Gossypol Inhibits Non-small Cell Lung Cancer Cells Proliferation by Targeting EGFRL858R/T790M

Background: Overexpression of epidermal growth factor receptor (EGFR) has been reported to be implicated in the pathogenesis of non-small cell lung cancer (NSCLC). Several EGFR inhibitors have been used in clinical treatment of NSCLC, but the emergence of EGFRL858R/T790M resistant mutation has reduced the efficacy of the clinical used EGFR inhibitors. There is an urgent need to develop novel EGFRL858R/T790M inhibitors for better NSCLC treatment.Methods: By screening a natural product library, we have identified gossypol as a novel potent inhibitor targeting EGFRL858R/T790M. The activity of gossypol on NSCLC cells was evaluated by cell proliferation, cell apoptosis and cell migration assays. Kinase activity inhibition assay and molecular docking were used to study the inhibition mechanism of gossypol to EGFRL858R/T790M. Western blotting was performed to study the molecular mechanism of gossypol inhibiting the downstream pathways of EGFR.Results: Gossypol inhibited the cell proliferation and cell migration of NSCLC cells, and induced caspase-dependent cell apoptosis of NSCLC cells by upregulating the expression of pro-apoptotic protein BAD. Molecular docking revealed that gossypol could bind to the kinase domain of EGFRL858R/T790M with good binding affinity through hydrogen bonds and hydrophobic interactions. Gossypol inhibited the kinase activity of EGFRL858R/T790M with EC50 of 150.1 nM. Western blotting analysis demonstrated that gossypol inhibited the phosphorylation of EGFR and its downstream signal pathways in a dose-dependent manner.Conclusion: Gossypol inhibited cell proliferation and induced apoptosis of NSCLC cells by targeting EGFRL858R/T790M. Our findings provided a basis for developing novel EGFRL858R/T790M inhibitors for treatment of NSCLC

Core-sheath nanofibers as drug delivery system for thermoresponsive controlled release

In this work, a smart drug delivery system of core–sheath nanofiber is reported. The core-sheath nanofibers were prepared with thermoresponsive poly-(N-isopropylacrylamide) (PNIPAAm) (as core) and hydrophobic ethylcellulose (EC) (as sheath) by coaxial electrospinning. Analogous medicated fibers were prepared by loading with a model drug ketoprofen (KET). The fibers were cylindrical without phase separation and have visible core-sheath structure as shown by scanning and transmission electron microscopy. X-ray diffraction patterns demonstrated the drug with the amorphous physical form was present in the fiber matrix. Fourier transform infrared spectroscopy analysis was conducted, finding that there were significant intermolecular interactions between KET and the polymers. Water contact angle measurements proved that the core-sheath fibers from hydrophobic transformed into hydrophobic when the temperature reached the lower critical solution temperature. In vitro drug-release study of nanofibers with KET displayed that the coaxial nanofibers were able to synergistically combine the characteristics of the two polymers producing a temperature-sensitive drug delivery system with sustained release properties. In addition, they were established to be non-toxic and suitable for cell growth. These findings show that the core–sheath nanofiber is a potential candidate for controlling drug delivery system

Regional Crustal Vertical Deformation Driven by Terrestrial Water Load Depending on CORS Network and Environmental Loading Data: A Case Study of Southeast Zhejiang

Monitoring regional terrestrial water load deformation is of great significance to the dynamic maintenance and hydrodynamic study of the regional benchmark framework. In view of the lack of a spatial interpolation method based on the GNSS (Global Navigation Satellite System) elevation time series for obtaining terrestrial water load deformation information, this paper proposes to employ a CORS (Continuously Operating Reference Stations) network combined with environmental loading data, such as ECMWF (European Centre for Medium-Range Weather Forecasts) atmospheric data, the GLDAS (Global Land Data Assimilation System) hydrological model, and MSLA (Mean Sea Level Anomaly) data. Based on the load deformation theory and spherical harmonic analysis method, we took 38 CORS stations in southeast Zhejiang province as an example and comprehensively determined the vertical deformation of the crust as caused by regional terrestrial water load changes from January 2015 to December 2017, and then compared these data with the GRACE (Gravity Recovery and Climate Experiment) satellite. The results show that the vertical deformation value of the terrestrial water load in southeast Zhejiang, as monitored by the CORS network, can reach a centimeter, and the amplitude changes from −1.8 cm to 2.4 cm. The seasonal change is obvious, and the spatial distribution takes a ladder form from inland to coastal regions. The surface vertical deformation caused by groundwater load changes in the east–west–south–north–central sub-regions show obvious fluctuations from 2015 to 2017, and the trends of the five sub-regions are consistent. The amplitude of surface vertical deformation caused by groundwater load change in the west is higher than that in the east. We tested the use of GRACE for the verification of CORS network monitoring results and found a relatively consistent temporal distribution between both data sets after phase delay correction on GRACE, except for in three months—November in 2015, and January and February in 2016. The results show that the comprehensive solution based on the CORS network can effectively improve the monitoring of crustal vertical deformation during regional terrestrial water load change

