scholarly journals Metformin and FTY720 Synergistically Induce Apoptosis in Multiple Myeloma Cells

2018 ◽  
Vol 48 (2) ◽  
pp. 785-800 ◽  
Author(s):  
Yi Zhao ◽  
Enfan Zhang ◽  
Ning Lv ◽  
Liang Ma ◽  
Shunnan Yao ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Er Stress ◽  
Resistance Mechanisms ◽  
Ros Generation ◽  
Dependent Manner ◽  
Mtor Signaling Pathway ◽  
Resistant Disease ◽  
Ros Scavenger ◽  
Anti Tumor Activity ◽  
Mtt Assays

Background/Aims: Patients with multiple myeloma (MM) invariably relapse with chemotherapy-resistant disease, underscoring the need for new therapeutic options that bypass these resistance mechanisms. Metformin is a widely prescribed antidiabetic drug with direct antitumor activity against various tumor cell lines. FTY720, also known as fingolimod, is an immune-modulating agent approved by the FDA as oral medication to treat the relapsing form of multiple sclerosis (MS). In recent years, FTY720 has attracted attention due to its anti-tumor activity. To explore an optimized combinational therapy, interactions between metformin and FTY720 were examined in MM cells. Methods: MTT assays were employed to assess the viability of MM cells. An apoptotic nucleosome assay was employed to measure apoptosis. Loss of mitochondrial membrane potential (MMP, ΔΨm) and cellular levels of ROS were measured by flow cytometry. qRT-PCR was used to analyze the expression of mRNAs. Western blotting assays were applied to measure the levels of proteins involved in different signaling pathways. Results: Coadministration of metformin and FTY720 synergistically inhibited the proliferation of MM cells. Increased levels of apoptosis, activation of caspase-3 and cleavage of PARP were detected after cotreatment with metformin and FTY720. These events were associated with modulation of Bcl-2 proteins, loss of MMP, ER stress induction, and inhibition of the PI3K/AKT/mTOR signaling pathway. The metformin/FTY720 regimen markedly induced ROS generation; moreover, apoptosis, ER stress and inhibition of PI3K/AKT/ mTOR were attenuated by the ROS scavenger NAC. Conclusions: Exposure to metformin in combination with FTY720 potently induces apoptosis in MM cells in a ROS-dependent manner, suggesting that a strategy combining these agents warrants further investigation in MM.

scholarly journals Anti-tumor activity of a novel proteasome inhibitor D395 against multiple myeloma and its lower cardiotoxicity compared with carfilzomib

2021 ◽  
Vol 12 (5) ◽  
Author(s):  
Xuxing Shen ◽  
Chao Wu ◽  
Meng Lei ◽  
Qing Yan ◽  
Haoyang Zhang ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Proteasome Inhibitor ◽  
Osteoclast Differentiation ◽  
Dependent Manner ◽  
Serum Enzyme ◽  
Herg Channels ◽  
Anti Tumor Activity ◽  
Dose Dependent

AbstractCarfilzomib, a second-generation proteasome inhibitor, has significantly improved the survival rate of multiple myeloma (MM) patients, but its clinical application is still restricted by drug resistance and cardiotoxicity. Here, we identified a novel proteasome inhibitor, D395, and assessed its efficacy in treating MM as well as its cardiotoxicity at the preclinical level. The activities of purified and intracellular proteasomes were measured to determine the effect of D395 on the proteasome. CCK-8 and flow cytometry experiments were designed to evaluate the effects of D395 on cell growth and apoptosis. The effects of D395 and carfilzomib on serum enzyme activity, echocardiography features, cardiomyocyte morphology, and hERG channels were also compared. In our study, D395 was highly cytotoxic to MM cell lines and primary MM cells but not normal cells, and it was well tolerated in vivo. Similar to carfilzomib, D395 inhibited osteoclast differentiation in a dose-dependent manner. In particular, D395 exhibited lower cardiotoxicity than carfilzomib in all experiments. In conclusion, D395 is a novel irreversible proteasome inhibitor that has remarkable anti-MM activity and mild cardiotoxicity in vitro and in vivo.

