Innsbruck Medical University View Institution's Website 14 articles published in JoVE Cancer Research A Data Integration Workflow to Identify Drug Combinations Targeting Synthetic Lethal Interactions Maximilian Marhold1, Andreas Heinzel2, Almas Merchant1, Paul Perco3, Michael Krainer1 1Department of Internal Medicine I - Division of Oncology, Comprehensive Cancer Center, Medical University of Vienna, 2Department of Nephrology, Medical University of Vienna, 3Department of Internal Medicine IV, Medical University Innsbruck Large genetic screens in model organisms have led to the identification of negative genetic interactions. Here, we describe a data integration workflow using data from genetic screens in model organisms to delineate drug combinations targeting synthetic lethal interactions in cancer. Neuroscience Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy Paul V. Ritschl*1,2,3, Lena Hofhansel*2,4, Bernhard Flörchinger5, Rupert Oberhuber2, Robert Öllinger1, Johann Pratschke1, Katja Kotsch6 1Department of Surgery Campus Charité Mitte/Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 2Department of Visceral, Transplant and Thoracic Surgery, Medical University of Innsbruck, 3Charité Clinician Scientist Program, Berlin Institute of Health, 4Department of Psychiatry, Psychotherapy and Psychosomatics, RWTH Aachen University, 5Department of Cardiothoracic Surgery, University Medical Center Regensburg, 6Department of General, Visceral and Vascular Surgery, Charité - Universitätsmedizin Berlin We present a murine model of brain death induction in order to evaluate the influence of its pathophysiological effects on organs as well as on consecutive grafts in the context of solid organ transplantation. Biochemistry Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay Ingo Bauer1, Angelo Pidroni1, Özgür Bayram2,3, Gerald Brosch1, Stefan Graessle1 1Division of Molecular Biology, Biocenter, Medical University of Innsbruck, 2Biology Department, Maynooth University, 3Maynooth University Human Health Research Institute Class 1 histone deacetylases (HDACs) like RpdA have gained importance as potential targets to treat fungal infections. Here we present a protocol for the specific enrichment of TAP-tagged RpdA combined with an HDAC activity assay that allows in vitro efficacy testing of histone deacetylase inhibitors. Immunology and Infection Simultaneous Quantification of Anti-vector and Anti-transgene-Specific CD8+ T Cells Via MHC I Tetramer Staining After Vaccination with a Viral Vector Sarah Wilmschen1, Zoltan Banki1, Dorothee von Laer1, Janine Kimpel1 1Division of Virology, Department of Hygiene, Microbiology, Social Medicine, Medical University of Innsbruck Here, we present a protocol for the ex vivo qualitative detection of antigen-specific CD8+ T cells. Analysis is possible with single cell suspensions from organs or from small amounts of blood. A broad range of studies require the analysis of cytotoxic T cell responses (vaccination and cancer immunotherapy studies). Medicine Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique Benno Cardini*1, Rupert Oberhuber*1, Sven R Hein1, Rebecca Eiter1, Martin Hermann2, Markus Kofler1,3, Stefan Schneeberger1, Gerald Brandacher1,4, Manuel Maglione1 1Center of Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Medical University Innsbruck, 2Department of Anesthesiology and Critical Care Medicine, Medical University Innsbruck, 4Department of Cardiac Surgery, Medical University Innsbruck, 3Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages. Developmental Biology Detection of Residual Donor Erythroid Progenitor Cells after Hematopoietic Stem Cell Transplantation for Patients with Hemoglobinopathies Roman Crazzolara1, Gabriele Kropshofer1, Michael Steurer2, Sieghart Sopper2,3, Wolfgang Schwinger4 1Department of Pediatrics, Medical University Innsbruck, 2Department of Internal Medicine V (Hematology & Oncology), Medical University Innsbruck, 3Tyrolean Cancer Research Institute, 4Department of Pediatrics, Medical University Graz Quantification of donor-derived cells is required to monitor engraftment after stem cell transplantation in patients with hemoglobinopathies. A combination of flow cytometry-based cell sorting, colony formation assay, and subsequent analysis of short tandem repeats may be used to assess the proliferation and differentiation of progenitors in the erythroid compartment. Cancer Research Harnessing the DNA Dye-triggered Side Population Phenotype to Detect and Purify Cancer Stem Cells from Biological Samples Maximilian Boesch*1,2,3, Elisabeth Hoflehner2,3, Dominik Wolf2,3,4, Guenther Gastl2, Sieghart Sopper*2,3 1Institute of Immunobiology, Kantonsspital St. Gallen, 2Internal