The circadian protein BMAL1 in myeloid cells is a negative regulator of allergic asthma

Asthma is of particular relevance to the area of circadian control of immunity, since it is a disease with very strong clinical evidence demonstrating regulation by circadian variation. Airway hypersensitivity and asthma attacks are more common at night in humans. The molecular basis for this is unknown and no model of asthma in animals with genetic distortion of the molecular clock exists.

Asthma is under strong circadian variation. Asthma symptoms worsen at night, particularly in the early hours of the morning. Lung function fluctuates in healthy individuals over 24 h period and these fluctuations are even more pronounced in asthmatics.

In this study, we showed that mice lacking the main clock transcription factor BMAL1 in myeloid cells have increased lung inflammation demonstrated by higher numbers of eosinophils and increased IL-5 (key pathogenic cytokine in asthma that recruits eosinophils).This suggests that Bmal1 is a potent negative regulator, in myeloid cells in the context of allergic asthma. Our findings might explain the increase in asthma incidents during the night in humans when BMAL1 expression is low.

Dr. Zbigniew Zaslona from TCD (pictured here) was the lead author on the study. Both Dr. Annie Curtis (MCT) and Prof. Luke O’Neill (TCD) were joint senior authors on the paper.

The circadian protein BMAL1 in myeloid cells is a negative regulator of allergic asthma.

Zaslona Z, Case S, Early JO, Lalor SJ, McLoughlin RM, Curtis AM*, O’Neill LA* – Both authors contributed equally to this study.

Am J Physiol Lung Cell Mol Physiol. 2017 Mar 23:ajplung.00072.2017. doi: 10.1152/ajplung.00072.2017. [Epub ahead of print]

International Research and Education

Prof Tracy Robson (MCT), Prof Jochen Prehn (Physiology & Medical Physics) and Dr Darran O’Connor (MCT) have recently returned from 1 week at the College of Pharmaceutical Sciences, Soochow University, Suzhou, China where they participated in a workshop with faculty to explore research collaborations and future joint funding applications under the newly announced SFI-NSF Partnerships for International Research and Education. Supported by an Erasmus+ programme coordinated by Prof Marc Devocelle (Department of Pharmaceutical and Medicinal Chemistry), the workshop involved presentations from RCSI and Soochow investigators describing their work and discussion to identify areas of synergy. Afternoon lectures by RCSI faculty were opened to postgraduate and postdoctoral researchers from Soochow, leading to a vigorous and stimulating discussion and Prof Xinliang Mao from Soochow will visit RCSI next month to further strengthen future collaborative research opportunities. 

Left to Right: Prof Tracy Robson (MCT), Prof Jochen Prehn (Physiology & Medical Physics) and Dr Darran O’Connor (MCT)

At the invitation of the President of the British Pharmacological Society, Professor John Waddington (Emeritus, RCSI) has been elected to Fellowship of the Society; this is in recognition of his career contributions to research, education and service in the discipline of pharmacology, not just in Ireland but globally. He has recently returned from 3 weeks at the College of Pharmaceutical Sciences, Soochow University, China, under his joint appointment as a Professor of Pharmacology. While there, he continued collaborative research, gave undergraduate lectures and fostered further joint endeavours between RCSI and Soochow University, which is in the top 5% of Chinese research universities.   

Tracy Robson

The Time Evolution of Shear-Induced Particle Margination and Migration in Flowing Blood

MCT Research Talks – 24th March 2017

Prof. Eric S. G. Shaqfeh, Qin M. Qi, Departments of Chemical and of Mechanical Engineering, Stanford University

The inhomogeneous center-of-mass distribution of red blood cells and platelets normal to the flow direction in small vessels plays a significant role in hemostasis, drug delivery and microfluidics. Under pressure-driven flow in channels, the migration of deformable red blood cells at steady state is characterised by a concentration peak at the channel center and a cell-free layer or Fahraeus-Lindqvist layer near the vessel wall.

