Chimeric antigen receptor T (CART) cells targeting CD19 have shown promising results in the treatment of chronic lymphocytic leukemia (CLL). However, efficacy seems to be inferior compared to diffuse ...large B‐cell lymphoma or acute lymphoblastic leukemia. Impaired T‐cell fitness of CLL patients may be involved in treatment failure. Less‐differentiated naïve‐like T cells play an important role in CART expansion and long‐term persistence in vivo. These cells are sparse in CLL patients. Therefore, optimization of CART cell production protocols enriching less differentiated T cell subsets may overcome treatment resistance. The B‐cell receptor inhibitor ibrutinib targeting Bruton's tyrosine kinase (BTK) is approved for the treatment of CLL. Besides BTK, ibrutinib additionally inhibits interleukin‐2‐inducible T‐cell kinase (ITK) which is involved in T‐cell differentiation. To evaluate the effect of ibrutinib on CART cell production, peripheral blood mononuclear cells from nine healthy donors and eight CLL patients were used to generate CART cells. T‐cell expansion and phenotype, expression of homing and exhaustion makers as well as functionality of CART cells were evaluated. CART cell generation in the presence of ibrutinib resulted in increased cell viability and expansion of CLL patient‐derived CART cells. Furthermore, ibrutinib enriched CART cells with less‐differentiated naïve‐like phenotype and decreased expression of exhaustion markers including PD‐1, TIM‐3 and LAG‐3. In addition, ibrutinib increased the cytokine release capacity of CLL patient‐derived CART cells. In summary, BTK/ITK inhibition with ibrutinib during CART cell culture can improve yield and function of CLL patient‐derived CART cell products.
What's new?
Chimeric antigen receptor T (CART) cells targeting CD19 have shown promising results in chronic lymphocytic leukemia (CLL). However, naïve‐like T cells play an important role in CART expansion and long‐term persistence in vivo, and these cells are sparse in CLL patients. Here, the authors show that BTK/ITK inhibition with Ibrutinib during CART cell generation may improve CLL patient‐derived CART cell products and enhance CART cell function. Supplementing CART cell production with ibrutinib increases CART cell yields and enriches CART cells with less‐differentiated phenotypes and lower expression of exhaustion markers, representing a potential avenue to improve the clinical outcome of patients.
•Tumor cells constitute the direct targets of anti-VEGF therapy;•Tumor cells possess the capacity to independently develop resistance to VEGF inhibition;•Acquired resistance to VEGF inhibition within ...tumor cells is reversible.
Vascular endothelial growth factor (VEGF) inhibition is an essential targeted strategy for malignant tumors, but its efficacy is severely constrained by drug resistance. The traditional view holds that the target of VEGF inhibition is endothelial cells, and thus compensatory angiogenesis is considered the main mechanism of drug resistance. In this study, we found that tumor cells themselves could develop acquired resistance to VEGF therapy, indicating an independent resistance mechanism apart from angiogenesis. Notably, this acquired resistance was temporary, disappearing completely four days after discontinuing exposure to the drug in vitro. Our findings suggest that tumor cells may also be targets of VEGF inhibition, and their response to treatment should not be overlooked in contributing to drug resistance.
Breast cancer has become one of the top five commonest causes of cancer death. The use of ferroptosis to induce the generation of reactive oxygen species (ROS) in cancer cells presents a promising ...and potential strategy for cancer treatment. Herein, a series of facile bimetallic nanoparticles (
% Fe-doped ZIF-8) were synthesized and tested, and doxorubicin (DOX), a classic drug for breast cancer therapy, was encapsulated. After comparing the ratios of Fe
/(Fe
+ Zn
), 7% Fe-doped ZIF-8 (7FZ) was found to be the most suitable particle for medical application. The drug loading efficiency of DOX@7FZ was 58.01 ± 0.02%. The pH-sensitive DOX@7FZ was degraded and DOX was released in lysosomes once internalized. Both the intracellular content of iron and ROS increased significantly. Meanwhile, the cell viability declined to 13.98% in 24 h at a concentration of 60 μg mL
and the IC
was 42.68 μg mL
. Moreover, the expression of Bcl-2 and GPX-4 proteins decreased in a time-dependent manner, indicating that DOX@7FZ was able to enhance the ROS level in cancer cells
a synergistic effect between apoptosis and ferroptosis. The mechanism of action of DOX@7FZ was further verified using hematoxylin and eosin staining and immunohistochemical staining of Bcl-2 and GPX-4. These remarkable characteristics of DOX@7FZ may inspire further advancements in the treatment of breast cancer.
