2013 Projects (31-60)

32. The incidence of thromboembolic events in patients with acute myeloid leukemia: a retrospective analysis

Song L1, Lee YG2, Kim I2

1UCD School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin 4.

2Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea. 

Patients with acute myeloid leukemia (AML) are disposed to hemostatic abnormalities, a potentially fatal clinical entity. Thrombosis is known to complicate the course of cancer management, and is triggered by various risk factors including age, chronic comorbities, and the use of central venous device1. The aim of this study was to investigate the incidence of thromboembolic events (TEEs) in AML patients.

A retrospective analysis was performed of 811 adult patients diagnosed with AML from January 2007 to December 2011. TEE occurrence was confirmed by throroughly reviewing all results of the imaging studies and their relevant electrical medical records.

Among 811 patients, 3 patients had TEE at AML diagnosis; the remaining 808 patients were entered for further analyses. Of the 808 patients, the 3-month, 6-month, and 1-year cumulative incidences of TEE were 1.8%, 2.7%, and 3.5%, respectively. A total of 23 (2.9%) TEEs occurred with a median time to thrombosis of 54 days (range 5-280 days). No significant predictors for TEE occurrence were found in univariate analysis. In multivariate analysis however, the intermediate and poor cytogenetic risk (vs. better risk) were an independent risk factor for TEE occurrence (hazard ratio 7.55; 95% CI 1.01-56.16; P= 0.048).

Cytogenetic risk factors proved to be the strongest independent predictor for increased risk of TEE in AML patients. Patients with intermediate or poor cytogenetic risk were found to have 7.55 times higher risk of TEE occurrence compared to patients with a better risk.

Reference:

1. Ku G, White R, Chew H, Harvey D, Zhou H, Wun T. Venous thromboembolism in patients with acute leukemia: incidence, risk factors, and effect on survival. Blood. 2009;113(17):3911-3917

  • Presenting Author: Mr Leo Song
  • Supervisor: Dr Inho Kim
  • Co-Supervisor: Dr Yun-Gyoo Lee

34. 3-lead ecg screening for prolonged qt-interval in methadone maintenance therapy clients: a pilot study

O'Carroll L, Bury G1

1UCD School of Medicine and Medical Science, University College Dublin, Belfield,Dublin 4.

Prolonged QT interval is a cardiac arrhythmia which may trigger episodes of ventricular fibrillation in certain individuals. Ventricular fibrillation is potentially deadly, as it prevents the efficient circulation of blood around the body. This study was designed to assess the feasibility of using three lead ECG to monitor the heart rhythms of clients attending clinics for methadone maintenance therapy. Regular methadone consumption demonstrably prolongs QT and QTc intervals1. It is difficult to establish whether this effect is dose-dependent, as prolongation of QT interval may be due to a diverse array of causes. Many risk factors may co-exist in a single client. These factors may interact synergistically or antagonise each other, producing an unpredictable effect on QTc. Notably, many drugs, both prescribed and non-prescribed, affect QTc. A large variety of psychiatric medicines, in particular SSRIs and atypical antipsychotics can provoke prolongation of QTc. This is important because methadone maintenance therapy (MMT) clients are frequently exposed to polypharmaceutical treatment of psychiatric co-morbidities with their substance misuse disorder.

It has been estimated that the rate of deaths due to prolonged QTc in MMT programmes is 0.06 per 100 treatment-years2. This is a significant burden of iatrogenic mortality considering there are around twelve thousand MMT patients in Ireland alone. This study aims to demonstrate the suitability and convenience of 3-lead ECG for the screening of prolonged QT interval in the MMT population. Convenience greatly improves client uptake of screening; and so a fast, reliable screening technology should make an appreciable difference to client safety in the future.

This project focuses on the planning and design of a pilot study which will be conducted to refine methods and instruments for use in the main study. Some key issues addressed over the course of the project related to: site selection, pilot power, sampling methodologies, instrument design, QT screening procedure, operational permissions and the logistics of providing sufficient defibrillators. 

Key outputs of the project included the identification of a pilot site, an agreed pilot sample size, pilot sample selection method, revised study instruments, and a pilot method designed to choose between two distinct client feedback generation modalities for use in the main study. The pilot site was selected after the total number of study sites were recruited in order to select a centre which we felt adequately reflected the demographics of the population, but which was small enough to use without sacrificing too much power from the main study. Potential sites were contacted, and if they expressed interest, they were sent follow up correspondence two to three times weekly, and were also given site visits by researchers, and took part in teleconferences discussing study design. Study instruments were revised constantly in response to clinician feedback arising from the site visits and teleconferences. Operational permissions were sought from the HSE for some state operated clinics to participate. This necessitated modification and resubmission of our application for ethical approval. Pilot screening is expected to begin at the end of September.

