SYK inhibition targets acute myeloid leukemia stem cells by blocking their oxidative metabolism

Spleen tyrosine kinase (SYK) is an important oncogene and signaling mediator activated by cell surface receptors crucial for acute myeloid leukemia (AML) maintenance and progression. Genetic or pharmacologic inhibition of SYK in AML cells leads to increased differentiation, reduced proliferation, and cellular apoptosis. Herein, we addressed the consequences of SYK inhibition to leukemia stem-cell (LSC) function and assessed SYK-associated pathways in AML cell biology. Using gain-of-function MEK kinase mutant and constitutively active STAT5A, we demonstrate that R406, the active metabolite of a small-molecule SYK inhibitor fostamatinib, induces differentiation and blocks clonogenic potential of AML cells through the MEK/ERK1/2 pathway and STAT5A transcription factor, respectively. Pharmacological inhibition of SYK with R406 reduced LSC compartment defined as CD34+CD38-CD123+ and CD34+CD38-CD25+ in vitro, and decreased viability of LSCs identified by a low abundance of reactive oxygen species. Primary leukemic blasts treated ex vivo with R406 exhibited lower engraftment potential when xenotransplanted to immunodeficient NSG/J mice. Mechanistically, these effects are mediated by disturbed mitochondrial biogenesis and suppression of oxidative metabolism (OXPHOS) in LSCs. These mechanisms appear to be partially dependent on inhibition of STAT5 and its target gene MYC, a well-defined inducer of mitochondrial biogenesis. In addition, inhibition of SYK increases the sensitivity of LSCs to cytarabine (AraC), a standard of AML induction therapy. Taken together, our findings indicate that SYK fosters OXPHOS and participates in metabolic reprogramming of AML LSCs in a mechanism that at least partially involves STAT5, and that SYK inhibition targets LSCs in AML. Since active SYK is expressed in a majority of AML patients and confers inferior prognosis, the combination of SYK inhibitors with standard chemotherapeutics such as AraC constitutes a new therapeutic modality that should be evaluated in future clinical trials

The Effect of the Parameters of Robotic TIG Welding on the Microstructure of 17-4PH Stainless Steel Welded Joint

17-4PH stainless steel finds application in the aerospace industry owing to its good mechanical properties and corrosion resistance. In the literature, this steel is described as good for welding, but research shows that it may be problematic due to the formation of defects. In this study, the welded joints were made by the robotic TIG welding method with various welding speeds (2 and 3 mm/s). The joints were subjected to non-destructive testing and were free from defects. The microstructure was observed by light microscopy and scanning electron microscopy. Changes in the microstructure of the heat affected zone were observed and discussed. Based on the observation of the microstructure and the change in the hardness profile, the heat affected zone was divided into 4 characteristic regions. δ-ferrite and NbC were observed in the martensite matrix. The welded joints were subjected to heat treatment consisting of solution and aging in 550°C for 4 h. The microstructure of the heat affected zone become homogenized as a result of the heat treatment. The content of stable austenite in the welded joint after the heat treatment was about 3%

The Influence of Efflux Pump Inhibitors on the Activity of Non-Antibiotic NSAIDS against Gram-Negative Rods.

