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Berlin Brandenburg

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  • 1
    Language: English
    In: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, April 2017, Vol.31(4), pp.1421-1433
    Description: CD63 is a ubiquitously expressed member of the tetraspanin superfamily. Using a mating-based split-ubiquitin-yeast 2-hybrid system, pull-down experiments, total internal reflection fluorescence microscopy, Förster resonance energy transfer, and biotinylation assays, we found that CD63 interacts with human organic cation transporter 2 (hOCT2), which transports endogenous and exogenous substrates, such as neurotransmitters and drugs in several epithelial cells. CD63 overexpression affects cellular localization of hOCT2 expressed in human embryonic kidney (HEK)293 cells. Studies with CD63-knockout mice indicate that in renal proximal tubules, CD63 determines the insertion of the mouse ortholog of the transporter into the proper membrane domain and mediates transporter regulation by trafficking processes. In polarized Madin-Darby kidney canine kidney (MDCK) epithelial cells, CD63 and hOCT2 colocalize with the small GTPase Rab4, which controls the rapid recycling from sorting endosomes back to the cell surface. Suitable negative and positive control experiments were performed for each experimental approach. Empty vector transfected cells and wild-type mice were used as control. CD63 seems to play a role in the recycling of hOCT2 from endosomes to the basolateral membrane in polarized epithelia. These data indicate that CD63 has a previously uncharacterized function in regulating trafficking of specific membrane proteins in polarized cells.-Schulze, U., Brast, S., Grabner, A., Albiker, C., Snieder, B., Holle, S., Schlatter, E., Schröter, R., Pavenstädt, H., Herrmann, E., Lambert, C., Spoden, G. A., Florin, L., Saftig, P., Ciarimboli, G. Tetraspanin CD63 controls basolateral sorting of organic cation transporter 2 in renal proximal tubules.
    Keywords: Regulation ; Trafficking ; Transport ; Kidney Tubules, Proximal -- Metabolism ; Organic Cation Transport Proteins -- Metabolism ; Tetraspanin 30 -- Metabolism
    ISSN: 08926638
    E-ISSN: 1530-6860
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  • 2
    Language: English
    In: PLoS ONE, 2011, Vol.6(9), p.e25787
    Description: MicroRNA expression is altered in cancer cells, and microRNAs could serve as diagnostic/prognostic biomarker for cancer patients. Our study was designed to analyze circulating serum microRNAs in patients with renal cell carcinoma (RCC). ; We first explored microRNA expression profiles in tissue and serum using TaqMan Low Density Arrays in each six malignant and benign samples: Although 109 microRNAs were circulating at higher levels in cancer patients' serum, we identified only 36 microRNAs with up-regulation in RCC tissue and serum of RCC patients. Seven candidate microRNAs were selected for verification based on the finding of up-regulation in serum and tissue of RCC patients: miR-7-1*, miR-93, miR-106b*, miR-210, miR-320b, miR-1233 and miR-1290 levels in serum of healthy controls (n = 30) and RCC (n = 33) patients were determined using quantitative real-time PCR (TaqMan MicroRNA Assays). miR-1233 was increased in RCC patients, and thus validated in a multicentre cohort of 84 RCC patients and 93 healthy controls using quantitative real-time PCR (sensitivity 77.4%, specificity 37.6%, AUC 0.588). We also studied 13 samples of patients with angiomyolipoma or oncocytoma, whose serum miR-1233 levels were similar to RCC patients. Circulating microRNAs were not correlated with clinical-pathological parameters. ; MicroRNA levels are distinctly increased in cancer patients, although only a small subset of circulating microRNAs has a tumor-specific origin. We identify circulating miR-1233 as a potential biomarker for RCC patients. Larger-scaled studies are warranted to fully explore the role of circulating microRNAs in RCC.
