Decrease of Bcl-xL and augmentation of thymocyte apoptosis in GILZ overexpressing transgenic mice

Blood ◽  
2004 ◽  
Vol 104 (13) ◽  
pp. 4134-4141 ◽  
Author(s):  
Domenico Vittorio Delfino ◽  
Massimiliano Agostini ◽  
Stefania Spinicelli ◽  
Pasquale Vito ◽  
Carlo Riccardi
Keyword(s):  
Transgenic Mice ◽  
Time Course ◽  
Leucine Zipper ◽  
Caspase 3 ◽  
Ex Vivo ◽  
Cell Lineage ◽  
Caspase 8 ◽  
Apoptotic Pathway ◽  
Cell Counts ◽  
Thymocyte Apoptosis

Abstract Glucocorticoids promote thymocyte apoptosis and modulate transcription of numerous genes. GILZ (glucocorticoid-induced leucine zipper), being one of them, is strongly up-regulated in the thymus. To elucidate its function we generated transgenic mice overexpressing it specifically in the T-cell lineage and characterized its influence on thymus function. In young adult transgenic mice CD4+CD8+ thymocyte number was significantly decreased and ex vivo thymocyte apoptosis was increased. Apoptotic pathway analysis detected reduced antiapoptotic B-cell leukemia XL (Bcl-xL) expression and increased activation of caspase-8 and caspase-3. Time-course experiments showed that in wild-type (WT) thymocytes GILZ up-regulation was followed by sequential Bcl-xL decreased expression and activation of caspase-8 and of caspase-3. Moreover, GILZ delivered inside WT thymocytes by a fusion protein with the transactivator of transcription (TAT) peptide decreased Bcl-xL and promoted their apoptosis. In aged mice perturbation of thymic subset numbers was amplified over time, as demonstrated by a further decrease in CD4+CD8+ cells and increases in CD4+CD8-, CD4-CD8-, and CD8+CD4- cell counts. These results support the hypothesis that GILZ participates in the regulation of thymocyte apoptosis by glucocorticoids. (Blood. 2004;104:4134-4141)

scholarly journals LIGHT, a Member of the Tumor Necrosis Factor Ligand Superfamily, Prevents Tumor Necrosis Factor-α-mediated Human Primary Hepatocyte Apoptosis, but Not Fas-mediated Apoptosis

2002 ◽  
Vol 277 (51) ◽  
pp. 50054-50061 ◽  
Author(s):  
Hideki Matsui ◽  
Yukiko Hikichi ◽  
Isamu Tsuji ◽  
Takao Yamada ◽  
Yasushi Shintani
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Time Course ◽  
Caspase 3 ◽  
Nuclear Factor Κb ◽  
Caspase 8 ◽  
Death Domain ◽  
Human Primary Hepatocytes ◽  
Induced Apoptosis ◽  
Necrosis Factor

LIGHT is a member of tumor necrosis factor (TNF) superfamily, and its receptors have been identified as lymphotoxin-β receptor (LTβR) and the herpesvirus entry mediator (HVEM)/ATAR/TR2, both of which lack the cytoplasmic sequence termed the “death domain.” The present study has demonstrated that LIGHT inhibits TNFα-mediated apoptosis of human primary hepatocytes sensitized by actinomycin D (ActD), but not Fas- or TRAIL-mediated apoptosis. Furthermore, LIGHT does not prevent some cell lines such as HepG2 or HeLa from undergoing ActD/TNFα-induced apoptosis. This protective effect requires LIGHT pretreatment at least 3 h prior to ActD sensitization. LIGHT stimulates nuclear factor-κB (NF-κB)-dependent transcriptional activity in human hepatocytes like TNFα. The time course of NF-κB activation after LIGHT administration is similar to that of the pretreatment required for the anti-apoptotic effect of LIGHT. LIGHT inhibits caspase-3 processing on the apoptotic protease cascade in TNFα-mediated apoptosis but not Fas-mediated apoptosis. In addition, increased caspase-3 and caspase-8 activities in ActD/TNFα-treated cells are effectively blocked by LIGHT pretreatment. However, LIGHT does not change the expression of TNFRp55, TNFRp75, and Fas. These results indicate that LIGHT may act as an anti-apoptotic agent against TNFα-mediated liver injury by blocking the activation of both caspase-3 and caspase-8.

