Diagnostic, prognostic and predictive value of cell-free miRNAs in prostate cancer : A systematic review

Publisher Copyright: © 2016 Endzeliņš et al.Prostate cancer, the second most frequently diagnosed cancer in males worldwide, is estimated to be diagnosed in 1.1 million men per year. Introduction of PSA testing substantially improved early detection of prostate cancer, however it also led to overdiagnosis and subsequent overtreatment of patients with an indolent disease. Treatment outcome and management of prostate cancer could be improved by the development of non-invasive biomarker assays that aid in increasing the sensitivity and specificity of prostate cancer screening, help to distinguish aggressive from indolent disease and guide therapeutic decisions. Prostate cancer cells release miRNAs into the bloodstream, where they exist incorporated into ribonucleoprotein complexes or extracellular vesicles. Later, cell-free miRNAs have been found in various other biofluids. The initial RNA sequencing studies suggested that most of the circulating cell-free miRNAs in healthy individuals are derived from blood cells, while specific disease-associated miRNA signatures may appear in the circulation of patients affected with various diseases, including cancer. This raised a hope that cell-free miRNAs may serve as non-invasive biomarkers for prostate cancer. Indeed, a number of cell-free miRNAs that potentially may serve as diagnostic, prognostic or predictive biomarkers have been discovered in blood or other biofluids of prostate cancer patients and need to be validated in appropriately designed longitudinal studies and clinical trials. In this review, we systematically summarise studies investigating cell-free miRNAs in biofluids of prostate cancer patients and discuss the utility of the identified biomarkers in various clinical scenarios. Furthermore, we discuss the possible mechanisms of miRNA release into biofluids and outline the biological questions and technical challenges that have arisen from these studies.publishersversionPeer reviewe

Nano-Sized α-Tricalcium Phosphate for Bone Cement

A new bottom-up approach that makes pure α-TCP has the potential to improve the field of calcium phosphate bone cements. This research takes on the challenge, with attention to microstructural characteristics, to develop a nano-sized building block of resorbable bone cements – by heating amorphous calcium phosphate (ACP) precursor – that upon mixing with water transforms into calcium-deficient apatite

Formation of Calcium-Deficient Hydroxyapatite via Hydrolysis of Nano-Sized Pure α-Tricalcium Phosphate

Nano-sized pure α-tricalcium phosphate (α-TCP) fabricated by a novel synthesis approach shows great potential for a faster transformation into calcium-deficient hydroxyapatite (CDHA) than conventionally prepared α-TCP. In this work, amorphous tricalcium phosphate precursors were precipitated and treated with a solvent (water or ethanol), and dried (freeze-dried and oven-dried) before heating at 775 °C. Nano-sized α-TCP powders were investigated for their phase composition and crystallinity, particle shape and size, reactivity and cellular biocompatibility. Reaction with water showed faster CDHA formation for freeze-dried powder, at 6 hours, compared to ethanol treated powders, whereas a higher biocompatibility was found for pure α-TCP

Formation of Calcium-Deficient Hydroxyapatite via Hydrolysis of Nano-Sized Pure α-Tricalcium Phosphate

Nano-sized pure α-tricalcium phosphate (α-TCP) fabricated by a novel synthesis approach shows great potential for a faster transformation into calcium-deficient hydroxyapatite (CDHA) than conventionally prepared α-TCP. In this work, amorphous tricalcium phosphate precursors were precipitated and treated with a solvent (water or ethanol), and dried (freeze-dried and oven-dried) before heating between 650 and 800 °C. Nano-sized α-TCP powders were investigated for their phase composition and crystallinity, particle shape and size, reactivity and cellular biocompatibility. Reaction with water showed faster CDHA formation for freeze-dried powder, at 6 hours, compared to ethanol treated powders whereas a higher biocompatibility was found in a pure α-TCP

Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages

Abstract. Background Macrophages are one of the most important players in the tumor microenvironment. The polarization status of tumor associated macrophages into a pro-inflammatory type M1 or anti-inflammatory type M2 may influence cancer progression and patient survival. Extracellular vesicles (EVs) are membrane-bound vesicles containing different biomolecules that are involved in cell to cell signal transfer. Accumulating evidence suggests that cancer-derived EVs are taken up by macrophages and modulate their phenotype and cytokine profile. However, the interactions of cancer-derived EVs with monocytes and macrophages at various differentiation and polarization states are poorly understood. In the current study, we have analyzed the uptake and functional effects of primary (SW480) and metastatic (SW620) isogenic colorectal cancer (CRC) cell line-derived EVs on monocytes (M), inactive macrophages (M0) and M1 and M2 polarized macrophages. Methods THP-1 monocytes were differentiated into M0 macrophages by addition of phorbol-12-myristate-13-acetate. Then M0 macrophages were further polarized into M1 and M2 macrophages in the presence of LPS, IFN- γ, IL-4, and IL-13 respectively. Internalization of SW480 and SW620-derived EVs was analyzed by flow cytometry and fluorescence microscopy. Changes in monocyte and macrophage immunophenotype and secretory profile upon EV exposure were analyzed by flow cytometry, quantitative PCR and Luminex assays. Results THP-1 monocytes and M0 macrophages efficiently take up SW480 and SW620-derived EVs, and our results indicate that dynamin-dependent endocytic pathways may be implicated. Interestingly, SW480 and SW620-derived EVs increased CD14 expression in M0 macrophages whereas SW480-derived EVs decreased HLA-DR expression in M1 and M2 polarized macrophages. Moreover, SW480-derived EVs significantly increased CXCL10 expression in monocytes and M0 macrophages. In contrast, SW620-derived EVs induced secretion of IL-6, CXCL10, IL-23 and IL-10 in M0 macrophages. However, addition of CRC cell line-derived EVs together with LPS, IFN- γ (M1) and IL-4, IL-13 (M2) stimuli during macrophage polarization had no additional effect on cytokine expression in M1 and M2 macrophages. Conclusion Our results suggest that CRC cell line-derived EVs are internalized and reprogram the immunophenotype and secretory profile in monocytes and inactive macrophages inducing mixed M1 and M2 cytokine response. Although CRC EVs decreased HLA-DR expression in M1, M2 polarized macrophages, their effect on the secretory profile of M1 and M2 polarized macrophages was negligible

Additional file 4: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages

TNFα, IL-23, IL-6, IL-1 β, CXCL10, CCL22, IL-10 and MMP9 secretion profile at different monocyte-macrophage differentiation stages. The graphs represent average biomolecule concentrations SEM (n = 3). Statistical analysis carried out with one-way ANOVA test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and **** ≤ 0.0001 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 63 kb

Additional file 3: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages

Effect of temperature on the SW480 EV uptake in THP-1 monocytes. Flow cytometry histograms showing Syto RNA Select fluorescence intensities of untreated (left) and Syto RNA Select-labeled SW480 EV-treated THP-1 monocytes following incubation at 4 °C (middle) and 37 °C (right). Histogram markers show the percentage of Syto RNA Select-positive cells. (PDF 53 kb

Additional file 2: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages

SW480 and SW620-derived EV effect on monocyte (M) and macrophage (M0, M1, M2) viability. a OD values at 450 nm which are in direct proportion of viable cell counts. b SW480 and SW620 EV cytotoxicity on THP-1 monocytes and M0, M1 and M2 macrophages. The graphs represent mean ± SEM (n = 3). Statistical analysis carried out with the t-test. *p ≤ 0.05, **p ≤ 0.01 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 50 kb