Membrane Lipid Biosynthesis As a Potential Therapeutic Target in Multiple Myeloma

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1836-1836
Author(s):  
Carolyne Bardeleben ◽  
Alan Lichtenstein
Keyword(s):  
Multiple Myeloma ◽  
Er Stress ◽  
Cell Lines ◽  
Cell Apoptosis ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Membrane Lipid ◽  
Dependent Manner ◽  
Kennedy Pathway ◽  
Phosphatidylcholine Synthesis

Abstract Abstract 1836 Phosphatidylcholine (PC) is the most prominent phospholipid in mammalian endoplasmic reticulum (ER) membranes. The rate-limiting step in PC synthesis through the Kennedy pathway is the conversion of phosphocholine + cytidine triphosphate (CTP) to cytidine diphosphocholine, (CDP)-choline, via the enzyme CTP:phosphocholine cytidylyltransferase (CCT) (see figure). Multiple myeloma (MM) cells may be particularly dependent on this biosynthetic reaction because of their high consistent level of ER stress and requirement to continuously replenish ER membranes. Indeed, CCT-null mice have a defect in differentiation of B lymphocytes to plasma cells and deficiencies in Ig synthesis. To test whether this pathway remains critical in survival of malignant MM cells, we exposed MM cell lines to an inhibitor shown to inhibit CCT activity, HexPC. HexPC induced apoptosis in all MM cell lines in a concentration- and time-dependent manner. The addition of lysophosphatidylcholine (LPC), presumably converted to PC independently of the Kennedy pathway, completely rescued MM cell apoptosis. In contrast, similar concentrations of LPC in the same cell lines could not rescue apoptosis induced by bortezomib. An additional intervention to inhibit phosphatidylcholine synthesis, namely inducing pyrimidine starvation, also resulted in MM cell apoptosis and down-regulation of CDP-choline levels. Apoptosis of MM cells induced by HexPC was associated with induction of ER stress as shown by enhanced phosphorylation of IRE1 and eIF-2alpha. This ER stress was also prevented when LPC was added to HexPC although LPC could not prevent similar ER stress induced by bortezomib. These results underscore the importance of this phosphatidylcholine synthesis pathway in MM cells and provide new targets for future therapy. Disclosures: No relevant conflicts of interest to declare.

Cadmium Inhibits Neurite Outgrowth in Differentiating Human SH-SY5Y Neuroblastoma Cells

2014 ◽  
Vol 33 (5) ◽  
pp. 412-418 ◽  
Author(s):  
Eun Joo Pak ◽  
Gi Dong Son ◽  
Byung Sun Yoo
Keyword(s):  
Neurite Outgrowth ◽  
Neuroblastoma Cells ◽  
Underlying Mechanism ◽  
Ros Generation ◽  
Dependent Manner ◽  
Cadmium Treatment ◽  
Trans Retinoic Acid ◽  
All Trans Retinoic Acid ◽  
Ros Scavenger ◽  
Dose Dependent

Cadmium, a highly ubiquitous heavy metal, is well known to induce neurotoxicity. However, the underlying mechanism of cadmium-mediated neurotoxicity remains unclear. We have studied cadmium inhibition of neurite outgrowth using human SH-SY5Y neuroblastoma cells induced to differentiate by all- trans-retinoic acid (RA). Cadmium, at a concentration of 3 μmol/L, had no significant effect on the viability of differentiating SH-SY5Y cells. However, the neurite outgrowth of the differentiating SH-SY5Y cells 48 hours after cadmium treatment (1-3 μmol/L cadmium) was significantly inhibited in a dose-dependent manner. Treatment of RA-stimulated differentiating SH-SY5Y cells with 1 to 3 μmol/L cadmium resulted in decreased level of cross-reactivities with 43-kDa growth-associated protein (GAP-43) in a dose-dependent manner. The reactive oxygen species (ROS) scavenger, NAC (N-acetyl-l-cysteine), recovered the expression of GAP-43 in cadmium-treated cells. The results indicate that cadmium is able to inhibit neurite outgrowth of differentiating SH-SY5Y cells and that this effect might result from ROS generation by cadmium.

