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The structurally unrelated antihistamine triprolidine does not induce cell death. (A) The chemical structures of cyproheptadine and triprolidine are shown. (B) LP-1 and KMS11 myeloma and OCI-AML2 and Jurkat leukemia cells were treated with increasing concentrations of cyproheptadine (CYP) and triprolidine (TPL). Seventy-two hours after incubation, cell viability was measured by MTS staining. Data represent the mean percentage plus or minus SD of cell viability relative to untreated controls.
Source publication
D-cyclins are regulators of cell division that act in a complex with cyclin-dependent kinases to commit cells to a program of DNA replication. D-cyclins are overexpressed in many tumors, including multiple myeloma and leukemia, and contribute to disease progression and chemoresistance. To better understand the role and impact of D-cyclins in hemato...
Context in source publication
Context 1
... further verify that cyproheptadine does not exert its cell- cycle and cytotoxic effects via the histamine receptor, we compared its activity with a structurally unrelated antihistamine, triprolidine. Unlike cyproheptadine, triprolidine did not reduce the viability of leukemia or myeloma cells ( Figure 6). Therefore, cyproheptadine's proapoptotic effects occur through a mechanism distinct from H1 receptor antagonism. ...
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The genetics of t(11;14)(q13;q32)/cyclin D1-negative mantle cell lymphoma (MCL) is poorly understood. We report here 8 MCL cases lacking t(11;14) or variant CCND1 rearrangement that showed expression of cyclin D1 (2 cases), D2 (2 cases), and D3 (3 cases). One case was cyclin D negative. Cytogenetics and fluorescence in situ hybridization detected t...
D-type cyclins form complexes with cyclin-dependent kinases (CDK4/6) and promote cell cycle progression. Although cyclin D functions appear largely tissue specific, we demonstrate that cyclin D3 has unique functions in lymphocyte development and cannot be replaced by cyclin D2, which is also expressed during blood differentiation. We show that only...
The relationship between activation-induced growth inhibition and regulation of the cell cycle progression was investigated in T cell hybridomas by studying the function of the cell cycle-regulating genes such as G1 cyclins and their associated kinases. Activation of T cell hybridomas by anti-T cell receptor antibody induces growth arrest at G1 pha...
Tumor progression in B-cell chronic lymphocytic leukemia (B-CLL) is thought to result from the gradual accumulation of small resting G0/G1 phase lymphoid cells rather than the proliferation of actively dividing cells. The recent identification of G1 cyclins that are likely to control both the progression through G0 and G1 phase and the G1/S transit...
Citations
... It binds to and stimulates CDK4/6, which cumulatively induces p-Rb phosphorylation, leading to cell cycle progression [34]. Several studies have shown that CCND3 might be a potential therapeutic target for T-ALL, AML, MCL and B-ALL [34][35][36][37][38]. We also found that the knockdown of CCND3 could reduce MCL-1, CDK4, and CDK6 proteins level, which indicates what was previously reported in the literature. ...
Introduction
B-cell acute lymphoblastic leukemia (B-ALL) is the most prevalent malignant tumor affecting children. While the majority of B-ALL patients (90%) experience successful recovery, early relapse cases of B-ALL continue to exhibit high mortality rates. MZ1, a novel inhibitor of Bromodomains and extra-terminal (BET) proteins, has demonstrated potent antitumor activity against hematological malignancies. The objective of this study was to examine the role and therapeutic potential of MZ1 in the treatment of B-ALL.
Methods
In order to ascertain the fundamental mechanism of MZ1, a sequence of in vitro assays was conducted on B-ALL cell lines, encompassing Cell Counting Kit 8 (CCK8) assay, Propidium iodide (PI) staining, and Annexin V/PI staining. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to examine protein and mRNA expression levels. Transcriptomic RNA sequencing (RNA-seq) was utilized to screen the target genes of MZ1, and lentiviral transfection was employed to establish stably-expressing/knockdown cell lines.
