TY - JOUR AB - Epidemiological studies established an association between chronic inflammation and higher risk of cancer. Inhibition of proteolytic enzymes represents a potential treatment strategy for cancer and prevention of cancer metastasis. Cathepsin C (CatC) is a highly conserved lysosomal cysteine dipeptidyl aminopeptidase required for the activation of pro-inflammatory neutrophil serine proteases (NSPs, elastase, proteinase 3, cathepsin G and NSP-4). NSPs are locally released by activated neutrophils in response to pathogens and non-infectious danger signals. Activated neutrophils also release neutrophil extracellular traps (NETs) that are decorated with several neutrophil proteins, including NSPs. NSPs are not only NETs constituents but also play a role in NET formation and release. Although immune cells harbor large amounts of CatC, additional cell sources for this protease exists. Upregulation of CatC expression was observed in different tissues during carcinogenesis and correlated with metastasis and poor patient survival. Recent mechanistic studies indicated an important interaction of tumor-associated CatC, NSPs, and NETs in cancer development and metastasis and suggested CatC as a therapeutic target in a several cancer types. Cancer cell-derived CatC promotes neutrophil recruitment in the inflammatory tumor microenvironment. Because the clinical consequences of genetic CatC deficiency in humans resulting in the elimination of NSPs are mild, small molecule inhibitors of CatC are assumed as safe drugs to reduce the NSP burden. Brensocatib, a nitrile CatC inhibitor is currently tested in a phase 3 clinical trial as a novel anti-inflammatory therapy for patients with bronchiectasis. However, recently developed CatC inhibitors possibly have protective effects beyond inflammation. In this review, we describe the pathophysiological function of CatC and discuss molecular mechanisms substantiating pharmacological CatC inhibition as a potential strategy for cancer treatment. AU - Korkmaz, B.* AU - Lamort, A.-S. AU - Domain, R.* AU - Beauvillain, C.* AU - Gieldon, A.* AU - Yildirim, A.Ö. AU - Stathopoulos, G.T. AU - Rhimi, M.* AU - Jenne, D. AU - Kettritz, R.* C1 - 63455 C2 - 51355 CY - The Boulevard, Langford Lane, Kidlington, Oxford Ox5 1gb, England SP - 114803 TI - Cathepsin C inhibition as a potential treatment strategy in cancer. JO - Biochem. Pharmacol. VL - 194 PB - Pergamon-elsevier Science Ltd PY - 2021 SN - 0006-2952 ER - TY - JOUR AB - Cathepsin C (CatC) is a tetrameric cysteine dipeptidyl aminopeptidase that plays a key role in activation of pro-inflammatory serine protease zymogens by removal of a N-terminal pro-dipeptide sequence. Loss of function mutations in the CatC gene is associated with lack of immune cell serine protease activities and cause Papillon-Lefevre syndrome (PLS). Also, only very low levels of elastase-like protease zymogens are detected by proteome analysis of neutrophils from PLS patients. Thus, CatC inhibitors represent new alternatives for the treatment of neutrophil protease-driven inflammatory or autoimmune diseases. We aimed to experimentally inactivate and lower neutrophil elastase-like proteases by pharmacological blocking of CatC-dependent maturation in cell-based assays and in vivo. Isolated, immature bone marrow cells from healthy donors pulse-chased in the presence of a new cell permeable cyclopropyl nitrile CatC inhibitor almost totally lack elastase. We confirmed the elimination of neutrophil elastase-like proteases by prolonged inhibition of CatC in a non-human primate. We also showed that neutrophils lacking elastase-like protease activities were still recruited to inflammatory sites. These preclinical results demonstrate that the disappearance of neutrophil elastase-like proteases as observed in PLS patients can be achieved by pharmacological inhibition of bone marrow CatC. Such a transitory inhibition of CatC might thus help to rebalance the protease load during chronic inflammatory diseases, which opens new perspectives for therapeutic applications in humans. AU - Guarino, C.* AU - Hamon, Y. AU - Croix, C.* AU - Lamort, A.-S. AU - Dallet-Choisy, S.* AU - Marchand-Adam, S.* AU - Lesner, A.* AU - Baranek, T.* AU - Viaud-Massuard, M.C.* AU - Lauritzen, C.* AU - Pedersen, J.H.* AU - Heuze-Vourc'h, N.