Unfolded Protein Response is Involved in Trans-Platinum (II) Complex-Induced Apoptosis in Prostate Cancer Cells via ROS Accumulation

2019 ◽  
Vol 19 (9) ◽  
pp. 1184-1195
Author(s):  
Didem Karakas ◽  
Buse Cevatemre ◽  
Arzu Y. Oral ◽  
Veysel T. Yilmaz ◽  
Engin Ulukaya
Keyword(s):  
Prostate Cancer ◽  
Cell Death ◽  
Cell Viability ◽  
Er Stress ◽  
Apoptotic Cell ◽  
Annexin V ◽  
Treatment Modalities ◽  
Drug Candidate ◽  
Ros Generation ◽  
Dependent Manner

Background:Prostate cancer is one of the most common cancer types and it is the sixth leading cause of cancer-related death in men worldwide. Even though novel treatment modalities have been developed, it still a lifethreatening disease. Therefore novel compounds are needed to improve the overall survival.Methods:In our study, it was aimed to evaluate the anti-cancer activity of newly synthesized Platinum (II) [Pt(II)] complex on DU145, LNCaP and PC-3 prostate cancer cell lines. The cytotoxic activity of Pt(II) complex was tested by SRB and ATP cell viability assays. To detect the mode of cell death; fluorescent staining, flow cytometry and western blot analyses were performed.Results:The Pt(II) complex treatment resulted in a decrease in cell viability and increasing levels of apoptotic markers (pyknotic nuclei, annexin-V, caspase 3/7 activity) and a decrease in mitochondrial membrane potential in a dose dependent manner. Among cell types, tested PC-3 cells were found to be more sensitive to Pt(II) complex, demonstrating elevation of DNA damage in this cell line. In addition, Pt(II) complex induced Endoplasmic Reticulum (ER) stress by triggering ROS generation. More importantly, pre-treatment with NAC alleviated Pt(II) complex-mediated ER stress and cell death in PC-3.Conclusion:These findings suggest an upstream role of ROS production in Pt(II) complex-induced ER stressmediated apoptotic cell death. Considering the ROS-mediated apoptosis inducing the effect of Pt(II) complex, it warrants further evaluation as a novel metal-containing anticancer drug candidate.

In Vivo Anti-Tumor Activity of Atiprimod on SCID Models of Multiple Myeloma.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 249-249
Author(s):  
Paola Neri ◽  
Pierfrancesco Tassone ◽  
Masood A. Shammas ◽  
Ramesh B. Batchu ◽  
Simona Blotta ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Cycle Control ◽  
Cell Signalling ◽  
Myeloma Cell ◽  
Scid Mice ◽  
Control Group ◽  
Dependent Manner ◽  
Growth And Survival ◽  
Anti Tumor Activity

