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Merkle cell

Josef Mang, Konstanze Merkle, Martina Heller, Julia Schüler, Yanis Tolstov, Jielin Li, Markus Hohenfellner, Stefan Duensing
BACKGROUND: Taxanes are routinely used to treat men with advanced prostate cancer, yet their molecular mode of action is poorly characterized. Taxanes stabilize microtubules and may hence interfere with a plethora of cellular processes, most notably mitosis. However, prostate cancer is typically a slowly growing tumor suggesting that additional processes play a role in the response to taxanes. METHODS: Here, we analyzed the potential effect of taxanes on microtubuli-dependent intracellular transport and signaling processes, specifically, nuclear translocation of the androgen receptor and modulation of the RAS-RAF-MEK-ERK signaling cascade...
September 28, 2016: Urologic Oncology
Michael Pilsl, Philipp E Merkl, Philipp Milkereit, Joachim Griesenbeck, Herbert Tschochner
RNA polymerase I (Pol I) activity is crucial to provide cells with sufficient amounts of ribosomal RNA (rRNA). Synthesis of rRNA takes place in the nucleolus, is tightly regulated and is coordinated with synthesis and assembly of ribosomal proteins, finally resulting in the formation of mature ribosomes. Many studies on Pol I mechanisms and regulation in the model organism S. cerevisiae were performed using either complex in vitro systems reconstituted from more or less purified fractions or genetic analyses...
2016: Methods in Molecular Biology
David P Santos, Evangelos Kiskinis, Kevin Eggan, Florian T Merkle
Genome editing of human pluripotent stem cells (hPSCs) with the CRISPR/Cas9 system has the potential to revolutionize hPSC-based disease modeling, drug screening, and transplantation therapy. Here, we aim to provide a single resource to enable groups, even those with limited experience with hPSC culture or the CRISPR/Cas9 system, to successfully perform genome editing. The methods are presented in detail and are supported by a theoretical framework to allow for the incorporation of inevitable improvements in the rapidly evolving gene-editing field...
2016: Current Protocols in Stem Cell Biology
Ruth Merkle, Bernhard Steiert, Florian Salopiata, Sofia Depner, Andreas Raue, Nao Iwamoto, Max Schelker, Helge Hass, Marvin Wäsch, Martin E Böhm, Oliver Mücke, Daniel B Lipka, Christoph Plass, Wolf D Lehmann, Clemens Kreutz, Jens Timmer, Marcel Schilling, Ursula Klingmüller
Lung cancer, with its most prevalent form non-small-cell lung carcinoma (NSCLC), is one of the leading causes of cancer-related deaths worldwide, and is commonly treated with chemotherapeutic drugs such as cisplatin. Lung cancer patients frequently suffer from chemotherapy-induced anemia, which can be treated with erythropoietin (EPO). However, studies have indicated that EPO not only promotes erythropoiesis in hematopoietic cells, but may also enhance survival of NSCLC cells. Here, we verified that the NSCLC cell line H838 expresses functional erythropoietin receptors (EPOR) and that treatment with EPO reduces cisplatin-induced apoptosis...
August 2016: PLoS Computational Biology
Sebastian Marwitz, Sofia Depner, Dmytro Dvornikov, Ruth Merkle, Magdalena Szczygieł, Karin Müller-Decker, Philippe Lucarelli, Marvin Wäsch, Heimo Mairbäurl, Klaus F Rabe, Christian Kugler, Ekkehard Vollmer, Martin Reck, Swetlana Scheufele, Maren Kröger, Ole Ammerpohl, Reiner Siebert, Torsten Goldmann, Ursula Klingmüller
Non-small cell lung cancer (NSCLC) is characterized by early metastasis and has the highest mortality rate among all solid tumors, with the majority of patients diagnosed at an advanced stage where curative therapeutic options are lacking. In this study, we identify a targetable mechanism involving TGFβ elevation that orchestrates tumor progression in this disease. Substantial activation of this pathway was detected in human lung cancer tissues with concomitant downregulation of BAMBI, a negative regulator of the TGFβ signaling pathway...
