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Zusammenfassung. Die Sarkoidose ist eine seltene Ursache der Hyperkalzämie. Als häufigste Ursachen kommen ein primärer Hyperparathyreoidismus sowie ein malignes Tumorleiden in Frage. Weitere Ursachen können eine Vitamin-D-Intoxikation, sonstige granulomatöse Erkrankungen (Tuberkulose, Pilzinfektionen etc.) oder ein multiples Myelom sein. Aufgrund der breiten Differenzialdiagnose ist eine systematische Abklärung entscheidend, um die korrekte Diagnose zu stellen. Zur Therapie einer Hyperkalzämie erfolgt eine Volumensubstitution und anschliessende Gabe von Bisphosphonaten. Bei einer Sarkoidose als Ursache der Hyperkalzämie ist zudem eine systemische Steroidtherapie mit Prednisolon indiziert.

Fibroblast specific protein 1 (FSP1/S100A4) is an intracellular calcium binding protein which has been linked to epithelial-mesenchymal transition, tissue fibrosis, pulmonary vascular disease, metastatic tumour development, increased tumour cell motility and invasiveness. This protein is reported to be uniquely expressed in newly formed and differentiated fibroblasts and has been used in various studies to demonstrate epithelial-mesenchymal transition (EMT). However, S100A4 immunoreactivity does not solely fit the distribution and shape of fibroblasts; therefore, we aimed to characterize S100A4 positive cells in different human tissue compartments. We found S100A4 expression in a wide range of cells, including cells of haematopoietic lineage, namely CD4 and CD8 positive T-lymphocytes, but not B-lymphocytes. All investigated monocytes, macrophages and specialised histiocytes were positive for S100A4. Only a limited number of fibroblasts and most smooth muscle actin positive myofibroblasts expressed S100A4. Even some epithelial cells of the kidney and bladder were positive for S100A4. Expression was also found in the vasculature. Here, cells of the subendothelial space, tunica adventitia and some smooth muscle cells of the tunica media were positive for S100A4. Expression was also detected in the stroma of the prostate, adipocytes and in skeletal muscle. In summary, S100A4 is expressed in various cell types of different lineage. It is therefore not possible to equate the expression of this protein to fibroblasts. Hence, studies that used this marker as evidence of transition from epithelia to fibroblasts must be revisited.

A Rare Cause of Hypercalcemia

Highlights • SalvGlandDx includes 27 genes known to be involved in salivary gland neoplasms • Can be applied to FFPE histological and cytological (cell block) specimen • Solely RNA extraction is needed • Gene fusions, hotspot mutations and gene expression levels can be detected • Guides diagnosis in uncertain cases and can detect therapeutically relevant fusions, Diagnosis of salivary gland neoplasms is often challenging due to their high morphological diversity and overlaps. Several recurrent molecular alterations have been described recently, which can serve as powerful diagnostic tools and potential therapeutic targets (e.g. NTRK or RET fusions). However, current sequential molecular testing can be expensive and time consuming. In order to facilitate the diagnosis of salivary gland neoplasms, we designed an all-in-one RNA-based next generation sequencing panel suitable for the detection of mutations, fusions and gene expression levels (including NR4A3) of 27 genes involved in salivary gland neoplasms. Here we present the validation of the “SalvGlandDx” panel on FFPE histological specimen including fine needle aspiration (FNA) cell block material, against the standard methods currently used at our institution. In a second part we describe selected unique cases in which the SalvGlandDx panel allowed proper diagnosis and new insights into special molecular characteristics of selected salivary gland tumors. We characterize a unique salivary gland adenocarcinoma harboring a ZCCHC7-NTRK2 fusion, a highly uncommon spindle cell and pseudoangiomatoid adenoid-cystic carcinoma with MYBL1-NFIB fusion, and a purely oncocytic mucoepidermoid carcinoma, whereas diagnosis could be made by detection of a CRTC3-MAML2 rearrangement on the cell block specimen of the FNA. Further, a rare case of a SS18-ZBTB7A rearranged low-grade adenocarcinoma previously described as potential spectrum of microsecretory adenocarcinoma, is reported. In addition, features of six cases within the spectrum of polymorphous adenocarcinoma / cribriform adenocarcinoma of salivary gland including PRKD1 p.E710D mutations and novel fusions involving PRKAR2A-PRKD1, SNX9-PRKD1 and ATL2-PRKD3, are described.

