Modeling fibrosis using fibroblasts isolated from scarred rat vocal folds.
Following injury, pathologically activated vocal fold fibroblasts (VFFs) can engage in disordered extracellular matrix (ECM) remodeling, leading to VF fibrosis and impaired voice function. Given the importance of scar VFFs to phenotypically appropriate in vitro modeling of VF fibrosis, we pursued detailed characterization of scar VFFs obtained from surgically injured rat VF mucosae, compared with those obtained from experimentally naïve, age-matched tissue. Scar VFFs initially exhibited a myofibroblast phenotype characterized by increased proliferation, increased Col1a1 transcription and collagen, type I synthesis, increased Acta2 transcription and α-smooth muscle actin synthesis, and enhanced contractile function. These features were most distinct at passage 1 (P1); we observed a coalescence of the scar and naïve VFF phenotypes at later passages. An empirical Bayes statistical analysis of the P1 cell transcriptome identified 421 genes that were differentially expressed by scar, compared with naïve, VFFs. These genes were primarily associated with the wound response, ECM regulation, and cell proliferation. Follow-up comparison of P1 scar VFFs and their in vivo tissue source showed substantial transcriptomic differences. Finally, P1 scar VFFs responded to treatment with hepatocyte growth factor and transforming growth factor-β3, two biologics with reported therapeutic value. Despite the practical limitations inherent to working with early passage cells, this experimental model is easily implemented in any suitably equipped laboratory and has the potential to improve the applicability of preclinical VF fibrosis research.
Department of Surgery, Division of Otolaryngology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Yo
Y
Kishimoto
Department of Surgery, Division of Otolaryngology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Ayami Ohno
AO
Kishimoto
Department of Biostatistics and Medical Informatics, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Shuyun
S
Ye
Department of Biostatistics and Medical Informatics, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Christina
C
Kendziorski
Department of Surgery, Division of Otolaryngology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
Nathan V
NV
Welham
D000818:Animals; D049109:Cell Proliferation; D002921:Cicatrix / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D000090602:Collagen Type I, alpha 1 Chain; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005355:Fibrosis; D020869:Gene Expression Profiling; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D058628:Myofibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051381:Rats; D011916:Rats, Inbred F344; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology; D014827:Vocal Cords / Q000293:injuries / Q000378:metabolism / Q000473:pathology*
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C509046:Tgfb3 protein, rat; D053782:Transforming Growth Factor beta3; D017228:Hepatocyte Growth Factor