9 results on '"Creemers, John W.M."'
Search Results
2. Functional and clinical relevance of novel and known PCSK1 variants for childhood obesity and glucose metabolism.
- Author
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Löffler, Dennis, Behrendt, Susanne, Creemers, John W.M., Klammt, Jürgen, Aust, Gabriela, Stanik, Juraj, Kiess, Wieland, Kovacs, Peter, and Körner, Antje
- Abstract
Objective Variants in Proprotein Convertase Subtilisin/Kexin Type 1 ( PCSK1 ) may be causative for obesity as suggested by monogenic cases and association studies. Here we assessed the functional relevance in experimental studies and the clinical relevance through detailed metabolic phenotyping of newly identified and known PCSK1 variants in children. Results In 52 obese children selected for elevated proinsulin levels and/or impaired glucose tolerance, we found eight known variants and two novel heterozygous variants (c.1095 + 1G > A and p.S24C) by sequencing the PCSK1 gene. Patients with the new variants presented with extreme obesity, impaired glucose tolerance, and PCOS. Functionally, c.1095 + 1G > A caused skipping of exon8 translation and a complete loss of enzymatic activity. The protein was retained within the endoplasmic reticulum (ER) causing ER stress. The p.S24C variant had no functional effect on protein size, cell trafficking, or enzymatic activity. The known variants rs6230, rs35753085, and rs725522 in the 5′ end did not affect PCSK1 promoter activity. In clinical association studies in 1673 lean and obese children, we confirmed associations of rs6232 and rs6234 with BMI-SDS and of rs725522 with glucose stimulated insulin secretion and Matsuda index. We did not find the new variants in any other subjects. Conclusions We identified and functionally characterized two rare novel PCSK1 variants of which c.1095 + 1G > A caused complete loss of protein function. In addition to confirming rs6232 and rs6234 in PCSK1 as polygenic risk variants for childhood obesity, we describe an association of rs725522 with insulin metabolism. Our results support the contribution of PCSK1 variants to obesity predisposition in children. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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- View/download PDF
3. Loss of hypothalamic Furin affects POMC to proACTH cleavage and feeding behavior in high-fat diet-fed mice.
- Author
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Coppola, Ilaria, Brouwers, Bas, Walker, Leanne, Alar, Ceyhun, Meulemans, Sandra, White, Anne, Ramos-Molina, Bruno, and Creemers, John W.M.
- Abstract
The hypothalamus regulates feeding and glucose homeostasis through the balanced action of different neuropeptides, which are cleaved and activated by the proprotein convertases PC1/3 and PC2. However, the recent association of polymorphisms in the proprotein convertase FURIN with type 2 diabetes, metabolic syndrome, and obesity, prompted us to investigate the role of FURIN in hypothalamic neurons controlling glucose and feeding. POMC-Cre
+/− mice were bred with Furinfl/fl mice to generate conditional knockout mice with Furin- deletion in neurons expressing proopiomelanocortin (POMC Fur KO), and Furinfl/fl mice were used as controls. POMC Fur KO and controls were periodically monitored on both normal chow diet and high fat diet (HFD) for body weight and glucose tolerance by established in-vivo procedures. Food intake was measured in HFD-fed Fur KO and controls. Hypothalamic Pomc mRNA was measured by RT-qPCR. ELISAs quantified POMC protein and resulting peptides in the hypothalamic extracts of POMC Fur KO mice and controls. The in-vitro processing of POMC was studied by biochemical techniques in HEK293T and CHO cell lines lacking FURIN. In control mice, Furin mRNA levels were significantly upregulated on HFD feeding, suggesting an increased demand for FURIN activity in obesogenic conditions. Under these conditions, the POMC Fur KO mice were hyperphagic and had increased body weight compared to Furinfl/fl mice. Moreover, protein levels of POMC were elevated and ACTH concentrations markedly reduced. Also, the ratio of α-MSH/POMC was decreased in POMC Fur KO mice compared to controls. This indicates that POMC processing was significantly reduced in the hypothalami of POMC Fur KO mice, highlighting for the first time the involvement of FURIN in the cleavage of POMC. Importantly, we found that in vitro , the first stage in processing where POMC is cleaved into proACTH was achieved by FURIN but not by PC1/3 or the other proprotein convertases in cell lines lacking a regulated secretory pathway. These results suggest that FURIN processes POMC into proACTH before sorting into the regulated secretory pathway, challenging the dogma that PC1/3 and PC2 are the only convertases responsible for POMC cleavage. Furthermore, its deletion affects feeding behaviors under obesogenic conditions. • Hypothalamic Furin mRNA levels were significantly upregulated on high-fat feeding. • High-fat diet fed mice lacking Furin in POMC neurons were hyperphagic and had increased body weight. • POMC processing to proACTH was significantly reduced in the hypothalami of mice lacking Furin in POMC neurons. • FURIN is a POMC-processing enzyme. [ABSTRACT FROM AUTHOR]- Published
- 2022
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4. Loss of the proprotein convertase Furin in T cells represses mammary tumorigenesis in oncogene-driven triple negative breast cancer.