In der vorliegenden Diplomarbeit wurden eine Strukturierung von Materialien auf GeoGebra und Unterrichtsplanungen für den Fachbereich Chemie erstellt. Zuerst werden theoretische Inhalte aufgezeigt, welche für die Erstellung einer passenden Strukturierung wichtig sind. Anschließend wird die Vorgangsweise für die Erstellung der Strukturierung beschrieben. Im Anschluss daran werden, zur Erweiterung der Materialiensammlung auf GeoGebra, Unterrichtsplanungen erstellt und frei verfügbar gemacht. Ziel dieser Arbeit ist es, eine Strukturierung von Materialien für den Fachbereich Chemie zu erstellen, die für verschiedenste Länder, Schultypen und Altersklassen geeignet ist, damit Lehrende bzw. Lernende entsprechende Materialien einfach und schnell finden können und des Weiteren, die Materialiensammlung auf GeoGebra zu erweitern. eingereicht von Stefanie Vogetseder Universität Linz 2021

Background Fos like 2 (Fosl-2) is a transcription factor belonging to the AP-1 transcription complex. We have recently described a Fosl-2 transgenic (tg) mouse model which develops a multi-organ inflammatory and autoimmune phenotype. In these mice, we have characterized a decrease in regulatory T cells (Tregs), which preceded the activation of T cells and the development of inflammation. Objectives To analyze how Fosl-2 reduces the Treg population and triggers autoimmunity in Fosl-2 tg mice. Methods We used previously generated Fosl-2 tg overexpressing and Fosl-2 T cell specific knock out (Fosl-2 ko) mice. For mixed bone marrow transfer experiments, lethally irradiated recipients received a one to one mix of Fosl-2 tg (CD45.2) and wt (CD45.1) bone marrow to create Fosl-2 tg-wt chimera. We addressed the contribution of Tregs to the inflammatory phenotype by co-transferring Fosl-2 tg CD4 T cells and wt Tregs to Rag2-/- recipients. Treg and T cell populations were analyzed by flow cytometry and inflammation was addressed by CD45 staining on paraffin-embedded sections. RNA-sequencing was used to compare wt, Fosl-2 tg and Fosl-2 ko CD4 T cells after 24 hours of stimulation with anti-CD3 (2μg/ml) and anti-CD28 (2μg/ml). Results We first addressed whether Fosl-2 affected Tregs in a cell intrinsic way using mixed bone marrow experiments. In these animals, the CD45.2 Fosl-2 tg CD4 T cells showed a much lower proportion of Tregs compared to the CD45.1 wt population, both in the spleen (0.73%±0.15 vs 31.6%±3.6, P=0.062) and thymus (1.3%±1.04 vs 3.23%±0.78, P=0.001). This demonstrates that Fosl-2 overexpression represses Treg development in a cell intrinsic way. In T cell transfer experiments, Rag2-/- mice receiving 106 Fosl-2 tg CD4 cells developed lung inflammation 5 weeks after transfer, confirming that T cells are inducers of inflammation in Fosl-2 tg