* AU - Si-Tahar, M.* AU - Firatli, E.* AU - Jenne, D. AU - Gauthier, F.* AU - Horwitz, M.S.* AU - Borregaard, N.* AU - Korkmaz, B.* C1 - 51029 C2 - 42902 CY - Oxford SP - 52-67 TI - Prolonged pharmacological inhibition of cathepsin C results in elimination of neutrophil serine proteases. JO - Biochem. Pharmacol. VL - 131 PB - Pergamon-elsevier Science Ltd PY - 2017 SN - 0006-2952 ER - TY - JOUR AU - Fimognari, C.* AU - Berti, F.* AU - Nüsse, M. AU - Cantelli-Forti, G.* AU - Hrelia, P.* C1 - 2861 C2 - 22693 SP - 2047-2056 TI - Induction of apoptosis in two human leukemia cell lines as well as differentiation in human promyelocytic cells by cyanidin-3-O-ß-glucopyranoside. JO - Biochem. Pharmacol. VL - 67 PY - 2004 SN - 0006-2952 ER - TY - JOUR AU - Fimognari, C.* AU - Nüsse, M. AU - Berti, F.* AU - Iori, R.* AU - Cantelli, Forti, G.* AU - Hrelia, P.* C1 - 2863 C2 - 22695 SP - 1133-1138 TI - Isothiocyanates as novel cytotoxic and cytostatic agents : molecular pathway on human transformed and non-transformed cells. JO - Biochem. Pharmacol. VL - 68 PY - 2004 SN - 0006-2952 ER - TY - JOUR AU - Neu, B.* AU - Puschmann, A.J.* AU - Mayerhofer, A.* AU - Hutzler, P. AU - Grossmann, J.* AU - Lippl, F.* AU - Schepp, W.* AU - Prinz, C.* C1 - 10336 C2 - 21094 SP - 1755-1760 TI - TNF-alpha induces apoptosis of parietal cells. JO - Biochem. Pharmacol. VL - 65 PY - 2003 SN - 0006-2952 ER - TY - JOUR AB - A protein capable of specifically binding polychlorinated biphenyls (PCB) was partially purified from rat liver cytosol. After labeling with [3H]2,2',4,4',5,5'-hexachlorobiphenyl (6-CB), protein enrichment was guided by monitoring the protein-bound radioactivity through a sequence of purification steps including ion exchange chromatography and preparative gel electrophoresis. In addition, specific binding tests of individual fractions were carried out. An average 100-fold enrichment of the 40 kDa protein was achieved. A variety of ligands were tested in competitive binding studies with 6-CB. Whereas penta- and hexachloro-PCB congeners are high affinity competitors, the 3,3',4,4'-tetrachlorobiphenyl congener does not compete for 6-CB binding. Studies on the species and tissue distribution suggest a prevalence of the binding protein in tissues of the rat. Since the natural physiological ligand of the protein has not yet been identified, the function of the protein can only be speculated on. AU - Bründl, A. AU - Buff, K. C1 - 20393 C2 - 13591 SP - 885-891 TI - Partial Purification and Characterization of a Rat Liver Polychlorinated Biphenyl (PCB) Binding Protein. JO - Biochem. Pharmacol. VL - 45 IS - 4 PY - 1993 SN - 0006-2952 ER - TY - JOUR AB - A protein capable of specifically binding polychlorinated biphenyls (PCB) was partially purified from rat liver cytosol. After labeling with [3H]2,2',4,4',5,5'-hexachlorobiphenyl (6-CB), protein enrichment was guided by monitoring the protein-bound radioactivity through a sequence of purification steps including ion exchange chromatography and preparative gel electrophoresis. In addition, specific binding tests of individual fractions were carried out. An average 100-fold enrichment of the 40 kDa protein was achieved. A variety of ligands were tested in competitive binding studies with 6-CB. Whereas penta- and hexachloro-PCB congeners are high affinity competitors, the 3,3',4,4'-tetrachlorobiphenyl congener does not compete for 6-CB binding. Studies on the species and tissue distribution suggest a prevalence of the binding protein in tissues of the rat. Since the natural physiological ligand of the protein has not yet been identified, the function of the protein can only be speculated on. AU - Bründl, A. AU - Buff, K. C1 - 40446 C2 - 0 SP - 885-891 TI - Partial purification and characterization of a rat liver polychlorinated biphenyl (PCB) binding protein. JO - Biochem. Pharmacol. VL - 45 IS - 4 PY - 1993 SN - 0006-2952 ER - TY - JOUR AB - Specific binding of polychlorinated biphenyls (PCBs) to rat liver cytosol protein has been detected using the 3H-labeled PCB probe 2,2',4,4',5,5'-hexachlorobiphenyl (6-CB). Binding of 6-CB to cytosol protein is displaced by its non-radioactive congener, is of high affinity (K(d) ~ 3nM) and is saturable (maximal binding capacity B(max) ~ 600 pmol/mg protein). 