Lymphedema-Associated Fibroblasts Are Related to Fibrosis and Stage Progression in Patients and a Murine Microsurgical Model

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Patrick Alessandro Marco Will Marks - , University Center for Orthopedics, Trauma and Plastic Surgery, Klinikum Ludwigshafen, Heidelberg University  (Author)
  • Katja Kilian - (Author)
  • Karen Bieback - (Author)
  • Fabia Fricke - (Author)
  • Juan Enrique Berner - (Author)
  • Ulrich Kneser - (Author)
  • Christoph Hirche - (Author)

Abstract

Background: The driver of secondary lymphedema (SL) progression is chronic inflammation, which promotes fibrosis. Despite advances in preclinical research, a specific effector cell subpopulation as a biomarker for therapy response or stage progression is still missing for SL. Methods: Whole skin samples of 35 murine subjects of a microsurgically induced SL model and 12 patients with SL were collected and their fibroblasts were isolated. These lymphedema-associated fibroblasts (LAFs) were cultured in a collagen I-poly-D-lysine 3-dimensional hydrogel to mimic skin conditions. Fibroblasts from non-lymphedema skin were used as negative control and transforming growth factor β (TGF-β)–stimulated fibroblasts were used to recreate profibrotic myofibroblasts. Quantitative immunocytofluorescence confocal microscopy analysis and invasion functional assays were performed in all subpopulations and statistically compared. Results: In contrast to normal skin fibroblasts, LAFs exhibit α–smooth muscle actin–positive stress fibers and a reduced number of tight junctions in 3-dimensional hydrogel conditions. The switch from normal E-cadherin high phenotype to an N-cadherin high-E-cadherin low morphology suggests epithelial-to-mesenchymal transition for expansion and proliferation. This pathologic behavior of LAF was confirmed by live cell imaging analysis of invasion assays. The significant activation of markers of the TGF-β receptor 2–Smad pathway and collagen synthesis (HSP-47 [heat shock protein 47]) in LAFs supports epithelial-to-mesenchymal transition phenotypic changes and previous findings relating to TGF-β1 and fibrosis with lymphedema. Conclusions: A characteristic SL myofibroblast subpopulation was identified and translationally related to fibrosis and TGF-β1–associated stage progression. This SL-related subpopulation was termed LAFs. A comprehensive stage-related characterization is required to validate LAFs as a reliable biomarker for SL disease progression.

Details

Original languageEnglish
Pages (from-to)688e-700e
Number of pages13
JournalPlastic and Reconstructive Surgery
Volume154
Issue number4
Early online date13 Oct 2023
Publication statusPublished - Oct 2024
Peer-reviewedYes

External IDs

Scopus 85204820025

Keywords

Library keywords