Evaluation of skin expression profiles of patients with vitiligo treated with narrow-band UVB therapy by targeted RNA-seq

Abstract

BackgroundVitiligo is characterized by a lack of pigmentation in the skin. To date, there are no studies that analyze the changes in gene expression in the skin of vitiligo patients in response to narrow-band ultraviolet B (nb-UVB) phototherapy treatment.ObjectiveExplore the usefulness of new generation RNA sequencing in the identification of gene expression changes in the skin of vitiligo patients treated with nb-UVB phototherapy.MethodsFour skin biopsies (4mm in diameter) were collected from 45 Mexican vitiligo vulgaris patients, 2 specimens before and 2 after treatment with nb-UVB phototherapy, obtained from pigmented and non-pigmented tissue. RNA extracted from the biopsies was analyzed using the Illumina TruSeq Targeted RNA Expression protocol to study the expression of genes that participate in pathways of skin homeostasis. The 2 groups were compared using Student's t-test and the Mann-Whitney U-test.ResultsThe expression analysis identified differences in 12 genes included in this study after comparing the samples obtained before and after treatment: 5 genes involved in skin pigmentation, 2 genes involved in apoptosis, 2 genes involved in cell survival, 2 genes involved in oxidative stress responses and 1 gene involved in signal transduction mechanisms (p<0.05).Study limitationsThe small size of skin biopsies limits the amount of RNA obtained, the number of genes to be analyzed and the use of conventional techniques such as RT-qPCR.ConclusionWe demonstrated usefulness of new generation RNA sequencing in the identification of gene expression changes, in addition to identifying new targets in the study of vitiligo.

Transcriptome Analysis and Emerging Driver Identification of CD8+ T Cells in Patients with Vitiligo

“…A cytotoxic role for activated CD8+ T cells mediating the destruction of melanocytes has been well established in vivo and in vitro [7, 10, 41, 42]. In order to decipher the intricate interplay of cellular processes and molecular mechanisms in the pathogenesis of vitiligo, previous studies using transcriptome sequencing have focused on the whole lesional skin and peripheral blood [43–45]. Nevertheless, despite the critical pathogenic role of CD8+ T cells in vitiligo, it is difficult to clarify their precise molecular mechanisms via transcriptome sequencing of whole lesional skin and peripheral blood due to the existence of other cells, such as keratinocytes, fibroblasts, CD4+ T cells, B cells, and macrophages.…”

Role of Isocitrate Dehydrogenase 2, Microphthalmia-Associated Transcription Factor and Alpha Melanocyte Stimulating Hormone in Patients with Non-Segmental Generalized Vitiligo

“…Techniques have been employed to promote repigmentation, such the ultraviolet B light (NB-UVB) narrow band, now regarded as the preferred therapy for vitiligo. This treatment utilises a 311nm light-spectrum that shows penetration in the epidermis' baseline, encourages melanocyte proliferation in the skin and on the external epithelial root sheath of the hair follicle, increases the melanogenesis and the generation of MSH [14].…”

“…Keratinocytes generate and release substances such as αmelanocyte stimulant (α-MSH) that control and accelerate melanocyte epidermal melanogenesis [17]. α-MSH interacts to encourage melanin formation and further specialised melanocyte activities with an unique cell surface receptor called the Melanocortin 1 Receptor (MC1R) [14].…”

TSH levels, overweight, BMI, and skin expression levels of DCT and CCBL2 genes are related to vitiligo treatment response with narrow band UVB phototherapy

“…Two skin biopsies (from vitiligo lesions and pigmented areas) of 4 mm in diameter were obtained before the administration of 48 nb‐UVB phototherapy sessions, performed two times a week for a period of 6 months with an initial dose of 150 mJ/cm 2 , and increments of 50 mJ/cm 2 every third session until the minimum dose of asymptomatic erythema was achieved. Each biopsy was used to isolate total RNA and to perform expression analysis with TruSeq Targeted RNA Expression analysis (RNA‐Seq, Illumina, Inc., San Diego, CA) (Ocampo‐Candiani et al, ), targeting 29 genes involved in skin pigmentation, apoptosis, cell survival, and signal transduction mechanisms.…”