RDS Primary Science Fair

Dr. Maria Morgan reports

On the 12th January 2017 I had the pleasure of attending the RDS Primary Science Fair which runs alongside the BT Young Scientist and Technology Exhibition. Although given the title of ‘Head Judge’ the Fair is non-competitive and provides a platform for showcasing STEM investigations (science, technology, engineering and maths) undertaken by primary school classes across Ireland.

Judges briefing at the RDS Primary Science Fair

Children exhibited their whole-class projects which included topics ranging from ‘How can we assist the declining bee population in our local area?’ to ‘Ambidextrous! Can I train my other hand?’. One of my favourites was ‘Why do we like Pink Lady apples so much?’ which demonstrated higher levels of sugar in Pink Ladies compared to other apple varieties. At another stand I was given the opportunity to have my lung capacity measured using a 5L water cooler bottle and some garden tubing (well how could I say no!). It’s such a privilege to attend the Fair each year where the positivity and enthusiasm for science means you come away with a feel-good scientific glow and a reassurance that the future of STEM in Ireland is in excellent little hands!
For more information on the RDS Primary Science Fair go to: www.rds.ie/Ireland-s-Philanthropic-Society/Our-Work/Projects/RDS-Primary-Science-Fair#sthash.z8tSvgG1.dpuf

Microcalcifications in breast cancer: Exploring their molecular formation and biological significance

MCT Research Talks – 23th January 2017

Survival rates for breast cancer have risen significantly over the past few decades, in large part due to a considerable increase in the number of tumours detected via mammography at an early, more easily-treated stage. The presence of microcalcifications on a mammogram constitutes an important diagnostic clue to radiographers, with approximately 30% of invasive breast tumours and up to 90% of cases of ductal carcinoma in situ (DCIS) being detected by the presence of calcifications. Some studies have also suggested that the presence of calcifications may act as a prognostic factor, as patients presenting with breast tumours with associated calcifications have a worse prognosis than those without.
Despite their importance in breast cancer diagnosis, the exact mechanism by which microcalcifications are formed remains largely unexplored. Our group previously established the first in vitro model of mammary cell microcalcification (1) which we have recently extended to the human the breast cancer cell line MDA-MB-231. When cultured with a cocktail of osteogenic-reagents for a prolonged period, these cells produce deposits of calcium phosphate.

Figure 1. Alizarin Red S stained MDA-MB-231 cell monolayer, grown in DMEM (Control) or DMEM supplemented with osteogenic cocktail and dexamethasone (OC+Dex). Red staining indicates presence of calcified deposits.

Using a combination of histological staining, quantitative measurement of calcium content, alkaline phosphatase activity and analysis of gene expression, we can monitor the changes in cell phenotype leading to onset of mineralisation. The nature of our model allows for easy manipulation of cell culturing conditions and by adding various inhibitory compounds or cytokines to our culture media, we can identify the key pathways and targets necessary for calcification production. In doing so, we hope to build up a comprehensive understanding of the cellular and molecular basis underlying the formation of these important diagnostic clues.

Recommended reading:

Cox RF, Hernandez-Santana A, Ramdass S, McMahon G, Harmey JH, Morgan MP.  Microcalcifications in breast cancer: novel insights into the molecular mechanism and functional consequence of mammary mineralisation. Br J Cancer. 106(3):525-37 PMID: 22233923 (Jan 2012)

Shane O’Grady, Maria Morgan

Irish Association of Pharmacologists Announcement

Dear IAP Members

I wanted to update you on developments over the past number of weeks.

I can report that we have made progress with an application for the IAP to join the European Association for Clinical Pharmacology & Therapeutics (EACPT -https://www.eacpt.eu/who-we-are-2/), who responded very positively and who have said that they will put our application to their executive committee at their next meeting in April. We look forward in hope to the last blank space in Western Europe being filled in blue very soon (see: https://www.eacpt.eu/members/national-society-affiliated-to-eacpt/).

We have also applied for membership of the Federation of European Pharmacological Societies (EPHAR -http://www.ephar.org/home.html), who have also responded very positively to our initial enquiries.

