A composite immune signature parallels disease progression across T1D subjects

At diagnosis, most people with type 1 diabetes (T1D) produce measurable levels of endogenous insulin, but the rate at which insulin secretion declines is heterogeneous. To explain this heterogeneity, we sought to identify a composite signature predictive of insulin secretion, using a collaborative assay evaluation and analysis pipeline that incorporated multiple cellular and serum measures reflecting beta cell health and immune system activity. The ability to predict decline in insulin secretion would be useful for patient stratification for clinical trial enrollment or therapeutic selection. Analytes from 12 qualified assays were measured in shared samples from subjects newly diagnosed with T1D. We developed a computational tool to identify a composite panel associated with decline in insulin secretion over 2 years after diagnosis. The tool employs multiple filtering steps to reduce data dimensionality, incorporates error-estimation techniques including cross-validation and sensitivity analysis, and is flexible to assay type, clinical outcome and disease setting. Using this novel analytical tool, we identified a panel of immune markers that, in combination, are highly associated with loss of insulin secretion. The methods used here represent a novel process for identifying combined immune signatures that predict outcomes relevant for complex and heterogeneous diseases like T1D

ZEB1 Is a Transcription Factor That Is Prognostic and Predictive in Diffuse Gliomas

Objective: To address the unmet medical need to better prognosticate patients with diffuse gliomas and to predict responses to chemotherapy regimens.Methods: ZEB1 alterations were retrospectively identified from a cohort of 1,160 diffuse glioma patients. Epigenome-wide association scans (EWAS) were performed on available data. We determined the utility of ZEB1 as a prognostic indicator of patient survival in diffuse gliomas and assessed the value of ZEB1 to predict the efficacy of treating diffuse glioma patients with procarbazine, CCNU, and vincristine along with radiation at diagnosis. Decision curve analysis (DCA) was used to determine if ZEB1 added benefit to clinical decision-making over and above conventional methods.Results: Fifteen percent of diffuse glioma patients had a ZEB1 deletion. ZEB1 deletion was associated with poor overall survival (OS) with and without adjustment for age and tumor grade (adjusted HR: 4.25; 95% CI: 2.35 to 7.66; P < 0.001). Decision curve analysis confirmed that ZEB1 status with or without IDH1 was more beneficial to clinical decision making than conventional information such as age and tumor grade. We showed that ZEB1 regulates TERT expression, and patients with ZEB1 deletions likely subsume patients with mutant TERT expression in diffuse gliomas. ZEB1 influenced clinical decision making to initiate procarbazine, CCNU, and vincristine treatment.Conclusion: We demonstrate the prognostic value of ZEB1 in diffuse glioma patients. We further determine ZEB1 to be a vital and influential molecular marker in clinical decisions that exceed conventional methods regarding whether to treat or not treat patients with diffuse glioma

Expression Signatures of Metastatic Capacity in a Genetic Mouse Model of Lung Adenocarcinoma

Background: Non-small cell lung cancer (NSCLC) is the foremost cause of cancer-related death in Western countries, which is due partly to the propensity of NSCLC cells to metastasize. The biologic basis for NSCLC metastasis is not well understood. Methodology/Principal Findings: Here we addressed this deficiency by transcriptionally profiling tumors from a genetic mouse model of human lung adenocarcinoma that develops metastatic disease owing to the expression of K-rasG12D and p53R172H. We identified 2,209 genes that were differentially expressed in distant metastases relative to matched lung tumors. Mining of publicly available data bases revealed this expression signature in a subset of NSCLC patients who had a poorer prognosis than those without the signature. Conclusions/Significance: These findings provide evidence that K-rasG12D; p53R172H mice recapitulate features of human NSCLC metastasis and will provide a useful platform on which to study the biologic basis for lung adenocarcinom

Effect of methionine sulfoximine on methylation of guanine residues in astroglial transfer ribonucleic acids

