Melanocyte Adhesion Research Articles

Role of Cytokines and Chemokines in Vitiligo and Their Therapeutic Implications.

Vitiligo is a persistent autoimmune disease characterized by progressive depigmentation of the skin caused by the selective destruction of melanocytes. Although its etiopathogenesis remains unclear, multiple factors are involved in the development of this disease, from genetic and metabolic factors to cellular oxidative stress, melanocyte adhesion defects, and innate and adaptive immunity. This review presents a comprehensive summary of the existing knowledge on the role of different cellular mechanisms, including cytokines and chemokines interactions, in the pathogenesis of vitiligo. Although there is no definitive cure for vitiligo, notable progress has been made, and several treatments have shown favorable results. A thorough understanding of the basis of the disease uncovers promising drug targets for future research, providing clinical researchers with valuable insights for developing improved treatment options.

Extraciliary OFD1 Is Involved in Melanocyte Survival through Cell Adhesion to ECM via Paxillin.

Primary cilia play a significant role in influencing cell fate, including apoptosis in multiple cell types. In the lesional epidermis of vitiligo patients, a reduced number of ciliated cells was observed. Our study also revealed a downregulation of oral-facial digital syndrome type 1 (OFD1) in the affected skin of vitiligo patients. However, it remains unknown whether primary cilia are involved in the control of melanocyte apoptosis. While both intraflagellar transport 88 (IFT88) and retinitis pigmentosa GTPase regulator-interacting protein-1 like (RPGRIP1L) are associated with ciliogenesis in melanocytes, only the knockdown of OFD1, but not IFT88 and RPGRIP1L, resulted in increased melanocyte apoptosis. OFD1 knockdown led to a decrease in the expression of proteins involved in cell-extracellular matrix (ECM) interactions, including paxillin. The OFD1 amino acid residues 601-1012 interacted with paxillin, while the amino acid residues 1-601 were associated with ciliogenesis, suggesting that the OFD1 domains responsible for paxillin binding are distinct from those involved in ciliogenesis. OFD1 knockdown, but not IFT88 knockdown, inhibited melanocyte adhesion to the ECM, a defect that was restored by paxillin overexpression. In summary, our findings indicate that the downregulation of OFD1 induces melanocyte apoptosis, independent of any impairment in ciliogenesis, by reducing melanocyte adhesion to the ECM via paxillin.

Melanocyte-keratinocyte cross-talk in vitiligo.

Vitiligo is a common acquired pigmentary disorder that presents as progressive loss of melanocytes from the skin. Epidermal melanocytes and keratinocytes are in close proximity to each other, forming a functional and structural unit where keratinocytes play a pivotal role in supporting melanocyte homeostasis and melanogenesis. This intimate relationship suggests that keratinocytes might contribute to ongoing melanocyte loss and subsequent depigmentation. In fact, keratinocyte dysfunction is a documented phenomenon in vitiligo. Keratinocyte apoptosis can deprive melanocytes from growth factors including stem cell factor (SCF) and other melanogenic stimulating factors which are essential for melanocyte function. Additionally, keratinocytes control the mobility/stability phases of melanocytes via matrix metalloproteinases and basement membrane remodeling. Hence keratinocyte dysfunction may be implicated in detachment of melanocytes from the basement membrane and subsequent loss from the epidermis, also potentially interfering with repigmentation in patients with stable disease. Furthermore, keratinocytes contribute to the autoimmune insult in vitiligo. Keratinocytes express MHC II in perilesional skin and may present melanosomal antigens in the context of MHC class II after the pigmented organelles have been transferred from melanocytes. Moreover, keratinocytes secrete cytokines and chemokines including CXCL-9, CXCL-10, and IL-15 that amplify the inflammatory circuit within vitiligo skin and recruit melanocyte-specific, skin-resident memory T cells. In summary, keratinocytes can influence vitiligo development by a combination of failing to produce survival factors, limiting melanocyte adhesion in lesional skin, presenting melanocyte antigens and enhancing the recruitment of pathogenic T cells.

Characteristics and pathogenesis of Koebner phenomenon.