6-CB binding is not found in liver cytosol of animals fed a PCB-supplemented diet (500 ppm PCB for 5 days). Binding is also in vitro inhibited by high concentrations of triglyceride. PCB congeners such as 3,3',4,4',5-pentachlorobiphenyl as well as the thyroid hormones 3,5,3',5'-tetraiodothyronine and 3,5,3'-triiodothyronine (the latter hormone with an order of magnitude lower affinity) compete for the PCB binding site. On the other hand, a number of biochemically important compounds including the PCB core compound biphenyl and the hormone ligands dexamethasone and estradiol, as well as 2,3,7,8-tetrachlorodibenzo-p-dioxin, do not compete for the 6-CB binding site. The data provide the first evidence of specific binding of unmetabolized PCB congeners to distinct binding sites in rat liver cytosol. AU - Buff, K. AU - Bründl, A. C1 - 40533 C2 - 12854 SP - 965-970 TI - Specific binding of polychlorinated biphenyls to rat liver cytosol protein. JO - Biochem. Pharmacol. VL - 43 IS - 5 PY - 1992 SN - 0006-2952 ER - TY - JOUR AB - Bile acid uptake, an important function of differentiated hepatocytes, is decreased in hepatocellular carcinomas and γ-glutamyltranspeptidase-positive, putatively preneoplastic hepatocytes. Whether hepatic uptake is also changed in carcinogen-induced diploid hepatocytes versus polyploid hepatocytes is unknown. The present study has determined whether the hepatic uptake of three model compounds, an anionic bile acid, an organic cation and a neutral organic compound, into diploid cells is different from that in polyploid hepatocytes. These two hepatocyte populations were separated from the parent freshly isolated hepatocyte suspension by centrifugal elutriation. Flow cytometric analysis indicated that the diploid fraction contained approximately 83% diploid cells and that the polyploid fraction had about 84% polyploid hepatocytes. Initial uptake velocity was determined for taurocholate (1-50 μM), ORG 9426 (20-400 μM), a vecuronium-like cation, and ouabain (20-500 μM). Apparent Km was not different between diploid and polyploid cells for the three tested substrates, whereas apparent Vmax was decreased in diploid hepatocytes for taurocholate and ouabain by 42 and 55%, respectively. There were no changes in the hepatic uptake of ORG 9426. These data indicate that uptake by the bile acid/multispecific carrier is compromised in carcinogen-induced diploid cells. AU - Schwarz, L.R. AU - Watkins, J.B.* C1 - 40687 C2 - 38897 SP - 1195-1201 TI - Uptake of taurocholate, a vecuronium-like organic cation, org 9426, and ouabain into carcinogen-induced diploid and polyploid hepatocytes obtained by centrifugal elutriation. JO - Biochem. Pharmacol. VL - 43 IS - 6 PY - 1992 SN - 0006-2952 ER - TY - JOUR AB - The metabolism of fluperlapine, a neuroleptic dibenzazepine derivative with a N-methyl-piperazinyl substituent, was investigated in continuous cultures of rat and human cells which express various cytochrome P450-dependent monooxygenase activities. The differentiated rat hepatoma cells H4IIEC3/G- and their variants 2sFou and FGC-5 metabolized fluperlapine predominantly by N-oxygenation and only to a minor degree by N-demethylation or glucuronidation of primary phenolic products. Total fluperlapine metabolism in dedifferentiated rat hepatoma cells H5 and partially differentiated human hepatoma cells HepG2 was much smaller than in the differentiated rat hepatoma lines. This was primarily attributable to their low capacity for N-oxygenation. Human lung adenocarcinoma lines NCI-H322 and NCI-H358 formed only trace amounts of fluperlapine N-oxide. Pretreatment of 2sFou cells with benz(a)anthracene, phenobarbital or dexamethasone markedly increased the formation of N-demethylated and glucuronidated products but did not affect the rate of N-oxide formation. Guanethidine and cysteamine, inhibitors of flavin-dependent monooxygenase activity, reduced fluperlapine N-oxidation more strongly than aldrin epoxidation, a marker for cytochrome P450 activity. In contrast, n-octylamine inhibited aldrin epoxidation but was without effect on fluperlapine N-oxygenation. The results suggest that certain cells in continuous culture are capable of expressing flavin-dependent monooxygenase(s) in addition to cytochrome P450-containing monooxygenases. Such cells may offer useful systems for studying the oxidative metabolism of a broad spectrum of xenobiotics and analysing the importance of the two oxygenation reactions for the biological effects of their substrates. AU - Fischer, V.