We are currently working on the IAP website and expect to be able to update you on developments shortly.

In the meantime, we would very much appreciate your promoting the newly constituted IAP to those who you think would be interested in becoming members. We anticipate that the benefits will soon include membership of European federations and the opportunities for contact with European colleagues.


Prof. Thomas Walther
Irish Association of Pharmacologists

The role of the anorectic neuropeptide CART in breast cancer

MCT Research Talks – 16th January 2017

Breast cancer currently affects 1 in 8 women in Ireland, with over 3000 reported cases each year. The most common subtype of breast cancer, known as Estrogen Receptor positive (ER+) breast cancer, accounts for roughly 70% of all breast cancers diagnosed. The most common drug used to treat this disease (Tamoxifen) works by preventing estrogen from driving the growth of the cancer cells, however, roughly 1 in 3 women will be resistant to tamoxifen treatment, highlighting the need for further research into this field. A number of years ago, though mining of publically available datasets, we identified a gene known as CART to be a marker of poor prognosis in ER+ breast cancer. CART (The Cocaine- and Amphetamine-Regulated Transcript) is a neuropeptide involved in processes such as feeding and drug reward. We have identified that high expression of CART in breast cancer patients correlates with poor overall survival, and also a poor response to tamoxifen. We also demonstrated that CART could influence the activity of ERα in a ligand-independent manner [1]. Our current research focuses on combining proteomic (mass-spectrometry) and transcriptomic (RNA-seq) approaches in order to fully understand the role CART plays in ER+ breast cancer. We aim to modulate the expression of these identified targets in order to investigate whether any of these targets could slow the growth of breast cancer cells in vitro. Combining these approaches, we hope to identify novel therapeutic opportunities for patients with ER+ breast cancer.

Recommended reading:

[1] DJ Brennan, DP O’Connor et al., The Cocaine- and Amphetamine-Regulated Transcript mediates ligand-independent activation of ERα, and is an independent prognostic factor in node-negative breast cancer. Oncogene 2012, 31, 3483–3494; doi:10.1038/onc.2011.519

Brian Mooney, Darran O’Connor

Recent MCT Papers In High Impact Journals With Commentaries

CNV and Schizophrenia Working Groups of the Psychiatric Genomics Consortium; Psychosis Endophenotypes International Consortium. [including Waddington J]. Contribution of copy number variants to schizophrenia from a genome-wide study of 41,321 subjects. Nat Genet. 2017 Jan;49(1):27-35. PMID: 27869829

John Waddington writes about his group’s participation in a global research endeavour that has produced a series of articles in Nature, Nature Genetics and Nature Neuroscience on the pathobiology of schizophrenia The Psychiatric Genomics Consortium (PGC) unites investigators around the world to conduct meta- and mega-analyses of genome-wide genomic data on psychiatric disorders, to achieve goals that cannot be reached by individual or indeed most national programmes. Its website (www.med.unc.edu/pgc) provides information about the organization, implementation and results of the PGC. This consortium began in early 2007 and has rapidly become a collaborative confederation of investigators from 38 countries. There are samples from more than 900,000 individuals currently in analysis, and this number is growing rapidly. The PGC is the largest consortium and the largest biological experiment in the history of psychiatry.
From 2007-11, the PGC focused on several disorders, from schizophrenia through to autism, attention-deficit hyperactivity disorder, bipolar disorder and major depressive disorder. These now extend to large studies of eating disorders, substance use disorders, obsessive-compulsive disorder/Tourette’s Syndrome and post-traumatic stress disorder. Initially, the PGC focused on common single nucleotide polymorphisms (SNPs) but has expanded to include copy number variation (CNVs) and uncommon/rare genetic variation. The PGC has received funding from many sources. It has relied heavily on the goodwill of its members and their donated effort, with the establishment andgenotyping of its primary studies funded by a wide range of national, international and commercial entities.