Abstract Atiprimod (N-N-diethl-8, 8-dipropyl-2-azaspiro [4.5] decane-2-propanamine) is an orally bio-available cationic amphiphilic agent previously studied for its in vivo anti-inflammatory activity. We previously demonstrated that Atiprimod, in a time- and dose-dependent manner, inhibits myeloma cell growth and survival in IL-6 independent (OPM1) as well as dependent (INA-6) MM cell-lines with an IC50 in the range of 0.5–2 μM. We have now further characterized the molecular changes in MM cells induced by Atiprimod, evaluating the gene expression profile of INA-6 and OPM-1 cells exposed to this drug. Following 24 h treatment, Atiprimod induced significant down-regulation of genes involved in growth and cell-cycle control (CDC14, CDC2-like 5, Cyclin-E binding protein 1 and MDM2), adhesion (ITGA-6, ITGA-X, ADAM-17, CDH-3, CDH-6, CTNND-1, PECAM-1 and MADCAM-1) cell-signalling pathways (PRKAB-1, Mapk-7, GPR-125 and GPR-32), up-regulation of genes implicated in apoptotic cascades (TNFAIP-3, TNFSF-10, TNFRSF-10c, CDKN2A and CDKN1A) and in negative regulation of signal transduction (RGS-4 and IGFBP-5). We next evaluated the in vivo activity of Atiprimod using three SCID mouse models of human MM: 1) To evaluate effects of Atiprimod directly on MM cells, we used SCID mice bearing subcutaneous OPM1 tumors and treated i.p. with Atiprimod or vehicle alone (PBS) on alternate days for 7 days. In this model 31%, 48% and 55% inhibition of tumor growth were observed in mice treated with Atiprimod at 20, 30 and 50 mg/kg, respectively, compared to control group. 2) To evaluate effects of Atiprimod on MM cells in the context of a human BM microenvironment, we used SCID mice implanted with a human fetal bone chip (SCID-hu) engrafted with IL-6-dependant INA-6 cells and treated i.p. with Atiprimod (40 mg/kg) or vehicle alone (PBS) for two weeks. The response was evaluated by detection of serum soluble human IL-6 receptor (shuIL-6R) released by MM cells in murine serum. We observed a 60% reduction in shuIL-6R level in mice treated with Atiprimod versus control group. 3) Finally, to evaluate effects of Atiprimod on primary patient cells in a human microenvironment, we used SCID-hu mice engrafted with patient MM cells (IgG λ) and treated i.p. with Atiprimod (50 mg/kg) or vehicle alone (PBS) for 4 weeks. Treatment with Atiprimod induced a reduction in both human IgG and λ light chain levels in mouse sera, whereas levels of both proteins continued to rise in mice treated with vehicle alone. Taken together these data demonstrate the in vivo anti-tumor activity of Atiprimod and provide a rational for its clinical evaluation in MM. Based on these data the drug is presently in a multi-site Phase I/IIa clinical trial in patients with relapsed or refractory multiple myeloma (MM).

scholarly journals Current State of the Art and Prospects of T Cell-Redirecting Bispecific Antibodies in Multiple Myeloma

2021 ◽  
Vol 10 (19) ◽  
pp. 4593
Author(s):  
Mashhour Hosny ◽  
Christie P. M. Verkleij ◽  
Jort van der Schans ◽  
Kristine A. Frerichs ◽  
Tuna Mutis ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
T Cells ◽  
T Cell ◽  
Phase 1 ◽  
Treatment Strategies ◽  
Resistance Mechanisms ◽  
Bispecific Antibodies ◽  
Great Promise ◽  
Resistant Disease ◽  
Novel Treatment Strategies

Multiple myeloma (MM) patients eventually develop multi-drug-resistant disease with poor survival. Hence, the development of novel treatment strategies is of great importance. Recently, different classes of immunotherapeutic agents have shown great promise in heavily pre-treated MM, including T cell-redirecting bispecific antibodies (BsAbs). These BsAbs simultaneously interact with CD3 on effector T cells and a tumor-associated antigen on MM cells, resulting in redirection of T cells to MM cells. This leads to the formation of an immunologic synapse, the release of granzymes/perforins, and subsequent tumor cell lysis. Several ongoing phase 1 studies show substantial activity and a favorable toxicity profile with BCMA-, GPRC5D-, or FcRH5-targeting BsAbs in heavily pre-treated MM patients. Resistance mechanisms against BsAbs include tumor-related features, T cell characteristics, and impact of components of the immunosuppressive tumor microenvironment. Various clinical trials are currently evaluating combination therapy with a BsAb and another agent, such as a CD38-targeting antibody or an immunomodulatory drug (e.g., pomalidomide), to further improve response depth and duration. Additionally, the combination of two BsAbs, simultaneously targeting two different antigens to prevent antigen escape, is being explored in clinical studies. The evaluation of BsAbs in earlier lines of therapy, including newly diagnosed MM, is warranted, based on the efficacy of BsAbs in advanced MM.

