Superior efficacy of co-targeting GFI1/KDM1A and BRD4 against AML and post-MPN secondary AML cells

There is an unmet need to overcome nongenetic therapy-resistance to improve outcomes in AML, especially post-myeloproliferative neoplasm (MPN) secondary (s) AML. Studies presented describe effects of genetic knockout, degradation or small molecule targeted-inhibition of GFI1/LSD1 on active enhancers, altering gene-expressions and inducing differentiation and lethality in AML and (MPN) sAML cells. A protein domain-focused CRISPR screen in LSD1 (KDM1A) inhibitor (i) treated AML cells, identified BRD4, MOZ, HDAC3 and DOT1L among the codependencies. Our findings demonstrate that co-targeting LSD1 and one of these co-dependencies exerted synergistic in vitro lethality in AML and post-MPN sAML cells. Co-treatment with LSD1i and the JAKi ruxolitinib was also synergistically lethal against post-MPN sAML cells. LSD1i pre-treatment induced GFI1, PU.1 and CEBPα but depleted c-Myc, overcoming nongenetic resistance to ruxolitinib, or to BETi in post-MPN sAML cells. Co-treatment with LSD1i and BETi or ruxolitinib exerted superior in vivo efficacy against post-MPN sAML cells. These findings highlight LSD1i-based combinations that merit testing for clinical efficacy, especially to overcome nongenetic therapy-resistance in AML and post-MPN sAML.Subject terms: Acute myeloid leukaemia, Targeted therapies     

Unraveling the impact of interval length between neoadjuvant chemoradiotherapy and surgery on perioperative and postoperative complications in rectal cancer patients

European Surgery - The optimal timing for surgery following neoadjuvant chemoradiotherapy (nCRT) in rectal cancer patients and its impact on perioperative and postoperative complications remain...     

The effects of black seed supplementation on cardiovascular risk factors in patients with nonalcoholic fatty liver disease: A randomized,double‐blind,placebo‐controlled clinical trial

Nonalcoholic fatty liver disease (NAFLD) is highly related to cardiovascular disorders risk factors. This study aimed to evaluate the effects of black seed (Nigella sativa) supplementation on cardiovascular disorders risk factors in patients with NAFLD. This randomized, double‐blind, placebo‐controlled clinical trial was conducted on 50 patients with NAFLD. Participants were assigned to receive a lifestyle modification plus 2 g/day of either N. sativa or placebo for 12 weeks. Compared with the placebo, N. sativa supplementation led to significant reductions in serum glucose (?7.95 vs. ?1.22; p = .041), serum insulin (?3.87 vs. ?1.07; p = .027), homeostatic model of assessment for insulin resistance (?1.02 vs. ?0.28; p = .021), and a significant increase in quantitative insulin sensitivity check index (0.03 vs. 0.006; p = .002). All of these changes were remained significant after adjusting for known confounding variables; however, there was no significant difference in lipid profile changes between the two groups (p = .05). N. sativa supplementation significantly decreased hepatic steatosis percentage compared with the placebo after adjustment for confounding variables (p = .005). In conclusion, our results indicate that daily intake of 2‐g N. sativa plus lifestyle modification is superior to lifestyle modification alone in amelioration of insulin resistance and hepatic steatosis in patients with NAFLD.     

Comparison of clinical characteristics of opium-induced and independent major depressive disorder

Psychiatric disorders and among them depression are common in substance dependent patients. The aim of this study is to compare the clinical characteristics of those that appear to have substance-induced depression and those that have independent major depression. One-hundred eighty-four independent and 187 opium-induced (OID) depressed male patients that met the DSM-IV criteria for major depressive disorder were randomly selected. Standard demographic data, including age, marital, employment and education status, were collected. The primary measure of depressive signs and symptoms was Hamilton Depression Scale (HAMD-21). The two groups were compared with each other for the HAMD total and subscales scores. The two groups were matched regarding age, educational level and marital status. Opium-induced depressed patients were more severely depressed and motor retarded and also they had more social and occupational problems. Gastrointestinal, sexual and somatic complaints were more common among them too. MDD patients had better insight than the other group. The results demonstrate that it is possible to differentiate between substance-induced and independent depression. Such differentiation might be important for establishing prognosis and optimal treatment.     

