A review on the interactions between the tumour microenvironment and androgen receptor signaling in prostate cancer

Prostate cancer growth is controlled by androgen receptor signaling via both androgen-dependent and androgen-independent pathways. Furthermore, the prostate is an immune competent organ with inflammatory changes both within the systemic and local environment contributing to the reprogramming of the prostatic epithelium with consistently elevated lymphocyte infiltration and pro-inflammatory cytokines being found in prostate cancer. The crosstalk between the tumour microenvironment and androgen receptor signaling is complex with both pro-tumorigenic and anti-tumourigenic roles observed. However, despite an increase in immune checkpoint inhibitors and inflammatory signaling blockades available for a range of cancer types, we are yet to see substantial progress in the treatment of prostate cancer. Therefore, this review aims to summarize the tumour microenvironment and its impact on androgen receptor signaling in prostate cancer

Stuttering, alcohol consumption and smoking

Purpose: Limited research has been published regarding the association between stuttering and substance use. An earlier study provided no evidence for such an association, but the authors called for further research to be conducted using a community sample. The present study used data from a community sample to investigate whether an association between stuttering and alcohol consumption or regular smoking exists in late adolescence and adulthood. Methods: Regression analyses were carried out on data from a birth cohort study, the National Child Development Study (NCDS), whose initial cohort included 18,558 participants who have since been followed up until age 55. In the analyses, the main predictor variable was parent-reported stuttering at age 16. Parental socio-economic group, cohort member’s sex and childhood behavioural problems were also included. The outcome variables related to alcohol consumption and smoking habits at ages 16, 23, 33, 41, 46, 50 and 55. Results: No significant association was found between stuttering and alcohol consumption or stuttering and smoking at any of the ages. It was speculated that the absence of significant associations might be due to avoidance of social situations on the part of many of the participants who stutter, or adoption of alternative coping strategies. Conclusion: Because of the association between anxiety and substance use, individuals who stutter and are anxious might be found to drink or smoke excessively, but as a group, people who stutter are not more likely than those who do not to have high levels of consumption of alcohol or nicotine

Drug screening of biopsy-derived multicellular spheroids using microfluidic technology

Performing drug screening, of physiologically relevant three-dimensional (3D) tumor models, for personalized treatment remains challenging, due to the small amount of tissue available from most biopsies. New microfluidic technologies, enabling greater control over cell positioning and fluid behavior at the micro-scale, allow extensive testing of anticancer agents on human tumor tissue preparations in 3D and offer new solutions for the development of anticancer compounds and personalized medicine. We have developed a microfluidic platform for extensive drug screening of tumor biopsies in a cost-effective manner and validated the system with tumor prostate patient samples. As a typical drug screening assay, up to 22 drug concentration-response curves could be generated from a single biopsy, within a time frame of up to 4 weeks. Biopsy tissue, grown as a heterogeneous co-culture from the primary sample, was prepared as cancer-cell enriched multicellular spheroids, cultured for 3 to 5 days prior to the application of a panel of standard-of-care drugs for prostate cancer. Readouts were obtained via bright-field and epifluorescence microscopy. The microfluidic platform was designed to be operated entirely without the need of external fluid actuation, with the microfluidic network capable of generating long-lasting, stable and repeatable drug concentration gradients across arrays of 240 spheroids. Outcomes were generated as 8-point drug concentration response curves per device, with each drug concentration tested on at least 24 spheroids. In-house developed software was used to analyze bright-field and fluorescent images to provide readouts of spheroid growth and viability, as well as information on drug penetration and drug efficacy over time. Following platform and assay validation using cancer cell lines, proof-of-concept screening was performed on prostate biopsies from 2 different patients. Results showed that biopsy-derived spheroids were more resistant to treatment than LNCaP spheroids, a prostate cancer cell line. For one biopsy, spheroids were sensitive to docetaxel, but resistant to enzalutamide, despite the presence of intact androgen receptors. This preliminary data outlines how this technology could become a useful tool to investigate patient-specific drug resistance and to test novel anticancer agents in a cost-effective manner, based on maximized screening of human tumor tissue in a 3D format

Inflammatory infiltration is associated with AR expression and poor prognosis in hormone naïve prostate cancer

