INTRODUCTION:Skin cancer is often fatal, which motivates new therapy avenues. Recent advances in cancer treatment are indicative of the importance of combination treatments in oncology. Previous studies have identified small molecule-based therapies and redox-based technologies, including photodynamic therapy or medical gas plasma, as promising candidates to target skin cancer. OBJECTIVE:We aimed to identify effective combinations of experimental small molecules with cold gas plasma for therapy in dermato-oncology. METHODS:Promising drug candidates were identified after screening an in-house 155-compound library using 3D skin cancer spheroids and high content imaging. Combination effects of selected drugs and cold gas plasma were investigated with respect to oxidative stress, invasion, and viability. Drugs that had combined well with cold gas plasma were further investigated in vascularized tumor organoids in ovo and a xenograft mouse melanoma model in vivo. RESULTS:The two chromone derivatives Sm837 and IS112 enhanced cold gas plasma-induced oxidative stress, including histone 2A.X phosphorylation, and further reduced proliferation and skin cancer cell viability. Combination treatments of tumor organoids grown in ovo confirmed the principal anti-cancer effect of the selected drugs. While one of the two compounds exerted severe toxicity in vivo, the other (Sm837) resulted in a significant synergistic anti-tumor toxicity at good tolerability. Principal component analysis of protein phosphorylation profiles confirmed profound combination treatment effects in contrast to the monotherapies. CONCLUSION:We identified a novel compound that, combined with topical cold gas plasma-induced oxidative stress, represents a novel and promising treatment approach to target skin cancer.

Skin cancers are the most common malignancies in the world. Among the most frequent skin cancer entities, squamous cell carcinoma (SCC) ranks second (~20%) after basal cell carcinoma (~77%). In early stages, a complete surgical removal of the affected tissue is carried out as standard therapy. To treat advanced and metastatic cancers, targeted therapies with small molecule inhibitors are gaining increasing attention. Small molecules are a heterogeneous group of protein regulators, which are produced by chemical synthesis or fermentation. The majority of them belong to the group of receptor tyrosine kinase inhibitors (RTKIs), which specifically bind to certain RTKs and directly influence the respective signaling pathway. Knowledge of characteristic molecular alterations in certain cancer entities, such as SCC, can help identify tumor-specific substances for targeted therapies. Most frequently, altered genes in SCC include TP53, NOTCH, EGFR, and CCND1. For example, the gene CCND1, which codes for cyclin D1 protein, is upregulated in nearly half of SCC cases and promotes proliferation of affected cells. A treatment with the small molecule 5′-nitroindirubin-monoxime (INO) leads to inhibition of cyclin D1 and thus inhibition of proliferation. As a component of Danggui Longhui Wan, a traditional Chinese medicine, indirubins are used to treat chronic diseases and have been shown to inhibit inflammatory reactions. Indirubins are pharmacologically relevant small molecules with proapoptotic and antiproliferative activity. In this review, we discuss the current literature on indirubin-based small molecules in cancer treatment. A special focus is on the molecular biology of squamous cell carcinomas, their alterations, and how these are rendered susceptible to indirubin-based small molecule inhibitors. The potential molecular mechanisms of the efficacy of indirubins in killing SCC cells will be discussed as well.

Loss-of-function mutations in the synaptosomal-associated protein 29 (SNAP29) lead to the rare autosomal recessive neurocutaneous cerebral dysgenesis, neuropathy, ichthyosis, and keratoderma (CEDNIK) syndrome. SNAP29 is a soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein. So far, it has been shown to be involved in membrane fusion, epidermal differentiation, formation of primary cilia, and autophagy. Recently, we reported the successful generation of two mouse models for the human CEDNIK syndrome. The aim of this investigation was the generation of a CRISPR/Cas9-mediated SNAP29 knockout (KO) in an immortalized human cell line to further investigate the role of SNAP29 in cellular homeostasis and signaling in humans independently of animal models. Comparison of different methods of delivery for CRISPR/Cas9 plasmids into the cell revealed that lentiviral transduction is more efficient than transfection methods. Here, we reported to the best of our knowledge the first successful generation of a CRISPR/Cas9-mediated SNAP29 KO in immortalized human MRC5Vi fibroblasts (c.169_196delinsTTCGT) via lentiviral transduction.