* AU - Wiebel, F.J. C1 - 42451 C2 - 40230 SP - 1327-1333 TI - Metabolism of fluperlapine by cytochrome p450-dependent and flavin-dependent monooxygenases in continuous cultures of rat and human cells. JO - Biochem. Pharmacol. VL - 39 IS - 8 PY - 1990 SN - 0006-2952 ER - TY - JOUR AB - Association of the PCB congener 2,2',4,4',5,5'-hexachlorobiphenyl (6-CB) with cell nuclei has been studied in cultured monolayer human Chang liver cells. Photo-induced formation of covalent bonds determined 6-CB binding to protein of cell nuclei and to DNA. Nuclear binding of 6-CB approached equilibrium after approximately 30 min of incubation. Photo-induced binding in vitro to purified Chang liver cell DNA substantiated direct interaction of the PCB congener with DNA. In monolayer cells, low levels of photo-induced 6-CB DNA adducts could be detected using the very sensitive 32P-postlabeling method. Adduct formation was dependent on 6-CB concentration as well as on incubation time. Highest adduct levels were in the range of 2 X 10(-8). Model reactions in vitro showed photo-induced binding of 6-CB to individual purine deoxyribonucleotide-3'-phosphates. The results demonstrate rapid intracellular movement of the PCB congener into the cell nucleus. The vast majority is associated with nuclear protein, minute amounts of 6-CB are found proximate to the DNA helix as evidenced by photo-induced adducts of purine nucleotides. AU - Buff, K. AU - Wegenke, M. AU - Bründl, A. C1 - 18067 C2 - 10910 SP - 2773-2779 TI - Photo-induced formation of DNA adducts of 2,2',4,4',5,5'-hexachlorobiphenyl in cultured human cells. JO - Biochem. Pharmacol. VL - 38 PY - 1989 SN - 0006-2952 ER - TY - JOUR AB - To examine the response of individual cytochrome P-450 species catalysing the epoxidation of aldrin (Wolff T and Guengerich FP, Biochem Pharmacol36: 2581–2588, 1987), monooxygenase systems reconstituted from these species were assayed in the presence of 5% (v/v) = 0.87 M ethanol. The activity of cytochromes P-450pb.b and P-450pb-d, two enzymes inducible by phenobarbital was increased seven-fold. The activity of two other P-450 enzymes purified from these animals was either inhibited by 50%, as observed for cytochrome P-450pb-c or remained unchanged, as noted with cytochrome P-450pcn-e. Two P-450 enzymes purified from untreated rats, cytochromes P-450ut-f and P-450ut-h, showed an inhibition by 50 and 20%, respectively, while the activity of cytochrome P-450ut-a was slightly increased by 50%. Indirect evidence that solvents enhance aldrin epoxidation by interacting with the hemoprotein was obtained by the finding that ethanol stimulated the activity of cytochrome P-450pb-b already, before addition of the lipid component, l-α-1,2-dilauroyl-in-glycero-3-phosphocholine. The Km of cytochrome P-450pb.b for NADPH cytochrome P-450 reductase was not altered by ethanol indicating that the interaction between the two enzymes was not affected by the solvent. Other results indicate that the stimulatory solvent binds to a site, apart from the type I or type II binding site. The potency of various hydrophylic solvents to modify aldrin epoxidase activity was assayed in microsomes of rats pretreated with phenobarbital and of untreated male rats. Ethanol, n-propranol, n-butanol, acetone and tetrahydrofuran enhanced enzyme activity of phenobarbital pretreated rats to a maximal extent of two-fold and, at similar concentrations, inhibited the enzyme activity of untreated rats by 50%. The potency of these solvents correlated with their lipophilicity. Methanol and dimethylsulfoxide only slightly modified the activity of induced and noninduced animals. In the presence of 0.5 M n-propranol as the modifying agent, microsomal epoxidase activity of rats pretreated with pregnenolone-16α-carbonitrile, dexamethasone, 3-methylcholanthrene and of control rats was inhibited by 60–80%, whereas the activity of animals pretreated with phenobarbital, DDT, or the polychlorinated biphenyl mixture, Clophen A 50, was stimulated between two- and three-fold. The results reveal that organic solvents frequently used to dissolve monooxygenase substrates may considerably