Results
MZ1 has been observed to induce the degradation of Bromodomain Containing 4 (BRD4), Bromodomain Containing 3 (BRD3), and Bromodomain Containing 2 (BRD2) in B-ALL cell strains, leading to inhibited cell growth and induction of cell apoptosis and cycle arrest in vitro. These findings suggest that MZ1 exhibits cytotoxic effects on two distinct molecular subtypes of B-ALL, namely 697 (TCF3/PBX1) and RS4;11 (MLL-AF4) B-ALL cell lines. Additionally, RNA-sequencing analysis revealed that MZ1 significantly downregulated the expression of Cyclin D3 (CCND3) gene in B-ALL cell lines, which in turn promoted cell apoptosis, blocked cell cycle, and caused cell proliferation inhibition.
Conclusion
Our results suggest that MZ1 has potential anti-B-ALL effects and might be a novel therapeutic target.
... Cyproheptadine inhibits In Vitro and xenograft growth of mantle lymphoma cells [34]. In mouse models of myeloma, cyproheptadine demonstrated inhibitory activity via cyclin D inhibition, inducing G0 arrest with subsequent apoptosis in the myeloma cells [35]. ...
Background:
Pharmacological targeting aberrant activation of epidermal growth factor receptor tyrosine kinase signaling is an established approach to treating lung adenocarcinoma. Osimertinib is a tyrosine kinase approved and effective in treating lung adenocarcinomas that have one of several common activating mutations in epidermal growth factor receptor. The emergence of resistance to osimertinib after a year or two is the rule. We developed a five-drug adjuvant regimen designed to increase osimertinib's growth inhibition and thereby delay the development of resistance. Areas of Uncertainty: Although the assembled preclinical data is strong, preclinical data and the following clinical trial results can be discrepant. The safety of OPALS drugs when used individually is excellent. We have no data from humans on their tolerability when used as an ensemble. That there is no data from the individual drugs to suspect problematic interaction does not exclude the possibility.
Data sources:
All relevant PubMed.org articles on the OPALS drugs and corresponding pathophysiology of lung adenocarcinoma and glioblastoma were reviewed. Therapeutic Opinion: The five drugs of OPALS are in wide use in general medicine for non-oncology indications. OPALS uses the anti-protozoal drug pyrimethamine, the antihistamine cyproheptadine, the antibiotic azithromycin, the antihistamine loratadine, and the potassium sparing diuretic spironolactone. We show how these inexpensive and generically available drugs intersect with and inhibit lung adenocarcinoma growth drive. We also review data showing that both OPALS adjuvant drugs and osimertinib have data showing they may be active in suppressing glioblastoma growth.
... Cyproheptadine (CPH) is a first-generation anti-histamine drug, which is often used to treat allergic reactions and common cold. CPH has also been demonstrated to have antitumor activity in multiple tumors, such as leukemia, myeloma, mantle cell lymphoma, and hepatocellular carcinoma [9][10][11]. Our previous study found that CPH exhibited anti-tumor activity in UC by targeting GSK3β signaling pathways [12]. ...
... In this regard, development of novel therapeutic strategy is an urgent issue. The serotonin antagonist and histamine H1 blocker CPH was recently reported to induce tumor cell apoptosis and inhibit tumor proliferation in myeloma, mantle-cell lymphoma, and hepatocellular carcinoma [9][10][11]. Our previous study also demonstrated that CPH exhibited antitumor activity in human UC cell lines and in vivo xenograft model [12]. ...
Background
Urothelial carcinoma (UC) is the second most common malignancy of the urinary system with high rate of recurrence, UC patients therefore needed to be treated with surgery followed by chemotherapy. Development of novel therapeutics with minimal side-effect is an urgent issue. Our previous study showed that cyproheptadine (CPH), an anti-histamine, exhibited antitumor activity in UC in vitro and in an xenograft model. However, the molecular mechanism of how CPH inhibits tumor progression is not fully understood.
