BACKGROUND: The mechanistic target of rapamycin (mTOR) is a crucial regulator of cell metabolic activity. It forms part of several distinct protein complexes, particularly mTORC1 and mTORC2. The lack of specific inhibitors still hampers the attribution of mTOR functions to these complexes. JR-AB2-011 has been reported as a specific mTORC2 inhibitor preventing mTOR binding to RICTOR, a unique component of mTORC2. We aimed to describe the effects of JR-AB2-011 in leukemia/lymphoma cells, where the mTOR pathway is often aberrantly activated. METHODS: The impact of JR-AB2-011 on leukemia/lymphoma cell metabolism was analyzed using the Seahorse platform. AKT phosphorylation at Ser473 was used as a marker of mTORC2 activity. mTOR binding to RICTOR was assessed by co-immunoprecipitation. RICTOR-null cells were derived from the Karpas-299 cell line using CRISPR/Cas9 gene editing. RESULTS: In leukemia/lymphoma cell lines, JR-AB2-011 induced a rapid drop in the cell respiration rate, which was variably compensated by an increased glycolytic rate. In contrast, an increase in the respiration rate due to JR-AB2-011 treatment was observed in primary leukemia cells. Unexpectedly, JR-AB2-011 did not affect AKT Ser473 phosphorylation. In addition, mTOR did not dissociate from RICTOR in cells treated with JR-AB2-011 under the experimental conditions used in this study. The effect of JR-AB2-011 on cell respiration was retained in RICTOR-null cells. CONCLUSION: JR-AB2-011 affects leukemia/lymphoma cell metabolism via a mechanism independent of mTORC2.
- MeSH
- fosforylace účinky léků MeSH
- leukemie * farmakoterapie metabolismus MeSH
- lidé MeSH
- mTOR inhibitory farmakologie MeSH
- mTORC2 * metabolismus MeSH
- nádorové buněčné linie MeSH
- protein RICTOR * metabolismus MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Kaurane-type diterpenoids, obtained from various natural sources, have shown many biological activities, including anti-inflammatory and antitumor effects. Caracasine, an ent-kaurane diterpenoid isolated from the flowers of Croton micans, was shown to induce apoptosis in leukaemia cell lines. OBJECTIVE: The present study aimed to ascertain the compound's mechanism of cell death induction using two leukaemia cell lines, Jurkat E6.1 (T cell) and HL-60 (promyeloblast cells). METHODS: Cell death in Jurkat and HL60 cells were evaluated by flow cytometry for apoptosis with annexin-V/PI, mitochondrial membrane potential disturbance, changes in cell cycle, CD95 expression, caspase activation, Nuclear Factor kappa B inhibition, and differentiation into a neutrophil-like cell (dHL60). RESULTS: Caracasine (10 μM) increased the G0/G1 phase in Jurkat and arrested the cell cycle in the S phase in HL60. Caracasine increased CD95 expression (p<0.01 in Jurkat and p<0.05 in HL60) and caspase-8 activation (p<0.001 in Jurkat and p<0.05 in HL60). Caspase-9 was activated in both cell lines (p<0.001) along with the decline in mitochondrial Δψm (p<0.05 in Jurkat and p<0.001 in HL60). In HL60 cells, the kaurane induced neutrophil differentiation was assessed by CD40 expression and reactive oxygen species production. In Jurkat cells, caracasine inhibited the NF-κB pathway in cells pretreated with PHA to activate the NF-κB pathway, suggesting a possible role in inflammatory diseases. CONCLUSION: Caracasine induced apoptosis through the intrinsic and extrinsic pathways in both cell lines were evaluated which could be the leading structure for new anti-leukemic and anti-inflammatory drugs.
- MeSH
- apoptóza MeSH
- diterpeny kauranové * farmakologie chemie MeSH
- diterpeny * farmakologie MeSH
- HL-60 buňky MeSH
- Jurkat buňky MeSH
- leukemie * farmakoterapie MeSH
- lidé MeSH
- NF-kappa B metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Leukemie jsou jedny z nejčastějších dětských onemocnění s maligním průběhem. Dnes naštěstí mají vysokou pravděpodobnost úspěšného vyléčení. Charakter onemocnění s sebou nese vysoké riziko iniciálního rozšíření, respektive pozdějšího relapsu onemocnění do CNS. Standardní léčba proto zahrnuje i intratekální aplikaci cytostatik, která toto riziko zásadně snižují, avšak mohou poškodit některé kognitivní funkce. Výzkum zabývající se touto problematikou je široký. Systematickou literární rešerší jsme vybrali 16 článků, které kognitivní funkce zkoumaly prostřednictvím neuropsychologických testů. Analýza výsledků svědčí o statisticky významném poklesu výkonu v testech pozornosti, rychlosti zpracování, pracovní paměti a exekutivních funkcí. Naopak celkový inteligenční kvocient, schopnost učení a dlouhodobá paměť se zdají být léčbou zasaženy jen mírně.