- Author
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He, Zongsheng, Khatib, Abdel-Majid, and Creemers, John W.M.
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TRIPLE-negative breast cancer , *T cells , *SUPPRESSOR cells , *TUMOR growth - Abstract
Immunotherapeutic interventions have become an important treatment for various cancer types including triple negative breast cancer (TNBC). Previous studies have shown that T cell-specific Furin deficient mice show regulatory CD4+ T cells (Tregs) malfunction phenotypes due to impaired cleavage of proTGF-β1. However, it is unknown how this phenotype influences tumor initiation and progression in TNBC. Here, we first show that there is a higher level of Furin expression in different immune cells compared to other proprotein convertase members, and its expression is clearly upregulated once immune cells are activated. Moreover, Furin expression levels negatively correlated with an abundance of different infiltrating immune cells in TNBC tumor samples. In an oncogene-induced TNBC mouse model, we demonstrate that Furin inactivation in T cells inhibits primary tumor growth and lung metastasis. Disruption of Furin in T cells in these mice led to a decreased peripheral and tumor-infiltrating Tregs. As a consequence, tumor-infiltrating CD8+ T cells showed a strong proliferative capacity and upregulated expression of IFN-γ and TNF-α. In these mice the repressed tumor growth was associated with induced apoptosis, which led to reduced lung metastases formation. Taken together, these finding revealed the potential therapeutic benefit of targeting Furin in cancer, particularly for immunotherapeutic interventions to treat TNBC. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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5. Amyloid precursor protein is not processed by furin, PACE 4, PC1/3, PC2, PC4 and PC5/6 of the furin family of proprotein processing enzymes
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De Strooper, Bart, Creemers, John W.M., Moechers, Dieder, Huylebroeck, Danny, Van De Ven, Wim J.M., Van Leuven, Fred, and Van den Berghe, Herman
- Published
- 1995
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6. Transgenic Artifacts Caused by Passenger Human Growth Hormone.
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de Faudeur, Geoffroy, Brouwers, Bas, Schuit, Frans, Creemers, John W.M., and Ramos-Molina, Bruno
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HUMAN growth hormone , *TRANSGENE expression , *TRANSGENIC mice , *PROLACTIN receptors , *NEUROENDOCRINOLOGY - Abstract
The minigene encoding human growth hormone (hGH) has been incorporated into over 300 transgenic mouse lines to improve transgene expression. However, unexpected and functional hGH expression can drastically alter physiology. We list here the mouse lines in which ectopic hGH has been confirmed, and we provide a wiki for lines awaiting analysis. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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7. Endosome to trans-Golgi network transport of Proprotein Convertase 7 is mediated by a cluster of basic amino acids and palmitoylated cysteines.
- Author
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Declercq, Jeroen, Ramos-Molina, Bruno, Sannerud, Ragna, Brouwers, Bas, Pruniau, Vincent P.E.G., Meulemans, Sandra, Plets, Evelyn, Annaert, Wim, and Creemers, John W.M.
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ENDOSOMES , *PROPROTEIN convertases , *FURIN protein , *AMINO acids , *CYTOPLASM - Abstract
Proprotein Convertase 7 (PC7) is a Furin-like endoprotease that cleaves precursor proteins at basic amino acids. PC7 is concentrated in the trans -Golgi network (TGN) but it shuttles between the plasma membrane and the TGN depending on sequences in the cytoplasmic tail. A short region containing a three amino acids motif, P 724 -L 725 -C 726 , is essential and sufficient for internalization of PC7 but not for TGN localization, which requires the additional presence of the juxtamembrane region. In this study we have investigated the contribution of a cluster of basic amino acids and two reversibly palmitoylated cysteine residues to endocytic trafficking. Stable cell lines overexpressing chimeric proteins (CD25 and CD46) containing the cytoplasmic domain of PC7 in which the basic cluster alone or together with both palmitoylated cysteines are mutated showed enhanced surface expression as demonstrated by immunofluorescence experiments and surface biotinylation. The mutant proteins no longer recycled to the TGN in antibody uptake experiments and accumulated in an endosomal compartment. Recycling of wild type PC7 to the TGN is blocked by nocodazole, suggesting that PC7 shuttles to the TGN via late endosomes, similar to Furin. Unlike furin, however, PC7 was found to recycle to a region within the TGN, which is deficient in sialyltransferase, as shown by resialylation experiments. In conclusion, a novel motif, composed of a basic amino acid cluster and two palmitoylated cysteines are essential for TGN localization and endocytic trafficking. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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8. Curcumin affects proprotein convertase activity: Elucidation of the molecular and subcellular mechanism.
