Targeting USP47 overcomes tyrosine kinase inhibitor resistance and eradicates leukemia stem/progenitor cells in chronic myelogenous leukemia.

Identifying novel drug targets to overcome resistance to tyrosine kinase inhibitors (TKIs) and eradicating leukemia stem/progenitor cells are required for the treatment of chronic myelogenous leukemia (CML). Here, we show that ubiquitin-specific peptidase 47 (USP47) is a potential target to overcome TKI resistance. Functional analysis shows that USP47 knockdown represses proliferation of CML cells sensitive or resistant to imatinib in vitro and in vivo. The knockout of Usp47 significantly inhibits BCR-ABL and BCR-ABLT315I-induced CML in mice with the reduction of Lin-Sca1+c-Kit+ CML stem/progenitor cells. Mechanistic studies show that stabilizing Y-box binding protein 1 contributes to USP47-mediated DNA damage repair in CML cells. Inhibiting USP47 by P22077 exerts cytotoxicity to CML cells with or without TKI resistance in vitro and in vivo. Moreover, P22077 eliminates leukemia stem/progenitor cells in CML mice. Together, targeting USP47 is a promising strategy to overcome TKI resistance and eradicate leukemia stem/progenitor cells in CML.

Wu-5, a novel USP10 inhibitor, enhances crenolanib-induced FLT3-ITD-positive AML cell death via inhibiting FLT3 and AMPK pathways.

The kinase FLT3 internal tandem duplication (FLT3-ITD) is related to poor clinical outcomes of acute myeloid leukemia (AML). FLT3 inhibitors have provided novel strategies for the treatment of FLT3-ITD-positive AML. But they are limited by rapid development of acquired resistance and refractory in monotherapy. Recent evidence shows that inducing the degradation of FLT3-mutated protein is an attractive strategy for the treatment of FLT3-ITD-positive AML, especially those with FLT3 inhibitor resistance. In this study we identified Wu-5 as a novel USP10 inhibitor inducing the degradation of FLT3-mutated protein. We showed that Wu-5 selectively inhibited the viability of FLT3 inhibitor-sensitive (MV4-11, Molm13) and -resistant (MV4-11R) FLT3-ITD-positive AML cells with IC50 of 3.794, 5.056, and 8.386 µM, respectively. Wu-5 (1-10 µM) dose-dependently induced apoptosis of MV4-11, Molm13, and MV4-11R cells through the proteasome-mediated degradation of FLT3-ITD. We further demonstrated that Wu-5 directly interacted with and inactivated USP10, the deubiquitinase for FLT3-ITD in vitro (IC50 value = 8.3 µM) and in FLT3-ITD-positive AML cells. Overexpression of USP10 abrogated Wu-5-induced FLT3-ITD degradation and cell death. Also, the combined treatment of Wu-5 and crenolanib produced synergistic cell death in FLT3-ITD-positive cells via the reduction of both FLT3 and AMPKα proteins. In support of this, AMPKα inhibitor compound C synergistically enhanced the anti-leukemia effect of crenolanib, while AMPKα activator metformin inhibited the anti-leukemia effect of crenolanib. In summary, we demonstrate that Wu-5, a novel USP10 inhibitor, can overcome FLT3 inhibitor resistance and synergistically enhance the anti-AML effect of crenolanib through targeting FLT3 and AMPKα pathway.

Characterization of naturally occurring pentacyclic triterpenes as novel inhibitors of deubiquitinating protease USP7 with anticancer activity in vitro.

Deubiquitinating protease USP7 is a promising therapeutic target for cancer treatment, and interest in developing USP7 inhibitors has greatly increased. In the present study, we reported a series of natural pentacyclic triterpenes with USP7 inhibitory activity in vitro. Among them, both the ursane triterpenes and oleanane triterpenes were more active than the lupine triterpenes, whereas ursolic acid was the most potent with IC50 of 7.0±1.5 µmol/L. Molecular docking studies showed that ursolic acid might occupy the ubiquitin binding pocket of USP7, with the 17-carboxyl group and 3-hydroxyl group playing a vital role in the USP7-ursolic acid interaction. Using the cellular thermal shift assay, we demonstrated that ursolic acid interacted with USP7 in RPMI8226 human myeloma cells. Ursolic acid dose-dependently inhibited the proliferation of the myeloma cells with IC50 of 6.56 µmol/L, accompanied by reductions in USP7 substrates such as MDM2, UHRF1 and DNMT1. Overexpression of USP7 partially, but significantly attenuated ursolic acid-induced cell death as well as downregulation of MDM2, UHRF1 and DNMT1. In conclusion, we demonstrate for the first time that pentacyclic triterpenes represent a novel scaffold for developing USP7 inhibitors and that USP7 inhibition contributes to the anti-cancer effect of ursolic acid.

[Study on the prediction of soil heavy metal elements content based on mid-infrared diffuse reflectance spectra].