scholarly journals Ricolinostat (ACY-1215), a Selective HDAC6 Inhibitor, Alone and in Combination with Bendamustine Is Effective in Preclinical Studies in Lymphoma Cell Lines

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2772-2772
Author(s):  
Maria Cosenza ◽  
Monica Civallero ◽  
Steven N Quayle ◽  
Stefano Sacchi ◽  
Samantha Pozzi
Keyword(s):  
Cell Cycle ◽  
Cell Viability ◽  
Er Stress ◽  
Cell Lines ◽  
Drug Combination ◽  
Histone Deacetylase Inhibitors ◽  
Epigenetic Therapy ◽  
Ros Generation ◽  
Preclinical Studies ◽  
Dependent Manner

Abstract Background. Histone deacetylase inhibitors (HDACi) have emerged as a new class of anticancer agents, targeting the biological processes including cell cycle and apoptosis. The discovery of isoform selective compounds may offer a therapeutic advantage by minimizing toxicity. Ricolinostat (ACY-1215) inhibits HDAC6, resulting in tubulin hyperacetylation. Previous studies have investigated the synergistic effects of the combination of HDACis and alkylating agents offering new therapeutic strategies. The aim of this in vitro study was to investigate the activity of ricolinostat alone and in combination with bendamustine in a panel of non-Hodgkin's lymphoma (NHL) cell lines. Methods. Ricolinostat was kindly provided by Acetylon Pharmaceuticals (Boston). NHL cell lines (WSU-NHL, RL, Granta-519, Jeko-1, Hut-78 and Karpas-299) were treated with ricolinostat and bendamustine alone and in combination. The interaction between the drugs was evaluated by the Chou-Talalay method and cell viability and clonogenicity were also evaluated. Apoptosis, ROS generation, Bcl-2 protein expression, cell cycle progression and tubulin expression were determined by flow cytometry. The effects of ricolinostat alone and in combination on caspases, PI3K/Akt, ER stress and UPR pathways were assessed by immunoblotting. Results. Ricolinostat was able to affect the cell viability in a dose dependent manner, with IC50 values ranging from 1.51 to 8.65 μM. Ricolinostat with bendamustine synergistically induced anti-proliferative and pro-apoptotic effects in lymphoma cells, even in the presence of the bone marrow microenvironment. Combination treatment decreased the percentage of viable peripheral blood mononuclear cells (PBMCs) from patients with lymphoma but had minimal or no cytotoxic effect on PBMCs from healthy donors. Clonogenic assay revealed that the drug combination significantly inhibited the colony formation compared with the drugs alone. Drug combination reduced the proportion of cells in the G0/G1 and S phases and caused an increase of "sub-G0/G1" peak with a decrease of cyclin D1 and cyclin E and an increase of p21 and p27 proteins. The synergistic effect was accompanied by increased reactive oxygen species (ROS) generation, which is linked to a decrease of thioredoxin-1 (Trx1) expression, activation of caspase -8, -9, and -3, the cleavage of PARP and modulation of the Bcl-2 protein family. Apoptosis was associated with increased hallmarks of ER stress and activation of UPR sensors and was mediated by dephosphorylation of the AKT. In addition, the exposure of ricolinostat induced the acetylation level of α-tubulin, the extent of which was not further modified by bendamustine. The direct cytotoxicity of ricolinostat/bendamustine may be mediated by an effect on microtubule stabilization. Finally, ricolinostat alone induced a significant down-regulation of IL-10 that was especially evident in WSU-NHL with a fold decrease of 6.6 compared to control. The drug combination affected IL-10 production in all three cell lines with a fold decrease of 5.77 in WSU-NL; 11.5 in Hut-78; and 10.9 in Jeko-1 cells compared with ricolinostat alone. Conclusion. These preclinical studies suggest that bendamustine in combination with epigenetic therapy, such as ricolinostat, may be a promising treatment regime for managing lymphomas. Disclosures Quayle: Acetylon Pharmaceuticals: Employment, Equity Ownership.