We have been involved in the Schizophrenia Working Group of the PGC for many years, from its progenitor organisation the International Schizophrenia Consortium, primarily through substantial funding from the Wellcome Trust for five centres across the island of Ireland (one led by us) to contribute DNA from well characterised patient populations to a national dataset curated at Trinity College Dublin. This dataset has then been shared with the PGC. The resultant studies have evolved from the first investigation of CNVs in schizophrenia (3,391 cases, 3,181 controls; Nature 2008; 455: 237-41), through to the largest GWAS study on schizophrenia undertaken to date (36,989 cases, 113,075 controls; Nature 2014; 511: 421-7), to this most recent publication (21,094 cases, 20,227 controls; Nature Genetics 2017; 49: 27-35), the largest study of CNVs in schizophrenia yet undertaken. This has been, and continues to be, an exemplary, rewarding journey of global collaboration to illuminate the pathobiology of the intractable human condition that is psychotic illness.


Chion A, O’Sullivan JM, Drakeford C, Bergsson G, Dalton N, Aguila S, Ward S, Fallon PG, Brophy TM, Preston RJ, Brady L, Sheils, O, Laffan M, McKinnon TA, O’Donnell JS. N-linked glycans within the A2 domain of von Willebrand factor modulate macrophage-mediated clearance. Blood. 2016 Oct 13;128(15):1959-1968. PMID: 27554083.

Jamie Marie O’Sullivan writes about this study:

Von Willebrand factor (VWF) is a large multimeric plasma sialoglycoprotein that plays two critical roles in normal haemostasis. First it mediates platelet adhesion to exposed subendothelial collagen at sites of vascular injury. Second, VWF acts as a carrier molecule for procoagulant factor VIII, thereby protecting it from premature proteolytic degradation and clearance. Deficiency of either VWF (von Willebrand disease, VWD) or FVIII (Haemophilia A) is associated with a significant bleeding phenotype. Conversely, elevated plasma levels of the VWF-FVIII complex constitute a dose dependent risk factor for both venous and arterial thrombosis. Consequently, understanding the factors that determine how plasma VWF levels are regulated is not only of basic scientific interest, but also direct clinical importance.

Despite the fact that type 1 von Willebrand disease constitutes the most common inherited bleeding disorder worldwide, the mechanisms underlying the reduced plasma VWF levels in these patients is not well established. However, emerging evidence suggests that reduced circulatory half-life may be important in a significant number of patients. However the biological mechanisms underlying VWF clearance from the plasma remains elusive. Work is on-going within the group of Professor James O’Donnell to investigate the importance of specific VWF domains in modulating half-life in vivo.

In the October issue of Blood the group published novel findings demonstrating a critical role for VWF glycosylation in determining its circulatory half-life. By examining the clearance of various truncated fragments of VWF, we observed that the A domains of VWF contain a receptor-recognition site important in mediating VWF binding to macrophages in vitro, and in regulating VWF clearance by macrophages in vivo. Furthermore site-directed mutagenesis demonstrated a key role for N-linked glycan structures at position N1515 and N1574 within the A domains in protecting VWF against macrophage-mediated clearance. More specifically, the protective effect of these large complex N-linked glycan structures may be due to steric hindrance, with shielding of cryptic binding sites for macrophage clearance receptor low density lipoprotein receptor-related protein 1 (LRP1).

Defining the mechanisms underpinning VWF clearance is not only of direct translational relevance, but may also have important implications for the development of novel therapeutic agents with significant commercial appeal. For example, the estimated cost of clotting factor concentrates to the global market is predicted to exceed €10 billion per annum by 2020. It is therefore perhaps unsurprising that major pharmaceutical companies (including Baxter®, Bayer®, Novo Nordisk® and Octapharma®) are actively pursuing development of longer-acting coagulation factor products (including VWF and FVIII) as a priority. Our novel findings further scientific understanding in relation to the molecular mechanisms involved in regulating coagulation glycoprotein clearance from plasma.

McGettrick AF, Corcoran SE, Barry PJ, McFarland J, Crès C, Curtis AM, Franklin E, Corr SC, Mok KH, Cummins EP, Taylor CT, O’Neill LA, Nolan DP. Trypanosoma brucei metabolite indolepyruvate decreases HIF-1α and glycolysis in macrophages as a mechanism of innate immune evasion. Proc Natl Acad Sci U S A. 2016 Nov 29;113(48):E7778-E7787. PMID: 27856732.