Inhibition of Netosis with PAD Inhibitor Attenuates Endotoxin Shock Induced Systemic Inflammation

Neutrophils play a pivotal role in innate immunity by releasing neutrophils extracellular traps (NETs). Excessive NETs are detrimental to the local tissue and further exacerbate inflammation. Protein arginine deiminases (PAD) mediate histone citrullination and NET formation that, in turn, exacerbate endotoxin shock damages. In this study, we further investigated the molecular mechanism underlying PAD and NETs in endotoxic stress in mice. The control group mice were injected with solvent, the LPS endotoxic shock group mice were intraperitoneally injected with LPS at 35 mg/kg only, while the LPS and PAD inhibitor YW3-56 treatment group mice were injected with YW3-56 at 10 mg/kg prior to the LPS injection. YW3-56 significantly prolonged the survival time of the LPS-treated mice. NETs, cfDNA, and inflammatory factors were detected by ELISA in serum, paitoneal cavity, and lung at 24 h after LPS administration. Lung injuries were detected by immunostaining, and lung tissue transcriptomes were analyzed by RNA-seq at 24 h after LPS administration. We found that YW3-56 altered neutrophil tissue homeostasis, inhibited NET formation, and significantly decreased cytokines (IL-6, TNFα and IL-1β) levels, cytokines gene expression, and lung tissue injury. In summary, NET formation inhibition offers a new avenue to manage inflammatory damages under endotoxic stress

The Crustal Vertical Deformation Driven by Terrestrial Water Load from 2010 to 2014 in Shaanxi–Gansu–Ningxia Region Based on GRACE and GNSS

The terrestrial water resources in Shaanxi–Gansu–Ningxia (SGN) region are relatively scarce, and its climate change is unstable. Research on the deformation driven by terrestrial water load is of great significance to the dynamic maintenance of reference station networks. In this paper, data derived from Gravity Recovery and Climate Experiment (GRACE) and Global Navigation Satellite System (GNSS) from 2010 to 2014 were combined to monitor the spatiotemporal characteristics of surface vertical deformation caused by terrestrial water load change. The single scale factor was calculated by comparing CPC, WGHM, and GLDAS hydrological model to restore filtering leakage signal. The singular spectrum analysis (SSA) method was used to extract the principal component of temporal vertical deformation, and its spatial distribution was analyzed. At the same time, in order to study the relationship between the terrestrial water load deformation from GRACE and that from GNSS, the first-order term correction, the Atmosphere and Ocean De-aliasing Level-1B product (GAC) correction, and the first-order load LOVE number correction for GRACE were adopted in this paper. In addition, a quantitative comparative analysis of both the monitoring results was carried out. The results show that the time-variable characteristics of surface vertical deformation characterized by the filtered three hydrological models were consistent with those of GRACE. The correlation coefficient and Nash–Sutcliffe efficiency coefficient (NSE) values were the highest in the GLDAS model and the GRACE model, respectively; the former index is 0.93, while the latter is 0.85. The crustal vertical deformation from terrestrial water load showed a declining rate from 2010 to 2014. Its spatial change rate showed an obvious ladder distribution, with the surface subsidence rate gradually decreasing from south to north. In addition, weighted root mean square (WRMS) contribution rate of the crustal vertical deformation resulting from GRACE with GAC correction between the different GNSS stations ranged from 18.52% to 54.82%. The correlation coefficient between them was close to 0.70. After deducting the mass load impact of GRACE only, the WRMS contribution rate of the corresponding stations decreased from −8.42% to 21.18%. The correlation coefficient between them reduced noticeably. Adding GAC back can increase the comparability with GRACE and GNSS in terms of monitoring the crustal vertical deformation. The annual amplitude and phase of surface vertical deformation resulting from GRACE with GAC correction were close to those of GNSS. The research results can help to explore the motion mechanism between water migration and surface deformation, which is of benefit in the protection of the water ecological environment in the region

