Репозиторий Университета

© 2020 Elsevier B.V. Neutrophils release neutrophil extracellular traps (NETs) in response to numerous pathogenic microbes as the last suicidal resource (NETosis) in the fight against infection. Apart from the host defense function, NETs play an essential role in the pathogenesis of various autoimmune and inflammatory diseases. Therefore, understanding the molecular mechanisms of NETosis is important for regulating aberrant NET release. The initiation of NETosis after the recognition of pathogens by specific receptors is mediated by an increase in intracellular Ca2+ concentration, therefore, the use of Ca2+ ionophore A23187 can be considered a semi-physiological model of NETosis. Induction of NETosis by various stimuli depends on reactive oxygen species (ROS) produced by NADPH oxidase, however, NETosis induced by Ca2+ ionophores was suggested to be mediated by ROS produced in mitochondria (mtROS). Using the mitochondria-targeted antioxidant SkQ1 and specific inhibitors of NADPH oxidase, we showed that both sources of ROS, mitochondria and NADPH oxidase, are involved in NETosis induced by A23187 in human neutrophils. In support of the critical role of mtROS, SkQ1-sensitive NETosis was demonstrated to be induced by A23187 in neutrophils from patients with chronic granulomatous disease (CGD). We assume that Ca2+-triggered mtROS production contributes to NETosis either directly (CGD neutrophils) or by stimulating NADPH oxidase. The opening of the mitochondrial permeability transition pore (mPTP) in neutrophils treated by A23187 was revealed using the electron transmission microscopy as a swelling of the mitochondrial matrix. Using specific inhibitors, we demonstrated that the mPTP is involved in mtROS production, NETosis, and the oxidative burst induced by A23187.

© 2020 Elsevier B.V. Cancer cells continuously secrete inflammatory biomolecules which play significant roles in disease progression and tumor metastasis toward secondary sites. Despite recent efforts to capture cancer cells' intercellular secretion heterogeneity using microfluidics, the challenges in operation of these systems as well as the complexity of designing a biosensing assay for long-term and real-time measurement of single cell secretions have become grand research barriers. Here, we present a new capillary-based microfluidic biosensing approach to easily and reliably capture ~500 single cells inside isolated dead-end nanoliter compartments using simple pipette injection, and quantify their individual secretion dynamics at the single cell resolution over a long period of culture (~16 h). We first present a detailed investigation of the fluid mechanics underlying the formation of nanoliter compartments in the microfluidic system. Based on the measurement of single cell capture efficiency, we employ a one-step FRET-based biosensor which monitors the single cancer cells' protease activity. The sensor reports the fluorescent signal as a product of amino acid chain cleavage and reduction in its quenching capability. Using the single cell protease secretion data, we identified modes of cell secretion dynamics in our cell sample. While most of the cells had low secretion levels, two other smaller and more aggressive secretion dynamics were cells with secretion modes that include sharp spikes or slow but progressive trend. The method presented here overcomes the difficulties associated with performing single cell secretion assays, enabling a feasible and reliable technique for high throughput measurement of metabolic activities in cancer cells.