July 1, 2016: Cancer Research
Rebecca Kesselring, Joachim Glaesner, Andreas Hiergeist, Elisabeth Naschberger, Helmut Neumann, Stefan M Brunner, Anja K Wege, Caroline Seebauer, Gudrun Köhl, Susanne Merkl, Roland S Croner, Christina Hackl, Michael Stürzl, Markus F Neurath, André Gessner, Hans-Juergen Schlitt, Edward K Geissler, Stefan Fichtner-Feigl
Colorectal cancer (CRC) is associated with loss of epithelial barrier integrity, which facilitates the interaction of the immunological microenvironment with the luminal microbiome, eliciting tumor-supportive inflammation. An important regulator of intestinal inflammatory responses is IRAK-M, a negative regulator of TLR signaling. Here we investigate the compartment-specific impact of IRAK-M on colorectal carcinogenesis using a mouse model. We demonstrate that IRAK-M is expressed in tumor cells due to combined TLR and Wnt activation...
May 9, 2016: Cancer Cell
Philipp Blüm, Joachim Pircher, Monika Merkle, Thomas Czermak, Andrea Ribeiro, Hanna Mannell, Florian Krötz, Alexander Hennrich, Michael Spannagl, Simone Köppel, Erik Gaitzsch, Markus Wörnle
Hepatitis C virus (HCV) infection is a major problem worldwide. HCV is not limited to liver disease but is frequently complicated by immune-mediated extrahepatic manifestations such as glomerulonephritis or vasculitis. A fatal complication of HCV-associated vascular disease is thrombosis. Polyriboinosinic:polyribocytidylic acid (poly (I:C)), a synthetic analog of viral RNA, induces a Toll-like receptor 3 (TLR3)-dependent arteriolar thrombosis without significant thrombus formation in venules in vivo. These procoagulant effects are caused by increased endothelial synthesis of tissue factor and PAI-1 without platelet activation...
April 18, 2016: Cellular & Molecular Immunology
M Merkl, R Ertl, S Handschuh, C Aurich, S Schäfer-Somi
In the present study, we assessed the presence of the ATP-binding-cassette (ABC) transporter molecules ABCA1 in spermatozoa of adult stallions and in testicular and epididymal tissue of prepubertal and adult stallions. For this purpose, semen samples from six fertile Shetland pony stallions aged 4 to 19 years were collected. Semen was collected from each stallion on three consecutive days. Ejaculates were analyzed immediately after collection, and only ejaculates meeting minimal requirements for fertile stallions were further evaluated...
April 1, 2016: Theriogenology
Kaitlyn R Ammann, Katrina J DeCook, Phat L Tran, Valerie M Merkle, Pak K Wong, Marvin J Slepian
BACKGROUND: Cell migration is a vital process for growth and repair. In vitro migration assays, utilized to study cell migration, often rely on physical scraping of a cell monolayer to induce cell migration. The physical act of scrape injury results in numerous factors stimulating cell migration - some injury-related, some solely due to gap creation and loss of contact inhibition. Eliminating the effects of cell injury would be useful to examine the relative contribution of injury versus other mechanisms to cell migration...
2015: Journal of Biological Engineering
Andrea Ribeiro, Monika Merkle, Nasim Motamedi, Hans Nitschko, Simone Köppel, Markus Wörnle
Polyomavirus-associated nephropathy due to BK virus infection (BKVAN) is recognized as an important cause of significant kidney transplant dysfunction often leading to renal graft loss. The activation of innate immune defense mechanisms during BKVAN is still poorly understood and an altered regulation of inflammatory mediators by resident kidney cells upon viral infection can be expected to contribute to the onset and progression of disease. TNFα interacting with its receptors, TNF receptor 1 (TNFR1) and TNF receptor 2 (TNFR2), is largely accepted to be involved in viral responses, exhibiting both proinflammatory and immunosuppressive effects...
