Target selection for T-cell therapy in epithelial ovarian cancer: systematic prioritization of self-antigens

Adoptive T cell-receptor therapy (ACT) could represent a promising approach in the targeted treatment of epithelial ovarian cancer (EOC). However, the identification of suitable tumor-associated antigens (TAAs) as targets is challenging. We identified and prioritized TAAs for ACT and other immunotherapeutic interventions in EOC. A comprehensive list of pre-described TAAs was created and candidates were prioritized, using predefined weighted criteria. Highly ranked TAAs were immunohistochemically stained in a tissue microarray of 58 EOC samples to identify associations of TAA expression with grade, stage, response to platinum, and prognosis. Preselection based on expression data resulted in 38 TAAs, which were prioritized. Along with already published Cyclin A1, the TAAs KIF20A, CT45, and LY6K emerged as most promising targets, with high expression in EOC samples and several identified peptides in ligandome analysis. Expression of these TAAs showed prognostic relevance independent of molecular subtypes. By using a systematic vetting algorithm, we identified KIF20A, CT45, and LY6K to be promising candidates for immunotherapy in EOC. Results are supported by IHC and HLA-ligandome data. The described method might be helpful for the prioritization of TAAs in other tumor entities

Solution Studies of βCyclodextrin-Pyrene Complexes under Reversed-Phase Liquid Chromatographic Conditions: Effect of Alcohols as Mobile-Phase Comodifiers

Studies of pyrene complexes with β-cyclodextrln, using reversed-phase (C18) liquid chromatography require a relatively more nonpolar mobile phase than water (mixtures of methanol-water \u3e55% methanol) In order to achieve a reasonable retention time. Although methanol has a very low association constant with β-cyclodextrin, It becomes strongly competitive at high concentrations, resulting In very weak Interaction between pyrene and cyclodextrin. The presence of tert-butyl alcohol or cyclopentanol hi the medium Increases the strength of the β-cyclodextrin-pyrene complex by various orders of magnitude due to the formation of a ternary complex. In the presence of these alcohols as mobile-phase comodifiers, the Interaction between β-cyclodextrln and pyrene becomes evident at methanol concentrations In the range of practical use for HPLC. © 1991, American Chemical Society. All rights reserved

Axon guidance factor Slit2 inhibits neural invasion and metastasis in pancreatic cancer

Pancreatic ductal adenocarcinoma (PDAC) metastasizes by neural, vascular and local invasion routes, which limit patient survival. In nerves and vessels, Slit2 and its Robo receptors constitute repellent guidance cues that also direct epithelial branching. Thus, the Slit2-Robo system may represent a key pinch point to regulate PDAC spread. In this study, we examined the hypothesis that escaping from repellent Slit2-Robo signaling is essential to enable PDAC cells to appropriate their local stromal infrastructure for dissemination. Through immunohistochemical analysis, we detected Slit2 receptors Robo1 and Robo4 on epithelia, nerves and vessels in healthy pancreas and PDAC specimens, respectively. Slit2 mRNA expression was reduced in PDAC compared to non-transformed pancreatic tissues and cell lines, suggesting a reduction in Slit2-Robo pathway activity in PDAC. In support of this interpretation, restoring the Slit2 expression in Slit2-deficient PDAC cells inhibited their bidirectional chemoattraction with neural cells, and more specifically impaired unidirectional PDAC cell navigation along outgrowing neurites in models of neural invasion. Restoring autocrine/paracrine Slit2 signaling was also sufficient to inhibit the directed motility of PDAC cells, but not their random movement. Conversely, RNAi-mediated silencing of Robo1 stimulated the motility of Slit2-competent PDAC cells. Furthermore, culture supernatants from Slit2-competent PDAC cells impaired migration of endothelial cells (HUVEC) whereas an N-terminal Slit2 cleavage fragment stimulated such migration. In vivo investigations of orthotopic pancreatic tumors with restored Slit2 expression demonstrated reduced invasion, metastasis and vascularization, with opposing effects produced by Robo1 silencing in tumor cells or sequestration of endogenous Slit2. Analysis of clinical specimens of PDAC showed that those with low Slit2-mRNA expression exhibited a higher incidence and a higher fraction of tumor-infiltrated lymph nodes. Taken together, our findings argue that disrupting Slit2-Robo signaling in PDAC may enhance metastasis and predispose PDAC cells to neural invasion

Increased Activity of the Immunoregulatory Enzyme Indoleamine-2,3-Dioxygenase (IDO) with Consecutive Tryptophan Depletion Predicts Death in Patients with Neuroendocrine Neoplasia.

BACKGROUND/AIMS Data from a considerable number of malignancies demonstrate that depletion of the essential amino acid tryptophan via induction of the immuno-regulatory enzyme Indoleamine-2,3-dioxygenase (IDO) serves as an important tumour escape strategy and is of prognostic importance. Here we investigate the predictive value of the activity of IDO as well as levels of tryptophan and respective downstream catabolites in a large cohort of patients with neuroendocrine neoplasia (NEN). METHODS 142 consecutive Caucasian patients (62 male, aged 60.3 ± 11.9 years) with histologically confirmed NEN were systematically analysed in a retrospective blinded endpoint analysis. Patients were followed up for a mean period of about 3.9 ± 1.9 years. Clinical outcome, levels of established biomarkers, and tryptophan degradation markers (assessed using tandem mass spectrometry) including estimated IDO-activity were recorded. Cox-proportional hazards regression models were performed for the assessment of prognostic power. RESULTS We found that baseline tryptophan levels were significantly lower and IDO-activity was significantly increased in non-survivors. The risk for death inclined stepwise and was highest in patients in the upper tertile of IDO-activity. Cox-proportional regression models identified IDO-activity as an independent predictor for death. CONCLUSIONS In this retrospective analysis, we observed that baseline activity of the immunoregulatory enzyme IDO was significantly increased in non-survivors. IDO-activity was identified as an independent predictor for death in this cohort of NEN patients. Whether IDO-activity or tryptophan depletion serves to guide future therapeutic interventions in NEN remains to be established

Measurement modelling and mapping of arsenic bioaccessibility in Northampton, United Kingdom

The human ingestion bioaccessibility of As was measured on 50 representative samples of soils selected from a 281-soil-sample geochemical survey of Northampton. The major and trace element content, pH and near infrared (NIR) spectra of the 281 soils were determined. A multiple linear regression (MLR) model using total As, major element composition and pH identified total As, pH and P to be the significant predictor variables for bioaccessible As (R2 = 0.72, median standard error of prediction = 1.5 mg kg−1 bioaccessible As). When spectral components (SC) derived from chemometric analysis of the NIR spectra were also included in the MLR, total As, pH, Mg and two NIR spectral components were found to be significant predictor variables (R2 = 0.84, median standard error of prediction = 1.2 mg kg−1 bioaccessible As). Correlation analysis of the SC with major element data suggested that the two NIR SC in the second model were related to different forms of Fe oxides in the soil. When plotted over a geological map of Northampton interpolated predictions of bioaccessible As showed clear geological control. The median total As concentration of the soils in Northampton was 30.2 mg kg−1 and the median bioaccessible As was 3.0 mg kg−1