Vitamin D supplementation in systemic lupus erythematosus patients with vitamin D deficiency and insufficiency : the effect on disease activity, fatigue and interferon signature gene expression

Background: Vitamin D deficiency is highly prevalent among patients with systemic lupus erythematosus (SLE) [1]. Evidence from multiple studies has shown that vitamin D deficiency in SLE is associated with a higher disease activity [2]. There is conflicting evidence with regards to the relationship between fatigue and vitamin D level [3,4].peer-reviewe

Phospho-Akt expression is high in a subset of triple negative breast cancer patients

The most commonly used biomarkers to predict the response of breast cancer patients to therapy are oestrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor 2 (HER2). Patients positive for these biomarkers are eligible for specific therapies such as anti-oestrogen therapy in the event of ER and PgR positivity, and trastuzumab, a monoclonal antibody, in the case of HER2 positive patients. Patients who are negative for all these three biomarkers, the so-called triple negatives, however, derive little benefit from such therapies. The PI3K/Akt pathway is activated in triple negative breast cancer cases, providing a possible target for therapy. The activation of Akt was investigated in Maltese triple negative breast cancer cases using an antibody detecting Akt phosphorylated at serine 473 (anti-Akt pS473). The study showed that 26\% of triple negative breast cancer patients had an elevated level of Akt (pS473). Furthermore, FTY720, a pharmacological activator of the phosphatase PP2A, was shown to block Akt activation at a concentration of 1\textmu M, in HCC1937 cells subjected to insulin-like growth factor 1 (IGF-1). Our data defined a subset of triple negative breast cancer patients based on high activity of AKT (pS473). This subset would be eligible for treatment using therapies which target the PI3K/Akt pathway, such as kinase inhibitors or phosphatase activators. In support of this, the BRCA1 mutant cells (HCC1937) were sensitive to the PP2a activator, FTY720. This suggests that FTY720 is a potential drug for use in adjuvant therapy in breast cancer cases having a high Akt (pS473).peer-reviewe

The re-emergence of the B1 cell compartment : is this a pre-lymphoma stage?

Chronic Lymphocytic Leukemia (CLL) are in some cases stereotyped for immunoglobulin variants in different populations, suggesting emergence of B cell subsets following presentation of the same antigen. CLL cells may originate from CD5+ naïve cells and from CD5 memory cells. Gene expression studies characterized a common cell of origin of the two clinical categories of CLL; the unmutated aggressive type and the mutated indolent type. The aim of this study was to investigate the presence of CD5 positive B cells in the elderly and their potential stimulation with exosomes derived from tumor cells. The findings from this study is aimed to create a model to identify instigating carcinomatous factors that may stimulate B1 cells to transform into a CLL-like model. In this study we show that CD19\textsuperscript+ cells (B cells) in cord blood have a high expression of CD5. CD19/CD5 staining of blood samples from senior citizens showed the presence of B cells which also express the CD5 marker, though at a lower expression when compared to CLL cells (CD19+/CD5 dim B cells). Measurement of clonality using λ/Κ flow cytometry staining show a monoclonal origin of the human CD19+/CD5 dim B cells. Monoclonal B cell Lymphocytosis in the elderly is a potential cell compartment that represents the origin of B cell proliferative disorders. The origin of the B cell proliferative disease requires antigen stimulation. A preliminary experiment showed that sorted lymphocytes can be stimulated by exosomes isolated from 2 cancer cells lines, A549 (lung epithelial) and PC3 (prostate cell line). In comparison with phytohaemagglutinin (PHA) and phorbolmyristate acetate (PMA), known lymphocyte stimulators, the exosomes stimulated the proliferation of monocytic-like cells. Further characterization is required to know the origin of these cells. The result shows that one can speculate that exosomes present cancer-derived antigens and stimulate cell proliferation. Further studies are required to evaluate the potential transformation capacity of cancer-derived exosomes. In addition, various cytokines were measured in the sera of senior citizens to investigate a differential release of cytokines in the presence or absence of the CD19+/CD5 dim B cells. Cytokines examined were not significantly different between the 2 groups and further evaluation of cytokine levels is required.peer-reviewe

Molecular classification of breast cancer patients using formalin-fixed paraffin-embedded derived RNA samples

The use of archival formalin-fixed paraffin-embedded (FFPE) material to analyse gene expression is limited by the low quality of extracted RNA. In this paper, we utilised an RNA based assay to quantify expression of luminal and basal markers, together with ERBB2 probes, in FFPE archival tissue from 2009 to 2010, all of which had clinical and therapeutic information of more than 5 years. Receptor status of the patients was characterised using the QuantiGene® Plex assay with 100% concordance to immunohistochemical (IHC) and fluorescence in situ hybridisation (FISH) results. A panel of molecular markers known to classify luminal and basal tumours were used and correlated with receptor status of the tumours. As expected, the triple negative breast cancer (TNBC) samples were classified as basal and oestrogen receptor (ER) positive cases as luminal. In summary, the QuantiGene® Plex technology provides a platform to quantitate novel panels of biomarkers on archival material. Moreover, multiplex analysis allows the use of minimal amounts of material providing an opportunity to utilise laser micro-dissected material. FFPE tissue samples are an invaluable resource for retrospective studies to interrogate current novel biomarkers, particularly to generate disease free survival and overall survival graphs to measure predictive value using well annotated retrospective samples with full clinical and pharmacological outcomes.This study was funded by the Faculty of Medicine and Surgery, University of Malta and the Italia Malta Genome Breast Cancer Cross Border Risk Surveillance (ImaGenX) project financed under Operational Program Italia Malta 2007-2013 and co-financed by the University of Malta.peer-reviewe