scholarly journals Tula hantavirus infection of Vero E6 cells induces apoptosis involving caspase 8 activation

2004 ◽  
Vol 85 (11) ◽  
pp. 3261-3268 ◽  
Author(s):  
Xiao-Dong Li ◽  
Sami Kukkonen ◽  
Olli Vapalahti ◽  
Alexander Plyusnin ◽  
Hilkka Lankinen ◽  
...  
Keyword(s):  
Influenza A ◽  
Time Course ◽  
Caspase 3 ◽  
Hantavirus Infection ◽  
Caspase 8 ◽  
Type I ◽  
Tnf Receptor ◽  
Necrosis Factor Alpha ◽  
Factor Alpha ◽  
Necrosis Factor

Hantaviruses are known to cause two severe human diseases: haemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome. The mechanisms of pathogenesis of these two diseases are progressively becoming understood. Recently, two hantaviruses, Hantaan and Prospect Hill were reported to cause programmed cell death of Vero E6 cells. This study shows that Tula hantavirus (TULV) infection efficiently triggers an apoptotic programme in infected Vero E6 cells, and that the replication of TULV is required for the activation of caspase 3 and the cleavage of poly (ADP-ribose) polymerase, two molecular hallmarks of apoptosis. The enforced treatment of infected Vero E6 cells with tumour necrosis factor alpha (TNF-α), but not interferon alpha (IFN-α), advanced the time course of apoptosis. Furthermore, caspase 8 was activated on day 4 post-infection, the same day when caspase 3 was activated. TNF receptor 1 was induced during a late stage of TULV infection. These data suggest that, unlike during influenza A virus infection, TNF-α, but not type I IFN-α/β, may contribute significantly to apoptosis in a synergistic manner with TULV propagation. Interestingly, pretreatment with a broad-spectrum caspase inhibitor, z-VAD-fmk, efficiently inhibited apoptosis of TULV-infected Vero E6 cells. Taken together, these results suggest that TULV replication initiates a typical apoptotic programme involving caspase 8 activation.

scholarly journals Plant Food Anthocyanins Induced Platelet Apoptosis Via BCL-2/BCL-XL Pathway

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4988-4988
Author(s):  
Yang Yan ◽  
Ma Jing ◽  
Tian Jinju ◽  
Chen Liyi ◽  
Songmei Yin ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Caspase 3 ◽  
Caspase 8 ◽  
Plant Food ◽  
Intrinsic Apoptotic Pathway ◽  
Caspase 9 ◽  
Cytochrome C Release ◽  
Apoptotic Pathway ◽  
Platelet Apoptosis ◽  
Activated Platelets