mice. Moreover, co-transfer of either 3*105 or 106 wt Treg cells resulted in a dose dependent reduction of inflammation. These data indicated that the decrease in the Treg population in Fosl-2 tg mice is responsible for the induction of inflammation. We then analysed Fosl-2 transcriptional targets in T cells by RNA-seq. Using a fold change > 1.5 and False Discovery Rate (FDR) of 0.05, we identified 191 differentially expressed genes in both Fosl-2 tg and Fosl-2 ko cells compared to wt. Interestingly, one of the top target genes of Fosl-2 was FoxP3. This unbiased approach thus revealed that FoxP3 expression is repressed by Fosl-2, with a 6.5 fold reduction in Fosl-2 tg cells and a 2.5 fold increase in Fosl-2 ko cells. This effect was confirmed on the protein level with a reduction in FoxP3 induction in Fosl-2 tg cells treated with TGFβ. The repression of Treg development observed in Fosl-2 tg mice could thus be explained by a direct transcriptional control of FoxP3 expression. Additionally, we found that Fosl-2 repressed a set of genes important for Tregs and other T helper cells. This included Nr4a2, a transcription factor involved in Treg development, IRF8 and Eomes, two genes involved in the polarization of Th1 and Th17 cells, or Ccl1 a chemokine important for Treg homeostasis. Conclusion Fosl-2 is involved in the control of FoxP3 expression in T cells. Through this, overexpression of Fosl-2 represses Treg development and induces a Treg dependent autoimmune phenotype in mice. This mechanism could thus be involved in the pathogenesis of autoimmune diseases and might represent a therapeutic target to modulate the Treg population. Disclosure of Interests Florian Renoux: None declared, Mara Stellato: None declared, Alexander Vogetseder: None declared, Riyung Huang: None declared, Arun Subramaniam: None declared, Przemyslaw Blyszczuk: None declared, Jorg Distler: None declared, Gabriela Kania: None declared, Onur Boyman: None declared, Oliver Distler Grant/research support from: Prof. Distler received research funding from Actelion, Bayer, Boehringer Ingelheim and Mitsubishi Tanabe to investigate potential treatments of scleroderma and its complications, Consultant for: Prof. Distler has/had consultancy relationship within the last 3 years with Actelion, AnaMar, Bayer, Boehringer Ingelheim, ChemomAb, espeRare foundation, Genentech/Roche, GSK, Inventiva, Italfarmaco, iQvia, Lilly, medac, MedImmune, Mitsubishi Tanabe Pharma, Pharmacyclics, Novartis, Pfizer, Sanofi, Serodapharm and UCB in the area of potential treatments of scleroderma and its complications. In addition, he had/has consultancy relationship within the last 3 years with A. Menarini, Amgen, Abbvie, GSK, Mepha, MSD, Pfizer and UCB in the field of arthritides and related disorders

