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Senescent Progenitor Cells in the Skin of Patients with Cutaneous Lupus Erythematosus

2021; Elsevier BV; Volume: 142; Issue: 3 Linguagem: Inglês

10.1016/j.jid.2021.06.022

1523-1747

Xiaoyan Wang, Gilles F.H. Diercks, Wietske M. Lambers, Johanna Westra, Hendrika Bootsma, Frans G. M. Kroese, Karina de Leeuw, Sarah Pringle,

Atherosclerosis and Cardiovascular Diseases

Cutaneous lupus erythematosus (CLE) is an autoimmune disease that can occur as an isolated skin condition or as a skin manifestation secondary to systemic autoimmune diseases such as systemic lupus erythematosus and primary Sjögren's syndrome (Kuhn and Landmann, 2014Kuhn A. Landmann A. The classification and diagnosis of cutaneous lupus erythematosus.J Autoimmun. 2014; 48–49: 14-19Crossref PubMed Scopus (135) Google Scholar; Wenzel, 2019Wenzel J. Cutaneous lupus erythematosus: new insights into pathogenesis and therapeutic strategies.Nat Rev Rheumatol. 2019; 15: 519-532Crossref PubMed Scopus (69) Google Scholar). CLE is commonly associated with IFN-1– and IFN-3–driven inflammation, with chronic discoid lupus erythematosus (CDLE) and subacute CLE (SCLE) as frequent subtypes (Wenzel, 2019Wenzel J. Cutaneous lupus erythematosus: new insights into pathogenesis and therapeutic strategies.Nat Rev Rheumatol. 2019; 15: 519-532Crossref PubMed Scopus (69) Google Scholar). Both types of CLE lesions contain infiltrating lymphoid cells in the epithelium, in the dermis, and in particular, at the dermo‒epidermal junction (DEJ), albeit CDLE lesions comprise more infiltrating cells than SCLE lesions (Supplementary Figure S1a). In addition, scarring frequently occurs after CDLE lesion but not after SCLE lesion, causing aesthetic and psychosocial discomfort (McDaniel et al., 2021McDaniel B. Sukumaran S. Tanner L.S. Discoid lupus erythematosus.in: StatPearls [internet]. StatPearls Publishing, Treasure Island, FL2021Google Scholar). Skin homeostasis and wound healing are normally maintained by the epithelial progenitor cell self-renewal and differentiation, facilitated by transient amplifying (TA) cells (Krafts, 2010Krafts K.P. Tissue repair: the hidden drama.Organogenesis. 2010; 6: 225-233Crossref PubMed Scopus (145) Google Scholar). Senescence is an irreversible state of cell cycle arrest, where a lack of proliferation ability is accompanied by apoptosis resistance (Gorgoulis et al., 2019Gorgoulis V. Adams P.D. Alimonti A. Bennett D.C. Bischof O. Bishop C. et al.Cellular senescence: defining a path forward.Cell. 2019; 179: 813-827Abstract Full Text Full Text PDF PubMed Scopus (662) Google Scholar). Senescent cells secrete a panel of proinflammatory cytokines and chemokines, termed the senescence-associated secretory phenotype, factors mitigating the spread of senescence to adjacent cells and potentially augmenting general inflammation (Hernandez-Segura et al., 2018Hernandez-Segura A. Nehme J. Demaria M. Hallmarks of cellular senescence.Trends Cell Biol. 2018; 28: 436-453Abstract Full Text Full Text PDF PubMed Scopus (724) Google Scholar). p16 and p21 are commonly used as senescence markers (Bernardes de Jesus and Blasco, 2012Bernardes de Jesus B. Blasco M.A. Assessing cell and organ senescence biomarkers.Circ Res. 2012; 111: 97-109Crossref PubMed Scopus (116) Google Scholar). We recently showed increased p16+ cells in a progenitor cell niche in the salivary gland of patients with the autoimmune disease primary Sjögren's syndrome, the extent of which correlated with hyposalivation and increased salivary gland lymphocytic infiltration (Wang et al., 2020Wang X. Bootsma H. Terpstra J. Vissink A. van der Vegt B. Spijkervet F.K.L. et al.Progenitor cell niche senescence reflects pathology of the parotid salivary gland in primary Sjögren's syndrome.Rheumatology (Oxford). 