Integrative analysis of spatial and single-cell transcriptome data from human pancreatic cancer reveals an intermediate cancer cell population associated with poor prognosis.

BACKGROUND: Recent studies using single-cell transcriptomic analysis have reported several distinct clusters of neoplastic epithelial cells and cancer-associated fibroblasts in the pancreatic cancer tumor microenvironment. However, their molecular characteristics and biological significance have not been clearly elucidated due to intra- and inter-tumoral heterogeneity. METHODS: We performed single-cell RNA sequencing using enriched non-immune cell populations from 17 pancreatic tumor tissues (16 pancreatic cancer and one high-grade dysplasia) and generated paired spatial transcriptomic data from seven patient samples. RESULTS: We identified five distinct functional subclusters of pancreatic cancer cells and six distinct cancer-associated fibroblast subclusters. We deeply profiled their characteristics, and we found that these subclusters successfully deconvoluted most of the features suggested in bulk transcriptome analysis of pancreatic cancer. Among those subclusters, we identified a novel cancer cell subcluster, Ep_VGLL1, showing intermediate characteristics between the extremities of basal-like and classical dichotomy, despite its prognostic value. Molecular features of Ep_VGLL1 suggest its transitional properties between basal-like and classical subtypes, which is supported by spatial transcriptomic data. CONCLUSIONS: This integrative analysis not only provides a comprehensive landscape of pancreatic cancer and fibroblast population, but also suggests a novel insight to the dynamic states of pancreatic cancer cells and unveils potential therapeutic targets.

Sequential ubiquitination of p53 by TRIM28, RLIM, and MDM2 in lung tumorigenesis.

Tripartite motif-containing 28 (TRIM28) is an E3 ubiquitin ligase harboring multiple cellular functions. We found that the TRIM28 protein is frequently overexpressed in patients with lung cancer. The stable overexpression of TRIM28 in lung cancer cells and xenograft models significantly increased the proliferation, migration, and invasiveness, whereas knockdown of TRIM28 had the opposite effect. We further observed that TRIM28 regulates the ubiquitin ligases RLIM and MDM2 to target the p53 levels during lung tumorigenesis. These data provide new insights into lung cancer development and potential new therapeutic targets for this disease.

CGP74514A enhances TRAIL-induced apoptosis in breast cancer cells by reducing X-linked inhibitor of apoptosis protein.

BACKGROUND: Despite the selectivity of Tumor necrosis factor Related Apoptosis-Inducing Ligand (TRAIL) for cancer cell killing activity, breast cancer cells are resistant to TRAIL-induced apoptosis for various reasons. MATERIALS AND METHODS: From a functionally-characterized small-molecule dataset, CGP74514A was identified as a TRAIL sensitizer in MCF-7 breast cancer cells. Combination of sub-toxic dose of TRAIL with CGP74514A was evaluated in three TRAIL-resistant breast cancer cells, MCF-7, T47D and SK-BR-3. RESULTS: In all tested cells, CGP74514A enhanced TRAIL sensitivity. Combination treatment triggered apoptotic events faster than single treatment. Regarding its mechanism of action, CGP74514A reduced cytosolic X-linked inhibitor of apoptosis protein (XIAP). Small interfering RNA-mediated knockdown experiments showed that reduction of XIAP is the reason of sensitization. CONCLUSION: CGP74514A sensitized breast cancer cells to TRAIL via reduction of XIAP expression level.

Cryptotanshinone induces G1 cell cycle arrest and autophagic cell death by activating the AMP-activated protein kinase signal pathway in HepG2 hepatoma.

AMP-activated protein kinase (AMPK) performs a pivotal function in energy homeostasis via the monitoring of intracellular energy status. Once activated under the various metabolic stress conditions, AMPK regulates a multitude of metabolic pathways to balance cellular energy. In addition, AMPK also induces cell cycle arrest or apoptosis through several tumor suppressors including LKB1, TSC2, and p53. LKB1 is a direct upstream kinase of AMPK, while TSC2 and p53 are direct substrates of AMPK. Therefore, it is expected that activators of AMPK signal pathway might be useful for treatment or prevention of cancer. In the present study, we report that cryptotanshinone, a natural compound isolated from Salvia miltiorrhiza, robustly activated AMPK signaling pathway, including LKB1, p53, TSC2, thereby leading to suppression of mTORC1 in a number of LKB1-expressing cancer cells including HepG2 human hepatoma, but not in LKB1-deficient cancer cells. Cryptotanshinone induced HepG2 cell cycle arrest at the G1 phase in an AMPK-dependent manner, and a portion of cells underwent apoptosis as a result of long-term treatment. It also induced autophagic HepG2 cell death in an AMPK-dependent manner. Cryptotanshinone significantly attenuated tumor growth in an HCT116 cancer xenograft in vivo model, with a substantial activation of AMPK signal pathways. Collectively, we demonstrate for the first time that cryptotanshinone harbors the therapeutic potential for the treatment of cancer through AMPK activation.