Methods
Genes that were upregulated after treatment with CPH in UC cells, were examined by RNA-Seq. Real-time quantitative PCR (RT-qPCR) was employed to detect IRF6 expression while COBRA assay and bisulphite pyrosequencing were used to examine promoter methylation of IRF6 . Enrichment of total H3K27 acetylation and H3K4 mono-methylation were detected by western blotting. Colony formation and flow cytometry were used to examine proliferation and apoptosis in UC cells overexpressed or depleted with IRF6. Nude mice xenograft model was used to examine the effect of IRF6 in UC.
Results
Our result showed that several genes, including IRF6 were upregulated after treatment with CPH in BFTC905 UC cells. Further experiments found that treatment of CPH could restore the expression of IRF6 in several other UC cell lines, probably due to promoter hypomethylation and enrichment of H3K27 acetylation and H3K4 mono-methylation. These results may be due to the fact that CPH could alter the activity, but not the expression of epigenetic modifiers. Finally, re-expression of IRF6 in UC inhibited tumor growth in vitro and in an xenograft mouse model, by inducing apoptosis.
Conclusion
In conclusion, our results suggested that CPH may be an epigenetic modifier, modulating the expression of the potential tumor suppressor IRF6 , in inhibiting tumor growth in UC.
... Although the patient samples and cancer cells belong to different stage, it should be reasonable to assume that there are some characteristics shared between different stages of the prostate cancer. In addition, it is found from some previous literatures (Mao et al., 2008;Kim et al., 2012;Barillari et al., 2014;Hsieh et al., 2016;Sun et al., 2016;Takemoto et al., 2016;Maksimovic-Ivanic et al., 2017;Van Eijk et al., 2019) that the three drugs (i.e., indinavir, imatinib, and cyproheptadine) predicted as one component of the drug combination exhibited anticancer impact on various cancers, including prostate cancer, which also to some extent supports the rationality of using perturbational gene-expression data on PC-3 cell lines to predict drug combinations for the prostate cancer. However, more in vitro and in vivo experiments will be needed to further validate the therapeutic efficacy of our predictive drug combinations for the prostate cancer. ...
Prostate cancer (PRAD) is a major cause of cancer-related deaths. Current monotherapies show limited efficacy due to often rapidly emerging resistance. Combination therapies could provide an alternative solution to address this problem with enhanced therapeutic effect, reduced cytotoxicity, and delayed the appearance of drug resistance. However, it is prohibitively cost and labor-intensive for the experimental approaches to pick out synergistic combinations from the millions of possibilities. Thus, it is highly desired to explore other efficient strategies to assist experimental researches. Inspired by the challenge, we construct the transcriptomics-based and network-based prediction models to quickly screen the potential drug combination for Prostate cancer, and further assess their performance by in vitro assays. The transcriptomics-based method screens nine possible combinations. However, the network-based method gives discrepancies for at least three drug pairs. Further experimental results indicate the dose-dependent effects of the three docetaxel-containing combinations, and confirm the synergistic effects of the other six combinations predicted by the transcriptomics-based model. For the network-based predictions, in vitro tests give opposite results to the two combinations (i.e. mitoxantrone-cyproheptadine and cabazitaxel-cyproheptadine). Namely, the transcriptomics-based method outperforms the network-based one for the specific disease like Prostate cancer, which provide guideline for selection of the computational methods in the drug combination screening. More importantly, six combinations (the three mitoxantrone-containing and the three cabazitaxel-containing combinations) are found to be promising candidates to synergistically conquer Prostate cancer.
... 6 A growing number of studies have shown that inhibition of the PI3K/Akt/mTOR signaling pathway triggers apoptosis in MM cells. 7,8 In the recent years advances in treatments, including immunomodulatory drugs (eg, thalidomide and lenalidomide), 9 proteasome inhibitors (eg, bortezomib and carfilzomib), 10 and autologous transplantation, 11 and so on, has changed outcome of MM patients tremendously. However, MM remains incurable, and there is a demand for novel therapeutic compounds. ...