2020; 59: 3003-3013Crossref PubMed Scopus (15) Google Scholar). We hypothesized that skin progenitor cells of patients with the autoimmune disease CLE may be senescent. Using immunohistochemistry, we first examined the expression of p16 and p21 in age-matched skin biopsies of patients with CLE lesions (n = 20, comprising 8 CDLE and 12 SCLE) compared with the expression in unaffected skin of patients with CLE as controls (n = 3) (Supplementary Figure S1b and c). Skin biopsies were taken during a routine biopsy for diagnosis and during the establishment of a local pathology database (METc201800245). According to national regulations in the Netherlands (Overheid.nl, 2018Overheid.nlMedical research involving humans act.https://wetten.overheid.nl/BWBR0009408/2018-08-01#Date: 2018Date accessed: June 24, 2021Google Scholar), this type of retrospective noninterventional study with leftover materials for diagnostic purposes does not require approval from the local medical ethical committee. For the analysis of p16+ and p21+ cells, the epidermis was divided into three areas: the basal layer (the progenitor cell niche), the suprabasal layer (the TA cell niche), and the further differentiated (FD) area (terminally differentiated keratinocytes) (Figure 1a). In unaffected skin, small numbers of scattered p16+ or p21+ cells were detected throughout the whole epidermis (Figure 1b, d, e, and g). In CLE lesions, significantly more p16+ cells per mm2 were detected in all the three layers, whereas for p21+ cells, this was only significant for the basal and suprabasal layer and as a trend for the FD area (Figure 1b‒g). Within CLE subtypes, there was a trend for more p16+ cells per mm2 in CDLE than in SCLE lesions in all the three layers of epidermis examined; however, this trend was seen for p21+ cells per mm2 only in the basal and suprabasal layers (Supplementary Figure S1d‒i). Next, to explore the possible influence of inflammation on skin stem cell senescence, we compared (i) CLE with 16 other dermatoses, with varying amounts and distribution of infiltrating cells in the skin. These dermatoses were split into four groups categorized into (ii) other dermatoses with DEJ infiltrates (non-CLE lichenoid dermatoses, n = 22) and dermatoses without DEJ infiltrates, including (iii) non-CLE sunlight induced, n = 7; (iv) spongiotic and psoriasiform, n = 6; and (v) superficial and deep, n = 14. The 18 individual dermatoses comprising a total of five pathological groups are listed in Supplementary Table S1. Skin lesions of non-CLE lichenoid dermatoses contained similar numbers of p16+ and p21+ cells per mm2 as the CLE group in all epidermal layers (Figure 2i and j). The p16+ and p21+ cells per mm2 were higher in CLE and non-CLE lichenoid groups than in all other dermatoses in the basal and suprabasal layers (Figure 2a‒j). In the FD area, CLE and non-CLE lichenoid groups also contain higher numbers of p16+ cells per mm2 than other groups, whereas the spongiotic and psoriasiform group contained the most p21+ cells per mm2 than all other groups (Figure 2i and j). When dermatoses were grouped according to the presence or absence of DEJ infiltration (see Supplementary Table S1), significantly more p16+ and p21+ cells per mm2 in the basal and suprabasal layers were found in dermatoses with DEJ infiltration than in those without DEJ infiltration and in the unaffected skin (Figure 2k and l). In the FD area, more p16+ cells per mm2 were observed in dermatoses with DEJ infiltration than in those without DEJ infiltration and in unaffected skin (Figure 2k and l). p21+ cells per mm2 in the FD area were similar with and without DEJ infiltration, both were higher than in unaffected skin (Figure 2k and l). p16+ and p21+ cells per mm2 in different epidermal layers in individual dermatosis are shown in Supplementary Figure S1d‒i. Across all dermatoses, p16+ and p21+ cells per mm2 were significantly positively correlated in both basal (R = 0.45, P < 0.0001) and suprabasal (R = 0.47, P < 0.0001) layers (Figure 2m). This was not the case in the FD area (R = 0.04, P = 0.76; Supplementary Figure S1j) and in total epidermis (R = 0.21, P = 0.07; Supplementary Figure S1k). Further probing revealed that p16+ cells were more likely in basal layer than in suprabasal layer (P < 0.0001), whereas p21 was conversely enriched in the suprabasal layer compared with that in the basal layer (P < 0.0001) (Figure 2n and o). Interestingly, the amount of basal p16+ cells per mm2 and suprabasal p21+ cells per mm2 were also significantly correlated (R = 0.46, P < 0.0001; Supplementary Figure S1l). Our findings reveal that dermatoses, including the autoimmune disease CLE, with infiltration spatially close to progenitor/TA niches (the DEJ) contain higher numbers of p16+ and p21+ progenitor/TA cells than dermatoses without these infiltrates. In line with this difference, within the CLE group, CDLE lesions, containing denser DEJ infiltration than SCLE lesions, contain more p16+ and p21+ cells. These findings suggest that inflammation and senescence (and senescence-associated secretory phenotype) are at least associated with each other, as has also been shown for other epithelial organs (e.g., salivary gland and intestine) (Coppé et al., 2010Coppé J.P. Desprez P.Y. Krtolica A. Campisi J. The senescence-associated secretory phenotype: the dark side of tumor suppression.Annu Rev Pathol. 2010; 5: 99-118Crossref PubMed Scopus (2385) Google Scholar; Freund et al., 2010Freund A. Orjalo A.V. Desprez P.Y. Campisi J. Inflammatory networks during cellular senescence: causes and consequences.Trends Mol Med. 2010; 16: 238-246Abstract Full Text Full Text PDF PubMed Scopus (832) Google Scholar; Parrinello et al., 2005Parrinello S. Coppe J.P. Krtolica A. Campisi J. Stromal-epithelial interactions in aging and cancer: senescent fibroblasts alter epithelial cell differentiation.J Cell Sci. 2005; 118: 485-496Crossref PubMed Scopus (437) Google Scholar; Pringle et al., 2018Pringle S. Wang X. Verstappen G.M.P.J. Terpstra J.H. Zhang C.K. He A. et al.Salivary gland stem cells age prematurely in primary Sjögren's syndrome.Arthritis Rheumatol. 2018; 71: 133-142Crossref Scopus (28) Google Scholar; Wang et al., 2020Wang X. Bootsma H. Terpstra J. Vissink A. van der Vegt B. Spijkervet F.K.L. et al.Progenitor cell niche senescence reflects pathology of the parotid salivary gland in primary Sjögren's syndrome.Rheumatology (Oxford). 2020; 59: 3003-3013Crossref PubMed Scopus (15) Google Scholar). Considering the proinflammatory nature of the components of the senescence-associated secretory phenotype, it is also possible that inflammation and senescence may augment each other's presence, also an interesting phenomenon to be explored in future studies. The nature of the infiltration in different dermatoses, in terms of immune cell types and their respective secretomes, may also play a significant role in the sum effect of the inflammation on progenitor and TA compartments. Inflammation has also been associated with impaired clearance of apoptotic epithelial cells in CLE and likely contributes significantly to CLE disease development (Kuhn and Bijl, 2008Kuhn A. Bijl M. Pathogenesis of cutaneous lupus erythematosus.Lupus. 2008; 17: 389-393Crossref PubMed Scopus (41) Google Scholar). Furthermore, the extent of inflammatory infiltration is also associated with the likelihood of scarring (Marshall et al., 2018Marshall C.D. Hu M.S. Leavitt T. Barnes L.A. Lorenz H.P. Longaker M.T. Cutaneous scarring: basic science, current treatments, and future directions.Adv Wound Care (New Rochelle). 