Acknowledgement:

I would like to thank the UCD Centre for Emergency Medical Science for allowing me to participate in this project.

References:

1. Roy AK, McCarthy C, Kiernan G, et al. Increased incidence of QT interval prolongation in a population receiving lower doses of methadone maintenance Therapy. Addiction 2011; 107; p1132-1139

2. Anchersen K, Clausen T, Gossop M, et al. Prevalence and clinical relevance of corrected QT interval prolongation during methadone and buprenorphine treatment: a mortality assessment study. Addiction 2009; 104; p993-999

 

  • Presenting Author: Mr Lorcan O'Carroll
  • Supervisor: Prof Gerard Bury
  • Co-Supervisor: Ms Helen Tobin

35. Novel therapy induction in a patient with high risk multiple myeloma followed by autologous stem cell transplantation (asct): a case report

Toolan S1, Bugler J1, Lane W1, Maglio M1, Cowzer D2, O’Gorman P2, Richardson PG1

1Jerome Lipper Multiple Myeloma Centre, Dana Farber Cancer Institute (DFCI), Boston                         

Department of Haematology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

We present the case of a 62-year-old woman with multiple myeloma (MM) who underwent autologous stem cell transplantation (ASCT), following novel therapy induction1 and whose post ASCT course was complicated by platelet refractory thrombocytopenia.

The patient was otherwise well when she presented with progressive fatigue and malaise. Preliminary evaluation revealed significant renal insufficiency. Serum protein electrophoresis with immunofixation revealed immunoglobulin G (IgG)-lambda monoclonal gammopathy and her disease was ISS stage 3, reflecting her high risk features2. Bone marrow aspiration and biopsy demonstrated an extensive plasmacytosis (90% plasma cells), with normal metaphase cytogenetics3. Skeletal survey was unremarkable apart from osteopenia.

She initially received a combination of bortezomib (Velcade) + thalidomide + dexamethasone (VTD), with cyclophosphamide then added (VTD+C) which resulted in a very good partial response (VGPR). She proceeded to stem cell mobilization with high dose cyclophosphamide shortly thereafter. Her pre-transplant course was complicated by a persistent left-sided otitis media and chronic sinusitis and a generalized tonic-clonic seizure which was attributed to her underlying seizure disorder. The patient proceeded with ASCT utilizing high dose melphalan, however her post-transplant course was complicated by the development of platelet-refractory thrombocytopenia; this was attributed to alloimmunisation4 and tissue typing for platelets was performed on day 10. The patient received HLA matched platelets over 3 days and her platelet count reached 50 x 109 by day 14, with subsequent further hematologic recovery allowing post-ASCT maintenance to follow utilizing bortezomib and lenalidomide.

Six months later, she is in sustained complete remission (CR) and is clinically well.

References:

1. Cavo M, Rajkumar SV, Palumbo A, Lonial S et al. International Myeloma Working Group consensus approach to the treatment of multiple myeloma patients who are candidates for autologous stem cell transplantation. Blood. 2011; 117 (23): 6063-73.

2. Greipp PR, San Miguel J, Durie BG Westin J et al. International staging system for multiple myeloma. J Clin Oncol. 2005; 23 (15): 3412-20.

3. Rajkumar SV. Multiple myeloma: 2012 update on diagnosis, risk stratification and management. Am J hematol. 2012; 87 (1): 78-88.

4. Hod E, Schwartz J. Platelet transfusion refractoriness. Br J Haematol. 2008; 142 (3): 348-60.

Acknowledgment:

Financial support for this project was provided by the Gerry Kelly Fellowship.

  • Presenting Author: Mr Shane Toolan
  • Supervisor: Dr Paul G Richardson
  • Co-Supervisor: Dr Peter O’Gorman

36. Retrospective analysis of extra-medullary disease in patients initially treated for multiple myeloma

Bugler J1,2, Toolan S1, Xie W1, Maglio M1, Mitsiades CS1, Laubach J1, O’Gorman P2, Richardson PG1

1Jerome Lipper Multiple Myeloma Center, Dana Farber Cancer Institute (DFCI), Boston, MA, USA

Department of Haematology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

Extramedullary disease (EMD) is the proliferation of neoplastic plasma cells within soft tissues and parenchymal organs, outside of the bone marrow. In multiple myeloma (MM), 6%-20% of patients develop EMD at relapse (1). Research suggests novel therapeutic agents decrease expression of adhesion molecules, impairing the adherence of myeloma cells to the bone marrow microenvironment, and so might increase the possibility of neoplastic plasma cells proliferating in the extra-medullary compartment (2). Combination-therapy approaches may overcome this effect. Our objective was to examine the relationship between various novel agents used as initial therapy and the prevalence of EMD at first relapse.

