ABSTRACT
Vitiligo is an autoimmune skin disease, characterized by depigmentation and epidermal melanocytes loss. The specific mechanisms underlying vitiligo have not been fully understood. As a result, treating vitiligo is a dermatological challenge. Recently, much attention has been paid to the dysfunction and interaction of organelles under environmental stress. The impaired organelles could generate misfolded proteins, particularly accumulated toxic premelanosome protein (PMEL) amyloid oligomers, activating the autoimmune system and cause melanocyte damage. Unfolded protein response (UPR) dysfunction accelerates toxic PMEL accumulation. Herein, we presented a narrative review on UPR's role in vitiligo, the misfolded PMEL-induced attack of the autoimmune system under autophagy dysfunction caused by abnormal activation of transient receptor potential (TRP) channels and the background of UPR system defects in melanocytes. All of these mechanisms were integrated to form UPR/PMEL-TRP channels/autophagy axis, providing a new understanding of vitiligo pathogenesis.
Subject(s)
Apoptosis , Autophagy , Melanocytes/metabolism , Melanosomes/metabolism , Transient Receptor Potential Channels/metabolism , Vitiligo/metabolism , gp100 Melanoma Antigen/chemistry , gp100 Melanoma Antigen/metabolism , Animals , Humans , Melanocytes/pathology , Melanosomes/pathology , Protein Conformation , Protein Unfolding , Signal Transduction , Skin Pigmentation , Structure-Activity Relationship , Vitiligo/pathologyABSTRACT
Melanosomes, lipofuscin, and melanolipofuscin are the three principal types of pigmented granules found in retinal pigment epithelium (RPE) cells. Changes in the density of melanosomes and lipofuscin in RPE cells are considered hallmarks of various retinal diseases, including Stargardt disease and age-related macular degeneration (AMD). Herein, we report the potential of an in vivo multimodal imaging technique based on directional back-scattering and short-wavelength fundus autofluorescence (SW-FAF) to study disease-related changes in the density of melanosomes and lipofuscin granules in RPE cells. Changes in the concentration of these granules in Abca4-/- mice (a model of Stargardt disease) relative to age-matched wild-type (WT) controls were investigated. Directional optical coherence tomography (dOCT) was used to assess melanosome density in vivo, whereas the autofluorescence (AF) images and emission spectra acquired with a spectrometer-integrated scanning laser ophthalmoscope (SLO) were used to characterize lipofuscin and melanolipofuscin granules in the same RPE region. Subcellular-resolution ex vivo imaging using confocal fluorescence microscopy and electron microscopy was performed on the same tissue region to visualize and quantify melanosomes, lipofuscin, and melanolipofuscin granules. Comparisons between in vivo and ex vivo results confirmed an increased concentration of lipofuscin granules and decreased concentration of melanosomes in the RPE of Abca4-/- mice, and provided an explanation for the differences in fluorescence and directionality of RPE scattering observed in vivo between the two mouse strains.
Subject(s)
ATP-Binding Cassette Transporters/physiology , Melanins/metabolism , Melanosomes/pathology , Multimodal Imaging/methods , Retinal Pigment Epithelium/pathology , Stargardt Disease/pathology , Animals , Mice , Mice, Knockout , Retinal Pigment Epithelium/diagnostic imaging , Retinal Pigment Epithelium/metabolism , Stargardt Disease/diagnostic imagingABSTRACT
Cutaneous melanoma tumors are heterogeneous and show diverse responses to treatment. Identification of robust molecular biomarkers for classifying melanoma tumors into clinically distinct and homogenous subtypes is crucial for improving the diagnosis and treatment of the disease. In this study, we present a classification of melanoma tumors into four subtypes with different survival profiles based on three distinct gene expression signatures: keratin, immune, and melanogenesis. The melanogenesis expression pattern includes several genes that are characteristic of the melanosome organelle and correlates with worse survival, suggesting the involvement of melanosomes in melanoma aggression. We experimentally validated the secretion of melanosomes into surrounding tissues by melanoma tumors, which potentially affects the lethality of metastasis. We propose a simple molecular decision tree classifier for predicting a tumor's subtype based on representative genes from the three identified signatures. Key predictor genes were experimentally validated on melanoma samples taken from patients with varying survival outcomes. Our three-pattern approach for classifying melanoma tumors can contribute to advancing the understanding of melanoma variability and promote accurate diagnosis, prognostication, and treatment.