The Koebner phenomenon, also known as isomorphic reaction, refers to the development of secondary lesions with the same clinical manifestations and histopathological characteristics as the primary lesions in normal skin after trauma or other stimuli. The triggering factors of Koebner phenomenon include physical trauma, chemical stimulation, mechanical stress, iatrogenic stimulation and pathogenic infection. Vitiligo, psoriasis and lichen planus are considered true Koebner phenomenon. Recent studies have shown that immunological disorders, oxidative stress, defective melanocyte adhesion and growth factor deficiency are the main pathological mechanisms of vitiligo Koebner phenomenon. In psoriasis, triggers may drive skin inflammation to induce a psoriatic phenotype through multiple signalling pathways and thereby cause Koebner phenomenon in susceptible individuals. Significantly, keratinocytes mediate the occurrence of Koebner phenomenon in psoriasis through mechano-induced signalling pathways after sensing mechanical signals and explains the high frequency of psoriasis lesions on the extensor side of the elbow and knee joints. On the contrary, TRPA1-driven mechano-transduction, autoimmunity and actinic damage are the underlying mechanisms of Koebner phenomenon in lichen planus. In this review, we have summarized the current understanding of the characteristics and pathogenesis of Koebner phenomenon.

32053 Vitiligo koebnerising to varicose veins of the lower limbs

A 29-year-old woman with Fitzpatrick skin type II noticed new white patches on her lower limbs after reporting a sudden onset of pain behind her knees along with the sudden appearance of bilateral varicose veins extending from the thighs down to the calves and shins. Subsequently she developed symmetrically distributed white patches in the submammary and groin areas. Examination of the affected areas revealed depigmented macular patches without scale, atrophy or induration in keeping with vitiligo. Strikingly, the white patches on the legs were confined to the prominent varicosities, consistent with koebnerisation of vitiligo to the skin overlying the varicose veins. Investigation of the pain behind the knees was unremarkable, leading to a likely diagnosis of symptomatic varicose veins. Koebner phenomenon, also called the isomorphic response, describes the development of lesions at sites of traumatised previously uninvolved skin, and is well-known to occur in vitiligo. The pathogenesis of this is thought to be an integration of immunologic mechanisms, deficiency of melanocyte growth factors, defective melanocyte adhesion, and increased oxidative stress. To our knowledge, this is only the second case reported in the literature of vitiligo koebnerising to areas of varicosities.

Inducible Nitric Oxide Synthase iNOS-954-G>C and Ex16+14-C>T Gene Polymorphisms and Susceptibility to Vitiligo in the Saudi Population.

ObjectiveVitiligo is an acquired pigmentary skin disorder with regional disappearance of melanocytes. Multigenic inheritance has been proposed in the pathogenesis of vitiligo. The present study aimed to investigate the possible association of inducible nitric oxide synthase polymorphisms iNOS-954-G/C (rs1800482 G>C) and iNOS-Ex16+14-C/T (rs2297518 C>T) with vitiligo in the Saudi population, if any.MethodsWe included 120 vitiligo cases and an equal number of age matched healthy controls. Polymerase chain reaction with restriction fragment length polymorphism method was used for the analysis of genetic polymorphisms.ResultsThe heterozygous (GC), (GC + CC) combined genotype and variant allele; C allele of rs1800482 G>C were associated significantly (p < 0.005, after Bonferroni correction) with increased risk of vitiligo (OR = 3.46, 95% CI = 1.99–6.01, p = 0.001), (OR = 3.30, 95% CI= 1.93–5.65, p = 0.001) and (OR = 1.94, 95% CI = 1.31–2.87, p = 0.001) respectively. When GC genotype of rs1800482 G>C was co-inherited with common genotype (CC) and heterozygous genotype (CT) of rs2297518 C>T, the risk of vitiligo was significantly increased ((OR = 4.51, 95% CI = 2.18–9.33, p = 0.001) and (OR = 3.60, 95% CI = 1.61–8.01, p = 0.001)) respectively. None of the rs1800482 G>C and rs2297518 C>T genotypes and alleles have been associated with non-segmental vitiligo in terms of gender, age of onset, and types of vitiligo.ConclusionThe heterozygous (GC), (GC+CC) combined genotype and variants allele; C allele of rs1800482 G>C, may cause overproduction of NO, which has been linked to melanocyte loss by increasing oxidative stress and decreasing melanocyte adhesion to the extracellular matrix components, and thus could be an associative risk factor for vitiligo.