Culture-grown astrocytes derived from 3-day-old rat brain were incubated in the presence of [ 3 H]guanosine and of the convulsant agent l -methionine- dl -sulfoximine (MSO). The resulting [ 3 H]tRNA was purified from control and MSO-exposed cells at several time points during the incubation and was hydrolyzed to [ 3 H]guanine and four [ 3 H]methyl guanines which were separated by high pressure liquid chromatography. Three of the four [ 3 H]methyl guanines were more highly labeled in the [ 3 H]tRNA of the MSO-exposed cells, relative to that of the control cells throughout the entire incubation period. The findings extend to cultured astrocytes, the stimulatory effect of MSO on the methylation of neural tRNA guanines, previouly observed both in vitro using [ 14 C] S -adenosyl- l -methionine and in vivo using [ methyl 3 -H] l -methionine.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45427/1/11064_2004_Article_BF00964832.pd

Late onset of necrotizing enterocolitis in the full-term infant is associated with increased mortality: Results from a two-center analysis

Purpose: The effect of timing of onset of necrotizing enterocolitis (NEC) on outcomes has not been determined for the full-term infant. In this study we aimed to characterize the full-term NEC population and to evaluate onset of NEC. Methods: We performed a two-center retrospective review of all full-term infants (≥ 37 weeks) with a diagnosis of NEC between 1990 and 2012. Patients were identified by ICD-9 and age. Early onset for NEC was ≤ 7 days and late onset after 7 days of life. Demographics, comorbidities, maternal factors, clinical factors, surgical intervention, complications, and mortality were evaluated. Wilcoxon's test was performed on continuous variables and Fisher's exact test on categorical data. A p-value b 0.05 was considered significant. Univariate outcomes with a p-value b 0.1 were selected for multivariable analysis. Results: Thirty-nine patients (24 boys, 15 girls) with median EGA of 39 weeks were identified. Overall mortality was 18%. Univariate predictors of mortality included congenital heart disease and placement of an umbilical artery (UA) catheter. Multivariate analysis revealed late onset of NEC to be an independent predictor of mortality (OR 90.8, 95% CI 2.6-3121). Conclusion: Full-term infants who develop NEC after 7 days of life, have congenital heart disease, and/or need UA catheterization have increased mortality. © 2014 Elsevier Inc. All rights reserved. The pathogenesis of necrotizing enterocolitis (NEC) in infants remains incompletely understood. Most commonly, NEC occurs in the premature infant, with less than 10% occurring in full-term neonates It is not known whether the underlying pathophysiology of NEC in the term infant is distinctly different from that in the premature infant. In addition to the frequent presence of significant co-morbidities, term infants are frequently reported to present with NEC earlier in postnatal life Methods Patient Population We performed a two-center retrospective review of all full-term infants (≥ 37 weeks) with a diagnosis of NEC between 1990 and 2012. Charts were identified by including ICD-9 codes (777.5-777.6) and were reviewed to verify diagnosis of NEC and gestational age. Term infants determined to have Bell's stage 2 or 3 NEC were included. Term infants with possible Bell's stage I NEC were excluded from analysis. Infants who had an estimated gestational age less than 37 weeks were also excluded from the study. The subject met inclusion criteria for NEC if he/she had evidence of Bell's stage II or greater and had diagnosis of NEC documented in the chart with signs and symptoms that included temperature instability, apnea, bradycardia, lethargy, pneumatosis, metabolic acidosis, peritonitis, and/or pneumoperitoneum Similar to a previous publication, early-onset NEC was defined as development of NEC during the 1st week of life (≤ 7 days) and lateonset NEC as any time after day 7 of lif

A Comparison of Ku0063794, a Dual mTORC1 and mTORC2 Inhibitor, and Temsirolimus in Preclinical Renal Cell Carcinoma Models