Subject(s)
Immunity/genetics , Melanins/genetics , Melanoma/genetics , Neoplasm Proteins/genetics , Carcinogenesis/genetics , Female , Gene Expression Regulation, Neoplastic/genetics , Humans , Kallikreins/genetics , Male , Melanins/biosynthesis , Melanoma/classification , Melanoma/pathology , Melanosomes/genetics , Melanosomes/pathology , Muscle Proteins/genetics , Neoplasm Metastasis/genetics , RNA-Seq , Receptors, Immunologic/genetics , Survival Analysis , Transcriptome/genetics , Tripartite Motif Proteins/genetics , Ubiquitin-Protein Ligases/geneticsABSTRACT
Aging may significantly modify antioxidant and photoprotective properties of melanin in retinal pigment epithelium (RPE). Here, photoreactivity of melanosomes (MS), isolated from younger and older human donors with and without added zeaxanthin and α-tocopherol, was analyzed by electron paramagnetic resonance oximetry, time-resolved singlet oxygen phosphorescence, and protein oxidation assay. The phototoxic potential of ingested melanosomes was examined in ARPE-19 cells exposed to blue light. Phagocytosis of FITC-labeled photoreceptor outer segments (POS) isolated from bovine retinas was determined by flow cytometry. Irradiation of cells fed MS induced significant inhibition of the specific phagocytosis with the effect being stronger for melanosomes from older than from younger human cohorts, and enrichment of the melanosomes with antioxidants reduced the inhibitory effect. Cellular protein photooxidation was more pronounced in samples containing older melanosomes, and it was diminished by antioxidants. This study suggests that blue light irradiated RPE melanosomes could induce substantial inhibition of the key function of the cells-their specific phagocytosis. The data indicate that while photoreactivity of MS and their phototoxic potential increase with age, they could be reduced by selected natural antioxidants.
Subject(s)
Antioxidants/pharmacology , Cellular Senescence/radiation effects , Light , Melanosomes/pathology , Melanosomes/radiation effects , Adolescent , Adult , Cell Death/drug effects , Cell Death/radiation effects , Cell Line , Cellular Senescence/drug effects , Humans , Luminescence , Melanosomes/drug effects , Middle Aged , Oxidation-Reduction/radiation effects , Oxygen/metabolism , Phagocytosis/drug effects , Phagocytosis/radiation effects , Stress, Physiological/drug effects , Stress, Physiological/radiation effects , Tissue Donors , Young AdultABSTRACT
A new form of open-angle glaucoma has been identified, in which calcification and silicification of the trabecular meshwork is a potentially significant component of outflow obstruction. It is noted that the mineralization of this area is promoted by various disturbances in the acid-base balance in the tissue. The role of melanosomal enzymes in the initiation of the formation of mineral calcium phosphate in trabecular tissue in open-angle glaucoma is considered.
Subject(s)
Calcinosis/pathology , Glaucoma, Open-Angle/pathology , Trabecular Meshwork/pathology , Acid-Base Imbalance/metabolism , Acid-Base Imbalance/pathology , Biomineralization , Calcinosis/metabolism , Glaucoma, Open-Angle/metabolism , Humans , Melanosomes/metabolism , Melanosomes/pathology , Microscopy, Electron, Scanning/methods , Trabecular Meshwork/metabolismABSTRACT
Melanosomes are specialized membrane-bound organelles that are involved in melanin synthesis. Unlike melanosome biogenesis, the melanosome degradation pathway is poorly understood. Among the cellular processes, autophagy controls degradation of intracellular components by cooperating with lysosomes. In this study, we showed that ursolic acid inhibits skin pigmentation by promoting melanosomal autophagy, or melanophagy, in melanocytes. We found that B16F1 cells treated with ursolic acid suppressed alpha-melanocyte stimulating hormone (α-MSH) stimulated increase in melanin content and activated autophagy. In addition, we found that treatment with ursolic acid promotes melanosomal degradation, and bafilomycin A1 inhibition of autophagosome-lysosome fusion blocked the removal of melanosomes in α-MSH-stimulated B16F1 cells. Furthermore, depletion of the autophagy-related gene 5 (ATG5) resulted in significant suppression of ursolic acid-mediated anti-pigmentation activity and autophagy in α-MSH-treated B16F1 cells. Taken together, our results suggest that ursolic acid inhibits skin pigmentation by increasing melanosomal degradation in melanocytes.
