Clinical and laboratory characterization of patients with localized scleroderma and response to UVA-1 phototherapy: In vivo and in vitro skin models.

BACKGROUND/PURPOSE: Localized scleroderma (LS) is a rare disease leading to progressive hardening and induration of the skin and subcutaneous tissues. LS is responsive to UVA-1 phototherapy, though its exact mechanism of action dermal fibrosis is yet to be fully elucidated. We aimed to investigate the molecular changes induced by UVA-1 rays in human primary fibroblasts cultures. METHODS: A total of 16 LS patients were treated with medium-dose UVA-1 phototherapy. At baseline, during and after therapy, Localized Scleroderma Assessment Tool, Dermatology Life Quality Index and lesions' staging and mapping were performed along with high-frequency ultrasound (HFUS) examination for dermal thickness assessment. Gene expression analysis for 23 mRNA transcripts, in vitro UVA-1 irradiation and viability tests were realized on lesional fibroblasts' primary cultures, before and 3 months after therapy. RESULTS: The dermal thickness, the LoSCAT and the DLQI progressively decreased starting from the last phototherapy session up to the 6 and 9 month follow-ups (-57% and -60%, respectively). Molecular gene analysis (rt-PCR) revealed that UVA-1 phototherapy exerts multiple effects: the activation of specific anti-fibrotic pathways (e.g., overexpression of CTHRC1 and metalloproteases 1, 2, 7, 8, 9, 12, suppression of TIMP-1), the downregulation of peculiar pro-fibrotic pathways (e.g., downregulation of TGF-ß, TGF-ßrII, Grb2, SMAD 2/3, TNRSF12A, CTGF) through a significant overexpression of IL-1ß; the stabilization of collagen synthesis acting on genes COL1A1, COL3A1, COL8A1, COL10A1, COL12A1. CONCLUSION: UVA-1 phototherapy adds significant benefits in local tissue remodeling, rebalancing the alteration between pro-fibrotic and anti-fibrotic pathways; these changes can be well monitored by HFUS.

UVA-1 phototherapy as adjuvant treatment for eosinophilic fasciitis: in vitro and in vivo functional characterization.

INTRODUCTION: Eosinophilic fasciitis (EF) is a rare autoimmune disease causing progressive induration of dermal, hypodermal, and muscularis fascia. The exact pathogenesis is yet to be fully understood, and a validated therapy protocol still lacks. We here aimed to realize a clinical-functional characterization of these patients. MATERIALS AND METHODS: A total of eight patients (five males, 45 years average) were treated with adjuvant high-dose UVA-1 phototherapy (90 J/cm), after having received the standard systemic immunosuppressive protocol (oral methylprednisolone switched to methotrexate). Body lesion mapping, Localized Scleroderma Assessment Tool (LoSCAT), Dermatology Life Quality Index (DLQI), High-Resolution Ultrasound (HRUS) (13-17MHz), and ultra HRUS (55-70 MHz) were performed at each examination time taking specific anatomical points. Gene expression analysis at a molecular level and in vitro UVA-1 irradiation was realized on lesional fibroblasts primary cultures. RESULTS: The LoSCAT and the DLQI showed to decrease significantly starting from the last UVA-1 session. A significant reduction in muscularis fascia thickness (-50% on average) was estimated starting from 3 months after the last UVA-1 session and maintained up to 12 months follow-up. Tissues was detected by HRUS. The UVA-1 in vitro irradiation of lesional skin sites cells appeared not to affect their viability. Molecular genes analysis revealed a significant reduction of IL-1ß and of TGF-ß genes after phototherapy, while MMPs 1,2,9 gene expression was enhanced. COMMENT: These preliminary in vivo and in vitro findings suggest that UVA-1 phototherapy is a safe and useful adjuvant therapy able to elicit anti-inflammatory effects and stimulate tissue matrix digestion and remodeling at lesional sites.

Identification of a novel, alpha2-fucosylation-dependent uptake system in highly proliferative cells.

In this paper we describe a new structure present in highly proliferative cells and absent in cells with normal growth potential. We used cultured bovine venular endothelial cells (CVEC) as examples of high proliferation, and dermal fibroblasts of a primary culture as examples of normal proliferation. The structure, consisting of tubules radiating from the nuclear region to the tips of cell protrusions, was revealed by its strong positivity to the fucose-binding lectin from Lotus (LTL) that prefers glycans with alpha-1,2-linked fucose. Another fucose-binding lectin that prefers glycans with alpha-1,6-linked fucose was instead found to localize glycans exclusively in Golgi complexes. LTL binding sites were also found at the surface of CVEC in a restricted region close to the nucleus. The role of alpha-1,2-linked fucose in forming or maintaining the tubules was confirmed by the fact that down-regulation of the fucosyltransferases FUT1 and FUT2 resulted in disappearance of the tubular structure. LTL also proved able to penetrate the cells through the tubular structures up to the nuclear region and to inhibit proliferation. Endostatin was also found to massively penetrate the cells in the tubular structures in control cells but not in FUT1/2 depleted cells. In cells of a first passage primary culture of dermal fibroblasts the tubular LTL-positive structure was absent as well as the LTL-positive sites at the external surface, and both fucose-binding lectins were found to exclusively localize glycans in Golgi complexes. Tubules were again found progressively in fibroblasts derived from repeated passages, where faster growing cells predominate. Disappearance of LTL-positivity in Golgi complexes paralleled appearance of LTL-positive tubules. The role of Golgi complexes in forming the tubules is discussed.