modify the activity of cytochrome P-450 dependent reactions, in particular when purified enzymes are assayed. AU - Wolff, T. AU - Wanders, H. AU - Guengerich, F.P. C1 - 18118 C2 - 10972 SP - 4217-4223 TI - Organic Solvents as Modifyers of Aldrin Epoxidase in Reconstituted Monooxygenase Systems in Microsomes. JO - Biochem. Pharmacol. VL - 38 IS - 23 PY - 1989 SN - 0006-2952 ER - TY - JOUR AB - Chinese hamster ovary (CHO) cells obtain a high capacity to utilize cystine from the growth medium by exposure to cysteamine (2-mercaptoethylamine, MEA) or N-acetylcysteine (NAC). For uptake studies a modified McCoy's 5A medium supplemented with 0.1 mM [35S]cystine was used. The uptake of cystine was dependent on the time of exposure (0-60 min) and the concentrations of MEA or NAC (0-8 mM). At high concentrations of MEA or NAC, the uptake of cystine became saturated. Half-maximal uptake of cysteine was observed at concentrations of 0.12 mM MEA and 0.66 mM NAC, respectively. Increase in temperature (37-44°) or pH (6.0-8.0) during MEA or NAC exposure further increased the cystine uptake. The increased uptake of cystine was not affected in the presence of glutamate or homocysteate which both inhibited the cystine uptake of control cells. Determination of both reduced (GSH) and oxidized (GSSG) cellular glutathione showed a twofold increase in MEA- or NAC-treated CHO cells. DL-buthionine-S,R-sulfoximine (BSO), an inhibitor of GSH biosynthesis completely blocked the promotion of cystine uptake by MEA and NAC. By further analysis using reversed-phase HPLC of cell extracts, more than 90% of the [35S] radioactive cystine taken up by the cells could be recovered within the pool of GSH. The results demonstrate that exposure of CHO cells with MEA and NAC leads to a promoted uptake of cystine from the culture medium and its rapid utilization for cellular GSH biosynthesis. AU - Issels, R.D. AU - Nagele, A.* AU - Eckert, K.G.* AU - Wllmanns, W.* C1 - 41148 C2 - 36175 SP - 881-888 TI - Promotion of cystine uptake and its utilization for glutathione biosynthesis induced by cysteamine and N-acetylcysteine. JO - Biochem. Pharmacol. VL - 37 IS - 5 PY - 1988 SN - 0006-2952 ER - TY - JOUR AB - Uptake of the persistent environmental chemicals 2,2',4,4',5,5'-hexachlorobiphenyl and 1,1,1-trichloro-2,2-di-(4- chlorophenyl)ethane (the insecticide DDT) by Chang liver cells, an established human cell line, has been investigated. Monolayer cells were incubated with culture medium to which the lipophilic model compounds had been added. The time course of uptake of either compound was biphasic, reaching equilibrium after about 5 hr of incubation. The ratio of DDT:hexachlorobiphenyl uptake was dependent on the presence of serum proteins. Increasing concentrations of serum proteins in the culture medium progressively inhibited uptake. Efflux from the cells was not entirely reversible: 10-20% of the chemicals were not released. Uptake was a linear function of the external concentration of the compounds. Absorptive binding to the outer cell plasma membrane could be determined by removing bound chemicals with fetal calf serum ('back exchange'). With this method, temperature-dependent translocation through the cell plasma membrane could directly be demonstrated. The effect of low temperature as well as the influence of metabolic inhibitors point out the contributin of energy-driven uptake pathways. Demonstration of LDL receptor-like binding protein on Chang liver cells facilitated estimation of the role of receptor-mediated uptake. This route of uptake proved to be of minor importance only, as was transport of the protein- bound chemicals via fluid pinocytosis. The results demonstrate that cellular endocytosis of plasma membrane-bound chemicals constitutes a major uptake pathway for lipophilic chemicals. AU - Mangelsdorf, I. AU - Buff, K. AU - Berndt, J. C1 - 41700 C2 - 36232 SP - 2071-2078 TI - Uptake of persistent environmental chemicals by cultured human cells. JO - Biochem. Pharmacol. VL - 36 IS - 13 PY - 1987 SN - 0006-2952 ER - TY - JOUR AU - Summer, K.H. AU - Greim, H.A. C1 - 41485 C2 - 38567 SP - 1719-1720 TI - Hepatic glutathione S-transferases: activities and cellular localization in rat, rhesus monkey, chimpanzee and man. JO - Biochem. Pharmacol. VL - 30 IS - 12 PY - 1981 SN - 0006-2952 ER -