Background
We aimed to investigate the anti-multiple myeloma (MM) activity of the new small molecular compound AE-848 (5-bromo-2-hydroxyisophthalaldehyde bis[(1-methyl-1H-benzimidazol-2-yl)hydrazone]) and its underlying anti-MM mechanism.
Methods
Cell viability and apoptosis were detected and quantified by using MTT and flow cytometry, respectively. JC-1 dye-related techniques were used to assess mitochondrial membrane potential (MMP). Western blotting was applied to detect the expression of NF-κB and PI3K/Akt/mTOR pathway-associated proteins. The in vivo activity of AE-848 against MM was evaluated in a MM mouse model.
Results
Application of AE-848 into the in vitro cell culture system significantly reduced the viability and induced apoptosis of the MM cell lines, RPMI-8226 and U266, in a dose- and time-dependent manner, respectively. JC-1 dye and Western blotting analysis revealed that AE-848 induced the cleavage of caspase-8, caspase-3, and poly ADP-ribose polymerase (PARP), resulting in loss of mitochondrial membrane potential (MMP). Both the NF-κB and PI3K/AKT/mTOR signaling pathways were involved in AE-848-induced apoptosis of U266 and RPMI8226 cells. Moreover, AE-848 leads to cell cycle arrest of MM cells. Its anti-MM efficacy was further confirmed in a xenograft model of MM. AE-848 administration significantly inhibited MM tumor progression and prolonged the survival of MM-bearing mice. More importantly, our results demonstrated that AE-848 markedly induced primary MM cell apoptosis.
Conclusion
Our results for the first time showed that the small compound AE-848 had potent in vitro and in vivo anti-myeloma activity, indicating that AE-848 may have great potential to be developed as a drug for MM treatment.
... The colony-forming unit assay was performed as described previously 33 . Briefly, bone marrow cells (6.25 × 10 5 /mL) were mixed with Nam-containing MethoCultGF H4434 medium (StemCell Technologies, Vancouver, Canada) followed by being seeded in 3.5-cm plates and were cultured for 14 d. ...
As a deubiqutinase Otub1 stabilizes and promotes the oncogenic activity of the transcription factor c-Maf in multiple myeloma (MM), a malignancy of plasma cells. In the screen for bioactive inhibitors of the Otub1/c-Maf axis for MM treatment, nanchangmycin (Nam), a polyketide antibiotic, was identified to suppress c-Maf activity in the presence of Otub1. By suppressing Otub1, Nam induces c-Maf polyubiquitination and subsequent degradation in proteasomes but does not alter its mRNA level. Consistently, Nam downregulates the expression of CCND2, ARK5, and ITGB7, the downstream genes regulated by c-Maf, and promotes MM cell apoptosis as evidenced by PARP and Caspase-3 cleavage, as well as Annexin V staining. In line with the hypothesis, overexpression of Otub1 partly rescues Nam-induced MM cell apoptosis, and interestingly, when Otub1 is knocked down, Nam-decreased MM cell survival is also partly ablated, suggesting Otub1 is essential for Nam anti-MM activity. Nam also displays potent anti-MM activity synergistically with Doxorubicin or lenalidomide. In the in vivo assays, Nam almost completely suppresses the growth of MM xenografts in nude mice at low dosages but it shows no toxicity. Given its safety and efficacy, Nam has a potential for MM treatment by targeting the Otub1/c-Maf axis.
... CY has been used for reducing all-cause deaths or deaths due to cancers in Taiwan [5]. CY is a firstgeneration anti-histamine, and it currently used to treat allergic reactions such as atopic dermatitis, anorexia, and migraines [6][7][8], and has been reported to be a novel therapeutic agent for treating multiple malignancies such as myeloma, leukemia and hepatocellular carcinoma (HCC) [9][10][11]. Other reported two advanced HCC cases with lung metastasis that experienced complete remission upon treatment with a combination of CY and thalidomide [10]. ...