Dieser Artikel bietet eine Ubersicht uber die neuen medikamentosen Behandlungsmoglichkeiten.

Background Fos-related antigen 2 (Fra2) is a transcription factor belonging to the Fos family proteins which is part of the AP-1 transcription complex. We recently described a Fra2 transgenic (tg) mouse model which develops a multi-organ inflammatory phenotype affecting skin, lungs, thymus, liver and salivary glands. We have observed abnormalities in the T cell compartment, particularly in regulatory T (Treg) cells, which led us to hypothesize that Fra2 tg mice develop a T cell driven autoimmune phenotype. Objectives To demonstrate the autoimmune phenotype of Fra2 tg mice and to characterize the mechanisms leading to Treg cell abnormality. Methods We used previously generated Fra2 tg overexpressing mice. T lymphocyte populations were analyzed by flow cytometry for expression of activation markers and secretion of cytokines. We transferred purified CD4+ T cells into Rag2-/- mice lacking T and B cells, and we generated Rag2-/-Fra2 tg mice. Bone marrow cells were transferred into lethally irradiated recipients to create Fra2-WT bone marrow chimeric mice. Results Fra2 tg mice backcrossed onto a Rag2-/- background did not develop inflammatory manifestations (n=6), demonstrating the dependence on T and/or B cells of the autoimmune phenotype. In line with this, the transfer of purified CD4+ cells from 16 week-old Fra2 tg mice into Rag2-/- recipients was sufficient to transfer the disease phenotype (n=3). Analysis of T cell populations from Fra2 tg mice showed the presence of activated CD4+ and CD8+ cells in the spleen and lungs. After in vitro stimulation, we found that CD4+ T cells from Fra2 tg mice produced the Th2 cytokines IL-4, IL-5 and IL-13. Thus, these data strongly suggest a T cell-driven autoimmune disease in these mice. We previously reported a striking decrease of Treg cells in Fra2 tg mice, which might explain the autoimmune phenotype observed. Supporting this idea, we found that 3 week-old mice were devoid of organ manifestations and of T cell activation, but presented the same defect in the Treg cell population (n=6, p Conclusions Our data suggest that Fra2 controls tTreg cell development, possibly by modulating IL-2 signaling in T cells, which leads to autoimmunity in this mouse model. This new pathway could be targeted in a translational approach to modulate the capacity of T cells to differentiate in Tregs during autoimmune disease. Disclosure of Interest F. Renoux Grant/research support from: Swisslife, M. Stellato: None declared, D. Impellizzieri: None declared, A. Vogetseder: None declared, R. Huang Employee of: Sanofi-Genzyme, A. Subramaniam Employee of: Sanofi-Genzyme, C. Dees: None declared, J. Distler Shareholder of: 4D Science, Grant/research support from: Anamar, Active Biotech, Array Biopharma, BMS, Bayer Pharma, Boehringer Ingelheim, Celgene, GSK, Novartis, Sanofi-Aventis, UCB, Consultant for: Actelion, Active Biotech, Anamar, Bayer Pharma, Boehringer Ingelheim, Celgene, Galapagos, GSK, Inventiva, JB Therapeutics, Medac, Pfizer, RuiYi and UCB, G. Kania: None declared, O. Boyman: None declared, O. Distler Grant/research support from: Actelion, Bayer, Boehringer Ingelheim, Pfizer, Sanofi, Consultant for: 4 D Science, Actelion, Active Biotec, Bayer, BiogenIdec, BMS, Boehringer Ingelheim, ChemomAb, EpiPharm, espeRare foundation, Genentech/Roche, GSK, Inventiva, Lilly, medac, Mepha, MedImmune, Mitsubishi Tanabe Pharma, Pharmacyclics, Pfizer, Sanofi, Serodapharm, Sinoxa, Speakers bureau: AbbVie, iQone Healthcare, Mepha

Cancer-testis antigens (CTAgs) play a major role in the immune response against cancer, but their biological functions in germ and cancer cells is still unclear. MAGE-C1 and MAGE-C2 are two CTAgs located at the Xq27 region of chromosome X and frequently expressed in multiple myeloma. Chromosomal rearrangements often occur in myeloma. We therefore investigated whether numerical and structural chromosomal aberrations correlate with their protein expression in primary multiple myelomas. To this aim, we designed new fluorescence in situ hybridization probes specific for the MAGE region in the Xq27 region and evaluated simultaneously aberrations of the X chromosome centromere. The comparison of MAGE copy number and chromosome X status revealed that MAGE copy number changes occurred in 6/43 (14%) cases, independent of concomitant X chromosome alterations. These numerical aberrations are less frequent than the expression of MAGE-C1 and MAGE-C2 (63% and 27% of patients, respectively) and do not always correlate with MAGE-C1 and MAGE-C2 expressions, suggesting alternative regulatory mechanisms in the expression of these genes. Copyright © 2014 John Wiley & Sons, Ltd.

To determine whether metastasis to brain is associated with altered expression patterns of integrins, we investigated the expression of αvβ3, αvβ5, αvβ6 and αvβ8 integrins in primary malignancies and metastases to brain of breast, lung and renal carcinomas and in malignant melanoma. Inhibitors of αv integrins are currently in clinical trials for glioblastoma. The role of integrins in the process of brain metastasis from other human tumors is unknown. Immunohistochemistry with novel integrin subtype specific rabbit monoclonal antibodies was performed on tissue microarrays of archival material of surgical biopsies taken from primary tumors and brain metastases. Integrin αvβ3 expression was increased in brain metastases compared to primary tumors of breast adenocarcinoma, non-small cell lung cancer, renal clear cell cancer and malignant cutaneous melanoma (all p