Cutaneous denervation of psoriasiform mouse skin improves acanthosis and inflammation in a sensory neuropeptide-dependent manner

Stephen M Ostrowski, Abdelmadjid Belkadi, Candace M Loyd, Doina Diaconu, Nicole L Ward, Stephen M Ostrowski, Abdelmadjid Belkadi, Candace M Loyd, Doina Diaconu, Nicole L Ward

Abstract

Nervous system involvement in psoriasis pathogenesis is supported by increases in nerve fiber numbers and neuropeptides in psoriatic skin and by reports detailing spontaneous plaque remission following nerve injury. Using the KC-Tie2 psoriasiform mouse model, we investigated the mechanisms by which nerve injury leads to inflammatory skin disease remission. Cutaneous nerves innervating dorsal skin of KC-Tie2 animals were surgically axotomized and beginning 1 day after denervation, CD11c(+) cell numbers decreased by 40% followed by a 30% improvement in acanthosis at 7 days and a 30% decrease in CD4(+) T-cell numbers by 10 days. Restoration of substance P (SP) signaling in denervated KC-Tie2 skin prevented decreases in CD11c(+) and CD4(+) cells, but had no effect on acanthosis; restoration of calcitonin gene-related peptide (CGRP) signaling reversed the improvement in acanthosis and prevented denervated-mediated decreases in CD4(+) cells. Under innervated conditions, small-molecule inhibition of SP in KC-Tie2 animals resulted in similar decreases to those observed following surgical denervation for cutaneous CD11c(+) and CD4(+) cell numbers; whereas small-molecule inhibition of CGRP resulted in significant reductions in CD4(+) cell numbers and acanthosis. These data demonstrate that sensory nerve-derived peptides mediate psoriasiform dendritic cell and T-cell infiltration and acanthosis and introduce targeting nerve-immunocyte/KC interactions as potential psoriasis therapeutic treatment strategies.

Conflict of interest statement

Conflict of Interest

The authors state no conflict of interest.

Figures

Figure 1. KC-Tie2 mice have increased expression…
Figure 1. KC-Tie2 mice have increased expression of neuropeptides in skin and DRG than control mice
Real time PCR was performed using primers targeting the neuropeptides somatostatin (SST), SP (SP), calcitonin gene related peptide (CGRP), vasoactive intestinal protein (VIP) and neuropeptide Y (NPY). (a) KC-Tie2 animals (n=4) have increased levels of SP and NPY expression in whole back skin as compared to control animals (n=4) and (b) increased level of SP and CGRP expression in their dorsal root ganglia (DRGs) as compared to control animals. p values are as indicated.
Figure 2. Surgical denervation of KC-Tie2 mouse…
Figure 2. Surgical denervation of KC-Tie2 mouse skin results in decreased acanthosis
Representative images of H&E staining of sections of (a) control, (b) KC-Tie2 sham operated and (c) KC-Tie2 denervated back skin. (d) Epidermal thickness (in µM) of the sham operated side and the denervated side of back skin is presented for individual animals (n=13). The hatched line represents average epidermal thickness levels in innervated control mouse back skin. Representative images of Ki67 stained sections of (e) KC-Tie2 sham operated and (f) KC-Tie2 denervated back skin. p values are as indicated. Scale bar = 50µM.
Figure 3. CD4 + T cell numbers…
Figure 3. CD4+ T cell numbers decrease in denervated skin while T cell derived cytokines remain unchanged
(a–c) Representative images of CD4 immunohistochemical staining in back skin sections of (a) control, (b) KC-Tie2 sham operated and (c) KC-Tie2 denervated back skin. (d) CD4+ T cell numbers in sham operated skin and denervated skin are presented for individual animals and are significantly decreased in KC-Tie2 denervated mouse skin compared to sham operated skin (n=10). The hatched line represents average number of CD4+ T cells in back skin of control mice. Western blotting (e) and ELISA (f–g) demonstrate no changes in IFNγ, IL-17 and IL-6 between sham operated and denervated KC-Tie2 mouse skin. p values are as indicated. Scale bar = 50µM.
Figure 4. Surgical denervation of KC-Tie2 mouse…
Figure 4. Surgical denervation of KC-Tie2 mouse skin reduces CD11c+ dendritic cell numbers back to control mouse levels and decreases IL-23 protein expression
(a–c) Representative images of CD11c+ immunohistochemical staining in back skin sections of (a) control, (b) KC-Tie2 sham operated and (c) KC-Tie2 denervated back skin. (d) CD11c+ cell numbers in sham operated skin and denervated skin are presented for individual animals (n=11). The hatched line represents average number of CD11c+ cells in back skin of control mice. (e) ELISA analysis of IL-23 protein expression demonstrates a significant decrease in denervated skin compared to sham operated skin in KC-Tie2 mice. p values are as indicated. p values are as indicated. Scale bar = 50µM.
Figure 5. Restoration of neuropeptide signaling under…
Figure 5. Restoration of neuropeptide signaling under denervated conditions returns CD11c+ and CD4+ cell numbers and acanthosis back to innervated conditions in a neuropeptide specific manner
Epidermal thickness (a), CD4+ cell number (b), and CD11c+ numbers (c) are presented for cohorts of animals (n=4 per group) that had PBS, SP receptor agonist (GR73632), or CGRP peptide injected intradermally into the denervated side of the back skin daily beginning one day after surgery. p values are as indicated.
Figure 6. Inhibition of SP or CGRP…
Figure 6. Inhibition of SP or CGRP activity under innervated conditions mimics denervated-mediated changes to acanthosis and CD11c+ and CD4+ cell numbers in a neuropeptide specific manner
Epidermal thickness (a), CD4+ cell number (b), and CD11c+ T cell number data (c) are presented for cohorts of animals (n=4–6 per group) prior to (pre-treatment) and after (post-treatment) 30 days of treatment with either PBS, the selective SP receptor NK-1R antagonist (RP67580), or the CGRP antagonist, CGRP8–37. p values are as indicated.