Subject(s)
Autophagy/drug effects , Melanoma, Experimental/pathology , Melanosomes/pathology , Skin Pigmentation/drug effects , Triterpenes/pharmacology , Animals , Cell Line, Tumor , Melanins/biosynthesis , Melanosomes/drug effects , Mice , Triterpenes/chemistry , alpha-MSH/pharmacology , Ursolic AcidABSTRACT
Although pityriasis alba is a common dermatologic condition, its pathogenesis is poorly understood, and there are many discrepancies in the literature. To assess the effect of the duration of disease on the histologic findings, a search of cases labeled "pityriasis alba" was performed on any cases submitted to our dermatopathology laboratory. Of 179 cases of pityriasis alba, five cases identified the duration of the disease, when the biopsy was taken. A biopsy for a lesion of only 1-month duration demonstrated groups of large, prominent melanocytes heaped up upon one another. Compared with biopsies from patients who had the lesions for increasingly longer periods of time, it was apparent that the melanocytes became progressively less abundant and smaller with less prominent dendritic processes. The time that the biopsy is taken may affect the histologic findings of pityriasis alba. Additionally, an abundance of melanosomes was observed between the melanocytes in all sections examined which may reflect a problem with the transfer of melanosomes into keratinocytes in this condition.
Subject(s)
Pityriasis/pathology , Skin/pathology , Adolescent , Biopsy , Child , Female , Humans , Keratinocytes/pathology , Male , Melanocytes/pathology , Melanosomes/pathology , Time FactorsABSTRACT
Cutaneous melanoma arises from melanocytes following genetic, epigenetic and allogenetic (i.e. other than epi/genetic) modifications. An estimated 10% of cutaneous melanoma cases are due to inherited variants or de novo mutations in approximately 20 genes, found using linkage, next-generation sequencing and association studies. Based on these studies, 3 classes of predisposing melanoma genes have been defined based on the frequency of the variants in the general population and lifetime risk of developing a melanoma: (i) ultra-rare variants with a high risk, (ii) rare with a moderate risk, and (iii) frequent variants with a low risk. Most of the proteins encoded by these genes have been shown to be involved in melanoma initiation, including proliferation and senescence bypass. This paper reviews the role(s) of these genes in the transformation of melanocytes into melanoma. It also describes their function in the establishment and renewal of melanocytes and the biology of pigment cells, if known.
Subject(s)
Biomarkers, Tumor/genetics , Melanocytes/pathology , Melanoma/genetics , Mutation , Skin Neoplasms/genetics , Animals , Cell Lineage , Genetic Predisposition to Disease , Humans , Melanins/metabolism , Melanocytes/metabolism , Melanoma/ethnology , Melanoma/metabolism , Melanoma/pathology , Melanosomes/metabolism , Melanosomes/pathology , Mutation Rate , Phenotype , Risk Assessment , Risk Factors , Skin Neoplasms/ethnology , Skin Neoplasms/metabolism , Skin Neoplasms/pathology , White People/geneticsABSTRACT
The pathogenesis of melasma is not fully understood, and the role of skin basement membrane zone (BMZ) alterations in disease development and the maintenance of hypermelanogenesis are also poorly known. We performed a comparative study to characterize the ultrastructural alterations that occur in BMZ in melasma and adjacent normal skin, as well as we discuss the implications of these changes in the physiopathology of the disease. Pairs of facial skin biopsies (2 mm) from 10 women with melasma and normal skin (< 2 cm apart) were processed by Transmission Electronic Microscopy or immunohistochemistry for Melan-A counterstained with Periodic acid-Schiff stain. Cytoplasmic organelles (from keratinocyte or melanocyte), BMZ damage were assessed and melanocyte counting (total and pendulous) was done. There was greater amount of cytoplasmic organelles inside basal keratinocytes and melanocytes in melasma, as well as structural damaged areas in the lamina densa (disruptions, gaps, lower density and thinning) and anchoring fibrils (lamina lucida), compared to healthy adjacent skin. Areas with pendulous melanocytes are characterized by discontinuity of BMZ ultrastructure. The prominence of cytoplasmic organelles from melanocytes and keratinocytes evidences the involvement of both cell groups in melasma. The damage in the lamina densa and lamina lucida suggest the role of upper dermis injury/repair process in the pathogenesis of the disease.