January 2016: Molecular and Cellular Biochemistry
Valerie M Merkle, Phat L Tran, Marcus Hutchinson, Kaitlyn R Ammann, Katrina DeCook, Xiaoyi Wu, Marvin J Slepian
UNLABELLED: Cardiovascular disease is the leading cause of death in the world. In this study, coaxial electrospinning is employed to fabricate fibers in a core-shell structure with polyvinyl alcohol (PVA) in the core and gelatin in the shell for evaluation as a potential vascular tissue engineering construct. PVA, a synthetic polymer, provides mechanical strength to the biocompatible and weak gelatin sheath. The HUVEC (human umbilical vein endothelial cells) and rSMC (rat smooth muscle cells) demonstrated a flattened morphology with multiple attachment sites on the gelatin and coaxial scaffolds, with an increase in cell spreading seen as mechanical stiffness of the scaffold increased...
November 2015: Acta Biomaterialia
Monika Merkle, Joachim Pircher, Hanna Mannell, Florian Krötz, Philipp Blüm, Thomas Czermak, Erik Gaitzsch, Christine Schneider, Simone Köppel, Andrea Ribeiro, Markus Wörnle
In viral infection, morbidity and mortality often result from extrahepatic disease manifestations such as vasculitis. We hereby show that human microvascular endothelial cells express viral receptors of the innate immune system which are induced upon ligand engagement. Furthermore, stimulation of endothelial cells with the synthetic analog of viral DNA, poly (dA:dT), human DNA and hepatitis B virus-containing immunoprecipitates from a patient with polyarteritis nodosa induces an inflammatory response including the upregulation of adhesion molecules, which is mediated exclusively by TLR9 and involves an IRF3-dependent pathway...
December 2015: Journal of Autoimmunity
Nisreen K Mesiwala, Nathan Shemonski, Michelle G Sandrian, Ryan Shelton, Hiroshi Ishikawa, Hussein A Tawbi, Joel S Schuman, Stephen A Boppart, Leanne T Labriola
BACKGROUND: The study aims to correlate Fourier-domain optical coherence tomography (FD-OCT) with Goldmann visual field (GVF) to show the photoreceptor (PR) structure and function relationship in the first described case of cancer-associated retinopathy (CAR) from Merkel cell carcinoma. FINDINGS: A case study of a patient with CAR who was imaged with serial GVF and FD-OCT over a 2-year period was carried out. En face images were created using a custom algorithm from the volumetric Fourier-domain OCT scans at the PR level...
December 2015: Journal of Ophthalmic Inflammation and Infection
Sarah H Merkling, Gijs J Overheul, Joël T van Mierlo, Daan Arends, Christian Gilissen, Ronald P van Rij
Innate immunity is the first line of defence against pathogens and is essential for survival of the infected host. The fruit fly Drosophila melanogaster is an emerging model to study viral pathogenesis, yet antiviral defence responses remain poorly understood. Here, we describe the heat shock response, a cellular mechanism that prevents proteotoxicity, as a component of the antiviral immune response in Drosophila. Transcriptome analyses of Drosophila S2 cells and adult flies revealed strong induction of the heat shock response upon RNA virus infection...
2015: Scientific Reports
Mayada Woriedh, Rainer Merkl, Thomas Dresselhaus
EMBRYO SAC1-4 (ES1-4) peptides belong to the defensin subgroup of cysteine-rich peptides known to mediate pollen tube burst in Zea mays (maize). ES1-4 are reported here to also be capable of inhibiting germination and growth of the maize fungal pathogens Fusarium graminearum and Ustilago maydis at higher concentrations. Dividing the peptides into smaller pieces showed that a 15-amino-acid peptide located in a highly variable loop region lacking similarity to other defensins or defensin-like peptides binds to maize pollen tube surfaces, causing swelling prior to burst...