Eric Stefan G. Shaqfeh

Rigid particles such as platelets, however, “marginate” and thus develop a near-wall excess concentration. This margination controls the time it takes for the initial stages of platelet binding and clotting in response to trauma.
In this talk, we investigate the time-dependent concentration distribution of red blood cells and platelets in pressure-driven flow by developing and solving a Boltzmann model, advection-diffusion equation for both species. From a fluid mechanics point of view, deformability-induced hydrodynamic lift and shear-induced diffusion are essential mechanisms for the cross-flow particle migration and margination. The governing equation for the distribution of red blood cells includes both lift flux away from the wall and shear-induced diffusion due to cell-cell “collisions”. On the other hand, the governing transport equation for platelets includes shear-induced diffusion from cell-platelet “collisions” and platelet-platelet “collisions”. We demonstrate that these predictions are in good agreement with full boundary element simulations of the margination process and we also compare directly to experimental results. We then examine, within this model and our full boundary element simulations, the time evolution and “entrance length” for red blood cell migration and platelet margination. The resulting complete model can serve as a fast and computationally efficient alternative to large-scale simulation with the application, for example, as a real-time computational tool for microfluidic blood assay systems.

A novel platelet function test (The Platelet Monitoring Biochip)

MCT Research Talks –13th March 2017 

Jonathan Cowman reports

Cardiovascular disease (CVD) is the leading cause of death and disability in the world (approx. 1.9 million deaths per year within the EU). Platelet’s play a key role in this process and hence is why antiplatelet therapy such as aspirin is effective in reducing its incidences. Platelet function testing has a role in identifying those that are at high risk of a CVD related event (example a heart attack) and also identifying those patients that do not respond to their medication. There are a number of platelet function tests on the market however these tests suffer from a number of disadvantages such as expense, high sample volume, requirement for trained lab personal and single drug test capability.

Jonathan Cowman

My current research under the supervision of Prof. Dermot Kenny (RCSI) and Dr. Niamh Gilmartin (DCU and DIT) is to work alongside our multi-disciplinary team to produce a cost effective, rapid, small sample volume platelet function assay which can detect the effect of multiple antiplatelet drugs in a single patient blood sample. The project is known as the Platelet Monitoring Biochip (PMB). The PMB device consists of 6 micron sized fibrinogen dots, which are micro contact printed to a Zeonor (plastic) surface, a bright-field imaging system and a custom designed platelet analysis software. Blood is added to the device and rocked for 30 minutes to allow platelets to adhere to the fibrinogen dots. The device has 3 channels, a control (no agonist well), an adenosine diphosphate (ADP) well and an Arachidonic acid (AA) well which can be used to detect P2Y12 platelet inhibition and aspirin effect simultaneously. The PMB device provides a fast, easy and low cost way to determine the effectively of antiplatelet therapy against multiple agonists in whole blood. The device is currently in operation in RCSI Beaumont.

MCT Awards at Research Day 2017

Dear all,

MCT was well represented in the award ceremony at the recent Research Day; our congratulations go to the following:

Dr Mark McCormack, for receiving prizes in the RCSI Author Citations Prizes in two categories – the 2011 Most Highly Cited RCSI Senior Authored Paper and the 2011-2015 Most Highly Cited RCSI Senior Authored Paper with International Collaboration – for his paper entitled “HLA-A*3101 and carbamazepine-induced hypersensitivity reactions in Europeans” published in the New England Journal of Medicine.

Dr Cathy Wyse, as part of Dr Annie Curtis’s group, was presented a prize for the front cover illustration of the RCSI Research Day abstract book, for a striking image of glial cells at the junction between the brain and pituitary gland.

Tony McHale, PhD student at the School of Pharmacy, & MCT, and the Irish Centre for Vascular Biology, RCSI received the prize for the best postgraduate oral presentation, for his talk on the topic of “First in Class Potential Novel Drug for the Treatment of Sepsis Caused by Urinary Tract Infections”.