    Keywords: Research Article ; Biology ; Medicine ; Urology ; Oncology ; Biochemistry
    E-ISSN: 1932-6203
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  • 3
    Language: English
    In: Cancer Research, 07/15/2016, Vol.76(14 Supplement), pp.476-476
    ISSN: 0008-5472
    E-ISSN: 1538-7445
    Source: CrossRef
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  • 4
    Language: English
    In: Kidney International, June 2014, Vol.85(6), pp.1369-1381
    Description: The role of calcium-activated chloride channels for renal function is unknown. By immunohistochemistry we demonstrate dominant expression of the recently identified calcium-activated chloride channels, Anoctamin 1 (Ano1, TMEM16A) in human and mouse proximal tubular epithelial (PTE) cells, with some expression in podocytes and other tubular segments. Ano1-null mice had proteinuria and numerous large reabsorption vesicles in PTE cells. Selective knockout of Ano1 in podocytes (Ano1-/-/Nphs2-Cre) did not impair renal function, whereas tubular knockout in Ano1-/-/Ksp-Cre mice increased urine protein excretion and decreased urine electrolyte concentrations. Purinergic stimulation activated calcium-dependent chloride currents in isolated proximal tubule epithelial cells from wild-type but not from Ano1-/-/Ksp-Cre mice. Ano1 currents were activated by acidic pH, suggesting parallel stimulation of Ano1 chloride secretion with activation of the proton–ATPase. Lack of calcium-dependent chloride secretion in cells from Ano1-/-/Ksp-Cre mice was paralleled by attenuated proton secretion and reduced endosomal acidification, which compromised proximal tubular albumin uptake. Tubular knockout of Ano1 enhanced serum renin and aldosterone concentrations, probably leading to enhanced compensatory distal tubular reabsorption, thus maintaining normal blood pressure levels. Thus, Ano1 has a role in proximal tubular proton secretion and protein reabsorption. The results correspond to regulation of the proton–ATPase by the Ano1-homolog Ist2 in yeast.
    Keywords: Ano1 ; Anoctamin 1 ; Ca2+-Activated Cl− Channels ; H+-Atpase ; Renal Proximal Tubules ; Tmem16a ; Medicine
    ISSN: 0085-2538
    E-ISSN: 1523-1755
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  • 5
  • 6
    Language: English
    In: Clinical cancer research : an official journal of the American Association for Cancer Research, 15 February 2012, Vol.18(4), pp.1101-8
    Description: Knowledge of transporters responsible for the renal secretion of creatinine is key to a proper interpretation of serum creatinine and/or creatinine clearance as markers of renal function in cancer patients receiving chemotherapeutic agents. Creatinine transport was studied in transfected HEK293 cells in vitro and in wild-type mice and age-matched organic cation transporter 1 and 2-deficient [Oct1/2(-/-)] mice ex vivo and in vivo. Clinical pharmacogenetic and transport inhibition studies were done in two separate cohorts of cancer patients. Compared with wild-type mice, creatinine clearance was significantly impaired in Oct1/2(-/-) mice. Furthermore, creatinine inhibited organic cation transport in freshly isolated proximal tubules from wild-type mice and humans, but not in those from Oct1/2(-/-) mice. In a genetic association analysis (n = 590), several polymorphisms around the OCT2/SLC22A2 gene locus, including rs2504954 (P = 0.000873), were significantly associated with age-adjusted creatinine levels. Furthermore, in cancer patients (n = 68), the OCT2 substrate cisplatin caused an acute elevation of serum creatinine (P = 0.0083), consistent with inhibition of an elimination pathway. Collectively, this study shows that OCT2 plays a decisive role in the renal secretion of creatinine. This process can be inhibited by OCT2 substrates, which impair the usefulness of creatinine as a marker of renal function.