Identification, Isolation, and Characterization of Melanocyte Precursor Cells in the Human Limbal Stroma.

Given their vital role in the homeostasis of the limbal stem cell niche, limbal melanocytes have emerged as promising candidates for tissue engineering applications. This study aimed to isolate and characterize a population of melanocyte precursors in the limbal stroma, compared with melanocytes originating from the limbal epithelium, using magnetic-activated cell sorting (MACS) with positive (CD117/c-Kit microbeads) or negative (CD326/EpCAM or anti-fibroblast microbeads) selection approaches. Both approaches enabled fast and easy isolation and cultivation of pure limbal epithelial and stromal melanocyte populations, which differed in phenotype and gene expression, but exhibited similar functional properties regarding proliferative potential, pigmentation, and support of clonal growth of limbal epithelial stem/progenitor cells (LEPCs). In both melanocyte populations, limbus-specific matrix (laminin 511-E8) and soluble factors (LEPC-derived conditioned medium) stimulated melanocyte adhesion, dendrite formation, melanogenesis, and expression of genes involved in UV protection and immune regulation. The findings provided not only a novel protocol for the enrichment of pure melanocyte populations from limbal tissue applying easy-to-use MACS technology, but also identified a population of stromal melanocyte precursors, which may serve as a reservoir for the replacement of damaged epithelial melanocytes and an alternative resource for tissue engineering applications.

Dopamine toxicity contributes to melanocyte loss via melanocytorrhagy: an invitro study.

Catecholamines (epinephrine, norepinephrine, and dopamine) have been proposed as a possible cause of melanocyte loss. Dopamine has been observed to cause apoptosis in melanocytes via reactive oxygen species development, but the effect on melanocyte adhesion and proliferation still remains to be elucidated. Thus, we explored the dose- and time-dependent toxicity of catecholamines and the effect of dopamine on the proliferation and adhesion potential of melanocytes. Primary culture of melanocytes was investigated in vitro for toxic effects of epinephrine, norepinephrine, and dopamine on metabolic activity via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide) assay. Cells were observed microscopically for any phenotypic changes. Further cell proliferation, cell adhesion, and cell death pathway were explored under dopamine toxicity in dose- and time-dependent manner with RNase/PI method of cell cycle analysis, cell adhesion assay, and Annexin V-FITC/PI assay, respectively. Altered gene expressions were confirmed with a real-time polymerase chain reaction. Metabolic activity of cells varied with time and different doses of epinephrine, norepinephrine, and dopamine. Dopamine was observed to be more toxic than epinephrine and norepinephrine. Melanocytes were observed to follow different cell death pathways at comparatively lower and higher concentrations of dopamine. Persistent exposure to dopamine resulted in decreased cell proliferation and adhesion potential with apoptotic changes. Gene expression changes also confirmed the weak adhesion and survival potential of cells under the toxic effects of dopamine. Dopamine can alter melanocytes' adhesion and survival potential, leading to apoptotic cell death or melanocytorrhagic loss.

Periorificial lesions associated with poor prognosis in patients with localized vitiligo: A retrospective study of 126 patients

Periorificial lesions associated with poor prognosis in patients with localized vitiligo: A retrospective study of 126 patients

Macrophage Migration Inhibitory Factor in Vitiligo: Pathogenesis and Potential Therapeutic Aspects

Vitiligo is an acquired skin disorder that is characterized by a gradual loss of skin pigmentation when melanocytes, the skin's pigment-producing cellsis lost. Pathogenicmechanisms are not well understood. Genetic, abnormal biochemical pathways, autoimmune, melanocyte adhesion deficits and nervous system imbalances are among the pathogenic triggers. Vitiligo lesions have also been shown to have macrophage infiltration.Macrophagemigration inhibitory factor (MIF) is a lymphokine that concentrates macrophages at inflammatory sites and is involved in cell-mediated immunity.MIF enhances chemotaxis and macrophage infiltration and upregulates inflammatory responses by inducing the expression of proinflammatory mediators such as TNF-α, nitric oxide and prostaglandin E2.Therapyfor vitiligo includes corticosteroids, immunomodulatory agents, vitamin D nalogues, antioxidants, phototherapy, laser and surgical therapy.However, no single treatment for vitiligo produces consistently good results and treatment response is variable. Narrow-band ultraviolet (NB-UVB, 311–313nm) phototherapy is viewed as backbone of treatment. Systemic therapies such as systemic corticosteroids and methotrexate were previously used to treat vitiligo which was assumed to be auto-immune nature.Pathogenic mechanisms, role of MIF and various treatment guidelines are discussed in this review

Translational Research in Vitiligo.