Abstract Background: Platelets are versatile cells and play important roles in hemostasis/thrombosis, inflammation, and atherosclerosis. The pathogenesis of cardiovascular diseases (CVDs) is linked to platelet hyperactivity which is considered an independent risk factor for CVDs. Platelets are critical for promoting the progression of CVDs, and platelet apoptosis have been reported to be involved in platelet activation. Anthocyanins are major phytochemicals abundant in plant food and have been shown to play a protective role against CVDs. Our previous studies demonstrated that anthocyanins from plant food significantly inhibited platelet activation, adhesion, aggregation and granule secretion, as well as attenuated thrombus growth at both arterial and venous shear stresses in vitro and in vivo, however, the effects of anthocyanin on platelet apoptosis and its mechanisms have not been explored. In the present study, we examined whether anthocyanin Cyanidin-3-glucoside (Cy-3-g) affect platelet apoptosis and the BCL-2/BCL-XL intrinsic apoptotic pathway. Methods: Cy-3-g, the predominant bioactive compound of anthocyanin preparations, was obtained from Polyphenol AS Company in Norway.Purified gel-filtered platelets from healthy volunteers were incubated at 37oC for 40 minutes with different concentrations of Cy-3-g (0.5、5、50μM) or PBS buffer as a control. the activated platelets were triggered with 0.5U thrombin for 15min to induce apoptosis. Mitochondria membrane potential (Δψm) and membrane phospholipid phosphatidylserine (PS) exposure in both activated and resting platelets were assessed by flow cytometry. Cytochrome C release, activation of caspase-3, caspase-8, caspase-9, cleavage of gelsolin, the levels of anti-apoptotic BCL-2 family proteins such as BCL-2, BCL-XL and proapoptotic BCL-2 family proteins Bax, Bak, Bad, Bid and tBid in both activated and resting platelets were measured by western blotting. Results: Cy-3-g at 5μM and 50μM directly induced significant ΔΨm dissipation in activated platelets dose dependently. Correspondingly, 50μM Cy-3-g increased cytochrome C release compared to control. The expression of pro-caspase-8 and pro-caspase-9 decreased, activation of caspase-3, caspase-8 and caspase-9 was induced in activated platelets in both 5μM and 50μM Cy-3-g groups. Both PS exposure and the cleavage of gelsolin increased in activated platelets, however these effects were only observed at Cy-3-g doses as high as 50μM. Cy-3-g did not induce the above changes in resting platelets. The intrinsic apoptotic pathway was initiated by Cy-3-g treatment in activated platelets; Cy-3-g significantly inhibited the expression of BCL-2, BCL-XL and increased the levels of Bax, Bak, Bad and Bid in activated platelets dose dependently. No significant difference was observed in resting platelets. Conclusions: Our data demonstrate for the first time that purified anthocyanin Cy-3-g directly accelerated apoptosis in activated platelets via the BCL-2/BCL-XL pathway. Anthocyanins may possess therapeutic potential for patients suffering from thrombotic conditions. Disclosures No relevant conflicts of interest to declare.

scholarly journals Moderate exercise attenuates caspase-3 activity, oxidative stress, and inhibits progression of diabetic renal disease in db/db mice

2009 ◽  
Vol 296 (4) ◽  
pp. F700-F708 ◽  
Author(s):  
S. Ghosh ◽  
M. Khazaei ◽  
F. Moien-Afshari ◽  
L. S. Ang ◽  
D. J. Granville ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Diabetic Nephropathy ◽  
Oxidative Damage ◽  
Renal Disease ◽  
Caspase 3 ◽  
Moderate Intensity ◽  
Caspase 8 ◽  
Apoptotic Pathway ◽  
Mitochondrial Apoptotic Pathway ◽  
Early Diabetic Nephropathy

Diabetic nephropathy, the leading cause of end-stage renal disease, is characterized by a proapoptotic and prooxidative environment. The mechanisms by which lifestyle interventions, such as exercise, benefit diabetic nephropathy are unknown. We hypothesized that exercise inhibits early diabetic nephropathy via attenuation of the mitochondrial apoptotic pathway and oxidative damage. Type 2 diabetic db/db and normoglycemic wild-type mice were exercised for an hour everyday at a moderate intensity for 7 wk, following which renal function, morphology, apoptotic signaling, and oxidative stress were evaluated. Exercise reduced body weight, albuminuria, and pathological glomerular expansion in db/db mice independent of hyperglycemic status. Changes in renal morphology were also related to reduced caspase-3 (main effector caspase in renal apoptosis), caspase-8 (main initiator caspase of the “extrinsic” pathway) activities, and TNF-α expression. A role for the mitochondrial apoptotic pathway was unlikely as both caspase-9 activity (initiator caspase of this pathway) and expression of regulatory proteins such as Bax and Bcl-2 were unchanged. Kidneys from db/db mice also produced higher levels of superoxides and had greater oxidative damage concurrent with downregulation of superoxide dismutase (SOD) 1 and 3. Interestingly, although exercise also increased superoxides, there was also upregulation of multiple SODs that likely inhibited lipid (hydroperoxides) and protein (carbonyls and nitrotyrosine) oxidation in db/db kidneys. In conclusion, exercise can inhibit progression of early diabetic nephropathy independent of hyperglycemia. Reductions in caspase-3 and caspase-8 activities, with parallel improvements in SOD expression and reduced oxidative damage, could underlie the beneficial effects of exercise in diabetic kidney disease.