Background: Tumor microenvironment inflammatory infiltration is proposed as a protumorigenic mechanism for prostate cancer with proinflammatory cytokines stimulating androgen receptor (AR) activity. However, association with patient prognosis remains unclear. This study derives an inflammatory gene signature associated with AR expression and investigates CD3+ and CD8+ T‐lymphocyte infiltration association with AR and prognosis. Methods: Gene profiling of inflammatory related genes was performed on 71 prostate biopsies. Immunohistochemistry on 243 hormone‐naïve prostate cancers was performed for CD3, CD8, AR, and phosphorylated AR tumor expression. Results: Multiple proinflammatory genes were differentially expressed in association with high AR expression compared with low AR expression including PI3KCA and MAKP8 (adjusted P < .05). High CD3+ and high CD8+ infiltration associated with reduced cancer‐specific survival (P = .018 and P = .020, respectively). High CD3+ infiltration correlated with high tumor cytoplasmic AR expression and if assessed together, they associated with reduced cancer‐specific and 5‐year survival from 90% to 56% (P = .000179). High CD8+ cytotoxic infiltration associated with high androgen‐independent tumor nuclear AR serine 213 phosphorylation (correlation coefficient = 0.227; P = .003) and when assessed together associated with poor clinico‐pathological features including perineural invasion (P = .001). Multiple genes involved in proinflammatory signaling pathways are upregulated in high AR expressing prostate samples. Conclusion: T‐lymphocyte infiltration in hormone‐naïve disease associates with androgen‐independent driven disease and provides possible therapeutic targets to reduce transformation from hormone‐naïve to castrate‐resistant disease

Androgen receptor phosphorylation at serine 81 and serine 213 in castrate-resistant prostate cancer

Background: Despite increases in diagnostics and effective treatments, over 300,000 men die from prostate cancer highlighting the need for specific and differentiating biomarkers. AR phosphorylation associates with castrate-resistance, with pARser213 promoting transcriptional activity. We hypothesise that combined pARser81 and pARser213 reduces survival and would benefit from dual-targeting androgen-dependent and Akt-driven disease. Methods: Immunohistochemistry and immunofluorescence were performed on matched hormone-naive and castrate-resistant prostate cancer samples. TempO-Seq gene profiling was analysed using DESeq2 package. LNCaP-AI cells were stimulated with DHT or EGF. WST-1 assays were performed to determine effects of Enzalutamide and BKM120 on cell viability. Results: Following the development of castrate-resistance, pARser81 expression reduced and pARser213 expression increased. Castrate-resistance pARser81 expression was not associated with survival but high pARser213 expression was associated with reduced survival from relapse. Combined high pARser81 and pARser213 was associated with reduced survival from relapse. pARser81 expression was induced by 10 nM DHT or 10 nM EGF and pARser213 expression was induced by treatment with 10 nM EGF in LNCaP-AI cells. Cell viability was reduced following treatment with 10 nM Enzalutamide and 10 nM BKM120. Eight genes were differentially expressed between hormone-naive and castrate-resistant tumours and twenty-five genes were differentially expressed between castrate-resistant tumours with high and low pARser213 expression. Conclusion: Combined pARser81 and pARser213 provides a novel prognostic biomarker for castrate-resistant disease and a potential predictive and therapeutic target for prostate cancer. Further studies will be required to investigate the combined effects of targeting AR and PI3K/AKT signalling

Efficacy and micro-characterization of pathophysiological events on caries-affected dentin treated with glass-ionomer cements