- Author
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Zhu, Jingjing, Bultynck, Geert, Luyten, Tomas, Parys, Jan B., Creemers, John W.M., Van de Ven, Wim J.M., and Vermorken, Alphons J.M.
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CURCUMIN , *PROPROTEIN convertases , *ENZYME activation , *MOLECULAR biology , *CANCER treatment , *VASCULAR endothelial growth factors - Abstract
Abstract: Proprotein convertases (PCs) form a group of serine endoproteases that are essential for the activation of proproteins into their active form. Some PCs have been proposed to be potential therapeutic targets for cancer intervention because elevated PC activity has been observed in many different cancer types and because many of the PC substrates, such as pro-IGF-1R, pro-TGF-beta, pro-VEGF, are involved in signaling pathways related to tumor development. Curcumin, reported to possess anticancer activity, also affects many of these pathways. We therefore investigated the effect of curcumin on PC activity. Our results show that curcumin inhibits PC activity in a cell lysate-based assay but not in vitro. PC zymogen maturation in the endoplasmic reticulum appears to be inhibited by curcumin. Treating cells with thapsigargin or cyclopiazonic acid, two structurally unrelated inhibitors of the sarco- and endoplasmic reticulum Ca2+ATPase (SERCA), also hampered both the PC zymogen maturation and the PC activity. Importantly, curcumin, like the SERCA inhibitors, impaired ATP-driven 45Ca2+ uptake in the endoplasmic reticulum. These results indicate that curcumin likely restrains PC activity by inhibiting SERCA-mediated Ca2+-uptake activity. Experiments in three colon cancer cell lines confirm that curcumin inhibits both the 45Ca2+ uptake and PC activity, notably the processing of pro-IGF-1R. Both curcumin and thapsigargin inhibit the anchorage-independent growth of these three colon carcinoma cell lines. In conclusion, our findings indicate that curcumin inhibits PC zymogen maturation and consequently PC activity and that its inhibitory effect on Ca2+ uptake into the ER allows and is sufficient to explain this phenomenon. [Copyright &y& Elsevier]
- Published
- 2013
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9. Proprotein convertases in human atherosclerotic plaques: The overexpression of FURIN and its substrate cytokines BAFF and APRIL
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Turpeinen, Hannu, Raitoharju, Emma, Oksanen, Anna, Oksala, Niku, Levula, Mari, Lyytikäinen, Leo-Pekka, Järvinen, Otso, Creemers, John W.M., Kähönen, Mika, Laaksonen, Reijo, Pelto-Huikko, Markku, Lehtimäki, Terho, and Pesu, Marko
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ATHEROSCLEROTIC plaque , *ENZYMES , *GENE expression , *CYTOKINES , *CHOLESTEROL metabolism , *ATHEROSCLEROSIS , *IMMUNOHISTOCHEMISTRY - Abstract
Abstract: Background: Proprotein convertase subtilisin/kexin (PCSK) enzymes cleave proproteins into mature end products. Previously, MBTPS1 and PCSK9 have been shown to regulate cholesterol metabolism and LDL receptor recycling, whereas FURIN and PCSK5 have been suggested to inactivate lipases and regulate inflammation in atherosclerosis. Here, we systematically analyzed the expression of PCSKs and their targets in advanced atherosclerotic plaques. Methods and results: Microarray and quantitative real-time PCR experiments showed that FURIN (42.86 median fold, p =2.1e−8), but no other PCSK, is universally overexpressed in the plaques of different vascular regions. The mRNA expression screen of PCSK target proteins in plaques identified many known factors, but it also identified the significant upregulation of the previously overlooked furin-processed B cell activating cytokines APRIL (TNFSF13, 2.52 median fold, p =3.0e−5) and BAFF (TNFSF13B, 2.97 median fold, p =7.6e−6). The dysregulation of FURIN did not associate with its htSNPs or the previously reported regulatory SNP (−229, rs4932178) in the promoter. Immunohistochemistry experiments showed the upregulation of FURIN in the plaque lymphocytes and macrophages where it was co-expressed with BAFF/TNFSF13B and APRIL/TNFSF13. Conclusions: Our data unequivocally show that FURIN is the primary PCSK that is dysregulated in the immune cells of advanced human atherosclerotic plaques, which implies a role for this enzyme in plaque pathology. Therefore, drugs that inhibit FURIN in arteries may modulate the course of this disease. [Copyright &y& Elsevier]
- Published
- 2011
- Full Text
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