In this work, an attempt was made to evaluate the effect of pesticides on growth pattern, surface morphology, cell viability and growth regulators of nitrogen fixing soil bacterium. Pesticide ...tolerant Azotobacter vinelandii strain AZ6 (Accession no. MG028654) was found to tolerate maximum level of pesticide and displayed multifarious PGP activities. At higher concentrations, pesticides triggered cellular/structural damage and reduced the cell viability as clearly shown under SEM and CLSM. With increase in concentration, pesticides exhibited a significant (p < 0.05) decrease in PGP traits of strain AZ6. Among all three groups of pesticides, herbicides glyphosate and atrazine were most toxic. Kitazin, hexaconazole, metalaxyl, glyphosate, quizalofop, atrazine, fipronil, monocrotophos and imidacloprid at 2400, 1800, 1500, 900, 1200, 900, 1800, 2100 and 2700 μg mL−1, respectively, decreased the production of IAA by 19.5 ± 1.9 (61%), 18.1 ± 1.2 (64%), 36.4 ± 3.4 (28%), 13.1 ± 0.8 (74%), 15.6 ± 1.0 (69%), 7.6 ± 0.5 (83%), 11.9 ± 0.8 (76%), 24.7 ± 1.7 (51%) and 32 ± 2.3 (37%) μg mL−1, respectively, over control (50.7 ± 3.6 μg mL−1). A maximum reduction of 8.4 ± 1.2 (46%), 5.8 ± 0.6 (62%) and 4 ± 0.2 (74%) μg mL−1 in 2, 3-DHBA at 300 (1×), 600 (2×) and 900 (3×) μg mL−1 glyphosate, respectively, While, 32.8 ± 2.7 (19%), 27.2 ± 2 (33%) and 21.5 ± 1.3 (47%) μg mL−1, respectively in the production of SA was observed at 300 (1×), 600 (2×) and 900 (3×) μg mL−1 atrazine, respectively. Likewise, with increase in concentration of pesticides, decrease in P solubilization ability and change in pH of broth was detected. The order of pesticide toxicity to PSE (percent decline over control) at highest concentration was: atrazine (45) > kitazin (44) > metalaxyl (43) > monocrotophos (43) > glyphosate (41) > hexaconazole (39) > quizalofop (33) > imidacloprid (31) > fipronil (25). The present study undoubtedly suggests that even at higher doses of pesticides, A. vinelandii maintained secreting plant growth regulators and this property makes this strain agronomically important microbe for enhancing the growth of plants.
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•Toxicity of pesticides to rhizosphere isolate varied under in vitro conditions.•Pesticides affects the growth pattern of bacterial strain A. vinelandii.•Pesticides alters the morphology and cell viability of strain AZ6.•Pesticides modulates the phytohormones and other active biomolecules secreted/produced by A. vinelandii.•Pesticide induced changes in amount of solubilized-P and drop in pH.
•Bioactive compounds of A. arguta leaves extracts are reported.•A. arguta leaves extracts strongly scavenged physiologically relevant ROS and RNS.•No adverse effects on Caco-2 and HT29-MTX ...cells.•Alcoholic extract was shown to have the strongest activities measured.
The present study reports for the first time the identification and quantification of phenolic compounds, the antioxidant and antimicrobial activities as well as the in vitro radical scavenging activity and intestinal cell effects of A. arguta leaves extracts. Extractions were carried out under water, water:ethanol (50:50) and ethanol. The highest antioxidant activity were obtained in alcoholic extract (IC50 = 53.95 ± 3.09 μg/mL for DPPH; 6628.42 ± 382.49 µmol/mg dry weight basis for FRAP) while the phenolic profile confirmed by HPLC analysis revealed highest amounts of phenolic acids (hydroxycinnamic acid derivatives) and flavonoids (flavan-3-ol and flavonols derivatives). An excellent scavenging activity against reactive oxygen and nitrogen species were determined for all extracts as well as no adverse effects on Caco-2 and HT29-MTX cells in concentrations below 100 μg/mL and 1000 μg/mL, respectively. These results highlight the potentialities of hardy kiwi leaves valorization.