August 2015: Journal of Experimental Botany
Dennis Doleschel, Anne Rix, Susanne Arns, Karin Palmowski, Felix Gremse, Ruth Merkle, Florian Salopiata, Ursula Klingmüller, Michael Jarsch, Fabian Kiessling, Wiltrud Lederle
Recombinant human erythropoietin (rhuEpo) is currently under debate for the treatment of chemotherapy-induced anemia due to clinical trials showing adverse effects in Epo-treated patients and the discovery of the erythropoietin-receptor (EpoR) in tumor and endothelial cells. Here, using Epo-Cy5.5 as theranostic near-infrared fluorescent probe we analyzed the effects of rhuEpo as co-medication to carboplatin in non-small-cell-lung-cancer (NSCLC)-xenografts with different tumor cell EpoR-expression (H838 ~8-fold higher than A549)...
2015: Theranostics
Narayan Yoganandan, Frank Pintar, Anjishnu Banerjee, Michael Schlick, Sajal Chirvi, Hermeeth Uppal, Andrew Merkle, Liming Voo, Michael Kleinberg
The objective of the present study was to derive injury probability curves applicable to the Hybrid III dummy (also termed the Anthropomorphic Test Device, ATD) lower leg under axial impacts for military applications. A matched-pair approach was used. Axial impacts were delivered to below knee foot-ankle complex preparations of the lower leg of the ATD using pendulum and custom vertical accelerator devices. Military boot was used in some tests. Post mortem human surrogate (PMHS) preparations were used as matched-pair tests for injury outcomes...
2015: Biomedical Sciences Instrumentation
Florian T Merkle, Werner M Neuhausser, David Santos, Eivind Valen, James A Gagnon, Kristi Maas, Jackson Sandoe, Alexander F Schier, Kevin Eggan
The CRISPR-Cas9 system has the potential to revolutionize genome editing in human pluripotent stem cells (hPSCs), but its advantages and pitfalls are still poorly understood. We systematically tested the ability of CRISPR-Cas9 to mediate reporter gene knockin at 16 distinct genomic sites in hPSCs. We observed efficient gene targeting but found that targeted clones carried an unexpectedly high frequency of insertion and deletion (indel) mutations at both alleles of the targeted gene. These indels were induced by Cas9 nuclease, as well as Cas9-D10A single or dual nickases, and often disrupted gene function...
May 12, 2015: Cell Reports
Justin Liu, Florian T Merkle, Avni V Gandhi, James A Gagnon, Ian G Woods, Cindy N Chiu, Tomomi Shimogori, Alexander F Schier, David A Prober
Loss of neurons that express the neuropeptide hypocretin (Hcrt) has been implicated in narcolepsy, a debilitating disorder characterized by excessive daytime sleepiness and cataplexy. Cell replacement therapy, using Hcrt-expressing neurons generated in vitro, is a potentially useful therapeutic approach, but factors sufficient to specify Hcrt neurons are unknown. Using zebrafish as a high-throughput system to screen for factors that can specify Hcrt neurons in vivo, we identified the LIM homeobox transcription factor Lhx9 as necessary and sufficient to specify Hcrt neurons...
March 15, 2015: Development
Florian T Merkle, Asif Maroof, Takafumi Wataya, Yoshiki Sasai, Lorenz Studer, Kevin Eggan, Alexander F Schier
Hypothalamic neurons orchestrate many essential physiological and behavioral processes via secreted neuropeptides, and are relevant to human diseases such as obesity, narcolepsy and infertility. We report the differentiation of human pluripotent stem cells into many of the major types of neuropeptidergic hypothalamic neurons, including those producing pro-opiolemelanocortin, agouti-related peptide, hypocretin/orexin, melanin-concentrating hormone, oxytocin, arginine vasopressin, corticotropin-releasing hormone (CRH) or thyrotropin-releasing hormone...
February 15, 2015: Development
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