This study was designed to compare the efficacy of Cyproheptadine (CY) in patients with bladder cancer (BC) who received different therapeutic modalities. We used the database from a hospital in Taiwan for analysis. We included patients diagnosed as having bladder cancer from January 1, 2008, to December 31, 2017. The patient cohort comprised those who received different treatments, and we compared patients who received CY with those who did not. In total, 627 patients were included, and the mean follow-up duration was 3.26 years. All data were filtered out by 230 million data and 119 patients had used CY. Among them, 32 patients were used over 3 months of CY. The CY treatment curve shown by Kaplan-Meier survival curves for patients treated is higher than that of the non-CY effect. The value of Chi-squared statistic was 4.138 with associated p-value less than 0.05. Two survival curves shown by the result of the log rank test differ significantly. The grouping variable different treatments for non-CY and CY has a significant influence on survival rate. These results suggest that the use of CY may improve the survival rate of patients with BC.
... The role of antihistamines in cancer has been controversial mainly due to the lack of consistency across different HRH1 inhibitors and the absence of a defined role of its well-described HRH1 molecular target on cancer cells [15]. In haematological neoplasias, several antihistamines have been shown to affect leukaemia cell viability to different extents [17,[68][69][70][71]. All of them display CAD-associated properties (high logD pH 7.4 and intermedium-low TPSA) compatible to the model proposed. ...
... All of them display CAD-associated properties (high logD pH 7.4 and intermedium-low TPSA) compatible to the model proposed. Interestingly, those antihistamines that presented no antileukaemic effect could be predicted by our model [17,68,69,71]. As most CADs, ANHAs are lysosomotropic drugs that induce irreversible damage to lysosomes [40,41]. ...
Background:
Despite great efforts to identify druggable molecular targets for AML, there remains an unmet need for more effective therapies.
Methods:
An in silico screening was performed using Connectivity Maps to identify FDA-approved drugs that may revert an early leukaemic transformation gene signature. Hit compounds were validated in AML cell lines. Cytotoxic effects were assessed both in primary AML patient samples and healthy donor blood cells. Xenotransplantation assays were undertaken to determine the effect on engraftment of hit compounds. The mechanism of action responsible for the antileukaemic effect was studied focussing on lysosomes and mitochondria.
Findings:
We identified a group of antihistamines (termed ANHAs) with distinct physicochemical properties associated with their cationic-amphiphilic nature, that selectively killed leukaemic cells. ANHAs behaved as antileukaemic agents against primary AML samples ex vivo, sparing healthy cells. Moreover, ANHAs severely impaired the in vivo leukaemia regeneration capacity. ANHAs' cytotoxicity relied on simultaneous mitochondrial and lysosomal disruption and induction of autophagy and apoptosis. The pharmacological effect was exerted based on their physicochemical properties that permitted the passive targeting of both organelles, without the involvement of active molecular recognition.
Interpretation:
Dual targeting of lysosomes and mitochondria constitutes a new promising therapeutic approach for leukaemia treatment, supporting the further clinical development. FUND: This work was funded by the Fundación Mutua Madrileña (RMR), CaixaImpulse (RMR), the Spanish Ministry of Economy (RMR), the Josep Carreras International Leukaemia Foundation (RMR), l'Obra Social "La Caixa" (RMR), and Generalitat de Catalunya (IJC).
... In particular, we observed that astemizole and terfenadine, both commercialized for their action against HRH1, were effective against AML LSC and bulk cell populations. There is growing evidence supporting the therapeutic potential of these two drugs as well as other H1-antihistamines for a variety of malignancies, including leukemia, myeloma, breast, prostate, colon, lung and liver cancers [29][30][31][32][33][34][35] . The evaluation of three additional molecules in this class, namely diphenhydramine and cetirizine (not predicted to target LSCs in our in silico analysis) and fexofenadine (not included in CMap), did not alter leukemia cell viability even at high micromolar concentrations. ...