References

    1. Arck PC, Handjiski B, Peters EM, Peter AS, Hagen E, Fischer A, et al. Stress inhibits hair growth in mice by induction of premature catagen development and deleterious perifollicular inflammatory events via neuropeptide substance P-dependent pathways. Am J Pathol. 2003;162:803–814.
    1. Arck PC, Slominski A, Theoharides TC, Peters EM, Paus R. Neuroimmunology of stress: skin takes center stage. J Invest Dermatol. 2006;126:1697–1704.
    1. Casasco A, Marchetti C, Calligaro A, Casasco M, Poggi P, Beolchini M. Immunocytochemical labelling of Merkel cells of human oral mucosa by means of antibodies to protein gene product 9.5. Bull Group Int Rech Sci Stomatol Odontol. 1990;33:61–64.
    1. Chamian F, Lowes MA, Lin SL, Lee E, Kikuchi T, Gilleaudeau P, et al. Alefacept reduces infiltrating T cells, activated dendritic cells, and inflammatory genes in psoriasis vulgaris. Proc Natl Acad Sci U S A. 2005;102:2075–2080.
    1. Chiang HY, Huang IT, Chen WP, Chien HF, Shun CT, Chang YC, et al. Regional difference in epidermal thinning after skin denervation. Exp Neurol. 1998;154:137–145.
    1. Costa R, Marotta DM, Manjavachi MN, Fernandes ES, Lima-Garcia JF, Paszcuk AF, et al. Evidence for the role of neurogenic inflammation components in trypsin-elicited scratching behaviour in mice. Br J Pharmacol. 2008;154:1094–1103.
    1. Costa SK, Yshii LM, Poston RN, Muscara MN, Brain SD. Pivotal role of endogenous tachykinins and the NK1 receptor in mediating leukocyte accumulation, in the absence of oedema formation, in response to TNFalpha in the cutaneous microvasculature. J Neuroimmunol. 2006;171:99–109.
    1. Dewing SB. Remission of psoriasis associated with cutaneous nerve section. Arch Dermatol. 1971;104:220–221.
    1. Ding W, Wagner JA, Granstein RD. CGRP, PACAP, and VIP modulate Langerhans cell function by inhibiting NF-kappaB activation. J Invest Dermatol. 2007;127:2357–2367.
    1. Farber EM, Lanigan SW, Boer J. The role of cutaneous sensory nerves in the maintenance of psoriasis. Int J Dermatol. 1990;29:418–420.
    1. Farber EM, Nickoloff BJ, Recht B, Fraki JE. Stress, symmetry, and psoriasis: possible role of neuropeptides. J Am Acad Dermatol. 1986;14:305–311.
    1. Fuller RW, Conradson TB, Dixon CM, Crossman DC, Barnes PJ. Sensory neuropeptide effects in human skin. Br J Pharmacol. 1987;92:781–788.
    1. Gottlieb AB, Chamian F, Masud S, Cardinale I, Abello MV, Lowes MA, et al. TNF inhibition rapidly down-regulates multiple proinflammatory pathways in psoriasis plaques. J Immunol. 2005;175:2721–2729.
    1. Goyarts E, Matsui M, Mammone T, Bender AM, Wagner JA, Maes D, et al. Norepinephrine modulates human dendritic cell activation by altering cytokine release. Exp Dermatol. 2008;17:188–196.
    1. Griffiths CE, Strober BE, van de Kerkhof P, Ho V, Fidelus-Gort R, Yeilding N, et al. Comparison of ustekinumab and etanercept for moderate-to-severe psoriasis. N Engl J Med. 2010;362:118–128.
    1. Gudjonsson JE, Johnston A, Dyson M, Valdimarsson H, Elder JT. Mouse models of psoriasis. J Invest Dermatol. 2007;127:1292–1308.
    1. Hosoi J, Murphy GF, Egan CL, Lerner EA, Grabbe S, Asahina A, et al. Regulation of Langerhans cell function by nerves containing calcitonin gene-related peptide. Nature. 1993;363:159–163.
    1. Hsieh ST, Lin WM. Modulation of keratinocyte proliferation by skin innervation. J Invest Dermatol. 1999;113:579–586.
    1. Huang IT, Lin WM, Shun CT, Hsieh ST. Influence of cutaneous nerves on keratinocyte proliferation and epidermal thickness in mice. Neuroscience. 1999;94:965–973.