Subject(s)
Basement Membrane/ultrastructure , Face/pathology , Keratinocytes/pathology , Melanocytes/pathology , Melanosis/pathology , Melanosomes/pathology , Skin/ultrastructure , Humans , Immunohistochemistry , Microscopy, Electron, TransmissionABSTRACT
Purpose: Mutations in the megalin-encoding gene, LRP2, cause high myopia as seen in patients suffering from Donnai-Barrow/facio-oculo-acoustico-renal syndrome. Megalin is present in both the nonpigmented epithelium of the ciliary body and in the RPE. In this study, we set out to establish an animal model to study the mechanisms underlying the ocular phenotype and to establish if high myopia/megaophthalmos is induced by postnatal megalin-deficiency in the RPE. Methods: Postnatal RPE-specific deletion of megalin was generated by crossing mice bearing a homozygous loxP-flanked Lrp2 allele with transgenic mice expressing the Cre recombinase driven by the BEST1 promotor. The model was investigated by immunohistologic techniques, and transmission electron microscopy. Results: Mice with postnatal RPE-specific loss of megalin developed a megaophthalmos phenotype with dramatic increase in ocular size and severe retinal thinning associated with compromised vision. This phenotype was present at postnatal day 14, indicating rapid development in the period from onset of BEST1 promotor activity at postnatal day 10. Additionally, RPE melanosomes exhibited abnormal size and morphology, suggested by electron tomography to be caused by fusion events between multiple melanosomes. Conclusions: Postnatal loss of megalin in the RPE induces dramatic and rapid ocular growth and retinal degeneration compatible with the high myopia observed in Donnai-Barrow patients. The morphologic changes of RPE melanosomes, believed to be largely inert and fully differentiated at birth, suggested a continued plasticity of mature melanosomes and a requirement for megalin to maintain their number and morphology.
Subject(s)
Eye Abnormalities/etiology , Low Density Lipoprotein Receptor-Related Protein-2/deficiency , Melanosomes/pathology , Retinal Degeneration/etiology , Retinal Pigment Epithelium/metabolism , Animals , Bestrophins/genetics , Eye Abnormalities/metabolism , Eye Abnormalities/pathology , Female , Integrases/metabolism , Male , Melanosomes/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Retinal Degeneration/metabolism , Retinal Pigment Epithelium/ultrastructureABSTRACT
There is a global trend of increase in the demand for three-dimensional electron microscopy with high resolution. The ultrastructural change and related functional studies are necessary to investigate biological phenomena. In this study, currently available 3D reconstruction techniques of electron microscopes (serial block-face scanning electron microscopy and focused ion beam-scanning electron microscopy) were used to investigate hyperpigmentary disorders in human skin. In the basal layer of the epidermis in the human skin, there are melanocytes that produce melanin and keratinocytes that act as a barrier against environmental damage. The 3D structure from serial images through scanning electron microscopy showed locations of melanosomes between melanocyte and keratinocyte in the hyperpigmentary disorder, in addition, the electron tomography showed pigment transfer through melanin instead melanosome. These results support the exocytosis-endocytosis theory of pigment in human skin.
Subject(s)
Endocytosis/physiology , Hyperpigmentation/pathology , Melanins/metabolism , Pigmentation/physiology , Skin/metabolism , Aged , Female , Humans , Imaging, Three-Dimensional , Keratinocytes/pathology , Male , Melanocytes/pathology , Melanosomes/pathology , Microscopy, Electron, Scanning , Middle Aged , Skin/ultrastructureABSTRACT
The chocolate plumage color in chickens is due to a sex-linked recessive mutation, choc, which dilutes eumelanin pigmentation. Because TYRP1 is sex-linked in chickens, and TYRP1 mutations determine brown coat color in mammals, TYRP1 appeared as the obvious candidate gene for the choc mutation. By combining gene mapping with gene capture, a complete association was identified between the chocolate phenotype and a missense mutation leading to a His214Asn change in the ZnA zinc-binding domain of the protein. A diagnostic test confirmed complete association by screening 428 non-chocolate chickens of various origins. This is the first TYRP1 mutation described in the chicken. Electron microscopy analysis showed that melanosomes were more numerous in feather follicles of chocolate chickens but exhibited an abnormal structure characterized by a granular content and an irregular shape. A similar altered morphology was observed on melanosomes of another TYRP1 mutant in birds, the roux mutation of the quail.