    Keywords: Creatinine -- Metabolism ; Kidney Tubules, Proximal -- Metabolism ; Organic Cation Transport Proteins -- Metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma -- Metabolism
    ISSN: 1078-0432
    E-ISSN: 15573265
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  • 7
    Language: English
    In: The Journal of Urology, April 2012, Vol.187(4), pp.1210-1214
    Description: Patients with stage pT3N0 urothelial bladder cancer vary in outcome after radical cystectomy. To improve prognosis estimation a model was recently developed that defines 3 risk groups for recurrence-free survival based on pT substaging, lymphovascular invasion and positive surgical margin. We present what is to our knowledge the first external validation of this risk model. Analogous to the risk model derivation cohort our study group comprised 472 patients with stage pT3, pN0, cM0 disease without perioperative chemotherapy and with a median followup of 42 months (IQR 20–75). The primary end point was recurrence-free survival. The effect of variables was determined by univariate and multivariate Cox regression analysis, and predictive accuracy was determined by ROC analysis. Stage pT3aN0 and pT3bN0 cases showed significantly different recurrence-free survival after 5 years (51% vs 29%, p 〈0.001). In the multivariate Cox model pT3 substage (HR 1.86, p 〈0.001), lymphovascular invasion (HR 1.48, p = 0.002), positive surgical margins (HR 1.90, p = 0.030) and patient age with a dichotomy at 70 years (HR 1.51, p = 0.001) had an independent effect on recurrence-free survival. In the low (221 patients or 47%), intermediate (184 or 39%) and high (67 or 14%) risk groups the 5-year recurrence-free survival rate was 55%, 45% and 13%, respectively (p 〈0.001). The concordance index of the risk model to predict recurrence-free survival was 0.64 (95% CI 0.59–0.69). This user friendly risk model can be recommended to estimate prognosis in patients with stage pT3N0 after radical cystectomy. Patients at high risk showed clearly compromised recurrence-free survival and should be included in adjuvant therapy studies.
    Keywords: Urinary Bladder ; Urothelium ; Carcinoma ; Mortality ; Germany ; Medicine
    ISSN: 0022-5347
    E-ISSN: 1527-3792
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  • 8
    Language: English
    In: Current Cancer Drug Targets, 2015, Vol.15(3), p.243-255
    Description: 〈i〉Introduction〈/i〉: Recently, an increasing number of systemic therapies with life extending capacity have become available in metastatic castration resistant prostate cancer (mCRPC) i.e. Abiraterone acetate, Enzalutamide, Sipuleucel-T, Docetaxel, Cabazitaxel and Radium-223. More compounds are currently being evaluated in promising pivotal trials (e.g. Tasquinimod, ARN-509, ODM-201, and more). Limitations of the currently available biomarkers make treatment decisions challenging. Considering the ever increasing complexity of treatment algorithms in mCRPC the current demand of research is to find and characterize biomarkers with prognostic, predictive and surrogate quality, allowing for information on clinically meaningful outcomes and on which therapy to offer patients in different and complex scenarios. 〈/p〉 〈p〉 〈i〉Methods〈/i〉: A comprehensive English-language literature review was performed through PubMed to identify articles and abstract presentations of the major conferences on cancer during December 2014. 〈/p〉 〈p〉 〈i〉Results〈/i〉: In this review we address established biomarkers like prostate specific antigen, lactate dehydrogenase and alkaline phosphatase. Emerging biomarkers like circulating tumor cells, androgen receptor splice variants, cancer stem cells and imaging biomarkers have also been reviewed and placed in the context of prognostic, predictive and surrogate implications in the current field of CRPC. We elaborate on the requirements of good biomarkers and discuss possible future developments of biomarkers in CRPC. Advances in knowledge of biomarkers in CRPC and thus biomarker driven therapy monitoring and up-front therapy decisions may help in the future to tailor treatment algorithms, give information on which therapy to offer and when to continue a given therapy in equivocal scenarios. This could maximize treatment benefit and minimize toxicity.
    Keywords: Biomarkers Castration Resistant Mcrpc Novel Therapeutics Predictive Prognostic Surrogate.
    ISSN: 1568-0096
    E-ISSN: 1873-5576
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  • 9
    Language: English
    In: Journal of Clinical Oncology, 02/20/2013, Vol.31(6_suppl), pp.264-264
    ISSN: 0732-183X
    E-ISSN: 1527-7755
    Source: CrossRef
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  • 10
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