    1. Joachim RA, Handjiski B, Blois SM, Hagen E, Paus R, Arck PC. Stress-induced neurogenic inflammation in murine skin skews dendritic cells towards maturation and migration: key role of intercellular adhesion molecule-1/leukocyte function-associated antigen interactions. Am J Pathol. 2008;173:1379–1388.
    1. Joachim RA, Kuhlmei A, Dinh QT, Handjiski B, Fischer T, Peters EM, et al. Neuronal plasticity of the "brain-skin connection": stress-triggered up-regulation of neuropeptides in dorsal root ganglia and skin via nerve growth factor-dependent pathways. J Mol Med. 2007;85:1369–1378.
    1. Joseph T, Kurian J, Warwick DJ, Friedmann PS. Unilateral remission of psoriasis following traumatic nerve palsy. Br J Dermatol. 2005;152:185–186.
    1. Kinnman E. Collateral sprouting of sensory axons in the hairy skin of the trunk: a morphological study in adult rats. Brain Res. 1987;414:385–389.
    1. Lee E, Trepicchio WL, Oestreicher JL, Pittman D, Wang F, Chamian F, et al. Increased expression of interleukin 23 p19 and p40 in lesional skin of patients with psoriasis vulgaris. J Exp Med. 2004;199:125–130.
    1. Legat FJ, Jaiani LT, Wolf P, Wang M, Lang R, Abraham T, et al. The role of calcitonin gene-related peptide in cutaneous immunosuppression induced by repeated subinflammatory ultraviolet irradiation exposure. Exp Dermatol. 2004;13:242–250.
    1. Lowes MA, Chamian F, Abello MV, Fuentes-Duculan J, Lin SL, Nussbaum R, et al. Increase in TNF-alpha and inducible nitric oxide synthase-expressing dendritic cells in psoriasis and reduction with efalizumab (anti-CD11a) Proc Natl Acad Sci U S A. 2005;102:19057–19062.
    1. Niizeki H, Kurimoto I, Streilein JW. A substance p agonist acts as an adjuvant to promote hapten-specific skin immunity. J Invest Dermatol. 1999;112:437–442.
    1. Papp KA, Langley RG, Lebwohl M, Krueger GG, Szapary P, Yeilding N, et al. Efficacy and safety of ustekinumab, a human interleukin-12/23 monoclonal antibody, in patients with psoriasis: 52-week results from a randomised, double-blind, placebo-controlled trial (PHOENIX 2) The Lancet. 2008;371:1675–1684.
    1. Pavlovic S, Daniltchenko M, Tobin DJ, Hagen E, Hunt SP, Klapp BF, et al. Further exploring the brain-skin connection: stress worsens dermatitis via substance P-dependent neurogenic inflammation in mice. J Invest Dermatol. 2008;128:434–446.
    1. Perlman HH. Remission of psoriasis vulgaris from the use of nerve-blocking agents. Arch Dermatol. 1972;105:128–129.
    1. Peters EM, Kuhlmei A, Tobin DJ, Muller-Rover S, Klapp BF, Arck PC. Stress exposure modulates peptidergic innervation and degranulates mast cells in murine skin. Brain Behav Immun. 2005;19:252–262.
    1. Peters EM, Liotiri S, Bodo E, Hagen E, Biro T, Arck PC, et al. Probing the effects of stress mediators on the human hair follicle: substance P holds central position. Am J Pathol. 2007;171:1872–1886.
    1. Raychaudhuri SP, Jiang WY, Raychaudhuri SK. Revisiting the Koebner phenomenon: role of NGF and its receptor system in the pathogenesis of psoriasis. Am J Pathol. 2008;172:961–971.
    1. Raychaudhuri SP, Raychaudhuri SK. Role of NGF and neurogenic inflammation in the pathogenesis of psoriasis. Prog Brain Res. 2004;146:433–437.
    1. Raychaudhuri SP, Sanyal M, Weltman H, Kundu-Raychaudhuri S. K252a, a High-Affinity Nerve Growth Factor Receptor Blocker, Improves Psoriasis: An In Vivo Study Using the Severe Combined Immunodeficient Mouse-Human Skin Model. Journal of Investigative Dermatology. 2004;122:812–819.