The aim of this study was to evaluate if mechanical cycling influences bioactivity and bond strength at the glass-ionomer cement-dentin interface, after load cycling. Microtensile bond strength (MTBS) was assessed with Ketac-Bond (conventional glass ionomer/GIC) or Vitrebond Plus (resin-modified/RMGIC), in sound dentin or in cariesaffected dentin (CAD). Debonded dentin surfaces were studied by field emission scanning electron microscopy (FESEM), and remineralization was evaluated through nanohardness (Hi) and Young’s modulus (Ei), Raman spectroscopy, and Masson's trichrome staining technique. Load cycling did not affect MTBS, except when Ketac- Bond was applied on sound dentin, which attained 100% pretesting failures. Minerals precipitated in porous platforms. GIC promoted total occlusion of tubules, and RMGIC originated empty or partial occluded tubules. In sound dentin, load cycling produced an increase of the relative presence of crystalline minerals after using Ketac-Bond (Phosphate peak, from 18.04 up to 81.29 cm-1 at hybrid layer, and from 19.28 up to 108.48 cm-1 at the bottom of the hybrid layer; Carbonate peak, from 8.06 up to 15.43 cm-1 at the hybrid layer, and from 7.22 up to 19.07 cm-1 at the bottom of the hybrid layer). Vitrebond Plus, in sound dentin, attained opposite outcomes. In CAD treated with Ketac- Bond, the highest Hi (1.11 GPa) and Ei (32.91 GPa) values were obtained at the hybrid layer after load cycling. This GIC showed increased and immature mineral components (an average of 25.82 up to 30.55 cm-1), higher frequencies of crosslinking (considering the pyridinium ring at hybrid layer, from 4.1 up to 6.86 cm-1; at bottom of the hybrid layer, from 7.55 up to 8.58 cm-1) and worst collagen quality (considering the ratio amide I/AGEs-pentosidine at the hybrid layer, from 0.89 up to 0.69 cm-1; at the bottom of the hybrid layer, from 1.39 up to 1.29 cm-1) after load cycling, at the interface of the CAD samples. Both Hi and Ei of CAD treated with RMGIC were not affected 4 after load cycling, though phosphates, carbonates and crystallinity increased. The organic components showed a dissimilar crosslinking and an improvement of the nature of collagen. Trichrome staining showed lower signs of demineralization or exposed proteins after mechanical loading, though Vitrebond Plus exhibited a slight increment in red intensity at the interface. The null hypothesis to be tested is that bond strength, chemical bonding and mechanical performance of the tested ionomer-based cements would not be influenced by the application of load cycling on restorations of sound and caries-affected dentin substrates.Project MAT2014-52036-P supported by the Ministery of Economy and Competitiveness (MINECO) and European Regional Development Fund (FEDER)

The investigation of androgen receptor phosphorylation and inflammation in prostate cancer

No abstract available

Drug screening of biopsy-derived spheroids using a self-generated microfluidic concentration gradient

Abstract Performing drug screening of tissue derived from cancer patient biopsies using physiologically relevant 3D tumour models presents challenges due to the limited amount of available cell material. Here, we present a microfluidic platform that enables drug screening of cancer cell-enriched multicellular spheroids derived from tumour biopsies, allowing extensive anticancer compound screening prior to treatment. This technology was validated using cell lines and then used to screen primary human prostate cancer cells, grown in 3D as a heterogeneous culture from biopsy-derived tissue. The technology enabled the formation of repeatable drug concentration gradients across an array of spheroids without external fluid actuation, delivering simultaneously a range of drug concentrations to multiple sized spheroids, as well as replicates for each concentration. As proof-of-concept screening, spheroids were generated from two patient biopsies and a panel of standard-of-care compounds for prostate cancer were tested. Brightfield and fluorescence images were analysed to provide readouts of spheroid growth and health, as well as drug efficacy over time. Overall, this technology could prove a useful tool for personalised medicine and future drug development, with the potential to provide cost- and time-reduction in the healthcare delivery

The Intestinal Microbiota in Colorectal Cancer Metastasis - Passive Observer or Key Player?

The association between colorectal cancer (CRC) and alterations in intestinal microbiota has been demonstrated by several studies, and there is increasing evidence that bacteria are an important component of the tumour microenvironment. Bacteria may contribute to the development of CRC metastasis by signalling through metabolites, promoting epithelial-mesenchymal transition, creating an immunosuppressive microenvironment and through the impairment of the gut-vascular barrier. Host immunity and intestinal microbiome symbiosis play a key role in determining innate and adaptive immune responses at the local and systemic level. How this gut-systemic axis might contribute to the development of CRC metastasis is however unclear. Several clinical trials are investigating the impact of microbiome-targeted interventions on the systemic inflammatory response, treatment-related complications, and side effects. This review examines pre-clinical and clinical studies which have examined the role of microbes in relation to CRC metastasis, the mechanisms which may contribute to tumour dissemination, and directions for future work