Crosstalk between adrenergic and toll-like receptors in human mesenchymal stem cells and keratinocytes: a recipe for impaired wound healing.
Previous studies demonstrate that skin wounds generate epinephrine (EPI) that can activate local adrenergic receptors (ARs), impairing healing. Bacterially derived activators of Toll-like receptors (TLRs) within the wound initiate inflammatory responses and can also impair healing. In this study, we examined the hypothesis that these two pathways crosstalk to one another, using EPI and macrophage-activating lipopeptide-2 (MALP2) to activate ARs and TLR2, respectively, in human bone marrow-derived mesenchymal stem cells (BM-MSCs) and neonatal keratinocytes (NHKs). BM-MSCs exposed to EPI significantly (p < .05) increased TLR2 message (sevenfold BM-MSCs), TLR2 protein (twofold), and myeloid differentiation factor 88 (MyD88) (fourfold). Conversely, activation of TLR2 by MALP2 in these cells increased β2-AR message (twofold in BM-MSCs, 2.7-fold in NHKs), β2-AR protein (2.5-fold), phosphorylation of β-AR-activated kinase (p-BARK, twofold), and induced release of EPI from both cell types (twofold). Treating cells with EPI and MALP2 together, as would be encountered in a wound, increased β2-AR and p-BARK protein expression (sixfold), impaired cell migration (BM-MSCs- 21%↓ and NHKs- 60%↓, p < .002), and resulted in a 10-fold (BM-MSCs) and 51-fold (NHKs) increase in release of IL-6 (p < .001) responses that were remarkably reduced by pretreatment with β2-AR antagonists. In vivo, EPI-stressed animals exhibited impaired healing, with elevated levels of TLR2, MyD88, and IL-6 in the wounds (p < .05) relative to nonstressed controls. Thus, our data describe a recipe for decreasing cell migration and exacerbating inflammation via novel crosstalk between the adrenergic and Toll-like receptor pathways in BM-MSCs and NHKs.
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA rrisseroff@ucdavis.edu rkdasu@ucdavis.edu rkmdasu@gmail.com.
Mohan R
MR
Dasu
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Sandra R
SR
Ramirez
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Thi Dinh
TD
La
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Farzam
F
Gorouhi
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Chuong
C
Nguyen
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Benjamin R
BR
Lin
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Chelcy
C
Mashburn
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Heather
H
Stewart
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Thomas R
TR
Peavy
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA.
Jan A
JA
Nolta
Department of Dermatology and Institute for Regenerative Cures, University of California, Davis, California, USA; Department of Biological Sciences, California State University, Sacramento, California, USA; Dermatology Service, Department of Veterans Affairs, Northern California Health Care System, Sacramento, California, USA rrisseroff@ucdavis.edu rkdasu@ucdavis.edu rkmdasu@gmail.com.
Roslyn R
RR
Isseroff
D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000319:Adrenergic beta-Antagonists / Q000097:blood; D000818:Animals; D002450:Cell Communication / Q000187:drug effects; D002465:Cell Movement; D002478:Cells, Cultured; D004837:Epinephrine / Q000378:metabolism / Q000494:pharmacology; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D055666:Lipopeptides / Q000494:pharmacology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D010766:Phosphorylation; D020239:Receptor Cross-Talk; D018343:Receptors, Adrenergic, beta-2 / Q000187:drug effects / Q000378:metabolism*; D012867:Skin / Q000293:injuries / Q000378:metabolism* / Q000473:pathology; D033581:Stem Cell Transplantation; D013997:Time Factors; D051195:Toll-Like Receptor 2 / Q000819:agonists / Q000378:metabolism*; D014945:Wound Healing / Q000187:drug effects
C553443:ADRB2 protein, human; D000318:Adrenergic beta-Agonists; D000319:Adrenergic beta-Antagonists; C508600:IL6 protein, human; D015850:Interleukin-6; D055666:Lipopeptides; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D018343:Receptors, Adrenergic, beta-2; C495342:TLR2 protein, human; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2; C577686:interleukin-6, mouse; C106335:macrophage stimulatory lipopeptide 2; C496567:GRK2 protein, human; D054769:G-Protein-Coupled Receptor Kinase 2; D004837:Epinephrine
Cell migration; Cell signaling; Mesenchymal stem cells; Stem cell-microenvironment interactions; Tissue regeneration