RCSI and ALMAC Discovery Partnership to Target the Root of Cancer: Cancer Stem Cells

A New Year…and a new challenge for MCT postdoctoral researcher Gillian Moore 

Between the post-Christmas blues, cold days and that painful wait for the next pay day, January can be a pretty long and gruelling month. This year, deviating from the norm, my January kicked off to a great start with my eagerly awaited move to RCSI. Before Christmas I was delighted to find out that I would be working alongside Prof. Tracy Robson in the Department of MCT and I’m really excited for 2017, and the new opportunities and challenges this postdoctoral research position has to offer.
An ongoing research collaboration between the Robson research group and leading oncology pharmaceutical company, ALMAC Discovery, resulted in the development of ALM201, an anti-cancer peptide-based drug currently in Phase I clinical trial for patients with solid tumours. ALM201 is structurally based on the naturally occurring protein, FKBPL. FKBPL and its peptide-derivative, ALM201, have demonstrated potent anti-angiogenic properties, and notably, a unique ability to target cancer stem cells. Targeting of cancer stem cells has arguably become the Holy Grail of cancer therapy in recent years. Within the mass of every tumour there is a subpopulation of cancer cells with the ability to self-regenerate. It is this cell population that are responsible for the initiation and propagation of a tumour, and recurrence of disease following resistance to chemo and/or radiotherapy. If we can robustly target the bulk of the tumour in addition to any residual cancer stem cells then we can potentially circumvent progression and indeed recurrence of disease.

Left to right: Dr Graham Cotton, Senior R&D Group Leader, Almac Discovery; Prof Tim Harrison, Vice President Discovery Chemistry, Almac Discovery; Prof Tracy Robson, Head of Molecular & Cellular Therapeutics, RCSI; Dr Gillian Moore, postdoctoral researcher (Robson Group); Seamus Browne, RCSI Head of Industry Partnerships; Dr Stephanie Annett, postdoctoral researcher (Robson Group).

Ovarian cancer is one of the top ten most common cancers in women and is associated with a poor prognosis, primarily due to the late presentation of disease. In the coming months, the next stage in the clinical trial of ALM201 will involve the treatment of a cohort of ovarian cancer patients. Recent, unpublished preclinical data in the Robson group has indicated promising anti-cancer stem cell efficacy of ALM201 in the ovarian cancer setting. I am interested in understanding the molecular mechanisms that underpin this observed anti-cancer stem cell activity of ALM201. A new phase of academic research funding from ALMAC Discovery will enable us to carry out this work. While the specific targeting of ovarian cancer stem cells is a relatively new research field, it has potential to provide much needed alternate treatment options for this aggressive tumour type and may have implications for other malignancies.

It’s great to be part of MCT at RCSI and I’m looking forward to sharing our research findings as the project develops.

The Immune-Clock laboratory of Dr. Annie Curtis, a recent recruit to RCSI

MCT Research Talks – 30th January 2017

Last week’s departmental talks encompassed a Deep Dive into Clock biology in Macrophages affecting the Inflammatory Response. This area is the focus of the Immune-Clock laboratory of Dr. Annie Curtis, a recent recruit to RCSI.

Jamie Early, my PhD student

Jamie Early (PhD student of the Curtis Lab) currently residing in the Luke O’Neill Laboratory presented his findings on the role of the circadian clock in suppressing inflammation in macrophages and if the anti-oxidant transcription factor and redox sensor NRF2 plays a role. His talk was titled ‘The macrophage clock is a key controller of the anti-oxidant and inflammatory response via the transcription factor Nrf2’.

Second up, we had Mariana Cervantes (PhD student and visiting scientist from the Instituto Politecnico Nacional (IPN) in Mexico) present her talk titled ‘The macrophage clock is having a profound impact on mitochondrial dynamics- what are the implications for inflammation?’

Mariana Cervantes

Mariana is interested in how mitochondria alter their morphology, either fusing together to form networks or fragmenting into smaller units termed fission. She is trying to uncover if the clock is regulating this process and if so what are the implications for the inflammatory response.

This work is part of a collaboration between RCSI and  Luke O’Neill laboratory at TCD and is funded through Science Foundation Ireland

A pharmacogenomic exploration of adverse drug reactions in epilepsy (PGXOME)

For most people with epilepsy, long term treatment with anti-epileptic drugs (AEDs) are necessary to prevent the seizure, and 40% do not respond to the first line of AED, leading to an often lifelong odyssey of trial and error towards effective treatment that is often not found. Epilepsy is primarily treated using AEDs, but these are associated with a considerable risk for adverse drug reactions (ADRs), some of which have been shown to have a genetic predisposition. For example, the genetic variant HLA-A*3101 is a common risk factor for rash and severe blistering skin reactions with the drug carbamazepine (Tegretol) in Europeans. However there are few other predictors of some more common ADRs.

Dr. Mark McCormack

The EpiPGX Consortium was established to identify genetic biomarkers of epilepsy treatment response from patient centres across Europe. The EpiPGX Consortium has generated genetic profiles on over 8000 patients with matching detailed drug response and medical histories. In order to investigate the links between genetic profiles and ADRs in epilepsy, Dr. Mark McCormack will travel to UMC Utrecht, the Netherlands for one year on a  Marie-Skłodowska-Curie Fellowship from the European Commission.

The aim of this fellowship is to identify clinically useful genetic variants to predict adverse reactions to AEDs. This will help optimize personalized treatment, limit the trial and error approach of AED choice, and thus improve medication safety and quality of life in epilepsy.

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

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

Follow me on Twitter @curtisannie

MCT student, Lisa Dwane, talks about her research and recent achievements

cropped-RCSI-logo-1.jpgFollowing completion of my Pharmacology degree in UCD, I began a PhD in breast cancer research under the supervision of Dr. Darran O’Connor, a career I have always been very determined to follow. My research is focused on endocrine-driven breast cancer and understanding the molecular mechanisms that drive this subtype of cancer. Currently, half of breast cancer patients that receive anti-endocrine therapies will relapse, so there is an urgent need for the identification of novel therapeutic targets. Our research is focused on the deubiquitinating enzyme USP11, which we believe plays a key role in driving endocrine-driven breast cancer. When we silence USP11 in vitro, we see a reduction in estrogen receptor activity and cell viability. During the final year of my PhD, I hope to elucidate the mechanism by which USP11 plays this role, and determine the prognostic relevance of USP11 in breast cancer. This could potentially lead to a better understanding of endocrine-driven breast cancer and with further validation, USP11 may represent a novel therapeutic target.

Lisa Dwane presents her research at the Irish Cancer Society’s Researcher of the Year Award. december, 1st, 2016. TCD
Lisa Dwane presents her research at the Irish Cancer Society’s Researcher of the Year Award. December, 1st, 2016. TCD

As a pharmacologist, I was thrilled to win best oral presentation at the Irish Association of Pharmacologists Annual Meeting! The standard of talks throughout the day were excellent, with a wide range of topics explored. I was also a finalist for the Irish Cancer Society’s Researcher of the Year Award, which took place 1st December at Trinity College Dublin. The purpose of the evening was to communicate our research to a lay audience, which proved more difficult than expected! Although I didn’t take home the award it was a very enjoyable evening, and the experience was invaluable. As scientists it is important for us to share and communicate our research with the general public and this was a skill I gained from the night!

Winners and finalists for the Irish Cancer Society’s Researcher of the Year Award
Winners and finalists for the Irish Cancer Society’s Researcher of the Year Award