Leukemia is one of the most common childhood diseases with a malignant course. Fortunately, today there is a high probability of its successful cure. The nature of the disease entails a high risk of initial spread, or later relapse of the disease into the central nervous system. Standard treatment including intrathecal application of cytostatics reduces this risk fundamentally; nevertheless it can damage some cognitive functions. There is extensive research dealing with this issue. Through a systematic literature research, we selected 16 articles that examined cognitive functions through neuropsychological tests. Result analysis indicates a statistically significant decrease in performance in tests of attention, processing speed, working memory and executive functions. Conversely, the overall intelligence quotient, learning ability and long-term memory seem to be only slightly affected by the treatment.
- MeSH
- diferenciální diagnóza MeSH
- hematologické nádory * diagnóza farmakoterapie terapie MeSH
- individualizovaná medicína metody MeSH
- leukemie diagnóza farmakoterapie terapie MeSH
- lidé MeSH
- lymfom diagnóza terapie MeSH
- příznaky a symptomy MeSH
- protinádorové látky farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
nestr.
Leukemie, které patří mezi maligní choroby, je nejčastější příčinou úmrtí v důsledku nádorových onemocnění u dětí. Stávající léčba již dosáhla kvůli vysoké toxicitě svých hranic, a tudíž je třeba nových strategií. Jeden z možných scénářů navrhuje vylepšit využití již zavedených terapeutik, zatímco druhý chce nalézt jejich nové možné cíle. Problémem této cílené terapie je však vysoká genetická heterogenita leukémií, což v praxi znamená, že konkrétní mutace musí být nejprve identifikovány, a až poté je možné začít s cílenou léčbou. Navíc, specifická inhibice konkrétního cíle může aktivovat zpětnou vazbu a zcela zmařit zamýšlený efekt. Tímto projektem navrhujeme obecnější metodu léčby, a to cílení na buněčný metabolismus, který je u nádorových buněk významně ovlivněn, a to zejména kvůli vyšší potřebě syntézy biomasy, nukleotidů a NADPH. Plánujeme podrobněji charakterizovat metabolismus leukemických buněk v průběhu indukce, progrese a léčby nemoci, abychom mohli navrhnout nové léčebné strategie pro pacienty nereagující na současnou terapii.; Leukemia is a malignant disease that ranks 1st in cancer-related deaths in children. Current treatment has reached its limits due to high toxicity, and there have been calls for new strategies. One of the possible scenarios is an improved use of the established drugs, while another involves introducing new druggable targets. Yet the bottle neck of such targeted therapy is the genetic heterogeneity of leukemia, meaning that specific driver mutations must first be identified before being targeted. Moreover, specific inhibition of one alteration can activate feedback loop pathways and impair the effect. We therefore propose a more generalized treatment strategy by targeting cellular metabolism, which is altered in cancer cells to meet the higher demands for biomass production, nucleotide synthesis, and NADPH. In the current project, we aim to characterize metabolism of leukemic cells during disease induction, progression and treatment to propose new therapeutic strategies for non-responsive patients.
- MeSH
- chemorezistence MeSH
- cílená molekulární terapie MeSH
- dítě MeSH
- leukemie farmakoterapie MeSH
- metabolismus MeSH
- nádorové procesy MeSH
- Check Tag
- dítě MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- onkologie
- hematologie a transfuzní lékařství
- pediatrie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
BACKGROUND: Recent discoveries in cancer therapeutics have proven combination therapies more effective than individual drugs. This study describes the efficacy of the combination of Cinnamomum zeylanicum and doxorubicin against benzene-induced leukemia. METHODS AND RESULTS: Brine shrimp assay was used to assess the cytotoxicity of C. zeylanicum, doxorubicin and their combination. After AML induction in Sprague Dawley rats, the same drugs were given to rat groups. Changes in organ weight, haematological profile, and hepatic enzymes were determined. Real-time PCR was used to elucidate the effect on the expression of STMN1, GAPDH, P53 and various TRAIL and NF-kappaB components. C. zeylanicum reduced the cytotoxicity of doxorubicin. The combination treatment showed better anti-leukemic results than any of the individual drugs as evident from STMN1 expression (p < 0.001). It was particularly effective in reducing total white blood cell counts and recovering lymphocytes, monocytes and eosinophils along with hepatic enzymes ALT and AST (p < 0.001). All doses recovered relative organ weights and improved blood parameters. The combination therapy was particularly effective in inducing apoptosis, inhibition of proliferation marker GAPDH (p < 0.001) and NF-kappaB pathway components Rel-A (p < 0.001) and Rel-B (p < 0.01). Expressions of TRAIL components c-FLIP (p < 0.001), TRAIL ligand (p < 0.001) and caspase 8 (p < 0.01) were also altered. CONCLUSION: Cinnamomum zeylanicum in combination with doxorubicin helps to counter benzene-induced cellular and hepatic toxicity and improves haematological profile. The anti-leukemic effects are potentially due to inhibition of GAPDH and NF-kappa B pathway, and through regulation of TRAIL pathway. Our data suggests the use of C. zeylanicum with doxorubicin to improve anti-leukemic therapeutic regimes.