The regulation of RNA Polymerase II mediated transcription by Caspase-8 in ovarian cancer.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e18082-e18082
Author(s):  
Sven Becker ◽  
Ranadip Mandal ◽  
Monika Raab ◽  
Mourad Sanhaji ◽  
Klaus Strebhardt
Keyword(s):  
Ovarian Cancer ◽  
Rna Polymerase Ii ◽  
Ex Vivo ◽  
New Drugs ◽  
Caspase 8 ◽  
Low Grade ◽  
Apoptotic Pathway ◽  
Rna Pol Ii ◽  
Pol Ii

e18082 Background: According to the WHO, in 2018, the number of new cases and mortality worldwide due to ovarian cancer, were 295,414 and 184,799, respectively, making it one of the most lethal gynaecological cancers in the world. The standard therapy of ovarian cancers is still comprehensive cytoreductive surgery followed by a combination of platinum-taxane based primary chemotherapy. Second line therapies include Carboplatin or Cisplatin in combination with Paclitaxel, PLD or Gemcitabine, with or without Bevacizumab. Unfortunately, late stage ovarian cancer cases are still incurable, however, several new drugs are under development or are undergoing clinical trials. At present, these new approaches can only stabilize the disease or delay its recurrence. Caspase-8, the predominant initiator of the extrinsic apoptotic pathway, also plays critical roles in a number of other non-apoptotic functions. It is also frequently down-regulated in ovarian cancer and we wanted to understand how this down-regulation could support the development of ovarian cancer. Methods: The analysis of the association between CASP8 expression and patient prognosis in ovarian cancer patients found that low CASP8 expression was significantly correlated with poor OS, with a median OS of 37.43 (low expression) and 49.97 (high expression) months, and higher clinical stages. CRISPR/Cas9 mediated CASP8 KO in multiple high- and low-grade ovarian cancer cell lines resulted in significant increases in their invasiveness. Results: Caspase-8 interactome analysis revealed that it can regulate the RNA Pol II mediated transcription, which was corroborated through multiple in vitro and ex vivo experiments. Global Transcriptomics and Proteomics analysis of the KO cells revealed significant upregulation of genes involved in metastasis, which was again verified in in vitro and ex vivo experiments. Additionally, the KO cells were significantly resistant towards standard chemotherapeutics like Carboplatin, a transcription inhibitor, either alone or in combination with Paclitaxel. Conclusions: Presently, we are working on orthotopic ovarian cancer mouse models to validate - the potential of Caspase-8 to regulate metastasis; and the targeting of RNA Pol II to sensitize them to standard chemotherapeutics. We are also developing organoid banks from patient derived ovarian cancer tissues, with low Caspase-8 expression, to validate the upregulation of genes identified in our ‘omics’ data and the potential of targeting them, along with RNA Pol II, to sensitize them to standard chemotherapeutics.

109. TROPHOBLAST DEPORTATION IS DEPENDENT UPON CASPASES 3, 8 AND ROCK

2009 ◽  
Vol 21 (9) ◽  
pp. 28
Author(s):  
K. J. Askelund ◽  
P. Stone ◽  
L. W. Chamley
Keyword(s):  
Caspase 3 ◽  
First Trimester ◽  
Normal Pregnancy ◽  
Maternal Blood ◽  
Caspase 8 ◽  
Caspase 9 ◽  
Apoptotic Pathway ◽  
Apoptosis Pathway ◽  
Villous Trophoblast