Dr. Anne McGettrick and Sarah Corcoran at Trinity College Dublin lead a team of researchers including Dr. Annie Curtis (now at MCT) on a study that uncovered how the African parasites trypanosomes evade the immune system in their hosts, The biochemists have unearthed a metabolic by-product of trypanosome activity known as indolepyruvate modulates the inflammatory response of the host and evades immune detection. This discovery may offer excellent possibilities for developing anti-trypanosome drugs and therapies because inhibiting the production of indolepyruvate may be key in fighting the parasite. Full details can be found here https://www.tcd.ie/news_events/articles/solving-a-putrid-camel-pee-riddle-may-aid-millions-affected-by-sleeping-sickness/7385

The cerebrovascular nature of neurological disorders

MCT Research Talks – 9th January 2017

Guest speaker: Dr. Matthew Cambell

At MCT, we are very excited to announce our visiting guest speaker Dr Matthew Campbell from the Smurfit Institute of Genetics in Trinity College Dublin. Dr Campbell is a prestigious and young emerging principal investigator in Ireland who has made significant contributions in the fields of Neuroscience, Inflammation and the Vasculature System, core research areas that are also conducted within the MCT department.

Dr. Matthew Cambell

Dr Campbell aims to elucidate and address questions associated with dysfunctional vasculature within neural tissues. Recently published in Nature Medicine, Dr Campbell made a significant discovery uncovering the role of the NLRP3 inflammasome in the development of one of the most common forms of central retinal blindness, AMD. His lab is now pursuing a range of novel therapeutic solutions for the treatment of AMD and recently reported on the translational potential of human IL-18 as an immunotherapy.

Dr Campbell interests also focus on the blood-brain barrier, where he recently reported for the first time on the auto-regulated diffusion of amyloid-β in Alzheimer’s disease. More recently, he has identified molecular mechanisms underlying the development of chronic traumatic encephalopathy (CTE) to concussive injuries in athletes and military personnel. He spearheads a project involving the use of RNA interference (RNAi) to modulate levels of distinct tight junction proteins at the blood-brain barrier. This led to a novel form of patented technology that was termed “Neural Barrier Modulation” which could have broad applications for a range of neurological conditions.

Dr Campbell is the recipient of Ireland’s most prestigious prize for young researchers, the “President of Ireland Young Researcher Award (PIYRA)“, in addition to the international Genentech/ARVO fellowship. He will be speaking today on ‘the cerebrovascular nature of neurological disorders’ at 12 pm in Tutorial Room 2/3. Lunch will be provided for all after the talk.

All welcome!!

Written by Dr Claire McCoy, Lecturer in Biochemistry, MCT, RCSI.

Immunometabolism, Is it under the eye of the clock

Annie Curtis reports

Our Immune-clock laboratory has a real interest in metabolism and how alterations in metabolic pathways termed “metabolic reprogramming” can shape the type of immune response. This area called “Immunometabolism” has exploded in the last 5 years, and the implications are massive. It appears that macrophages use one metabolic pathway to become highly proinflammatory and another metabolic pathway to resolve inflammation and promote wound healing.  So why is our laboratory so interested in this? Well, if you think about daily changes in our environment, the two biggest are the sleep/wake cycle and the other is feeding/fasting. It is now clear that clocks in metabolic tissues like the liver/pancreas/adipose tissue prepare the body to deal with this daily rhythm in feeding/fasting. Based on this, our interest is to figure out if the clock within macrophages is somehow altering its metabolism over the course of the day and is that leading to changes in macrophage function, particularly the inflammatory response.

Jamie Early, my PhD student

Jamie Early, my PhD who “lives” in Prof. Luke O’Neills laboratory at TCD and I were invited to submit this first ever review on circadian immunometabolism, and you can find it here.

Additional reading

  1. Early JO, Curtis AM Immunometabolism: Is it under the eye of the clock?Semin Immunol. 2016 Oct;28(5):478-490

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