Subject(s)
Hair Color/genetics , Melanosomes/pathology , Mutation, Missense , Oxidoreductases/genetics , Pigmentation Disorders/pathology , Pigmentation/genetics , Animals , Base Sequence , Chickens , Female , Male , Melanosomes/genetics , Phenotype , Pigmentation Disorders/genetics , Sequence HomologyABSTRACT
INTRODUCTION: The effect of melanogenesis intensity on melanoma biology remains an open question, and the biological differences between melanotic and amelanotic melanoma cells have not yet been satisfactorily documented. As a result, the melanization of melanoma cells in in vitro cultures is not considered among experimental procedures. The aim of this study was to investigate the effect of the medium used to culture Bomirski amelanotic Ab melanoma cells on the melanogenesis process. MATERIAL AND METHODS: Amelanotic melanoma cells (Ab) were cultured in two media recommended for in vitro melanoma cell cultures, RPMI and DMEM. The melanization was evaluated by determining the melanin and tyrosinase presence in the cells using spectrophotometrical and western blot methods, respectively. Changes in Ab melanoma cells' ultrastructure were determined using electron microscopy (EM). RESULTS: The medium with higher level of tyrosine (DMEM) induced significant melanization of amelanotic melanoma cells (Ab) after only 24 h, while the RPMI medium, with a lower level of tyrosine, weakly affected melanin production. Melanization of Ab cells was paralleled by an increase in the amount of tyrosinase protein. Induced melanization was easily observed on EM-micrographs in the form of newly formed melanosomes containing melanin pigment. Melanosomes at stages from one (I) to four (IV) were observed. CONCLUSIONS: Culture medium has an important effect on the in vitro biology of amelanotic melanoma cells, since it can affect the rate of cellular melanization. The appropriate medium should be carefully selected, taking into account the known biology of the melanoma cells being used.
Subject(s)
Culture Media/pharmacology , Melanins/biosynthesis , Melanoma/metabolism , Melanosomes/metabolism , Animals , Cell Culture Techniques/methods , Cell Line, Tumor , Cricetinae , Culture Media/chemistry , Melanoma/pathology , Melanosomes/pathologyABSTRACT
Purpose: Oral nitisinone has been shown to increase fur and ocular pigmentation in a mouse model of oculocutaneous albinism (OCA) due to hypomorphic mutations in tyrosinase (TYR), OCA1B. This study determines if nitisinone can improve ocular and/or fur pigmentation in a mouse model of OCA type 3 (OCA3), caused by mutation of the tyrosinase-related protein 1 (Tyrp1) gene. Methods: Mice homozygous for a null allele in the Tyrp1 gene (C57BL/6J-Tyrp1 b-J/J) were treated with 8 mg/kg nitisinone or vehicle every other day by oral gavage. Changes in fur and ocular melanin pigmentation were monitored. Mature ocular melanosome number and size were quantified in pigmented ocular structures by electron microscopy. Results: C57BL/6J-Tyrp1 b-J/J mice carry a novel c.403T>A; 404delG mutation in Tyrp1, predicted to result in premature truncation of the TYRP1 protein. Nitisinone treatment resulted in an approximately 7-fold increase in plasma tyrosine concentrations without overt toxicity. After 1 month of treatment, no change in the color of fur or pigmented ocular structures was observed. The distribution of melanosome cross-sectional area was unchanged in ocular tissues. There was no significant difference in the number of pigmented melanosomes in the RPE/choroid of nitisinone-treated and control groups. However, there was a significant difference in the number of pigmented melanosomes in the iris. Conclusions: Treatment of a mouse model of OCA3 with oral nitisinone did not have a favorable clinical effect on melanin production and minimally affected the number of pigmented melanosomes in the iris stroma. As such, treatment of OCA3 patients with nitisinone is unlikely to be therapeutic.