Medical student Jack Donohue [RSS student], was awarded the Dr Harry O’Flanagan Prize for Excellence in Undergraduate Research for the best undergraduate oral presentation for a project carried out as part of the RCSI Research Summer School entitled “Sanger Confirmation of Suspected Epilepsy-Related Pathogenic Variants Identified Through Next-Generation Sequencing”.

Very well done to all!

Tracy Robson

Irish Association For Cancer Research Meeting 2017

Irish Association for Cancer Research – Annual Meeting takes place at Newpark Hotel, Kilkenny on Thursday 23 and Friday 24 February 2017.

MCT cancer researchers secured oral presentations at different sessions. Prof Ray Stallings is a guest speaker at the Plenary Session focused on challenges in childhood cancers. He will be discussing ‘Modulation of neuroblastoma phenotype with epigenetically regulated miRNAs’.

Stephanie Annett will be giving a talk ‘FKBPL as a novel prognostic biomarker and therapeutic agent in high-grade serous ovarian cancer’ at Proffered Paper Session on Thursday morning. Two Irish Cancer Society funded PhD students will be discussing their findings at the Irish Cancer Society Scholar and Fellow Presentation session. Louise Walsh – ‘RNA sequencing identifies bromodomain proteins as a therapeutic strategy for invasive lobular carcinoma’ and Brian Mooney – ‘Expression of the cocaine- and amphetamine-regulated transcript recruits BAF chromatin remodelling complexes to the estrogen receptor’.

Good luck to our presenters!

Olga Piskareva

 

 

Diagnostic gene sequencing in adults with epilepsy and intellectual disability

MCT Research Talks – 20th February 2017

Sinead Heavin reports

Sinead Heavin, PhD Post-Doctoral Researcher

Epilepsy is a common neurological disorder that affects ~40,000 people in Ireland. There are many different types of seizures which are caused by uncontrolled electrical impulses in the brain. Anti-epileptic drugs control seizures for ~50% of people with epilepsy but up to ~30% of patients remain uncontrolled despite treatment with multiple drugs. Epilepsy is caused by a number of factors include stroke, trauma and infections. However, more recently we have learned that epilepsy can be caused by genetic mutations. Some epilepsies are heritable while others arise de-novo. Many patients with an intellectual disability (ID) also have epilepsy. Many of these patients lack a specific diagnosis due to limited testing and available investigations. We sequenced a cohort of 99 adult patients with epilepsy and ID on a custom gene panel of ~150 genes. A likely pathogenic variant was identified in 20 patients in 19 different genes, including SCN1A, DCX and DEPDC5, well-known epilepsy genes. Furthermore, we identified copy number variants in two patients which are likely causative. Further work is needed to investigate the phenotype-genotype correlations identified in this study and any potential treatment options that may arise.

MCT researchers shed light on the ancestry of the Irish Travellers from the perspective of DNA

Edmund Gilbert reports

A new study, led by Prof. Gianpiero Cavalleri at MCT and Prof. Jim Wilson at the University of Edinburgh, has examined the population history of the Irish Travellers and has confirmed that the Irish Travellers share a common Irish origin with the settled Irish population. The work has also for the first time estimated the date which this divergence occurred.

