Pemphigus Vulgaris (PV) is a life-threatening autoimmune disease manifested with blisters in the skin and mucosa and caused by autoantibodies against adhesion protein desmoglein-3 (Dsg3) expressed in epithelial membrane linings of these tissues. Despite many studies, the pathogenesis of PV remains incompletely understood. Recently we have shown Dsg3 plays a role in regulating the yes-associated protein (YAP), a co-transcription factor and mechanical sensor, and constraining reactive oxygen species (ROS). This study investigated the effect of PV sera as well as the anti-Dsg3 antibody AK23 on these molecules. We detected elevated YAP steady-state protein levels in PV cells surrounding blisters and perilesional regions and in keratinocytes treated with PV sera and AK23 with concomitant transient ROS overproduction. Cells treated with hydrogen peroxide also exhibited augmented nuclear YAP accompanied by reduction of Dsg3 and α-catenin, a negative regulator of YAP. As expected, transfection of α-catenin-GFP plasmid rendered YAP export from the nucleus evoked by hydrogen peroxide. In addition, suppression of total YAP was observed in hydrogen peroxide treated cells exposed to antioxidants with enhanced cell-cell adhesion being confirmed by decreased fragmentation in the dispase assay compared to hydrogen peroxide treatment alone. On the other hand, the expression of exogenous YAP disrupted intercellular junction assembly. In contrast, YAP depletion resulted in an inverse effect with augmented expression of junction assembly proteins, including Dsg3 and α-catenin capable of abolishing the effect of AK23 on Dsg3 expression. Finally, inhibition of other kinase pathways, including p38MAPK, also demonstrated suppression of YAP induced by hydrogen peroxide. Furthermore, antioxidant treatment of keratinocytes suppressed PV sera-induced total YAP accumulation. In conclusion, this study suggests that oxidative stress coupled with YAP dysregulation attributes to PV blistering, implying antioxidants may be beneficial in the treatment of PV.

• MeSH
• adaptorové proteiny signální transdukční genetika   metabolismus   MeSH
• alfa-katenin metabolismus   MeSH
• antioxidancia farmakologie   terapeutické užití   MeSH
• autoprotilátky krev   imunologie   metabolismus   MeSH
• buněčná adheze účinky léků   imunologie   MeSH
• buněčné linie MeSH
• desmoglein 3 imunologie   metabolismus   MeSH
• genový knockdown MeSH
• keratinocyty MeSH
• lidé MeSH
• MAP kinasový signální systém účinky léků   imunologie   MeSH
• oxidační stres účinky léků   imunologie   MeSH
• pemfigus krev   farmakoterapie   imunologie   patologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• studie případů a kontrol MeSH
• transkripční faktory genetika   metabolismus   MeSH
• ústní sliznice imunologie   patologie   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH

To improve the treatment of psoriasiform inflammation, we developed actively targeted nanocarriers loaded with the phosphodiesterase 4 inhibitor AN2728. Methods: Phospholipid-poly(lactic-co-glycolic acid) nanohybrids were prepared and conjugated with monovalent anti-desmoglein 3 antibody to bind keratinocytes. Results: The actively targeted nanohybrids were 229 nm in mean size with a nearly neutral surface charge. Flow cytometry and confocal microscopy showed a 9-fold increase in keratinocyte uptake of targeted nanohybrids relative to non-targeted nanoparticles. The nanoparticles localized mainly in lysosomes after internalization. AN2728-loaded antibody-conjugated nanocarriers inhibited cytokine/chemokine overexpression in activated keratinocytes without affecting cell viability. The targeted nanohybrids also suppressed neutrophil migration by reducing CXCL1 and CXCL2 release from keratinocytes. Following subcutaneous administration in mice, the nanohybrids distributed to the epidermis and hair follicles. In a psoriasis-like skin mouse model, the actively targeted nanoparticles were superior to free drug and non-targeted nanoparticles in mitigating skin inflammation. Intervention with the targeted nanosystem reduced the epidermal thickness of the psoriasiform lesion from 191 to 42 µm, decreased the Psoriasis Area Severity Index by 74%, restored barrier function, and returned chemokine levels to baseline. Conclusions: Our developed nanosystem was safe and demonstrated efficient targeting properties for the treatment of cutaneous inflammation.