Subject(s)
Albinism, Oculocutaneous/drug therapy , Cyclohexanones/therapeutic use , Enzyme Inhibitors/therapeutic use , Nitrobenzoates/therapeutic use , Administration, Oral , Albinism, Oculocutaneous/blood , Albinism, Oculocutaneous/genetics , Albinism, Oculocutaneous/pathology , Animals , Blotting, Western , Disease Models, Animal , Genotyping Techniques , Melanins/metabolism , Melanosomes/pathology , Membrane Glycoproteins/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred DBA , Microscopy, Electron, Transmission , Oxidoreductases/genetics , Real-Time Polymerase Chain Reaction , Treatment Outcome , Tyrosine/bloodABSTRACT
Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic disorder that leads to premature aging. In this study, we used induced pluripotent stem cells to investigate the hypopigmentation phenotypes observed in patients with progeria. Accordingly, two iPS cell lines were derived from cells from HGPS patients and differentiated into melanocytes. Measurements of melanin content revealed a lower synthesis of melanin in HGPS melanocytes as compared to non-pathologic cells. Analysis of the melanosome maturation process by electron microscopy revealed a lower percentage of mature, fully pigmented melanosomes. Finally, a functional rescue experiment revealed the direct role of progerin in the regulation of melanogenesis. Overall, these results report a new dysregulated pathway in HGPS and open up novel perspectives in the study of pigmentation phenotypes that are associated with normal and pathological aging.
Subject(s)
Induced Pluripotent Stem Cells , Melanocytes , Melanosomes , Models, Biological , Pigmentation Disorders , Progeria , Humans , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/pathology , Melanocytes/metabolism , Melanocytes/pathology , Melanosomes/metabolism , Melanosomes/pathology , Pigmentation Disorders/metabolism , Pigmentation Disorders/pathology , Progeria/metabolism , Progeria/pathologyABSTRACT
Large-scale sequencing studies have revealed several genes that are recurrently mutated in melanomas. To annotate the melanoma genome, we have expressed tumor-associated variants of these genes in zebrafish and characterized their effects on melanocyte development and function. Here, we describe expression of tumor-associated variants of the recurrently mutated metabotropic glutamate receptor 3 (GRM3) gene. Unlike wild-type GRM3, tumor-associated GRM3 variants disrupted trafficking of melanosomes, causing their aggregation in the cell body. Melanosomes are trafficked in a cAMP-dependent manner, and drugs that directly or indirectly increased cAMP levels were able to suppress melanosome aggregation in mutant GRM3-expressing melanocytes. Our data show that oncogenic GRM3 variants dysregulate cAMP signaling, a heretofore unknown role for these oncogenes. cAMP signaling has been implicated in melanoma progression and drug resistance, and our data show that oncogenic properties of GRM3 could be mediated, at least in part, by alterations in cAMP signaling.
Subject(s)
Cyclic AMP/metabolism , Genetic Variation , Melanocytes/pathology , Melanoma/pathology , Melanosomes/pathology , Receptor, Metabotropic Glutamate 5/genetics , Zebrafish/metabolism , Animals , Humans , Melanocytes/metabolism , Melanoma/genetics , Melanoma/metabolism , Melanosomes/metabolism , Receptor, Metabotropic Glutamate 5/metabolism , Signal Transduction , Zebrafish/growth & developmentSubject(s)
Cerebral Ventricle Neoplasms/pathology , Neurocytoma/pathology , Humans , Male , Melanosomes/pathology , Pigmentation , Young AdultABSTRACT
In this review, I will discuss how careful scrutiny of genetic skin disorders could help us to understand human biology. Like other organs, the skin and its appendages, such as hairs and teeth, experience fundamental biological processes ranging from lipid metabolism to vesicular transport and cellular migration. However, in contrast to other organ systems, they are accessible and can be studied with relative ease. By visually revealing the functional consequences of single gene defects, genetic skin diseases offer a unique opportunity to study human biology. Here, I will illustrate this concept by discussing how human genetic disorders of skin pigmentation reflect the mechanisms underlying this complex and vital process.