Background: Trophoblast deportation is the process whereby multinucleated fragments of the syncytiotrophoblast are shed from the placenta into the maternal blood. It is estimated that 150,000 are shed from the placenta and deported daily in normal pregnancy and that more are shed during preeclampsia1. In normal pregnancy deported trophoblasts are thought to die by apoptosis, which is also increased in villous trophoblast in preeclampsia2. However, experimental confirmation that apoptosis leads to trophoblast shedding is required and it is not clear which components of the apoptotic pathway are involved in trophoblast shedding. Objectives: To determine the effect of inhibiting caspase 3 (executioner), caspases 8 and 9 (initiators), and Rho-associated kinase (ROCK; bleb formation) on the number of trophoblasts shed from first trimester human placentae. Methods : Using an in vitro placental explant model of trophoblast deportation, first trimester placentae were cultured for 72 hours in media containing specific inhibitors of ROCK, caspases 3, 8 or 9. Trophoblasts shed from quintuple explants/inhibitor from five placentae were depleted of contaminating leucocytes and erythrocytes, labelled with trypan blue and the sizes and numbers of shed trophoblasts quantified using a Nexcelom automated counter. Results: The number of trophoblasts that were shed from the explants was significantly increased (p=0.04) when caspase 3 (2.4 fold) and caspase 8 (2.7 fold) were inhibited. There was no significant change following caspase 9 inhibition. The number of shed trophoblasts was significantly decreased when ROCK was inhibited. None of the inhibitors significantly altered the size of the shed trophoblasts. Conclusion: Our data suggest that the apoptosis pathway is involved in trophoblast shedding in vitro from first trimester placentae. That caspase 8 but not caspase 9 affected shedding suggests trophoblasts from normal placentae are induced to die via the extrinsic apoptosis pathway. Aberrant regulation of the apoptosis pathway may contribute to pregnancy pathology.

scholarly journals AAV8 vector expressing IL24 efficiently suppresses tumor growth mediated by specific mechanisms in MLL/AF4-positive ALL model mice

Blood ◽  
2012 ◽  
Vol 119 (1) ◽  
pp. 64-71 ◽  
Author(s):  
Hayato Tamai ◽  
Koichi Miyake ◽  
Hiroki Yamaguchi ◽  
Miyuki Takatori ◽  
Kazuo Dan ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Caspase 3 ◽  
Lymphoblastic Leukemia ◽  
C2c12 Cell ◽  
Caspase 8 ◽  
Apoptotic Pathway ◽  
High Relapse Rate ◽  
Tnf Α

Abstract Mixed-lineage leukemia (MLL)/AF4-positive acute lymphoblastic leukemia (ALL) is a common type of leukemia in infants, which is associated with a high relapse rate and poor prognosis. IL24 selectively induces apoptosis in cancer cells and exerts immunomodulatory and antiangiogenic effects. We examined the effects of adeno-associated virus type 8 (AAV8) vector-mediated muscle-directed systemic gene therapy in MLL/AF4-positive ALL using IL24. In a series of in vitro studies, we examined the effects of AAV8-IL24–transduced C2C12 cell-conditioned medium. We also examined the effects of AAV8-IL24 in MLL/AF4 transgenic mice. The results revealed the effects of AAV8-IL24 in MLL/AF4-positive ALL both in vitro and in vivo. With regard to the mechanism of therapy using AAV8-IL24 in MLL/AF4-positive ALL, we demonstrated the antiangiogenicity and effects on the ER stress pathway and unreported pathways through inhibition of S100A6 and HOXA9, which is specific to MLL/AF4-positive ALL. Inhibition of S100A6 by IL24 was dependent on TNF-α and induced acetylation of p53 followed by activation of the caspase 8–caspase 3 apoptotic pathway. Inhibition of HOXA9 by IL24, which was independent of TNF-α, induced MEIS1 activation followed by activation of the caspase 8–caspase 3 apoptotic pathway. Thus, gene therapy using AAV8-IL24 is a promising treatment for MLL/AF4-positive ALL.

scholarly journals The MEK-ERK-MST1 Axis Potentiates the Activation of the Extrinsic Apoptotic Pathway during GDC-0941 Treatment in Jurkat T Cells