... Cell proliferation is another process regulated by Ca2 + -dependent signaling pathways, including expression of cell cycle regulator controlling of the G1/S cell cycle transition 39 . The H1-antihistamines terfenadine and cyproheptadine were shown to cause G0/ G1 cell cycle arrest and apoptosis in leukemia 29,40 . Our gene-set enrichment analysis of transcriptomic fingerprints of astemizole in cancer cells was consistent with a possible alteration of G1/S cell cycle progression. ...
Therapy for acute myeloid leukemia (AML) involves intense cytotoxic treatment and yet approximately 70% of AML are refractory to initial therapy or eventually relapse. This is at least partially driven by the chemo-resistant nature of the leukemic stem cells (LSCs) that sustain the disease, and therefore novel anti-LSC therapies could decrease relapses and improve survival. We performed in silico analysis of highly prognostic human AML LSC gene expression signatures using existing datasets of drug-gene interactions to identify compounds predicted to target LSC gene programs. Filtering against compounds that would inhibit a hematopoietic stem cell (HSC) gene signature resulted in a list of 151 anti-LSC candidates. Using a novel in vitro LSC assay, we screened 84 candidate compounds at multiple doses and confirmed 14 drugs that effectively eliminate human AML LSCs. Three drug families presenting with multiple hits, namely antihistamines (astemizole and terfenadine), cardiac glycosides (strophanthidin, digoxin and ouabain) and glucocorticoids (budesonide, halcinonide and mometasone), were validated for their activity against human primary AML samples. Our study demonstrates the efficacy of combining computational analysis of stem cell gene expression signatures with in vitro screening to identify novel compounds that target the therapy-resistant LSC at the root of relapse in AML.
... The use of primary bone marrow cells was approved by the Review Board and Ethical Committee of Soochow University, and each patient provided written informed consent to donate 2-5 ml of bone marrow for this study after diagnostic and clinical procedures in accordance with the Declaration of Helsinki. Mono-nuclear cells were isolated by Lympholyte® Cell Separation (Cedarlane, Canada) [13]. ...
Background
UBE2O is proposed as a ubiquitin-conjugating enzyme, but its function was largely unknown.
Methods
Mass spectrometry was applied to identify c-Maf ubiquitination-associated proteins. Immunoprecipitation was applied for c-Maf and UBE2O interaction. Immunoblotting was used for Maf protein stability. Luciferase assay was used for c-Maf transcriptional activity. Lentiviral infections were applied for UBE2O function in multiple myeloma (MM) cells. Flow cytometry and nude mice xenografts were applied for MM cell apoptosis and tumor growth assay, respectively.
Results
UBE2O was found to interact with c-Maf, a critical transcription factor in MM, by the affinity purification/tandem mass spectrometry assay and co-immunoprecipitation assays. Subsequent studies showed that UBE2O mediated c-Maf polyubiquitination and degradation. Moreover, UBE2O downregulated the transcriptional activity of c-Maf and the expression of cyclin D2, a typical gene modulated by c-Maf. DNA microarray revealed that UBE2O was expressed in normal bone marrow cells but downregulated in MGUS, smoldering MM and MM cells, which was confirmed by RT-PCR in primary MM cells, suggesting its potential role in myeloma pathophysiology. When UBE2O was restored, c-Maf protein in MM cells was significantly decreased and MM cells underwent apoptosis. Furthermore, the human MM xenograft in nude mice showed that re-expression of UBE2O delayed the growth of myeloma xenografts in nude mice in association with c-Maf downregulation and activation of the apoptotic pathway.
Conclusions
UBE2O mediates c-Maf polyubiquitination and degradation, induces MM cell apoptosis, and suppresses myeloma tumor growth, which provides a novel insight in understanding myelomagenesis and UBE2O biology.