<div><p>Rapamycin analogs, temsirolimus and everolimus, are approved for the treatment of advance renal cell carcinoma (RCC). Currently approved agents inhibit mechanistic target of rapamycin (mTOR) complex 1 (mTORC1). However, the mTOR kinase exists in two distinct multiprotein complexes, mTORC1 and mTORC2, and both complexes may be critical regulators of cell metabolism, growth and proliferation. Furthermore, it has been proposed that drug resistance develops due to compensatory activation of mTORC2 signaling during treatment with temsirolimus or everolimus. We evaluated Ku0063794, which is a small molecule that inhibits both mTOR complexes. Ku0063794 was compared to temsirolimus in preclinical models for renal cell carcinoma. Ku0063794 was effective in inhibiting the phosphorylation of signaling proteins downstream of both mTORC1 and mTORC2, including p70 S6K, 4E-BP1 and Akt. Ku0063794 was more effective than temsirolimus in decreasing the viability and growth of RCC cell lines, Caki-1 and 786-O, <em>in vitro</em> by inducing cell cycle arrest and autophagy, but not apoptosis. However, in a xenograft model there was no difference in the inhibition of tumor growth by Ku0063794 or temsirolimus. A potential explanation is that temsirolimus has additional effects on the tumor microenvironment. Consistent with this possibility, temsirolimus, but not Ku0063794, decreased tumor angiogenesis <em>in vivo,</em> and decreased the viability of HUVEC (Human Umbilical Vein Endothelial Cells) cells <em>in vitro</em> at pharmacologically relevant concentrations. Furthermore, expression levels of VEGF and PDGF were lower in Caki-1 and 786-O cells treated with temsirolimus than cells treated with Ku0063794.</p> </div

Ku0063794 and temsirolimus induced G1 cell cycle arrest and autophagy in RCC cells.

<p>Caki-1 (<b>A</b>) and 786-O (<b>B</b>) cells were plated and treated with 2 µM Ku0063794, 300 nM temsirolimus or DMSO (vehicle control) for 72 hours. The treated cells were subjected to cell cycle analysis (<b>left</b>). Cells were trypsinized and live cells were counted (<b>right</b>). The percent of cells at each cell cycle and the standard error of the mean are provided. *p<0.01 comparing either Ku0063794 or temsirolimus treatment to either DMSO treatment or untreated control. #p<0.01 comparing Ku0063794 to temsirolimus. Ku0063794 and temsirolimus induced autophagy in RCC cells as indicated by the increase in LC3-2/LC3-1 ratio. (<b>C</b>). Caki-1 (<b>left</b>) and 786-O (<b>right</b>) cells were treated with Ku0063794 or temsirolimus at the indicated concentration, or DMSO (vehicle control) for 24 hours with/without pepstatin A and E-64d. The figure is representative of triplicate experiments. (<b>D</b>) Ku0063794 and temsirolimus failed to induce apoptosis in RCC cells. Annexin-V and propidium iodide staining was performed following Ku0063794 or temsirolimus treatment at the indicated concentrations for 24 hours. As a positive control, 786-O cells were treated with 20 mM H₂O₂ for 30 minutes. Error bars indicate the standard error of the mean for triplicate experiments.</p

Ku0063794 and temsirolimus inhibited tumor growth in a xenograft model of RCC.

<p>(<b>A</b>) The treatment with Ku0063794 or temsirolimus significantly inhibited tumor growth in Nu/Nu nude mice. Following subcutaneous injection of 786-O cells, mice (5 mice in each group) were treated from days 33–78. The error bars indicate the standard error of the mean. *p<0.05 when the Ku0063794 or temsirolimus group was compared with control. (<b>B</b>) 90 minutes after drug administration, both Ku0063794 and temsirolimus inhibited the mTORC1 pathway <i>in vivo</i> as indicated by decrease in S6P phosphorylation in the tumor tissues, while only Ku0063794 inhibited the mTORC2 pathway <i>in vivo</i> as indicated by decrease in Akt phosphorylation on Ser473. Each lane represents a tumor from a different mouse.</p