Plasma medicine is gaining increasing attention and is moving from basic research into clinical practice. While areas of application are diverse, much research has been conducted assessing the use of cold atmospheric pressure plasma (CAP) in wound healing and cancer treatment—two applications with entirely different goals. In wound healing, a tissue-stimulating effect is intended, whereas cancer therapy aims at killing malignant cells. In this review, we provide an overview of the latest clinical and some preclinical research on the efficacy of CAP in wound healing and cancer therapy. Furthermore, we discuss the current understanding of molecular signaling mechanisms triggered by CAP that grant CAP its antiseptic and tissue regenerating or anti-proliferative and cell death-inducing properties. For the efficacy of CAP in wound healing, already substantial evidence from clinical studies is available, while evidence for therapeutic effects of CAP in oncology is mainly from in vitro and in vivo animal studies. Efforts to elucidate the mode of action of CAP suggest that different components, such as ultraviolet (UV) radiation, electromagnetic fields, and reactive species, may act synergistically, with reactive species being regarded as the major effector by modulating complex and concentration-dependent redox signaling pathways.

ZusammenfassungSeit Antritt von Prof. Dr. med. Steffen Emmert als Ordinarius der Klinik und Poliklinik für Dermatologie und Venerologie im Jahr 2015 konnte das dermatologische Forschungslabor sukzessive aufgebaut und erweitert werden. Im Einklang mit dem onkologischen Schwerpunkt der Universitätsmedizin Rostock sowie dem von der Landesregierung forcierten „Gesundheitsland Mecklenburg-Vorpommern“ wird grundlagenorientierten und translationalen Projekten nachgegangen. Das vorwiegend drittmittelfinanzierte und stetig wachsende Forschungsteam bearbeitet diverse Fragestellungen in den Bereichen der Dermato-Onkologie, Plasmamedizin und seltenen Hauterkrankungen. Inzwischen auf einem soliden Fundament stehend, befindet sich der Forschungsbereich weiterhin in einem dynamischen Entwicklungsprozess. Nicht nur personell, sondern auch thematisch und methodisch wird er derzeit durch die Integration weiterer Arbeitsgruppen unter der Leitung von Ärzten aus der Klinik ergänzt und ausgebaut. Diverse Kollaborationen an der Universitätsmedizin Rostock und im Land zeugen von einem freundlichen, unterstützenden und kollegialen Umfeld, das die Integration am Standort befördert hat.

Recently, the potential use of cold atmospheric pressure plasma (CAP) in cancer treatment has gained increasing interest. Especially the enhanced selective killing of tumor cells compared to normal cells has prompted researchers to elucidate the molecular mechanisms for the efficacy of CAP in cancer treatment. This review summarizes the current understanding of how CAP triggers intracellular pathways that induce growth inhibition or cell death. We discuss what factors may contribute to the potential selectivity of CAP towards cancer cells compared to their non-malignant counterparts. Furthermore, the potential of CAP to trigger an immune response is briefly discussed. Finally, this overview demonstrates how these concepts bear first fruits in clinical applications applying CAP treatment in head and neck squamous cell cancer as well as actinic keratosis. Although significant progress towards understanding the underlying mechanisms regarding the efficacy of CAP in cancer treatment has been made, much still needs to be done with respect to different treatment conditions and comparison of malignant and non-malignant cells of the same cell type and same donor. Furthermore, clinical pilot studies and the assessment of systemic effects will be of tremendous importance towards bringing this innovative technology into clinical practice.

Plasmamedizin hat sich zu einem innovativen Forschungsgebiet mit diversen Anwendungsmöglichkeiten insbesondere in der Dermatologie entwickelt. Kaltes Atmosphärendruckplasma (KAP) kann sowohl gewebeschonend als auch zur Destruktion von malignem Gewebe eingesetzt werden. KAP besteht aus einer komplexen Mixtur aus verschiedenen biologisch aktiven Agenzien, die zum Teil synergistisch auf das zu behandelnde Material oder Gewebe wirken. Dargestellt werden die Anwendungsgebiete für KAP in der Dermatologie sowie der aktuelle Stand der Forschung. Literatur zur Anwendung von KAP in der Dermatologie wurde gesichtet und als Übersichtsarbeit zusammengefasst. Mit einer KAP-Behandlung können beispielsweise antimikrobielle, gewebestimulierende, durchblutungsfördernde, aber auch proapoptotische Effekte erreicht werden. Unter Ausnutzung dieser Effekte kommt KAP in der Dermatologie erfolgreich zur Desinfektion und Wundbehandlung zum Einsatz. Des Weiteren zeigen Untersuchungen zur Anwendung von KAP bei der Behandlung von Tumoren, aktinischen Keratosen, Narben, Ichthyose, atopischen Ekzemen sowie zur Linderung von Schmerz und Juckreiz positive Effekte. Während die Anwendung von KAP zur Desinfektion und zur Wundbehandlung vereinzelt bereits den Einzug in die klinische Praxis gefunden hat, befindet sich die Anwendung in weiteren Bereichen, beispielsweise zur Tumortherapie, noch im Forschungskontext.