Interferon-γ Induces Apoptosis and Augments the Expression of Fas and Fas Ligand by Microglia in Vitro

Activation of microglia by interferon-γ (IFN-γ) has been implicated in a number of central nervous system (CNS) inflammatory disease processes. Because IFN-γ has also been shown to play a role in programmed cell death, we investigated its cytotoxicity and its effect on the Fas apoptotic pathway in microglia. Flow cytometry was used to quantify the IFN-γ-mediated apoptotic response and Fas and Fas ligand (FasL) expression in two well-characterized murine microglia cell lines (BV-2 and N9). Nuclear fragmentation, suggestive of apoptosis, was noted within 24 h of incubation of microglia with IFN-γ (10 U/ml). After a 72-h incubation, almost every BV-2 and N9 microglia, but not GL261 glioma cells, underwent cell death and detached from the culture plates. This cytotoxicity occurred even at low IFN-γ concentrations (1 U/ml) and was inhibited by BAF, a pan-caspase inhibitor. Incubation of BV-2 and N9 microglia, but not GL261 glioma cells, with IFN-γ also potentiated the expression of Fas and FasL in a similar dose–response and time-course manner, as seen for the apoptotic response. Whereas Fas expression increased by 100% in both microglia cells, FasL upregulation was more pronounced and increased by as much as 200% in the N9 cells. These findings suggest that in addition to its role as a microglia activator, IFN-γ may also induce apoptosis of microglia, possibly through simultaneous upregulation of Fas and FasL. Interferon-γ modulation of the Fas pathway and apoptosis in microglia may be important in the pathogenesis of inflammatory CNS disease processes.     

Molecular dynamics simulation on the low sensitivity of mutants of NEDD-8 activating enzyme for MLN4924 inhibitor as a cancer drug

MLN4924 is an experimental cancer drug known as inhibitor of NEDD8-activating enzyme (NAE). This anti-tumor candidate is a selective small-molecule inhibitor of NAE which is conjugated to cullin protein on Cullin-RING ligases (CRLs). This covalent modification actives cullin complex to recruit an ubiquitin-charged E2 and leads to downstream target protein polyubiquitination and proteasomal degradation. MLN4924, which can form a covalent adduct with NEDD8, and block NAE at the first step in this pathway, has shown anti-tumor activity in many kinds of cancer cell lines and also xenograft models, including lung cancer, colon cancer, melanoma and lymphoma. The anti-tumor activity of MLN4924 results from inactivation of CLRs, which causes DNA re-replication and inhibition of nuclear factor (NF)-κB signaling, thus leading to cancer cell death. A mutation can reduce the enzyme’s sensitivity to MLN4924. Verma et al. in 2013 studied on molecular dynamics simulation of a mutant A171T and consequently found out that this mutation reduce MLN4924 interaction with DNA Binding site of enzyme as a result of reduction of enzyme affinity to ATP. One year later, in 2014, Wei Xu et al. carried out a research on inhibitor resistant cell lines and revealed that a couple of mutations so called Y352H and I310N leads to enzyme resistance to MLN4924 inhibitor, interestingly, the cause reported was the increase of enzyme affinity to ATP. As in Wei Xu et al. experiment the molecular dynamics simulation was not considered, present study is conducted to identify enzyme mutation mechanism by molecular dynamics approach using advantages of Gromacs software version 4.5.6.     

Locomotor sensitization to cocaine is associated with distinct pattern of glutamate receptor trafficking to the postsynaptic density in prefrontal cortex: Early versus late withdrawal effects

Glutamatergic neurotransmission plays an important role in the behavioral and molecular plasticity observed in cocaine mediated locomotor sensitization. Recent studies show that glutamatergic signaling is regulated by receptor trafficking, synaptic localization, and association with scaffolding proteins. The trafficking of the glutamate receptors was investigated in the dorsal and ventral prefrontal cortex at 1 and 21 days after repeated cocaine administration which produced robust locomotor sensitization. A subcellular fractionation technique was used to isolate the cellular synaptosomal fraction containing the postsynaptic density. At early withdrawal, the prefrontal cortex displayed a reduction in the synaptosomal content of the AMPA and NMDA receptor subunits. In contrast, after extended withdrawal, there was a significant increase in the trafficking of the receptors into the synaptosomal compartment. These changes were accompanied by corresponding trafficking of the postsynaptic glutamatergic scaffolding proteins. Thus, enhanced trafficking of glutamate receptors from cytosolic to synaptosomal compartment is associated with prolonged withdrawal from repeated exposure to cocaine and may have functional consequences for the synaptic and behavioral plasticity.