The present paper analyzed the possibility of mid-infrared diffuse reflectance spectra for quick assessment of heavy metal element content in soil quickly. Soil samples were collected from Jiangning District and Baguazhou Island, and the numbers of sample were 103 and 58 separately. Jiangning District samples were used as calibration set while Baguazhou Island samples as validation set. To assess the utility of different pre-treatment process of MIR spectroscopy for soil heavy metal element content analysis, we used PLSR method to develop the calibration between spectral data and soil elements content. Three spectral pretreating techniques such as smooth, log(1/N), baseline correction, multiplicative scatter correction were used for promotion of predicting performance. The result showed that the progress of (log-BC-MSC) in turn achieved optimal calibration of MIR spectra and better prediction for ex-situ soils. Though the calibration data were treated by different pre-treating schema, the R2 of the 8 elements followed the same law: Ni > 0.8 > Cr, Cu, Zn, Pb, Hg > 0.6 > As, Cd. When we applied these calibrations to Baguazhou Island soils, (log-BC-MSC) treated data results in the smallest RMSEp-BGZ. We used the same calibration method to compare the predictive ability of MIR spectra to VNIR spectra. The R2 of 8 elements developed by VNIR spectral calibration are sometime larger than MIR's, but after we applied these calibrations to validation set, the RSME of MIR data for prediction of BGZ soil samples is 21% to 73% of VNIR's. This result showed us that for predicting ex-situ soils, MIR analysis substantially outperformed VNIR These results indicated that MIR spectra can be used to predict soil heavy metal content quickly and non-destructively.

[Research on spectrum denoising methods based on the combination of wavelet package transformation and mathematical morphology].

The present study introduced the generalized morphological filter into the denoising of visible and near infrared spectra for the first time, and provided a new method for denoising the reflectance spectra by combining mathematical morphology methods with the wavelet packet transformation. The authors used vegetable spectra from USGS spectral library as the reference spectra, and obtained the noised spectra by adding noises with different signal-to-noise ratios to the referenced spectra. The results were evaluated by signal-to-noise ratio (SNR), root mean squared error (RMSE), normalized correlation coefficient (NCC) and smoothness ratio (SR) of the denoised spectra. The authors' results showed that both the thresholding on wavelet packet decomposition best bases method and the generalized morphological filter method could maintain the spectral shape and the spectral smoothness after denoising. The generalized morphological filter method can remove larger amplitude random noise whereas the continuous small amplitude random noise could not be removed well. Hence, the denoised spectra were not smooth. Nevertheless, the denoised spectra using the thresholding on the best base groups of wavelet packet decomposition method were smooth, but the larger amplitude noise could not be removed completely. The authors' method by combining the two methods has the merits of the two methods but removing their defects. The results showed that both large and small amplitude noise could be removed completely, meanwhile the normalized correlation coefficient (NCC) and smoothness ratio (SR) were improved, which indicated that the authors' method is superior to other methods in denoising visible and near infrared spectra.

[A study of environmental changes on new snail habitat though multi-temporal relative radiometric normalized TM images].

OBJECTIVE: To study the environmental changes of new snail habitual by using multi-temporal relative radiometric normalized TM images taken in Wuzhoutou of Nanchang, Jiangxi province. METHODS: TM images of Poyang Lake were collected on 3rd April 1998, 16th April 2000 and 5th May 2004. Taking images in 2004 as a reference, the TM images in 1998 and 2000 were radiometric normalized. The images were analyzed to trace the vegetation background and snail historical information. RESULTS: The vegetation coverage rate of Wuzhaoutou on April 3rd in 1998 was 4.76%, but the vegetation coverage rates of Wuzhoutou on April 16th in 2000 and May 5th in 2004 were above 80%; the NDVI values of Wuzhoutou on April 3rd in 1998 were around -0.4 to 0.2, but the NDVI values of Wuzhoutou on April 16th in 2000 and May 5th in 2004 were mainly around 0.2 to 0.6, which had statistical difference with that in 1998 (q = 162.285321, P < 0.01 and q = 161.669725, P < 0.01). The greenness of three images have statistical differences(q = 178.679245, P < 0.01; q = 130.320755, P < 0.01 and q = - 48.358491, P < 0.01). CONCLUSION: Combining with the snail survey, the TM images could be an effective measure for setting up models to forecast the areas where snails might live to control the Schistosomiasis epidemic.

Feasibility of reflectance spectroscopy for the assessment of soil mercury contamination.

Conventional methods for investigating soil Hg contamination based on raster sampling and chemical analysis are time-consuming and relatively expensive. The objective of this study was to develop a rapid method for investigating Hg concentration in suburban agricultural soils of the Nanjing region using reflectance spectra within the visible-near-infrared (VNIR) region. Several spectral pretreatments (absorbance, Kubelka-Munk transformations and their derivatives) were applied to the reflectance spectra to optimize the accuracy of prediction. The prediction of Hg concentration was achieved by univariate regression and principal component regression (PCR) approaches. The optimal model (R= 0.69, RMSEP = 0.15) for predicting Hg was achieved using the PCR method with the Kubelka-Munktransformation asthe spectral predictor. Comparison of three wavelength ranges (0.38-1.1, 1.0-2.5, and 0.38-2.5 microm) on the effect of prediction accuracy showed that the best results were acquired using the 1.0-2.5 microm spectral region. Correlation analysis revealed that Hg concentration was negatively correlated with soil reflectance while positively correlated with the absorption depths of goethite at 0.496 microm and clay minerals at 2.21 microm, suggesting that Hg-sorption by clay-size mineral assemblages in soils was the mechanism by which to predict spectrally featureless Hg. These results indicate that it is feasible to predict Hg levels in agricultural soils using the rapid and cost-effective reflectance spectroscopy. Future study with operational remote sensing techniques and field measurements is strongly recommended.