a gene expression profile did not correspond to any dermal fibroblast subpopulation identified by single-cell RNAseq of freshly isolated human skin cells. In skin reconstitution assays, c64a fibroblasts did not support epidermal stratification as effectively as other lines tested. In fibroblast lines generated in-house, shRNA-mediated knockdown of IGF1 increased α-SMA expression without affecting epidermal stratification. Conversely, WNT4 knockdown had no consistent effect on α-SMA expression, but increased the ability of fibroblasts to support epidermal stratification. Thus, by comparing the properties of different lines of cultured dermal fibroblasts, we have identified IGF1 and WNT4 as candidate mediators of two distinct dermal functions: myofibroblast formation and epidermal maintenance.

UR - http://www.scopus.com/inward/record.url?scp=85104679483&partnerID=8YFLogxK

U2 - 10.3389/fcell.2021.628039

DO - 10.3389/fcell.2021.628039

M3 - Article

C2 - 33889572

SN - 2296-634X

VL - 9

JO - Frontiers in Cell and Developmental Biology

JF - Frontiers in Cell and Developmental Biology

M1 - 628039

ER -