scholarly journals Homocysteine Induces Apoptosis of Human Umbilical Vein Endothelial Cells via Mitochondrial Dysfunction and Endoplasmic Reticulum Stress

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Zhimin Zhang ◽  
Congying Wei ◽  
Yanfen Zhou ◽  
Tao Yan ◽  
Zhengqiang Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Er Stress ◽  
Mitochondrial Dysfunction ◽  
Umbilical Vein ◽  
Dependent Manner ◽  
Human Umbilical Vein ◽  
Homologous Protein ◽  
Intracellular Ros ◽  
Underlying Mechanisms ◽  
Umbilical Vein Endothelial Cells

Homocysteine- (Hcy-) induced endothelial cell apoptosis has been suggested as a cause of Hcy-dependent vascular injury, while the proposed molecular pathways underlying this process are unclear. In this study, we investigated the adverse effects of Hcy on human umbilical vein endothelial cells (HUVEC) and the underlying mechanisms. Our results demonstrated that moderate-dose Hcy treatment induced HUVEC apoptosis in a time-dependent manner. Furthermore, prolonged Hcy treatment increased the expression of NOX4 and the production of intracellular ROS but decreased the ratio of Bcl-2/Bax and mitochondrial membrane potential (MMP), resulting in the leakage of cytochrome c and activation of caspase-3. Prolonged Hcy treatment also upregulated glucose-regulated protein 78 (GRP78), activated protein kinase RNA-like ER kinase (PERK), and induced the expression of C/EBP homologous protein (CHOP) and the phosphorylation of NF-κb. The inhibition of NOX4 decreased the production of ROS and alleviated the Hcy-induced HUVEC apoptosis and ER stress. Blocking the PERK pathway partly alleviated Hcy-induced HUVEC apoptosis and the activation of NF-κb. Taken together, our results suggest that Hcy-induced mitochondrial dysfunction crucially modulated apoptosis and contributed to the activation of ER stress in HUVEC. The excessive activation of the PERK pathway partly contributed to Hcy-induced HUVEC apoptosis and the phosphorylation of NF-κb.

scholarly journals Crosstalk between endoplasmic reticulum stress and oxidative stress: a dynamic duo in multiple myeloma

2021 ◽  
Author(s):  
Sinan Xiong ◽  
Wee-Joo Chng ◽  
Jianbiao Zhou
Keyword(s):  
Oxidative Stress ◽  
Multiple Myeloma ◽  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Cell Fate ◽  
Stress Responses ◽  
Plasma Cells ◽  
Reactive Oxygen Species Generation ◽  
Future Research ◽  
And Oxidative Stress

AbstractUnder physiological and pathological conditions, cells activate the unfolded protein response (UPR) to deal with the accumulation of unfolded or misfolded proteins in the endoplasmic reticulum. Multiple myeloma (MM) is a hematological malignancy arising from immunoglobulin-secreting plasma cells. MM cells are subject to continual ER stress and highly dependent on the UPR signaling activation due to overproduction of paraproteins. Mounting evidence suggests the close linkage between ER stress and oxidative stress, demonstrated by overlapping signaling pathways and inter-organelle communication pivotal to cell fate decision. Imbalance of intracellular homeostasis can lead to deranged control of cellular functions and engage apoptosis due to mutual activation between ER stress and reactive oxygen species generation through a self-perpetuating cycle. Here, we present accumulating evidence showing the interactive roles of redox homeostasis and proteostasis in MM pathogenesis and drug resistance, which would be helpful in elucidating the still underdefined molecular pathways linking ER stress and oxidative stress in MM. Lastly, we highlight future research directions in the development of anti-myeloma therapy, focusing particularly on targeting redox signaling and ER stress responses.

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