Novel method of producing nanoparticles for gadolinium-scintillator-based digital radiography.

Radiation image sensor properties affect the dose of radiation that patients are exposed to in a clinical setting. Numerous radiation imaging systems use scintillators as materials that absorb radiation. Rare-earth scintillators produced from elements such as gadolinium, yttrium, lutetium, and lanthanum have been investigated to improve the properties of radiation imaging systems. Although such rare-earth scintillators are manufactured with a bulk structure, they exhibit low resolution and low efficiency when they are used as conversion devices. Nanoscintillators have been proposed and researched as a possible solution to these problems. According to the research, the optical properties and size of fine scintillators are affected by the sintering temperature used to produce nanoscintillators instead of the existing bulk-structured scintillators. Therefore, the main purpose of this research is to develop radiation-imaging sensors based on nanoscintillators in order to evaluate the quantitative properties of various scintillators produced under various conditions such as sintering temperature. This is accomplished by measuring acquired phantom images, and modulation transfer functions (MTFs) for complementary-symmetry metal-oxide-semiconductor (CMOS) image sensors under the same X-ray conditions. Low-temperature solution combustion was used to produce fine scintillators consisting of 5 wt% of europium as an activator dopant in a Gd2O3 scintillator host. Variations in the characteristics of the fine scintillators were investigated. The characteristics of fine scintillators produced at various sintering temperatures (i.e., 600, 800, or 1000 degrees C) and with a europium concentration of 0.5 wt% were also analyzed to determine the optimal conditions for synthesizing the fine scintillators.

Optimization of nano-phosphor synthesis by including sensitizer doping for medical X-ray imaging.

Medical radiation imaging systems employ phosphors such as CaWO4 as X-ray receptor materials. Unfortunately, the conversion efficiencies of these materials are rather low (approx. 5%). Alternatives that comprise a bulk structure have been fabricated from rare earth metals, but they are not efficient enough to produce high quality images. Nano-phosphors do not suffer from the limitations inherent to the bulk structures of conventional phosphors. We examined the effects of sensitizer doping conditions on the optical characteristics and morphology of the rare earth phosphor Gd2O3:Eu to fabricate a novel type of nano-phosphor. We optimized a temperature solution-combustion procedure for producing phosphors doped with 5 wt% Eu. Scanning electron microscopy images showed that the phosphors were 20-30 nm in diameter and X-ray diffraction analysis revealed that they underwent polycrystalline growth upon the addition of a sensitizer, similar to the polycrystalline growth of bulk phosphors. In addition, the phosphors exhibited a strong peak at 613 nm and luminescence similar to conventional phosphors. Phosphors that were produced using citric acid as a sensitizer showed more than double the level of luminescence and could be used to produce higher quality images compared to non-sensitized phosphors. The phosphors also exhibited a high degree of luminescence stability.

Development of quantification software using model-based segmentation of left ventricular myocardium in gated myocardial SPECT.

Gated myocardial single photon emission computed tomography (SPECT) is being used for the diagnosis of coronary artery diseases. In this study, we developed new software for the quantification of volumes and ejection fraction (EF) on the gated myocardial SPECT data using a cylindrical model. Volumes and EF by developed software were validated by comparing with those quantified by quantitative gated SPECT (QGS) software. Cylinder model for left ventricular myocardium was used to eliminate background activity and count profiles across the myocardium were fitted to the Gaussian curve to determine the endocardial and epicardial boundary. End-diastolic volume (EDV), end-systolic volume (ESV) and EF were calculated using this boundary information. Gated myocardial SPECT was performed in 83 patients. EDV, ESV and EF values estimated using present method were compared to those obtained using the commercialized software QGS, and reproducibility in the parameter estimation was assessed. EF, EDV and ESV obtained using two methods were correlated well (correlation coefficients = 0.96, 0.96 and 0.98). The correlation between the parameters repetitively estimated from the same data set by an operator was very high (correlation coefficients = 0.96, 0.99 and 0.99 for EF, EDV and ESV). On the repeated acquisition, reproducibility was also high with correlation coefficients of 0.89, 0.97 and 0.98. The present software will be useful for the development of new parameters for describing the perfusion and function of the LV.

Multi-resolution wavelet-transformed image analysis of histological sections of breast carcinomas.

Multi-resolution images of histological sections of breast cancer tissue were analyzed using texture features of Haar- and Daubechies transform wavelets. Tissue samples analyzed were from ductal regions of the breast and included benign ductal hyperplasia, ductal carcinoma in situ (DCIS), and invasive ductal carcinoma (CA). To assess the correlation between computerized image analysis and visual analysis by a pathologist, we created a two-step classification system based on feature extraction and classification. In the feature extraction step, we extracted texture features from wavelet-transformed images at 10x magnification. In the classification step, we applied two types of classifiers to the extracted features, namely a statistics-based multivariate (discriminant) analysis and a neural network. Using features from second-level Haar transform wavelet images in combination with discriminant analysis, we obtained classification accuracies of 96.67 and 87.78% for the training and testing set (90 images each), respectively. We conclude that the best classifier of carcinomas in histological sections of breast tissue are the texture features from the second-level Haar transform wavelet images used in a discriminant function.