Medicine V, Medical University of Innsbruck, 3Tyrolean Cancer Research Institute (TKFI), 4Medical Clinic III, Oncology, Hematology, Immunoncology and Rheumatology, University Clinic Bonn (UKB) Methods allowing the characterization and isolation of stem cell populations from biological samples are critical for the advance of stem cell-targeted treatments in cancer and beyond. Here, we provide a detailed protocol for cancer stem cell isolation using the dye-triggered side population phenotype. Immunology and Infection Generation of Human Monocyte-derived Dendritic Cells from Whole Blood Wilfried Posch1, Cornelia Lass-Flörl1, Doris Wilflingseder1 1Division of Hygiene and Medical Microbiology, Medical University of Innsbruck Here, we demonstrate how monocytes are isolated by magnetic bead separation from peripheral blood mononuclear cells after density gradient centrifugation of human anti-coagulated blood. Following incubation for 5 days, human monocytes are differentiated into immature dendritic cells and are ready for experimental procedures in a non-clinical setting. Neuroscience Combined Optogenetic and Freeze-fracture Replica Immunolabeling to Examine Input-specific Arrangement of Glutamate Receptors in the Mouse Amygdala Sabine Schönherr1, Anna Seewald1, Yu Kasugai1, Daniel Bosch2, Ingrid Ehrlich2, Francesco Ferraguti1 1Department of Pharmacology, Medical University of Innsbruck, 2Hertie Institute for Clinical Brain Research and Centre for Integrative Neuroscience, University of Tübingen This article illustrates how the expression of neurotransmitter receptors can be quantified and the pattern analyzed at synapses with identified pre and postsynaptic elements using a combination of viral transduction of optogenetic tools and the freeze-fracture replica immunolabeling technique. Medicine Orthotopic Hind Limb Transplantation in the Mouse Georg J. Furtmüller*1, Byoungchol Oh*1, Johanna Grahammer*2, Cheng-Hung Lin3, Robert Sucher4, Madeline L. Fryer1, Giorgio Raimondi1, W.P. Andrew Lee1, Gerald Brandacher1 1Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine, 2Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 3Center for Vascularized Composite Allotransplantation, Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital and School of Medicine, 4Department of General, Visceral and Transplant Surgery, Charite Berlin This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA). Medicine Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis Agnieszka Martowicz*1,4, Johann Kern*1,2, Eberhard Gunsilius1, Gerold Untergasser1,3 1Department of Internal Medicine V, Innsbruck Medical University, 2Oncotyrol GmbH, 3Tyrolean Cancer Research Institute, 4Division of Vascular Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute Human multiple myeloma (MM) cells require the supportive microenvironment of mesenchymal cells and extracellular matrix components for survival and proliferation. We established an in vivo chicken embryo model with engrafted human myeloma and mesenchymal cells to study effects of cancer drugs on tumor growth, invasion and angiogenesis. Medicine Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique Rupert Oberhuber*1, Benno Cardini*1, Markus Kofler1, Paul Ritschl1, Robert Oellinger1, Felix Aigner1, Robert Sucher1, Stefan Schneeberger1, Johann Pratschke1, Gerald Brandacher2, Manuel Maglione1 1Center of Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 2Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach. Herein, we provide additional details of this technique to supplement the video. Bioengineering Shock Wave Application to Cell Cultures Johannes Holfeld1, Can Tepeköylü1, Radoslaw Kozaryn1, Wolfgang Mathes1, Michael Grimm1, Patrick Paulus2 1Department of Cardiac Surgery, Innsbruck Medical University, 2Clinic of Anesthesiology, Intensive Care Medicine and Pain Therapy, Goethe-University Hospital Shock waves nowadays are well known for their regenerative effects. Therefore in vitro experiments are of increasing interest. We therefore developed a model for in vitro shock wave trials (IVSWT) that enables us to mimic in vivo conditions thereby avoiding distracting physical effects. Immunology and Infection Orthotopic Hind-Limb Transplantation in Rats Robert Sucher*1, Rupert Oberhuber*1, Christian Margreiter1, Guido Rumberg1, Rishi Jindal2, WP Andrew Lee2, Raimund Margreiter1, Johann Pratschke1, Stefan Schneeberger1, Gerald Brandacher1 1Department of Visceral, Transplant, and Thoracic Surgery, Daniel Swarovski Research Laboratory, Innsbruck Medical University, 2Department of Surgery, Division of Plastic and Reconstructive Surgery, University of Pittsburgh Medical Center Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.