National strategy for health research and innovation

In 2011, the Malta Council for Science and Technology (MCST) commissioned the Development of a dedicated strategy for health research and innovation in line with its mandate from Government to identify areas of national priority and design and to also implement strategic approaches to enhance economic competitiveness and quality of life. The Strategy was drawn up by a steering group which also included people from outside the health sector, to ensure that it also keeps note of the economic side of things.peer-reviewe

Thr300Ala ATG16L1 polymorphisms and bone strength in Crohn’s disease patients

Introduction: Studies on the effect of deletion of ATG5 and ATG7 proteins on bone cell function and bone strength in laboratory mice have revealed an association between autophagy and osteoporosis. The effect on bone strength of the Thr300Ala variant (rs2241880 polymorphism) of the ATG16l1 gene, a Crohn’s disease susceptibility gene essential in macroautophagy, has not yet been explored. Methods: 101 Crohn’s disease patients underwent DEXA bone density scanning. Real time PCR, high resolution melt (HRM) and restriction fragment length polymorphism (RFLP) were made use of as to assess for the rs2241880 polymorphism of the ATG16L1 gene in these patients. Results: HRM and RFLP demonstrated that 39.6% had the wild type rs2241880 (Thr300Ala) polymorphism while 7.9% were homozygous and 52.5% were heterozygous for the polymorphism. Mean DEXA bone mineral density scores in these patients showed lower T scores at the hip (1.74) among patients with the homozygous polymorphism than among patients with the heterozygous polymorphism (mean T score hip: -1.29). The highest mean T scores were found in patients with the wild type polymorphism (-1.04). Discussion: This study demonstrates the first evidence that polymorphisms in the ATG16L1 gene may play a role in bone metabolism.peer-reviewe

Biomarkers predicting sensitivity to PP2A activation therapy in cancer

The invention relates to the treatment and classification of cancer, in particular breast cancer. It relates to identifying patients who are likely to respond to cancer therapy with a PP2 A activator. Background: The provision of new treatments for cancer is of high importance, including for cancers resistant to known treatments. There is also a need for identification of markers that allow for detection, prognosis and classification of cancer, and which can predict responsiveness of a patient to a given therapy. The deregulation of the protein phosphatase 2A (PP2A) complex is known to be a common event in cancer [Grech et al; Tumor Biol; DOl 10.1007/s13277-016-5145-4 (2016)].peer-reviewe

Deregulation of the phosphatase, PP2A is a common event in breast cancer, predicting sensitivity to FTY720

We would like to thank Professor Andrew Hanby, Professor Valerie Speirs and Dr Thomas A Hughes for the breast cancer cell lines used in this study. This research was supported by the Faculty of Medicine and Surgery, University of Malta.Background: The most commonly used biomarkers to predict the response of breast cancer patients to therapy are the oestrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor 2 (HER2). Patients positive for these biomarkers are eligible for specific therapies such as endocrine treatment in the event of ER and PgR positivity, and the monoclonal antibody, trastuzumab, in the case of HER2-positive patients. Patients who are negative for these three biomarkers, the so-called triple negatives, however, derive little benefit from such therapies and are associated with a worse prognosis. Deregulation of the protein serine/threonine phosphatase type 2A (PP2A) and its regulatory subunits is a common event in breast cancer, providing a possible target for therapy. Methods: The data portal, cBioPortal for Cancer Genomics was used to investigate the incidence of conditions that are associated with low phosphatase activity. Four (4) adherent human breast cancer cell lines, MDA-MB-468, MDA-MB-436, Hs578T and BT-20 were cultured to assess their viability when exposed to various dosages of rapamycin or FTY720. In addition, RNA was extracted and cDNA was synthesised to amplify the coding sequence of PPP2CA. Amplification was followed by high-resolution melting to identify variations. Results and conclusion: The sequence of PPP2CA was found to be conserved across a diverse panel of solid tumour and haematological cell lines, suggesting that low expression of PPP2CA and differential binding of inhibitory PPP2CA regulators are the main mechanisms of PP2A deregulation. Interestingly, the cBioPortal for Cancer Genomics shows that PP2A is deregulated in 59.6% of basal breast tumours. Viability assays performed to determine the sensitivity of a panel of breast cancer cell lines to FTY720, a PP2A activator, indicated that cell lines associated with ER loss are sensitive to lower doses of FTY720. The subset of patients with suppressed PP2A activity is potentially eligible for treatment using therapies which target the PI3K/AKT/mTOR pathway, such as phosphatase activators.peer-reviewe