The tyrosine kinase inhibitor lenvatinib is used to treat advanced hepatocellular carcinoma (HCC). Ferroptosis is a type of cell death characterized by the iron‐dependent accumulation of lethal lipid ...reactive oxygen species (ROS). Nuclear factor erythroid‐derived 2‐like 2 (Nrf2) protects HCC cells against ferroptosis. However, the mechanism of lenvatinib‐induced cytotoxicity and the relationships between lenvatinib resistance and Nrf2 are unclear. Thus, we investigated the relationship between lenvatinib and ferroptosis and clarified the involvement of Nrf2 in lenvatinib‐induced cytotoxicity. Cell viability, lipid ROS levels, and protein expression were measured using Hep3B and HuH7 cells treated with lenvatinib or erastin. We examined these variables after silencing fibroblast growth factor receptor‐4 (FGFR4) or Nrf2 and overexpressing‐Nrf2. We immunohistochemically evaluated FGFR4 expression in recurrent lesions after resection and clarified the relationship between FGFR4 expression and lenvatinib efficacy. Lenvatinib suppressed system Xc− (xCT) and glutathione peroxidase 4 (GPX4) expression. Inhibition of the cystine import activity of xCT and GPX4 resulted in the accumulation of lipid ROS. Silencing‐FGFR4 suppressed xCT and GPX4 expression and increased lipid ROS levels. Nrf2‐silenced HCC cells displayed sensitivity to lenvatinib and high lipid ROS levels. In contrast, Nrf2‐overexpressing HCC cells displayed resistance to lenvatinib and low lipid ROS levels. The efficacy of lenvatinib was significantly lower in recurrent HCC lesions with low‐FGFR4 expression than in those with high‐FGFR4 expression. Patients with FGFR4‐positive HCC displayed significantly longer progression‐free survival than those with FGFR4‐negative HCC. Lenvatinib induced ferroptosis by inhibiting FGFR4. Nrf2 is involved in the sensitivity of HCC to lenvatinib.
Regulation of ferroptosis by fibroblast growth factor receptor 4 (FGFR4) and nuclear factor erythroid‐derived 2‐like 2 (Nrf2) in hepatocellular carcinoma GPX4, glutathione peroxidase 4.
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Recently, polysaccharides based microparticles have been found to offer an attractive potential as a carrier in drug delivery field. In this study, bare gellan gum microparticles (GG ...MPs) and methotrexate (MTX) loaded gellan gum microparticles (MTX-GG MPs) prepared by using simple water-in-oil (W/O) emulsion solvent diffusion method. The developed microparticles (MPs) were found discretely distributed in a spherical shape. MTX has been encapsulated in microparticles with 84.8 ± 1.68% encapsulation efficiency (%EE) and 6.45 ± 0.07% loading capacity (%LC). The Fourier Transform Infrared Spectroscopy (FTIR) characterization of the MPs clearly indicated the physical encapsulation of MTX into polymeric matrix of MPs. Thermogravimetric analysis (TGA) characterization showed slightly higher thermal stability of MTX-GG MPs in comparison to the GG MPs. In vitro release study of MTX-GG MPs showed 84% drug release within 24 h. The hemolysis study of GG MPs and MTX-GG MPs on human red blood cells (RBCs) showed <1.0% hemolysis. The cell viability studies on L929 showed GG MPs, and MTX-GG MPs are biocompatible.
Islet amyloid deposition and reduced ß-cell mass are pathologic hallmarks of type 2 diabetes. Amyloid deposits contain the 37 amino acid hIAPP, aggregation of which is toxic to ß cells. The 20-29 ...amino acid region of the peptide is known to confer its amyloidogenic potential. We previously reported that plasmin, a fibrinolytic protease, cleaves hIAPP thereby inhibiting fibril formation. We now sought to determine whether hIAPP fragments arising from plasmin-mediated cleavage retain the potential to aggregate and induce cytotoxicity. The ß-cell line INS-1 was treated for 24 h with full-length hIAPP (20 µM) alone or in the presence of plasmin (0.4 µM); plasmin alone or buffer were used as controls. Exposure to hIAPP alone decreased cell viability (53±1.4% of buffer, p<0.05, n=5). Addition of plasmin to hIAPP restored cell viability (90±4.3% of buffer, p=0.3 vs. buffer and p<0.05 vs. hIAPP alone, n=5), while treatment with plasmin alone had no effect (102±2.9% of buffer, p=0.7, n=5). LC/MS analysis of hIAPP cleavage products showed that plasmin induced a rapid decrease in the abundance of full-length hIAPP and appearance of the hIAPP 1-11 and 12-37 fragments. Since the hIAPP 12-37 contains the amyloidogenic region, we compared the amyloidogenic and cytotoxic properties of this fragment to full-length hIAPP. In a thioflavin T assay, hIAPP 12-37 was less amyloidogenic than full-length hIAPP. As expected, treatment of INS-1 cells for 24 h with full-length hIAPP (0-60 µM) reduced cell viability in a dose-dependent manner (67±1.3%, 30±0.4% and 16±0.7% of buffer, at 20, 40 and 60 µM respectively; p<0.05 vs. buffer for all, n=2), while hIAPP 12-37 was not toxic at any dose tested (90±3.2%, 90±3.3% and 108±3.5% of buffer, at 20, 40 and 60 µM respectively; n=2).