2018; 7: 29-45Crossref PubMed Scopus (99) Google Scholar; Wilgus, 2020Wilgus T.A. Inflammation as an orchestrator of cutaneous scar formation: a review of the literature.Plast Aesthet Res. 2020; 7: 54PubMed Google Scholar). Whether DEJ infiltration, progenitor cell senescence, or epithelial cell apoptosis contributes equally or not to scar formation in CDLE lesions remains to be established and may dictate the most appropriate treatment regime. In particular, timely removal of senescent cells using senolytics, paired with blockade of chronic inflammation, may promote optimal CDLE lesion resolution. The preferable expression of p16 in the progenitor niche and p21 in the TA niche were also observed in another fast-turnover organ in an autoimmune disease context, namely intestines, from patients with Crohn disease (Wang et al., 2019Wang X. Bootsma H. Kroese F. Dijkstra G. Pringle S. Senescent stem and transient amplifying cells in Crohn's disease intestine.Inflamm Bowel Dis. 2019; 26: e8-e9Crossref Scopus (3) Google Scholar). In our study of the salivary gland (a slow turnover organ) in primary Sjögren's syndrome, only p16 was observed (Wang et al., 2020Wang X. Bootsma H. Terpstra J. Vissink A. van der Vegt B. Spijkervet F.K.L. et al.Progenitor cell niche senescence reflects pathology of the parotid salivary gland in primary Sjögren's syndrome.Rheumatology (Oxford). 2020; 59: 3003-3013Crossref PubMed Scopus (15) Google Scholar). We hypothesize that p21-induced senescence is more likely to occur in epithelial organs containing TA cells, usually fast-turnover tissues, whereas p16 drives a more general epithelial senescence. The different p16/p21 expression profiles in the FD area (this study) among dermatoses may also underpin the differences in p16- or p21-mediated senescence pathway activation, of which the mechanisms remain to be investigated. In conclusion, we propose that dysfunctional epidermal progenitor and TA cells in CLE and other dermatoses with DEJ infiltration may be senescent as a result of close proximity to inflammatory infiltrate and may be hindered in their ability to repair epidermal damage. No datasets were generated or analyzed during this study. Xiaoyan Wang: http://orcid.org/0000-0002-6778-727X Gilles Diercks: http://orcid.org/0000-0001-8053-216X Wietske M. Lambers: http://orcid.org/0000-0002-2026-5191 Johanna Westra: http://orcid.org/0000-0002-6581-6508 Hendrika Bootsma: http://orcid.org/0000-0001-7126-9785 Frans G.M. Kroese: http://orcid.org/0000-0003-3660-0617 Karina de Leeuw http://orcid.org/0000-0003-0218-9364 Sarah Pringle: http://orcid.org/0000-0002-0779-1680 The authors state no conflict of interest. The authors gratefully acknowledge the patients who took part in this research and agreed to provide their skin biopsies for this study. This research was funded by a China Scholarship Council grant (201606220074), a Dutch Arthritis Foundation Translational Research Grant (T015-052), and a Dutch Arthritis Foundation Long Term Project Grant (LLP-29). Conceptualization: SP, XW, GD, KDL; Data Curation: XW; Formal analysis: XW, SP, GD, KDL, FGMK; Methodology: SP, XW, GD, KDL; Writing - Original Draft Preparation: XW, GD, WML, JW, HB, FGMK, KDL, SP; Writing - Review and Editing: XW, GD, WML, JW, HB, FGMK, KDL, SP Supplementary Table S1Grouping of DermatosesDermatosesGroupsMain Infiltration in the DEJ AreaCDLECLEYesSCLEBenign lichenoid keratosisNon-CLE lichenoidEEMLichen planusLichenoid drug eruptionGVHDFixed drug reactionPLEVARosaceaNon-CLE sunlight inducedNoPorphyriaPMLESuperficial and deepUrticariaInsect bitePerniosisEACEczemaSpongiotic and psoriasiformPsoriasisAbbreviations: CDLE, chronic discoid lupus erythematosus; CLE, cutaneous lupus erythematosus; DEJ, dermo‒epidermal junction; EAC, erythema annulare centrifugum; EEM, erythema multiforme; GVHD, graft-versus-host disease; PLEVA, pityriasis lichenoides et varioliformis acuta; PMLE, polymorphous light eruption; SCLE, subacute cutaneous lupus erythematosus. Open table in a new tab Abbreviations: CDLE, chronic discoid lupus erythematosus; CLE, cutaneous lupus erythematosus; DEJ, dermo‒epidermal junction; EAC, erythema annulare centrifugum; EEM, erythema multiforme; GVHD, graft-versus-host disease; PLEVA, pityriasis lichenoides et varioliformis acuta; PMLE, polymorphous light eruption; SCLE, subacute cutaneous lupus erythematosus.