Vitiligo is a disease of the skin characterized by the appearance of white spots. Significant progress has been made in understanding vitiligo pathogenesis over the past 30 years, but only through perseverance, collaboration, and open-minded discussion. Early hypotheses considered roles for innervation, microvascular anomalies, oxidative stress, defects in melanocyte adhesion, autoimmunity, somatic mosaicism, and genetics. Because theories about pathogenesis drive experimental design, focus, and even therapeutic approach, it is important to consider their impact on our current understanding about vitiligo. Animal models allow researchers to perform mechanistic studies, and the development of improved patient sample collection methods provides a platform for translational studies in vitiligo that can also be applied to understand other autoimmune diseases that are more difficult to study in human samples. Here we discuss the history of vitiligo translational research, recent advances, and their implications for new treatment approaches.

Type-1 cytokines regulate MMP-9 production and E-cadherin disruption to promote melanocyte loss in vitiligo.

Loss of melanocytes is the pathological hallmark of vitiligo, a chronic inflammatory skin depigmenting disorder induced by exaggerated immune response, including autoreactive CD8 T cells producing high levels of type 1 cytokines. However, the interplay between this inflammatory response and melanocyte disappearance remains to be fully characterized. Here, we demonstrate that vitiligo skin contains a significant proportion of suprabasal melanocytes, associated with disruption of E-cadherin expression, a major protein involved in melanocyte adhesion. This phenomenon is also observed in lesional psoriatic skin. Importantly, apoptotic melanocytes were mainly observed once cells were detached from the basal layer of the epidermis, suggesting that additional mechanism(s) could be involved in melanocyte loss. The type 1 cytokines IFN-γ and TNF-α induce melanocyte detachment through E-cadherin disruption and the release of its soluble form, partly due to MMP-9. The levels of MMP-9 are increased in the skin and sera of patients with vitiligo, and MMP-9 is produced by keratinocytes in response to IFN-γ and TNF-α. Inhibition of MMP-9 or the JAK/STAT signaling pathway prevents melanocyte detachment in vitro and in vivo. Therefore, stabilization of melanocytes in the basal layer of the epidermis by preventing E-cadherin disruption appears promising for the prevention of depigmentation occurring in vitiligo and during chronic skin inflammation.

533 Type-1 cytokines induce depigmentation through JAK/STAT signaling, MMP9 production and disruption of E-cadherin expression

Depigmentation observed in vitiligo results from the loss of epidermal melanocytes, involving the combination of several factors, including exaggerated inflammatory and immune response with overproduction of the type-1 cytokines interferon (IFN)-g and tumor necrosis factor (TNF)-a. Nonetheless, the precise mechanism leading to such disappearance remains controversial. We used ex vivo, in vitro and in vivo approaches to decipher the interplay between type-1 cytokines and melanocyte loss. We show that basal melanocytes from vitiligo patients skin are not apoptotic but suffer from a disruption of E-cadherin expression, the main protein involved in melanocyte adhesion to keratinocytes, explaining the presence suprabasal melanocytes. Importantly, we found that type-1 cytokines IFNg and TNFa reproduced this phenotype both in vitro in a 3D model of human pigmented epidermis, and vivo in mice. Such effect was mediated through the promotion of epidermal production of matrix metalloprotease MMP9, resulting in disruption of E-cadherin expression and subsequent destabilization of melanocytes from the basal layer of the epidermis. In addition, inhibition of MMP9 or of JAK signaling prevented such detachment of melanocytes. Therefore, preventing E-cadherin disruption appears as an attractive therapeutic approach to stabilize melanocytes in the basal layer of the epidermis and prevent depigmentation.