We examined patients with MM treated on protocol with various combinations of lenalidmide (R), bortezomib (V), dexamethasone (D) and cyclophosphamide (C) versus bortezomib-monotherapy. Patient’s records were analysed to assess whether EMD was noted on diagnosis or relapse, and if so, whether it was intra- or extra-osseous in nature; histological confirmation of EMD was necessary to distinguish this from other soft-tissue neoplasms.

We evaluated 98 patient records involved in DFCI treatment protocols. For 27 patients treated with CRVD,19% of patients relapsed with EMD; for 32 patients who received RVD therapy,16% relapsed with EMD; while for 39 patients treated with bortezomib-monotherapy, 51% of patients relapsed with EMD, suggesting a lower rate of EMD recurrence for the combination regimens studied (p=0.002). Prevalence of EMD at diagnosis was similar among the three treatment groups (33%, 28%, 28%, respectively, p=0.886).

The data suggests that there may be a significantly higher rate of EMD at relapse when single-agent therapy is used as initial treatment, compared to multi-agent combinations, and further studies of this important phenomenology are warranted.

References:

1. Chen HF, Wu TQ, Li ZY, Shen HS, Tang JQ, Fu WJ, et al. Extramedullary Plasmacytoma in the presence of Multiple Myeloma: Clinical Correlates and prognostic Relevance. Onco Targets and Therapy. 2012;5:329-34

2. Roodman GD. Role of the bone Marrow Microenvironment in Multiple Myeloma. Journal of Bone and Mineral Research. 2002;17(11):1921-1925

Acknowledgement:

This project was supported by the Gerry Kelly Fellowship.

  • Presenting Author: Ms Jane Bugler
  • Supervisor: Dr Paul G Richardson
  • Co-Supervisors: Dr Jacob Laubach and Dr Peter O’Gorman

37. Incidence of picc line-associated venous thrombosis in haematology patients at mater misericordiae university hospital

Iglesias J1, MacMahon P2, Fay M3, Ní Ainle F3, O’Gorman P3, Bacon L3, Fortune A3.

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2Department of Radiology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

3Department of Haematology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

Peripherally inserted central catheters (PICCs) provide a reliable means of venous access for treatment of haematological disease. However, development of venous thrombosis is regarded as a common and potentially serious complication. The reported incidence ranges from 1-22%. This study aimed to determine the incidence of PICC-associated venous thrombosis as a basis for PICC removal or exchange in haematology patients at Mater Misericordiae University Hospital (MMUH).

This observational retrospective cohort study reviewed consecutive haematology patients who underwent placement of one or more PICC lines between September 2011 and July 2013. Occurrence of upper limb venous thrombosis was confirmed by Doppler sonography. Presence and classification of concurrent line-associated bacterial infection was determined by review of blood culture results. Clinical data pertaining to medical history, haematological diagnosis and chemotherapy regimen was obtained from the medical notes.

Data in relation to 101 consecutive PICC placements was collated. The cohort comprised 54 patients, 29 male and 25 female, age range 18-81 years. Twenty one (38.9%) patients received more than one line insertion. Eight individual thrombotic events occurred in total (male n=2, female n=7) representing an overall incidence of 7.9%. Of these 8 patients 6 had a diagnosis of relapsed lymphoma, and 2 acute leukaemia. One patient had a concurrent line infection.

While the incidence of PICC line-associated venous thrombosis in our cohort compares to that reported in recent literature for cancer patients it continues to be cause of considerable morbidity. The apparent association with relapsed lymphoma requires further study.

References:

1. Chopra V, Anand S, Krein SL, Chenoweth C, Saint S. Bloodstream infection, venous thrombosis, and peripherally inserted central catheters: reappraising the evidence. Am J Med. 2012 Aug;125(8):733-41.

2. Yi XL, Chen J, Li J, Feng L, Wang Y, Zhu JA, Shen E, Hu B. Risk factors associated with PICC-related upper extremity venous thrombosis in cancer patients. J Clin Nurs. 2013 May 28.