A roadside camp in County Mayo 1972. Courtesy of George Gmelch

The Irish Travellers are a small nomadic population, making up about 0.6% of the total population on the island of Ireland, or between 29,000 and 40,000 individuals. Within the population cousin marriages (consanguineous marriages) are common, and the population is socially isolated from the surrounding settled Irish population.
The researchers, who also include MCT PhD student Edmund Gilbert, Shai Carmi of the Hebrew University of Jerusalem, and Sean Ennis of University College Dublin, used SNP-array based genotype data to compare the population genetics of the Irish Travellers to neighbouring Irish and British populations, as well as world-wide groups and European Roma Gypsies.
The study found that although the Irish Travellers were genetic closest to the settled Irish population, they showed significant differences. The study also confirmed the lack of recent shared genetic ancestry between the Irish Travellers and Roma Gypsies. The Irish Travellers, therefore, represent a subset of Irish genetic diversity, and the significant differences can be attributed to genetic drift, brought on by hundreds of years of genetic isolation and a decreasing population size. The analysis showed Irish Travellers also exhibit within-population sub-structure with four apparently distinct groups emerging, and interestingly these groups mirror different forms of the Shelta language and sociological groups within the Irish Travellers.

Galway John Ward making tinware and Galway 1971. Courtesy of George Gmelch

The dating of the origin of the Irish Travellers is of considerable interest, but this is a distinct date from the genetic origins of each population. This study has estimated a time of genetic divergence of the Irish Travellers and the settled Irish population using genomic tracts of shared identity. This method estimated the divergence to about 12 generations (360 years) ago, which is far older than common belief that the Irish Traveller population arose from the time of the Great Famine. The size of the dataset limited the authors to exploring the relatively simple model of one divergence event, future work is required to expand the study to explore more complex demographic models. The Irish Traveller population was shown to have high proportions of the genome where both maternal and paternal copies are identical, at similar levels to other consanguineous populations around the world.
The research was also welcomed by author and Traveller activist, Michael McDonagh said, “As a Traveller who has spoken on the history and identity of Irish Travellers to many groups ranging from children to academics, you sometimes rely on anecdotal information in trying to put across a serious message about Irish Traveller history. I am delighted that now we have qualified evidence that substantiates the argument I have made for many years, which is that Travellers did not descend from the Famine in Ireland. This research allows us to bring Irish Traveller history back many and gives us a factual identity.”

Kay McKeon

8 February 2017

An informal, private reception was held in the College for Kay McKeon on Wednesday, 8th February, to mark her retirement and contribution to RCSI after 39 years.

Kay joined Clinical Pharmacology in 1978 and with Prof. Kevin O’Malley was responsible for commissioning the then ‘new’ laboratories. She continued to play a fundamental role in developing Clinical Pharmacology’s laboratories and building the department’s reputation through the 1980s and 1990s into RCSI’s premier research department.

She played a central role in assisting Kevin O’Malley’s successor, Prof. Des Fitzgerald, in securing RCSI’s first large HEA-PRTLI and SFI grants, working long hours with the intricate details and logistics for such applications.

During this period, Kay was seconded to oversee the development of the RCSI Centre for Human Proteomics, before returning to base and seeing in another period of change on the departure of Prof. Fitzgerald and formation of Molecular & Cellular Therapeutics (MCT) from the Departments of Biochemistry and Clinical Pharmacology under Profs. David Croke and John Waddington as HODs; she played a key role in the management of MCT as a member of it’s Executive.

At a more personal level, Kay was someone who carried out her responsibilities in a genuinely supportive and politically astute manner; many appreciated her sensitivity, in assisting all ‘new staff’ settle in and in maintained balance and stability in overseeing laboratories with up to 100 staff.

Yet in addition to this, Kay also found the time to contribute more broadly to RCSI, particularly its philanthropic activities, for example, the Old Folks Christmas Lunch for those living in the vicinity of the College and related activities; an all-too-rare rare combination of professionalism and altruism.

An important part of what Clinical Pharmacology and MCT has achieved serves as her legacy to the College, in making MCT what it is today and the entity that Prof. Tracy Robson has recently inherited.

Hebrews 6 (10 … 19): ‘For God would not be so unjust as to forget all that you did for love of his name, when you rendered service to his people, as you still do … It is like an anchor for our lives, an anchor safe and sure’.

Kay has been an ‘anchor safe and sure’ across four decades. We thank you, Kay, for everything you’ve done for Clinical Pharmacology, MCT, RCSI and the community at large and wish you well for the future.

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.