Combining metaphase cytogenetics with single nucleotide polymorphism arrays can improve the diagnostic yield and identify prognosis more precisely in myelodysplastic syndromes

Myelodysplastic syndromes (MDS) encompass a group of heterogeneous haematopoietic stem cell malignancies characterised by ineffective haematopoiesis, cytological aberrations, and a propensity for progression to acute myeloid leukaemia. Diagnosis and disease prognostic stratification are much based on genomic abnormalities. The traditional metaphase cytogenetics analysis (MC) can detect about 40–60% aberrations. Single-nucleotide polymorphism arrays (SNP-A) karyotyping can detect copy number variations with a higher resolution and has a unique advantage in detection of copy number neutral loss of heterozygosity (CN-LOH). Combining these two methods may improve the diagnostic efficiency and accuracy for MDS. We retrospectively analysed the data of 110 MDS patients diagnosed from January 2012 to December 2019 to compare the detection yield of chromosomal abnormalities by MC with by SNP-A, and the relationship between chromosomal abnormalities and prognosis. Our results showed that SNP-A improved the detection yield of chromosomal aberrations compared with MC (74.5 vs. 55.5%, p 65 years, bone marrow blasts ≥5%, with acquired CN-LOH, new aberrations detected by SNP-A, TGA value > the median (81.435 Mb), higher risk by IPSS-R-MC, higher risk by IPSS-R-SNP-A all had poorer prognosis. More critically, multivariable analysis showed that age >65 years and higher risk by IPSS-R-SNP-A were independent predictors of inferior OS in MDS patients. The combination of MC and SNP-A based karyotyping can further improve the diagnostic yield and provide more precise prognostic stratification in MDS patients. However, SNP-A may not completely replace MC because of its inability to detect balanced translocation and to detect different clones. From a practical point of view, we recommend the concurrent use of SNP-A and MC in the initial karyotypic evaluation for MDS patients on diagnosis and prognosis stratification.KEY MESSAGESSNP-A based karyotyping can further improve the MDS diagnostic yield and provide more precise prognostic stratification in MDS patients.Acquired CN-LOH is a characteristic chromosomal aberration of MDS, which should be integrated to the diagnostic project of MDS.The concurrent use of SNP-A and MC in the initial karyotypic evaluation for MDS patients can be recommended. SNP-A based karyotyping can further improve the MDS diagnostic yield and provide more precise prognostic stratification in MDS patients. Acquired CN-LOH is a characteristic chromosomal aberration of MDS, which should be integrated to the diagnostic project of MDS. The concurrent use of SNP-A and MC in the initial karyotypic evaluation for MDS patients can be recommended.</p

Presentation_1_Gossypol Inhibits Non-small Cell Lung Cancer Cells Proliferation by Targeting EGFRL858R/T790M.pdf

<p>Background: Overexpression of epidermal growth factor receptor (EGFR) has been reported to be implicated in the pathogenesis of non-small cell lung cancer (NSCLC). Several EGFR inhibitors have been used in clinical treatment of NSCLC, but the emergence of EGFR^L858R/T790M resistant mutation has reduced the efficacy of the clinical used EGFR inhibitors. There is an urgent need to develop novel EGFR^L858R/T790M inhibitors for better NSCLC treatment.</p><p>Methods: By screening a natural product library, we have identified gossypol as a novel potent inhibitor targeting EGFR^L858R/T790M. The activity of gossypol on NSCLC cells was evaluated by cell proliferation, cell apoptosis and cell migration assays. Kinase activity inhibition assay and molecular docking were used to study the inhibition mechanism of gossypol to EGFR^L858R/T790M. Western blotting was performed to study the molecular mechanism of gossypol inhibiting the downstream pathways of EGFR.</p><p>Results: Gossypol inhibited the cell proliferation and cell migration of NSCLC cells, and induced caspase-dependent cell apoptosis of NSCLC cells by upregulating the expression of pro-apoptotic protein BAD. Molecular docking revealed that gossypol could bind to the kinase domain of EGFR^L858R/T790M with good binding affinity through hydrogen bonds and hydrophobic interactions. Gossypol inhibited the kinase activity of EGFR^L858R/T790M with EC₅₀ of 150.1 nM. Western blotting analysis demonstrated that gossypol inhibited the phosphorylation of EGFR and its downstream signal pathways in a dose-dependent manner.</p><p>Conclusion: Gossypol inhibited cell proliferation and induced apoptosis of NSCLC cells by targeting EGFR^L858R/T790M. Our findings provided a basis for developing novel EGFR^L858R/T790M inhibitors for treatment of NSCLC.</p