Neben akuten Wunden durch Traumen oder chirurgische Eingriffe stellen chronische Wunden eine relativ große, heterogene Erkrankungsgruppe dar. Dazu zählen Beinulzera, bei denen die venöse Genese die der arteriellen Genese deutlich übertrifft, aber auch das diabetische Fußsyndrom und Druckulzera. Aufgrund der Therapieresistenz v. a. chronischer Wunden werden dringend weitere, additiv wirksame Therapieoptionen benötigt. Die zusätzliche Behandlung mit kaltem Atmosphärendruckplasma (KAP) stellt eine derartige innovative Option dar. Betrachtet wird die Wirkung von Kaltplasma auf die Heilung akuter und chronischer Wunden, und es wird der aktuelle Stand der Forschung aufgezeigt. Literatur zur Anwendung von KAP zur Wundbehandlung wurde gesichtet und als Übersichtsarbeit zusammengefasst. Mit einer KAP-Behandlung können mehrere wundheilungsfördernde Effekte in einer Anwendung erzielt werden. Zum einen besitzt KAP eine starke und breite antimikrobielle Aktivität auch gegen Biofilm. Zum anderen mediiert der Plasmacocktail, der aus reaktiven Stickstoff- und Sauerstoffspezies, UV und geladenen Teilchen (Stromfluss) besteht, gewebestimulierende und durchblutungsfördernde sowie antiinflammatorische Effekte. Eine deutliche Reduktion der Keimbesiedelung und eine schnellere Wundheilung konnten in kontrollierten Studien bereits klar nachgewiesen werden. Die mittlerweile umfassende Studienlandschaft mit strukturierten Fallberichten und randomisierten Fall-Kontroll-Studien lässt den Schluss zu, dass KAP zur Wundbehandlung sicher, effektiv und einfach einsetzbar ist. Die Anwendung von KAP zur Wundbehandlung hat vereinzelt bereits den Einzug in die klinische Routine gefunden.

The ability to produce cold plasma at atmospheric pressure conditions was the basis for the rapid growth of plasma-related application areas in biomedicine. Plasma comprises a multitude of active components such as charged particles, electric current, UV radiation, and reactive gas species which can act synergistically. Anti-itch, antimicrobial, anti-inflammatory, tissue-stimulating, blood flow-enhancing, and proapoptotic effects were demonstrated in in vivo and in vitro experiments, and until now, no resistance of pathogens against plasma treatment was observed. The combination of the different active agents and their broad range of positive effects on various diseases, especially easily accessible skin diseases, renders plasma quite attractive for applications in medicine. For medical applications, two different types of cold plasma appear suitable: indirect (plasma jet) and direct (dielectric barrier discharge—DBD) plasma sources. The DBD device PlasmaDerm® VU-2010 (CINOGY Technologies GmbH), the atmospheric pressure plasma jet (APPJ) kINPen® MED (INP Greifswald/neoplas tools GmbH), and the SteriPlas (Adtec Ltd., London, United Kingdom) are CE-certified as a medical product to treat chronic wounds in humans and showed efficacy and a good tolerability. Recently, the use of plasma in cancer research and oncology is of particular interest. Plasma has been shown to induce proapoptotic effects more efficiently in tumor cells compared with the benign counterparts, leads to cellular senescence, and—as shown in vivo—reduces skin tumors. To this end, a world-wide first Leibniz professorship for plasmabiotechnology in dermatology has been introduced to establish a scientific network for the investigation of the efficacy and safety of cold atmospheric plasma in dermatooncology. Hence, plasma medicine especially in dermatology holds great promise.