Vitiligo: An Update on Pathophysiology and Treatment Options.

The pathophysiology of vitiligo is becoming increasingly clarified. In non-segmental vitiligo, early factors include activation of innate immunity, inflammasome activation, oxidative stress, and loss of melanocyte adhesion. Nonetheless, the main mechanism leading to non-segmental vitiligo involves an immune-mediated destruction of melanocytes. Anti-melanocyte-specific cytotoxic T cells exert a central role in the final effector stage. Genetic research revealed a multi-genetic inheritance displaying an overlap with other autoimmune disorders. However, some melanocyte-specific genes were also affected. Segmental vitiligo carries a different pathogenesis with most evidence indicating a mosaic skin disorder. Current management includes topical corticosteroids and immunomodulators. Narrow-band ultraviolet B can be used in patients not responding to topical treatment or in patients with extensive disease. Pigment cell transplantation offers an alternative for the treatment of segmental vitiligo or stable non-segmental lesions. Recent findings have revealed new targets for treatment that could lead to more efficient therapies. Targeted immunotherapy may halt the active immune pathways, although combination therapy may still be required to induce satisfying repigmentation. A recently established core set of outcome measures, new measurement instruments, and biomarker research pave the way for future standardized clinical trials.

Altered levels of LXR‐α: crucial implications in the pathogenesis of vitiligo

In recent times, the role of LXRs in skin physiology and pathology has evolved rapidly because of their role in proliferation, carcinogenesis, differentiation and permeability barrier function. LXRs were identified as promising drug targets for the treatment of many skin diseases. For this study, skin biopsies were taken from 15 patients with vitiligo and six controls to culture melanocytes from clinically active perilesional and normal skin. Gene expression was examined by reverse transcriptase-polymerase chain reaction analysis. Role of LXR-α in regulating the expression of MMPs was checked by gene knock-down, and its role in vitiligo pathogenesis was checked by treatment with LXR-α agonist 22(R)-hydroxycholesterol. After treatment adhesion assay, annexin V staining and proliferation assay were performed. The expression of LXR-α was relatively more in perilesional skin melanocytes as compared to uninvolved skin melanocytes of non-segmental vitiligo patient, and controls on the other hand, perilesional melanocytes were more prone to apoptosis. LXR-α gene knock-down significantly increases the expression of MMPs. LXR-α agonist 22(R)-hydroxycholesterol treatment significantly decreases melanocyte adhesion, apoptosis and proliferation. Higher expression of LXR-α in perilesional skin melanocytes significantly decreases the adhesion, proliferation and matrix metalloproteinases and increases apoptosis.

Membrane‐bound Kit ligand regulates melanocyte adhesion and survival, providing physical interaction with an intraepithelial niche

Morphogenesis, as illustrated by melanocyte migration and homing to the skin, requires cadherin adhesion, integrin-dependent migration and Kit-ligand growth factor signaling. However, it is not known how Kit ligand regulates integrin or cadherin-dependent intraepidermal melanocyte behavior. To answer this question, we developed specific 2-dimensional (2D) and 3D culture systems analyzing how soluble or immobilized Kit-ligand-regulated melanocyte migration on vitronectin and laminin, or within a monolayer of kidney epithelial cells. In a 2D system, soluble Kit ligand stimulated integrin-dependent melanoblast migration and chemotaxis and accelerated integrin turnover. In contrast, immobilized, but not soluble, Kit ligand, enhanced integrin-dependent melanocyte spreading on suboptimal laminin concentrations. In 3D, membrane-bound Kit ligand induced intraepithelial melanocyte proliferation, survival, and tight adhesion to epithelial cells, while cleavable Kit ligand was less effective. In contrast, melanocyte motility was independent of membrane-bound Kit ligand, but increased in the presence of the cleavable Kit-ligand isoform. Transmembrane-dimerization or basolateral-targeting mutants of Kit ligand altered intraepithelial melanocyte localization, survival, and adhesion to epithelial cells. These data and the identification of c-kit/Kit-ligand clusters at cell contacts suggest that membrane-bound Kit ligand captures cell surface-expressed c-kit, providing mechanical anchoring and survival signaling within intraepithelial niches, and thereby defining a new mechanism for melanocyte homeostasis and requirement for environmental niches.