- MeSH
- apoptóza MeSH
- benzen farmakologie MeSH
- doxorubicin farmakologie MeSH
- krysa rodu rattus MeSH
- leukemie * farmakoterapie MeSH
- NF-kappa B metabolismus MeSH
- oleje prchavé * farmakologie MeSH
- potkani Sprague-Dawley MeSH
- protein TRAIL metabolismus farmakologie MeSH
- skořicovník ceylonský metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The newly emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cause life-threatening diseases in millions of people worldwide, in particular, in patients with cancer, and there is an urgent need for antiviral agents against this infection. While in vitro activities of artemisinins against SARS-CoV-2 and cancer have recently been demonstrated, no study of artemisinin and/or synthetic peroxide-based hybrid compounds active against both cancer and SARS-CoV-2 has been reported yet. However, the hybrid drug's properties (e. g., activity and/or selectivity) can be improved compared to its parent compounds and effective new agents can be obtained by modification/hybridization of existing drugs or bioactive natural products. In this study, a series of new artesunic acid and synthetic peroxide based new hybrids were synthesized and analyzed in vitro for the first time for their inhibitory activity against SARS-CoV-2 and leukemia cell lines. Several artesunic acid-derived hybrids exerted a similar or stronger potency against K562 leukemia cells (81-83 % inhibition values) than the reference drug doxorubicin (78 % inhibition value) and they were also more efficient than their parent compounds artesunic acid (49.2 % inhibition value) and quinoline derivative (5.5 % inhibition value). Interestingly, the same artesunic acid-quinoline hybrids also show inhibitory activity against SARS-CoV-2 in vitro (EC50 13-19 μm) and no cytotoxic effects on Vero E6 cells (CC50 up to 110 μM). These results provide a valuable basis for design of further artemisinin-derived hybrids to treat both cancer and SARS-CoV-2 infections.
- MeSH
- antivirové látky farmakologie terapeutické užití MeSH
- artemisininy * farmakologie MeSH
- Cercopithecus aethiops MeSH
- chinoliny * terapeutické užití MeSH
- COVID-19 * MeSH
- farmakoterapie COVID-19 MeSH
- leukemie * farmakoterapie MeSH
- lidé MeSH
- nádory * farmakoterapie MeSH
- peroxidy MeSH
- SARS-CoV-2 MeSH
- Vero buňky MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
N-acetylcysteine (NAC), often used as an antioxidant-scavenging reactive oxygen species (ROS) in vitro, was recently shown to increase the cytotoxicity of other compounds through ROS-dependent and ROS-independent mechanisms. In this study, NAC itself was found to induce extensive ROS production in human leukemia HL-60 and U937 cells. The cytotoxicity depends on ROS-modulating enzyme expression. In HL-60 cells, NAC activated NOX2 to produce superoxide (O2•-). Its subsequent conversion into H2O2 by superoxide dismutase 1 and 3 (SOD1, SOD3) and production of ClO- from H2O2 by myeloperoxidase (MPO) was necessary for cell death induction. While the addition of extracellular SOD potentiated NAC-induced cell death, extracellular catalase (CAT) prevented cell death in HL-60 cells. The MPO inhibitor partially reduced the number of dying HL-60 cells. In U937 cells, the weak cytotoxicity of NAC is probably caused by lower expression of NOX2, SOD1, SOD3, and by the absence of MOP expression. However, even here, the addition of extracellular SOD induced cell death in U937 cells, and this effect could be reversed by extracellular CAT. NAC-induced cell death exhibited predominantly apoptotic features in both cell lines. Conclusions: NAC itself can induce extensive production of O2•- in HL-60 and U937 cell lines. The fate of the cells then depends on the expression of enzymes that control the formation and conversion of ROS: NOX, SOD, and MPO. The mode of cell death in response to NAC treatment bears apoptotic and apoptotic-like features in both cell lines.
- MeSH
- acetylcystein farmakologie MeSH
- HL-60 buňky MeSH
- katalasa genetika MeSH
- leukemie farmakoterapie genetika metabolismus MeSH
- lidé MeSH
- NADPH-oxidasa 2 genetika MeSH
- oxidační stres účinky léků MeSH
- peroxidasa genetika MeSH
- proliferace buněk účinky léků MeSH
- reaktivní formy kyslíku metabolismus MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- stanovení celkové genové exprese MeSH
- superoxiddismutasa genetika MeSH
- U937 buňky MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