© 2020 Elsevier B.V. Cancer cells continuously secrete inflammatory biomolecules which play significant roles in disease progression and tumor metastasis toward secondary sites. Despite recent efforts to capture cancer cells' intercellular secretion heterogeneity using microfluidics, the challenges in operation of these systems as well as the complexity of designing a biosensing assay for long-term and real-time measurement of single cell secretions have become grand research barriers. Here, we present a new capillary-based microfluidic biosensing approach to easily and reliably capture ~500 single cells inside isolated dead-end nanoliter compartments using simple pipette injection, and quantify their individual secretion dynamics at the single cell resolution over a long period of culture (~16 h). We first present a detailed investigation of the fluid mechanics underlying the formation of nanoliter compartments in the microfluidic system. Based on the measurement of single cell capture efficiency, we employ a one-step FRET-based biosensor which monitors the single cancer cells' protease activity. The sensor reports the fluorescent signal as a product of amino acid chain cleavage and reduction in its quenching capability. Using the single cell protease secretion data, we identified modes of cell secretion dynamics in our cell sample. While most of the cells had low secretion levels, two other smaller and more aggressive secretion dynamics were cells with secretion modes that include sharp spikes or slow but progressive trend. The method presented here overcomes the difficulties associated with performing single cell secretion assays, enabling a feasible and reliable technique for high throughput measurement of metabolic activities in cancer cells.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Elsevier Ltd Immunosuppressive chemoresistance is a major barrier in lung cancer treatment. Tumor immunosuppressive environments mediated by myeloid-derived suppressor cells (MDSCs) play a key role in chemotherapy induced MDSC development and differentiation but their mechanistic role has not been elucidated. Here, we define a role for carboplatin based chemotherapy in potentiating an MDSC-dependent pathway that triggers the chemoresistance mechanism. Findings reveal MDSC differentiation and activation of IL-13/IL-33-mediated pathway through VCAM/RANTES following carboplatin treatment. Furthemore, secretion of T regulatory IL-10-producing CD4+Foxp3+ cells was increased followed by expression of co-inhibitory receptor TIGIT on T cells, leading to a dysfunctional T cell phenotype. These cells were characterized by an immunosuppressive phenotype with impaired activation, proliferation and cytokine production. Lung cancer tissues expressed CD155, which bound TIGIT receptors and inactivated CD8 T cells. This TIGIT expression on tumor-infiltrating T cells was found to be associated with tumor progression and was linked to functional exhaustion of T cells. In addition, the presence of plasmacytoid dendritic cells (pDCs) exposed to tumor-derived factors further enhanced tumor progression through IL-10 production and up-regulation of the inducible co-stimulatory ligand (ICOS-L). Deciphering these deranged immune mechanisms and how they are impacted by chemotherapy induction is essential for incorporation of novel immune-based strategies in order to restore immunity and inhibit the immunosuppressive phenotype of metastatic lung cancer.

© 2019 Stem cells and the niche in which they reside feature a complex microenvironment with tightly regulated homeostasis, cell-cell interactions and dynamic regulation of metabolism. A significant number of organoid models has been described over the last decade, yet few methodologies can enable single cell level resolution analysis of the stem cell niche metabolic demands, in real-time and without perturbing integrity. Here, we studied the redox metabolism of Lgr5-GFP intestinal organoids by two emerging microscopy approaches based on luminescence lifetime measurement – fluorescence-based FLIM for NAD(P)H, and phosphorescence-based PLIM for real-time oxygenation. We found that exposure of stem (Lgr5-GFP) and differentiated (no GFP) cells to high and low glucose concentrations resulted in measurable shifts in oxygenation and redox status. NAD(P)H-FLIM and O2-PLIM both indicated that at high ‘basal’ glucose conditions, Lgr5-GFP cells had lower activity of oxidative phosphorylation when compared with cells lacking Lgr5. However, when exposed to low (0.5 mM) glucose, stem cells utilized oxidative metabolism more dynamically than non-stem cells. The high heterogeneity of complex 3D architecture and energy production pathways of Lgr5-GFP organoids were also confirmed by the extracellular flux (XF) analysis. Our data reveals that combined analysis of NAD(P)H-FLIM and organoid oxygenation by PLIM represents promising approach for studying stem cell niche metabolism in a live readout.

© Springer Science+Business Media, LLC, part of Springer Nature 2020. We describe here the Oncobox method for scoring efficiencies of anticancer target drugs (ATDs) using high throughput gene expression data. The method rationale, design, and validation are given along with the examples of its practical applications in biomedicine. The method is based on the analysis of intracellular molecular pathways activation and measuring expressions of molecular target genes for every ATD under consideration. Using Oncobox method requires collection of normal (control) expression profiles and annotated databases of molecular pathways and drug target genes. Both microarray and RNA sequencing profiles are acceptable, although the latter type of data prevails in the most recent applications of this technique.

© Springer Science+Business Media, LLC, part of Springer Nature 2020. Intracellular molecular pathways (IMPs) control all major events in the living cell. IMPs are considered hotspots in biomedical sciences and thousands of IMPs have been discovered for humans and model organisms. Knowledge of IMPs activation is essential for understanding biological functions and differences between the biological objects at the molecular level. Here we describe the Oncobox system for accurate quantitative scoring activities of up to several thousand molecular pathways based on high throughput molecular data. Although initially designed for gene expression and mainly RNA sequencing data, Oncobox is now also applicable for quantitative proteomics, microRNA and transcription factor binding sites mapping data. The Oncobox system includes modules of gene expression data harmonization, aggregation and comparison and a recursive algorithm for automatic annotation of molecular pathways. The universal rationale of Oncobox enables scoring of signaling, metabolic, cytoskeleton, immunity, DNA repair, and other pathways in a multitude of biological objects. The Oncobox system can be helpful to all those working in the fields of genetics, biochemistry, interactomics, and big data analytics in molecular biomedicine.