Study of CCN3 (NOV) and DDR1 in normal melanocytes and vitiligo skin

We have hypothesised that melanocytes disappear in vitiligo because they are weakly attached to the epidermal basal membrane (melanocytorrhagy). In the epidermis, attachment of melanocytes to collagen IV is mediated through DDR1, which is under the control of CCN3. DDR1 genetic variants have been associated with vitiligo in patients of different ethnic origin. In vitro studies have shown that inhibition of CCN3 induces the detachment of melanocytes. We have studied in parallel the expression of CCN3 and DDR1 in lesional and perilesional skin of patients with vitiligo and the impact of the silencing of CCN3 and DDR1 in normal human melanocytes on their behaviour in epidermal reconstructs. Our in vivo study provides evidence of a dysregulation of the DDR1-CCN3 interaction in vitiligo skin as melanocytes remaining in perilesional skin did not express CCN3. Expression of DDR1 was decreased in lesional versus perilesional vitiligo skin in the majority of patients, and the expression of collagen IV was found decreased in all patients. Silencing of CCN3 in melanocytes induced a significant inhibition of cell adhesion to collagen IV whereas melanocytes transduced with shDDR1 still adhered well on collagen IV and did not increase melanocyte loss in epidermal reconstructs as compared with normal melanocytes. Melanocyte detachment was observed but not in all reconstructs using CCN3 silenced melanocytes. Overall, our study confirms that a downregulation of CCN3 is implicated in melanocyte adhesion in part through DDR1. In vitiligo skin, the interaction of CCN3 with other molecules, such as TGFβ and CCN2, needs to be addressed.

Role of apoptosis and melanocytorrhagy: a comparative study of melanocyte adhesion in stable and unstable vitiligo

Role of apoptosis and melanocytorrhagy: a comparative study of melanocyte adhesion in stable and unstable vitiligo

Integrins: versatile receptors controlling melanocyte adhesion, migration and proliferation

From the onset of melanocyte specification from the neural crest, throughout their migration during embryogenesis and until they reside in their niche in the basal keratinocyte layer, melanocytes interact in dynamic ways with the extracellular environment of the growing embryo. To recognize and to adhere to their environment, melanocytes depend on heterodimeric cell surface receptors of the family of integrins. In addition to the control of adhesive interactions between melanocytes and the extracellular matrix scaffold secreted by fibroblasts and keratinocytes, the integrin receptors allow cells also to sense the mechanical condition of the extracellular environment, responding by intracellular signaling, triggering cell survival, proliferation or migration events. In this review, we summarize the recently emerged concepts that explain integrin-dependent adhesion and how this adhesion system interfaces with integrin-dependent signaling events. The gained information will help to understand melanocyte behavior in pathological situations such as melanoma growth and metastasis formation.

Role of apoptosis and melanocytorrhagy: a comparative study of melanocyte adhesion in stable and unstable vitiligo

Apoptosis and melanocytorrhagy have been proposed as mechanisms of melanocyte disappearance although there are few controlled studies. We undertook this project to study melanocyte morphology and adhesion defects in patients with stable and unstable disease in controls. In this comparative study we included seven patients with stable disease and seven patients with unstable vitiligo. We cultured perilesional skin melanocytes from these patients with stable and unstable vitiligo and studied for morphological changes, adhesion to collagen type IV and caspase 3 expression. Melanocytes were also treated with okadaic acid and annexin V expression was then checked and compared between controls and patients with stable and unstable vitiligo. Perilesional skin melanocytes from patients with unstable vitiligo revealed some significant morphological changes. Melanocytes from unstable vitiligo showed significantly low adhesion to collagen type IV compared with control and stable vitiligo melanocytes. Our results showed that caspase 3 and annexin V staining was significantly greater in melanocytes cultured from unstable vitiligo compared with the control. In this study we demonstrated that melanocytes in the patients with unstable vitiligo were in their detachment phase, which ultimately leads to apoptosis of these cells, whereas melanocytes cultured from controls and from patients with stable vitiligo were morphologically normal without any adhesion defects. These morphological and adhesion findings support the theory of melanocytorrhagy as the primary defect underlying melanocyte loss in unstable vitiligo.