    1. Saint-Mezard P, Chavagnac C, Bosset S, Ionescu M, Peyron E, Kaiserlian D, et al. Psychological stress exerts an adjuvant effect on skin dendritic cell functions in vivo. J Immunol. 2003;171:4073–4080.
    1. Saraceno R, Kleyn CE, Terenghi G, Griffiths CE. The role of neuropeptides in psoriasis. Br J Dermatol. 2006;155:876–882.
    1. Seiffert K, Hosoi J, Torii H, Ozawa H, Ding W, Campton K, et al. Catecholamines inhibit the antigen-presenting capability of epidermal Langerhans cells. J Immunol. 2002;168:6128–6135.
    1. Seike M, Ikeda M, Morimoto A, Matsumoto M, Kodama H. Increased synthesis of calcitonin gene-related peptide stimulates keratinocyte proliferation in murine UVB-irradiated skin. J Dermatol Sci. 2002;28:135–143.
    1. Seville RH. Psoriasis and stress. Br J Dermatol. 1977;97:297–302.
    1. Siebenhaar F, Magerl M, Peters EM, Hendrix S, Metz M, Maurer M. Mast cell-driven skin inflammation is impaired in the absence of sensory nerves. J Allergy Clin Immunol. 2008;121:955–961.
    1. Stankovic N, Johansson O, Hildebrand C. Increased occurrence of PGP 9.5-immunoreactive epidermal Langerhans cells in rat plantar skin after sciatic nerve injury. Cell Tissue Res. 1999;298:255–260.
    1. Stratigos AJ, Katoulis AK, Stavrianeas NG. Spontaneous clearing of psoriasis after stroke. J Am Acad Dermatol. 1998;38:768–770.
    1. Swindell WR, Johnston A, Elder JT, Ward NL, Gudjonsson JE. Microarray-based comparison of gene expression in human psoriasis and a mouse model driven by overexpression of Tie2 in keratinocytes. J Invest Dermatol. 2010;130:S93.
    1. Talme T, Liu Z, Sundqvist KG. The neuropeptide calcitonin gene-related peptide (CGRP) stimulates T cell migration into collagen matrices. J Neuroimmunol. 2008;196:60–66.
    1. Torii H, Tamaki K, Granstein RD. The effect of neuropeptides/hormones on Langerhans cells. J Dermatol Sci. 1998;20:21–28.
    1. van der Fits L, Mourits S, Voerman JS, Kant M, Boon L, Laman JD, et al. Imiquimod-induced psoriasis-like skin inflammation in mice is mediated via the IL-23/IL-17 axis. J Immunol. 2009;182:5836–5845.
    1. Ward NL, Hatala DA, Wolfram JA, Knutsen DA, Loyd CM. Cutaneous manipulation of vascular growth factors leads to alterations in immunocytes, blood vessels and nerves: Evidence for a cutaneous neurovascular unit. Journal of Dermatological Science. 2010a In Press, Corrected Proof.
    1. Ward NL, Loyd CM, Wolfram JA, Diaconu D, Michaels CM, McCormick TS. Depletion of antigen presenting cells by clodronate liposomes reverses the psoriatic skin phenotype in KC-Tie2 mice. British Journal of Dermatology. 2010b In Press.
    1. Wolfram JA, Diaconu D, Hatala DA, Rastegar J, Knutsen DA, Lowther A, et al. Keratinocyte but not endothelial cell specific overexpression of Tie2 leads to the development of psoriasis. Am J Pathol. 2009;174:1443–1458.
    1. Yu XJ, Li CY, Xu YH, Chen LM, Zhou CL. Calcitonin gene-related peptide increases proliferation of human HaCaT keratinocytes by activation of MAP kinases. Cell Biol Int. 2009
    1. Zaba LC, Cardinale I, Gilleaudeau P, Sullivan-Whalen M, Farinas MS, Fuentes-Duculan J, et al. Amelioration of epidermal hyperplasia by TNF inhibition is associated with reduced Th17 responses. J Exp Med. 2007 20071094.
    1. Zheng Y, Danilenko DM, Valdez P, Kasman I, Eastham-Anderson J, Wu J, et al. Interleukin-22, a T(H)17 cytokine, mediates IL-23-induced dermal inflammation and acanthosis. Nature. 2007;445:648–651.

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