BACKGROUND:Most patients with bacterial infections suffer from fever and various pains that require complex treatments with antibiotics, antipyretics, and analgaesics. The most common drugs used to relieve these symptoms are non-steroidal anti-inflammatory drugs (NSAIDs), which are not typically considered antibiotics. Here, we investigate the effects of NSAIDs on bacterial susceptibility to antibiotics and the modulation of bacterial efflux pumps. METHODOLOGY:The activity of 12 NSAID active substances, paracetamol (acetaminophen), and eight relevant medicinal products was analyzed with or without pump inhibitors against 89 strains of Gram-negative rods by determining the MICs. Furthermore, the effects of NSAIDs on the susceptibility of clinical strains to antimicrobial agents with or without PAβN (Phe-Arg-β-naphtylamide) were measured. RESULTS:The MICs of diclofenac, mefenamic acid, ibuprofen, and naproxen, in the presence of PAβN, were significantly (≥4-fold) reduced, decreasing to 25-1600 mg/L, against the majority of the studied strains. In the case of acetylsalicylic acid only for 5 and 7 out of 12 strains of P. mirabilis and E. coli, respectively, a 4-fold increase in susceptibility in the presence of PAβN was observed. The presence of Aspirin resulted in a 4-fold increase in the MIC of ofloxacin against only two strains of E. coli among 48 tested clinical strains, which included species such as E. coli, K. pneumoniae, P. aeruginosa, and S. maltophilia. Besides, the medicinal products containing the following NSAIDs, diclofenac, mefenamic acid, ibuprofen, and naproxen, did not cause the decrease of clinical strains' susceptibility to antibiotics. CONCLUSIONS:The effects of PAβN on the susceptibility of bacteria to NSAIDs indicate that some NSAIDs are substrates for efflux pumps in Gram-negative rods. Morever, Aspirin probably induced efflux-mediated resistance to fluoroquinolones in a few E. coli strains

The activity of NSAIDs and paracetamol (active substances and medicinal products) against clinical isolates of <i>Enterobacteriaceae</i> and non-fermentative Gram-negative rods.

<p>The activity of NSAIDs and paracetamol (active substances and medicinal products) against clinical isolates of <i>Enterobacteriaceae</i> and non-fermentative Gram-negative rods.</p

Effects of PAβN on the MIC values of tested NSAIDs against clinical isolates of <i>Enterobacteriaceae</i> and non-fermentative Gram-negative rods.

<p>Effects of PAβN on the MIC values of tested NSAIDs against clinical isolates of <i>Enterobacteriaceae</i> and non-fermentative Gram-negative rods.</p

Effects of acetylsalicylic acid and its metabolites on the susceptibility of Gram-negative clinical strains (12 isolates of each species) to quinolones in the presence or absence of PAβN.

<p>Effects of acetylsalicylic acid and its metabolites on the susceptibility of Gram-negative clinical strains (12 isolates of each species) to quinolones in the presence or absence of PAβN.</p

The activity of NSAIDs (active substances and medicinal products) with and without the efflux pump inhibitor PAβN against standard Gram-negative strains.

<p>The activity of NSAIDs (active substances and medicinal products) with and without the efflux pump inhibitor PAβN against standard Gram-negative strains.</p

Susceptibility of clinical strains of Gram-negative rods to selected antimicrobial agents in the presence or absence of PAβN.

<p>Susceptibility of clinical strains of Gram-negative rods to selected antimicrobial agents in the presence or absence of PAβN.</p

Targeting the thioredoxin system as a novel strategy against B cell acute lymphoblastic leukemia.

B cell precursor acute lymphoblastic leukemia (BCP-ALL) is a genetically heterogeneous blood cancer characterized by abnormal expansion of immature B cells. Although intensive chemotherapy provides high cure rates in a majority of patients, subtypes harboring certain genetic lesions, such as MLL rearrangements or BCR-ABL1 fusion, remain clinically challenging, necessitating a search for other therapeutic approaches. Herein, we aimed to validate antioxidant enzymes of the thioredoxin system as potential therapeutic targets in BCP-ALL. We observed oxidative stress along with aberrant expression of the enzymes associated with the activity of thioredoxin antioxidant system in BCP-ALL cells. Moreover, we found that auranofin and adenanthin, inhibitors of the thioredoxin system antioxidant enzymes, effectively kill BCP-ALL cell lines and pediatric and adult BCP-ALL primary cells, including primary cells co-cultured with bone marrow-derived stem cells. Furthermore, auranofin delayed the progression of leukemia in MLL-rearranged patient-derived xenograft model and prolonged the survival of leukemic NSG mice. Our results unveil the thioredoxin system as a novel target for BCP-ALL therapy, and indicate that further studies assessing the anticancer efficacy of combinations of thioredoxin system inhibitors with conventional anti BCP-ALL drugs should be continued