Cells ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 191 ◽  
Author(s):  
Jana Nováková ◽  
Pavel Talacko ◽  
Petr Novák ◽  
Karel Vališ
Keyword(s):  
T Cells ◽  
Caspase 3 ◽  
Phosphorylation Site ◽  
Conditional Knockout ◽  
Caspase 8 ◽  
Pi3k Signaling ◽  
Hippo Signaling ◽  
Dependent Manner ◽  
Jurkat T Cells ◽  
Apoptotic Pathway

The discrete activation of individual caspases is essential during T-cell development, activation, and apoptosis. Humans carrying nonfunctional caspase-8 and caspase-8 conditional knockout mice exhibit several defects in the progression of naive CD4+ T cells to the effector stage. MST1, a key kinase of the Hippo signaling pathway, is often presented as a substrate of caspases, and its cleavage by caspases potentiates its activity. Several studies have focused on the involvement of MST1 in caspase activation and also reported several defects in the immune system function caused by MST1 deficiency. Here, we show the rapid activation of the MEK-ERK-MST1 axis together with the cleavage and activation of caspase-3, -6, -7, -8, and -9 after PI3K signaling blockade by the selective inhibitor GDC-0941 in Jurkat T cells. We determined the phosphorylation pattern of MST1 using a phosphoproteomic approach and identified two amino acid residues phosphorylated in an ERK-dependent manner after GDC-0941 treatment together with a novel phosphorylation site at S21 residue, which was extensively phosphorylated in an ERK-independent manner during PI3K signaling blockade. Using caspase inhibitors and the inhibition of MST1 expression using siRNA, we identified an exclusive role of the MEK-ERK-MST1 axis in the activation of initiator caspase-8, which in turn activates executive caspase-3/-7 that finally potentiate MST1 proteolytic cleavage. This mechanism forms a positive feed-back loop that amplifies the activation of MST1 together with apoptotic response in Jurkat T cells during PI3K inhibition. Altogether, we propose a novel MEK-ERK-MST1-CASP8-CASP3/7 apoptotic pathway in Jurkat T cells and believe that the regulation of this pathway can open novel possibilities in systemic and cancer therapies.

scholarly journals Human Embryonic Stem Cells Acquire Responsiveness to TRAIL upon Exposure to Cisplatin

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Lucie Pešková ◽  
Vladimír Vinarský ◽  
Tomáš Bárta ◽  
Aleš Hampl
Keyword(s):  
Stem Cells ◽  
Dna Damage ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Caspase 3 ◽  
Embryonic Stem ◽  
Caspase 8 ◽  
Apoptotic Pathway ◽  
Extrinsic Apoptotic Pathway ◽  
Induced Apoptosis

Tumor necrosis factor-related apoptosis-inducing ligand—TRAIL—is a protein operating as a ligand capable of inducing apoptosis particularly in cancerously transformed cells, while normal healthy cells are typically nonresponsive. We have previously demonstrated that pluripotent human embryonic stem cells (hESC) are also refractory to TRAIL, even though they express all canonical components of the death receptor-induced apoptosis pathway. In this study, we have examined a capacity of DNA damage to provoke sensitivity of hESC to TRAIL. The extent of DNA damage, behavior of molecules involved in apoptosis, and response of hESC to TRAIL were investigated. The exposure of hESC to 1 μM and 2 μM concentrations of cisplatin have led to the formation of 53BP1 and γH2AX foci, indicating the presence of double-strand breaks in DNA, without affecting the expression of proteins contributing to mitochondrial membrane integrity. Interestingly, cisplatin upregulated critical components of the extrinsic apoptotic pathway—initiator caspase 8, effector caspase 3, and the cell death receptors. The observed increase of expression of the extrinsic apoptotic pathway components was sufficient to sensitize hESC to TRAIL-induced apoptosis; immense cell dying accompanied by enhanced PARP cleavage, processing of caspase 8, and full activation of caspase 3 were all observed after the treatment combining cisplatin and TRAIL. Finally, we have demonstrated the central role of caspase 8 in this process, since its downregulation abrogated the sensitizing effect of cisplatin.

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