• MeSH
• bicyklické sloučeniny heterocyklické aplikace a dávkování   farmakologie   MeSH
• buněčné linie keratinocytů HaCaT MeSH
• chemokin CXCL1 účinky léků   imunologie   MeSH
• chemokin CXCL2 účinky léků   imunologie   MeSH
• chemotaxe účinky léků   MeSH
• desmoglein 3 imunologie   MeSH
• epidermis MeSH
• fosfolipidy * MeSH
• imunokonjugáty farmakologie   MeSH
• inhibitory fosfodiesterasy 4 aplikace a dávkování   farmakologie   MeSH
• keratinocyty účinky léků   imunologie   MeSH
• kopolymer kyseliny glykolové a mléčné * MeSH
• lékové transportní systémy MeSH
• lidé MeSH
• lyzozomy metabolismus   ultrastruktura   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• nanočástice * MeSH
• neutrofily účinky léků   MeSH
• nosiče léků MeSH
• protilátky imunologie   MeSH
• psoriáza imunologie   patologie   MeSH
• sloučeniny boru aplikace a dávkování   farmakologie   MeSH
• vlasový folikul MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Autoři popisují případ 18letého pacienta trpícího pemphigus vulgaris, u kterého pro protrahovaný průběh léčby, četné exacerbace onemocnění, rozvoj komplikací v rámci dlouhodobé kortikoterapie a neúčinnost předchozí adjuvantní terapie (azathioprin, cyklofosmamid, mykofenolát mofetil), byla zahájena adjuvantní terapie rituximabem. V současné době je popisovaný pacient již 21 měsíců v kompletní remisi onemocnění, jen s podpůrnou terapií osteoporózy. Rituximab je chimérická (myší/lidská) monoklonální protilátka namířená proti povrchovému receptoru CD20 B-lymfocytů, která je nově indikována v první linii celkové terapie pemfigu a která v klinických studiích prokázala dobrou terapeutickou odpověď a steroidy šetřící efekt. Autoři předkládají přehled současných poznatků o léčbě tohoto onemocnění rituximabem.

The authors describes a case of a 18-year-old patient suffering from pemphigus vulgaris in whom adjuvant therapy with rituximab has been initiated due to the protracted course of treatment, frequent exacerbations of the disease, development of multiple complications during long-term systemic corticosteroid therapy and ineffective prior adjuvant therapy (azathioprine, cyclophosphamide, mycophenolate mofetil). Currently, the described patient has been in complete remission of the disease for 21 months only with supportive osteoporosis therapy. Rituximab is a chimeric (murine/human) mono-clonal antibody targeting B cell surface receptor CD20 and has been newly introduced to the first line of systemic therapy of pemphigus and induced good therapeutic response and steroid-sparing effect in clinical trials. The authors present an overview of current knowledge about the treatment of this disease with rituximab.

• Klíčová slova
• CD19 pozitivní B-lymfocyty,
• MeSH
• desmoglein 1 MeSH
• desmoglein 3 MeSH
• hormony kůry nadledvin škodlivé účinky   MeSH
• lidé MeSH
• mladiství MeSH
• osteoporóza etiologie   terapie   MeSH
• pemfigus * terapie   MeSH
• rituximab terapeutické užití   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH

Desmoglein-3 (Dsg3), the Pemphigus Vulgaris (PV) antigen (PVA), plays an essential role in keratinocyte cell-cell adhesion and regulates various signaling pathways involved in the progression and metastasis of cancer where it is upregulated. We show here that expression of Dsg3 impacts on the expression and function of p53, a key transcription factor governing the responses to cellular stress. Dsg3 depletion increased p53 expression and activity, an effect enhanced by treating cells with UVB, mechanical stress and genotoxic drugs, whilst increased Dsg3 expression resulted in the opposite effects. Such a pathway in the negative regulation of p53 by Dsg3 was Dsg3 specific since neither E-cadherin nor desmoplakin knockdown caused similar effects. Analysis of Dsg3-/- mouse skin also indicated an increase of p53/p21WAF1/CIP1 and cleaved caspase-3 relative to Dsg3+/- controls. Finally, we evaluated whether this pathway was operational in the autoimmune disease PV in which Dsg3 serves as a major antigen involved in blistering pathogenesis. We uncovered increased p53 with diffuse cytoplasmic and/or nuclear staining in the oral mucosa of patients, including cells surrounding blisters and the pre-lesional regions. This finding was verified by in vitro studies where treatment of keratinocytes with PV sera, as well as a characterized pathogenic antibody specifically targeting Dsg3, evoked pronounced p53 expression and activity accompanied by disruption of cell-cell adhesion. Collectively, our findings suggest a novel role for Dsg3 as an anti-stress protein, via suppression of p53 function, and this pathway is disrupted in PV.

• MeSH
• desmoglein 3 nedostatek   metabolismus   MeSH
• desmozomy metabolismus   MeSH
• fyziologický stres * MeSH
• kaspasa 3 metabolismus   MeSH
• keratinocyty metabolismus   MeSH
• kultivované buňky MeSH
• kůže metabolismus   MeSH
• leupeptiny farmakologie   MeSH
• lidé MeSH
• myši MeSH
• nádorový supresorový protein p53 metabolismus   MeSH
• pemfigus krev   imunologie   patologie   MeSH
• proteolýza MeSH
• protilátky imunologie   MeSH
• psi MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH