PhD Scientific Days 2020

Molecular Sciences V. Lectures

Nonlinear Optical Microscopy as a Novel Tool for the Analysis of Cutaneous Alterations in Pseudoxanthoma Elasticum

Előadó neve

Dr. Fésüs, Luca

Előadó munkahelye

Department of Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary

Előadó e-mail címe

luca.fesus@gmail.com

Az előadás címe

Nonlinear Optical Microscopy as a Novel Tool for the Analysis of Cutaneous Alterations in Pseudoxanthoma Elasticum

Szerző(k) neve és munkahelye

Luca Fésűs1,2, Norbert Kiss1,2, Szabolcs Bozsányi1,2, Flóra Szeri3, Matthias Van Gils4,5, Róbert Szipőcs2,8, Ludovic Martin9, Olivier Vanakker4,5, Tamás Arányi10, Béla Merkely7, Norbert M Wikonkál1* and Márta Medvecz1*
1 Department of Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary
2 Wigner RCP, Institute for Solid State Physics and Optics, Hungarian Academy of Sciences, Budapest, Hungary
3 The PXE International Center of Excellence in Research and Clinical Care, Department of Dermatology and Cutaneous Biology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania
4 Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium
5 Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
7 Heart and Vascular Center, Semmelweis University, Budapest, Hungary
8 R&D Ultrafast Lasers Ltd, Budapest, Hungary
9 PXE Reference Center (MAGEC Nord), Angers University Hospital, Angers, France
10 Research Center for Natural Sciences, Institute of Enzymology, Hungarian Academy of Sciences, Budapest, Hungary

Szekció

Molecular Sciences V. Lectures

Language of the presentation

English

Section, first choice

Clinical Medicine

Section, second choice

Molecular Sciences

Összefoglaló szövege

Introduction: Pseudoxanthoma elasticum (PXE, OMIM#264800) is a rare, genetic disease with an autosomal recessive inheritance. PXE is caused by mutations in the ABCC6 gene, which result in decreased serum levels of inorganic pyrophosphate (PPi). As a consequence of low PPi levels, ectopic mineralization and fragmentation of elastic fibers develop in the skin, the endothelium of blood vessels and the Bruch membrane of the eye. PXE is often diagnosed only after severe ophthalmological and cardiovascular complications occur. Multidisciplinary clinical examination, histopathologic evaluation of skin biopsy, in addition to molecular genetic testing is needed for proper diagnosis. Nonlinear optical (NLO) microscopy is an innovative non-invasive imaging technology that is increasingly applied in life sciences. NLO processes, generated by ultrafast laser pulses provide stain-free imaging with submicron resolution and higher penetration depth compared to reflectance confocal microscopy. Fibrillar collagen, as being a highly non-centrosymmetric molecule emits strong second harmonic generation (SHG) signal. Elastin fibers and calcium deposits, as endogenous chromophores can be selectively visualized by two-photon fluorescence (TPF).
Methods: In five patients with PXE we performed deep phenotyping, molecular genetic testing, ophthalmological and cardiovascular assessment. In addition, plasma PPi measurements, quantitative ex vivo SHG and TPF imaging was carried out and compared to five healthy controls. Histological assessment was carried out on hematoxylin and eosin, van Gieson, Weigert’s elastic and von Kossa stained slides.
Results: Pathological mutations were found in the ABCC6 gene in all patients. Their plasma PPi level was found significantly lower compared to controls (0.232±0.063µM vs 0.947±0.108µM). Conventional histology confirmed characteristic histological alterations in PXE. NLO displayed significantly more (175.7±79.49 vs 97.56±29.30) and shorter (59d.49±0.66µm vs 91.64±5.6µm) elastin fibers, while collagen fibers were shorter (67.99±0.79µm vs 73.53±1.04µm) and thinner (6.8±0.06µm vs 7.33±0.07µm).
Conclusion: Our data suggest, that NLO enables high-resolution imaging and quantification of the specific features of PXE-affected skin. In vivo NLO may be a promising method in the assessment of PXE facilitating early diagnosis and follow-up.

Additional Information

Supervizor:
Prof.Dr. Wikonkál Norbert, Department of Dermatology, Venereology and Dermatooncology, Semmelweis University,

Bemutatás módja

Szóbeli

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

5031

Start

18:00

End

18:15

Authors (legacy)

Luca Fésűs1,2, Norbert Kiss1,2, Szabolcs Bozsányi1,2, Flóra Szeri3, Matthias Van Gils4,5, Róbert Szipőcs2,8, Ludovic Martin9, Olivier Vanakker4,5, Tamás Arányi10, Béla Merkely7, Norbert M Wikonkál1* and Márta Medvecz1*
1 Department of Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary
2 Wigner RCP, Institute for Solid State Physics and Optics, Hungarian Academy of Sciences, Budapest, Hungary
3 The PXE International Center of Excellence in Research and Clinical Care, Department of Dermatology and Cutaneous Biology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania
4 Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium
5 Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
7 Heart and Vascular Center, Semmelweis University, Budapest, Hungary
8 R&D Ultrafast Lasers Ltd, Budapest, Hungary
9 PXE Reference Center (MAGEC Nord), Angers University Hospital, Angers, France
10 Research Center for Natural Sciences, Institute of Enzymology, Hungarian Academy of Sciences, Budapest, Hungary