In summary, plasmin protects ß cells from hIAPP-induced toxicity by cleaving hIAPP between amino acids 11 and 12. Thus, increasing islet plasmin activity might be a strategy to limit ß-cell loss in type 2 diabetes.
Disclosure
N. Esser: None. M.F. Hogan: None. A.T. Templin: None. J.J. Castillo: None. D. Raleigh: None. J.S. Edgar: None. S. Zraika: Research Support; Self; Novartis Pharmaceuticals Corporation. R.L. Hull: None. S.E. Kahn: Advisory Panel; Self; Boehringer Ingelheim International GmbH, Eli Lilly and Company, Intarcia Therapeutics, Janssen Scientific Affairs, LLC., Merck & Co., Inc., Novo Nordisk A/S, Pfizer Inc.
Funding
U.S. Department of Veterans Affairs (I01BX001060)
Alzheimer's disease (AD) is a common neurodegenerative disease with an increasing incidence rate. Numerous microRNAs (miRNAs) have been found to be involved in AD progression. This study aimed to ...investigate the expression and diagnostic value of microRNA-331-3p (miR-331-3p) in AD patients and to explore the effects of miR-331-3p on neuronal viability and neuroinflammation.
This study recruited AD patients and used Aβ1–40 treated SH-SY5Y cells mimicking AD characteristics. The expression of miR-331-3p was estimated using reverse transcription quantitative PCR. A receiver operating characteristic (ROC) analysis was used to evaluate the diagnostic value of miR-331-3p, and the correlation of miR-331-3p with patients' Mini-Mental State Examination (MMSE) scores and serum proinflammatory cytokines were analyzed. The effects of miR-331-3p on neuronal viability and inflammatory response were explored in SH-SY5Y cells by in vitro analysis.
In AD patients and Aβ1–40 treated SH-SY5Y cells, the expression of miR-331-3p was significantly downregulated. Serum miR-331-3p had certain diagnostic potential and was correlated with the MMSE scores and serum proinflammatory cytokine levels of AD patients. In Aβ1–40-treated SH-SY5Y cells, the overexpression of miR-331-3p enhanced cell viability and inhibited inflammatory responses.
The data of this study indicated that serum expression of miR-331-3p is decreased in AD patients, and is correlated with the MMSE scores and proinflammatory cytokine levels of AD patients. In addition, miR-331-3p can regulate the cell viability and the expression of pro-inflammatory cytokines of Aβ1–40 treated SH-SY5Y cells, indicating the potential neuroprotective role of miR-331-3p.
•Alzheimer's disease patients have decreased serum miR-331-3p expression.•Serum miR-331-3p is correlated with patients' MMSE scores and inflammation.•miR-331-3p can promote cell viability and attenuate inflammation of SH-SY5Y cells.•Von Hippel-Lindau tumor suppressor serves as a target of miR-331-3p in SH-SY5Y cells.
OBJECTIVEIn the current research, lornoxicam-loaded in situ gels were developed, and their potential usage in ocular inflammation was evaluated.SIGNIFICANCELornoxicam cyclodextrin complex prepared ...with hydroxypropyl methylcellulose and poloxamer P407 because of the low viscosity of in situ gels to provide easy application. However, washing and removing it from the ocular surface becomes difficult due to the gelation formation with heat.METHODSA three-level factorial experimental design was used to evaluate the effects of poloxamer 407 concentration, polymer type, and polymer concentration on viscosity, pH, gelation capacity, gelation time, and gelation temperature, which were considered the optimal indicators of lornoxicam-containing formulations.RESULTSAs a result of the three-level factorial experimental design, the optimized formulation contained 15 (%w/v) poloxamer 407 and 1 (%w/v) hydroxypropyl methylcellulose. The optimize formulation viscosity 25 °C = 504 ± 49cP, viscosity 35 °C = 11247 ± 214cP, pH = 6.80 ± 0.01, gelation temprature = 35 ± 0.2 °C, and gelation time= 34 ± 0.2 s was obtained. In the in vitro release studies, 68% of lornoxicam was released with a burst effect in the first three hours; then, the release continued for eight hours with controlled release. Release kinetics of the formulations were modeled mathematically, and it was found to be compatible with the Korsemeyer-Peppas and Weibull models. In cell culture studies, cell viability at 100 µg/mL was 83% and 96% for NL6 and NL6-CD, respectively. In Draize's in vivo test, no negative conditions occurred in rats.CONCLUSIONSTherefore, the NL6-CD formulation has the potential to be a favorable option for treating ocular inflammation.
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