Effect of monitor luminance and ambient light on observer performance in soft-copy reading of digital chest radiographs.

PURPOSE: To examine the combined effects of monitor luminance and ambient light on observer performance for detecting abnormalities in a soft-copy interpretation of digital chest radiographs. MATERIALS AND METHODS: A total of 254 digital chest radiographs were displayed on a high-resolution cathode ray tube monitor at three luminance levels (25, 50, and 100 foot-lamberts) under three ambient light levels (0, 50, and 460 lux). Six chest radiologists reviewed each image in nine modes of combined luminance and ambient light. The observers were allowed to adjust the window width and level of the soft-copy images. The abnormalities included nodule, pneumothorax, and interstitial disease. Observer performance was analyzed in terms of the receiver operating characteristics. The observers reported their subjective level of visual fatigue with each viewing mode. A statistical test was conducted for each of the abnormalities and for fatigue score by using repeated-measures two-way analysis of variance with an interaction. RESULTS: The detection of nodules was the only reading that was affected by the ambient light with a statistically significant difference (P <.05). Otherwise, observer performance for detecting a nodule, pneumothorax, and interstitial disease was not significantly different in the nine-mode comparison. There was no evidence that the luminance of the monitors was related to the ambient light for any of the abnormalities. The fatigue score showed a statistically significant difference due to both the luminance and ambient light. CONCLUSION: When adequate window width and level are applied to soft-copy images, the primary diagnosis with chest radiographs on the monitor is unlikely to be affected under low ambient light and a monitor luminance of 25 foot-lamberts or more.

Ex vivo proton MR spectroscopy (1H-MRS) for evaluation of human gastric carcinoma.

The present study was performed to determine the characteristics of the biochemical metabolites related to gastric cancer using ex vivo (1)H magnetic resonance spectroscopy (MRS), and to assess the clinical usefulness. A total of 35 gastric specimens resected during surgery for gastric cancer were used to compare MR spectra. A 1.5-T (64-MHz) clinical MR imager equipped with facilities for spectroscopy was used to obtain MR spectra from 33 gastric specimens. High-resolution (1)H nuclear magnetic resonance (NMR) spectra of the remains of two specimens were also examined with a 9.4-T (400-MHz) NMR spectrometer. Localized spectroscopic measurements were performed in two layers of gastric tissue, the proper muscle layer and the composite mucosa/submucosa layer. T(2) FSE and 3D SPGR images were used to determine the voxel size and the location for MRS data collection. MR spectra were obtained using the single-voxel PRESS technique with parameters of TR/TE = 2000/30 ms, NA = 256, and voxel size = 3 x 3 x 3 mm(3) (27 microL). Cancerous and noncancerous gastric tissues in the voxel were determined by histopathological analysis. On 9.4-T ex vivo NMR spectroscopy, the following metabolite peaks were found: lipids at 0.9 ppm (CH(3)) and 1.3 ppm (CH(2)); alanine (beta-CH(3)) at 1.58 ppm; N-Acetyl neuraminic acid (NANA: sialic acid) at 2.03 ppm; and glutathione at 2.25 ppm in normal gastric tissue layers. In the 1.5-T MR system, broad and featureless spectral peaks of the various metabolites in normal human gastric tissue were observed at 0.9 ppm, 1.3 ppm, 2.0 ppm, and 2.2 ppm regardless of gastric tissue layer. In specimens (Borrmann type III) with tubular adenocarcinoma, resonance peaks were observed at 1.26 ppm, 1.36 ppm (doublet of lactate), and 3.22 ppm (choline). Cancer lesions showed decreased levels of lipid peaks, showing the significant lactate doublet peaks, and increased intensity of the choline peak as compared with noncancerous gastric tissue. We found that decreased levels of lipids and increases in lactate and choline peaks in gastric tissue were markers for malignancy in gastric lesions. Information provided by ex vivo (1)H MRS, together with the development of in vivo (1)H MRS with high field strength and high resolution, may be very useful for the diagnosis of gastric cancer in clinical situation.

Evaluation of automated and semi-automated skull-stripping algorithms using similarity index and segmentation error.

The skull-stripping in the MR brain image appears to be a key issue in neuroimage analysis. In this paper, we evaluated the accuracy and efficiency of both automated and semi-automated skull-stripping methods. The evaluation was performed on both simulated and real data with the ground truth in skull-stripping. Although automated method showed better efficient results, it should require additional intervention. In contrast to that, semi-automated method showed better accurate results, but it was time consuming and prone to operator bias. Therefore, it might be practical that the semi-automated method was used as the post-processing of the automated one.