  • Presenting Author: Mr Jan Iglesias
  • Supervisor: Dr Anne Fortune
  • Co-supervisor: Dr Peter O'Gorman

38. Reconstitution of cortical bone and healing of pathological fracture by targetted anti-myeloma and anabolic bone medical therapy in a patient with a pathological fracture due to multiple myeloma

Ho MZG1, Ting KR2, O’Gorman, P1,2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Department of Haematology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

Plasmacytoma is a discrete, solitary mass of neoplastic monoclonal plasma cells in either bone or soft tissue. We aim to review the effect of Bortezomib on bone formation in myeloma patients and the efficacy of biological therapy in treating Plasmacytoma.

A 70-year-old gentleman presented with sudden onset of persisting “horrendous, sharp, and immobilizing pain” in his right shoulder following a game of golf with a history of drenching night sweats and lethargy without weight-loss. Patient was referred for X-ray and tissue biopsy diagnosing IgG kappa Plasmacytoma. Investigations showed no evidence of Multiple Myeloma.

Baseline X-Ray showed pathological fracture of proximal right humerus with underlying lytic lesion. PET/CT revealed intense hypermetabolism (SUV: 6.1) extending over 7.6 cm.

Recommended treatment for plasmacytoma is radiotherapy. However, in light of the pathological fracture, patient was started on Bisphosphonate and six cycles of VTD (Bortezomib, Thalidomide, Dexamethasone) with a review after two cycles wherein En-Bloc Resection would be considered if there was neither radiological nor symptomatic improvement.

After treatment, shoulder X-Ray showed callus formation secondary to healing pathological fracture. PET/CT revealed significant reduction in metabolic activity (SUV: 2.7) and increased sclerosis.

Lytic bone lesions in Multiple Myeloma rarely heal1. VTD was chosen as unlike radiotherapy which kills osteoblasts2, Bortezomib stimulates bone anabolism. The need for invasive orthopedic surgery or radiotherapy was obviated through targeted therapy which facilitated the eradication of myeloma cells, concomitant stimulation of osteoblastogenesis, and suppression of osteoclastogenesis (CTX-1 fell from 0.447 to 0.070 µg/L after treatment). 

References:

1. Zangari M, Terpos E, Zhan F, Tricot G. Impact of bortezomib on bone health in myeloma: a review of current evidence. Cancer treatment reviews. 2012;38(8):968-80.

2.  Cunha SSD, Sarmento VA, Ramalho LMP, Freitas ACd, Almeida Dd, Tavares ME, et al. Effects of radiotherapy on bone tissues. Radiologia Brasileira. 2007;40(3):189-92.

  • Presenting Author: Mr Matthew Ho-Zhi-Guang
  • Supervisor: Dr Peter O'Gorman
  • Co-Supervisor: Dr Kay Reen Ting

39. The effect of additional antibiotic dosing on sepsis following transrectal ultrasound guided biopsy of the prostate - a retrospective study

Browne E1,2, Cham Boon Way A3, Hegarty N3

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2 Misericordiae University Hospital, Eccles Street, Dublin 7.

3Mater Private Hospital, Eccles St, Dublin 7.

Transrectal ultrasound guided (TRUS) biopsies are one of main tools for the diagnosis of prostate cancer of which sepsis is a major complication requiring hospitalisation. This study aims to investigate the difference in sepsis post TRUS biopsy of the prostate in patients who received different antibiotic regimens.

Patient data was collected from 155 patients under a single surgeon, from Rapid Access Prostate and Outpatients clinics between May 2011 to July 2013. Of the 162 biopsies performed the Rapid Access group (n=71) received antibiotic regimen 1 (750mg Ciprofloxacin pre-biopsy, 750mg post-biopsy), and the Outpatient group (n=90) received regimen 2 (2x750mg Ciprofloxacin pre-biopsy, 4x750mg post-biopsy).

The two groups were compared looking at sepsis rate post-biopsy. The incidence of sepsis was obtained by looking at patient and radiological follow up records.

Of the 162 procedures performed, 1 patient was diagnosed with definitive sepsis. This is a sepsis rate of 0.65%, lower than the estimated incidence of 3%1. This patient belonged to the Rapid Access group who received regimen 1, thereby giving this group a sepsis rate of 1.4%. The group which received regimen 2 had no reports of sepsis and therefore had a sepsis rate of 0%. These results give a p value of 0.3, a not statistically significant but demonstrable difference between the two antibiotic regimens.

Given the high cost of admission for sepsis and the risk of mortality to the patient a change in antibiotic prophylaxis may be considered for those patients under the Rapid Access Prostate Service.

Reference:

1. Ismail M, Saini A, Nigam R. Ciprofloxacin-resistant infection after transrectal ultrasonography-guided prostate biopsy: Should we reassess our practice? BJU Int. 2011 Aug;108(3):305-6.

  • Presenting Author: Ms Eva Browne
  • Supervisor: Mr Hegarty

40. Bioinformatics analysis to identify processed pseudogenes in breast cancer

Ew JV1, Freitas I2, Tarrant F2, O’ Gaora P2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2UCDConway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4.

Pseudogenes are mutated copies of genes which are generally considered non-functional. However, new evidence shows that pseudogenes play a role in tumour biology. Processed pseudogenes (PPGs) arise from reverse transcription of mRNA and transposition into the genome. In this pilot study, breast cancer genomic data were subjected to a bioinformatics workflow to detect potential PPGs.

Whole exome sequencing data from 5 normal breast tissue samples and 5 breast cancer tumour samples were downloaded from The Cancer Genome Atlas repository. Reads were aligned to the human genome using the splice-aware aligner STAR. Uniquely aligned reads which aligned across exon-exon junctions (splitreads) were identified for further processing. The splitread counts in tumour samples were compared to those in normal tissue to identify significant differences in potential PPGs.

Hierarchical clustering analysis showed clear differentiation of normal and tumour samples. From 23,230 genes in the annotated genome, 2,597 were found to harbour aligned splitreads. Using negative binomial distribution testing implemented in DESeq, 30 genes were found to show statistically significant elevations or depressions in the tumour samples (p<0.05).

This study identifies 30 genes as potentially important PPGs in breast cancer genomes. Nevertheless, further lab work needs to be performed to confirm the presence of pseudogenes. Furthermore, increased numbers of samples will provide greater statistical power for detection of PPGs. Pseudogene signature profiles may eventually be correlated to tumour subtypes and disease outcome. In conclusion, this study is the first step to understanding breast cancer pseudogene biology.

Acknowledgement:

With thanks to the Pathological Society of Great Britain and Ireland for funding this project.

  • Presenting Author: Ms Ju Vern Ew
  • Supervisor: Dr Peadar O’Gaora
  • Co-Supervisor: Mr Finbarr Tarrant

41. Is compression plating for humeral shaft fracture associated with lower iatrogenic radial nerve palsy as compared to intramedullary nailing technique? : a review and meta-analysis

Ng CH, Lyons F, McCarthy T 1and Hogan N1

1Trauma Orthopaedics. St James’s Hospital, Dublin 8

Open reduction and fixation with compression plate has advantage of accurate fracture reduction but has disadvantage of additional soft tissue, periosteal stripping and potential radial nerve damage. Intramedullary nail, however, is associated with more shoulder pain and reduced range of motion. Both intramedullary nail and compression plating can achieve similar functional result.

We evaluate the current evidence for using the compression plating (CP) vs. the traditional and gold standard intramedullary nailing (IMN) technique for fixation of the humeral shaft fracture with regards to iatrogenic radial nerve injury.

An electronic literature search was undertaken on the CP and IMN for management of humeral shaft fracture. The search term used was: (Humerus) AND (Fracture) OR (Humeral fracture) AND ((Shaft) OR (Diaphysis)). The authors involved in the study assessed the eligibility of trial for inclusion in the analysis.

A total of 10 studies were included with 436 patients for analysis with RevMan 5.1. In the included studies for IMN approach (n=10), the radial nerve palsy ranged from 0 to 15.4%. In the included studies for the CP approach (n=10), the radial nerve palsy ranged from 0 to 14.2%. The forest plot obtained favours IMN with odd ratios of 1.25 [0.58, 2.68] at 95% Cl.

Compression plating should be considered as a good alternative to intramedullary nail technique for management of humeral shaft fracture. We showed that in additional to having similar non-union and infection rates, compression plating is associated with low rates of iatrogenic radial nerve palsy.

References:

1. Heineman DJ, Poolman RW, Nork SE. Plate fixation or intramedullary fixation of humeral shaft fractures. Acta Orthop 2010; 81 (2): 216-223.

2. Kurup H, Hossain M, Andrew JG. Dynamic compression plating versus locked intramedullary nailing for humeral shaft fractures in adults. Cochrane Database Syst Rev 2011; 15 (6): CD005959.

  • Presenting Author: Mr Chee Hon Ng
  • Supervisor: Mr Niall Hogan
  • Co-supervisor: Mr Thomas McCarthy

42. Investigation of the apoptotic effect of histone deacetylase inhibitor saha (vorinostat) on htlv-1 infected cells

Narulla MS, McCabe Á, Hall WW1, Sheehy N1

1Centre for Research in Infectious Diseases, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Human T-cell lymphotropic virus, type 1 (HTLV-I) infects 10-20 million people worldwide and is the causative agent of adult T-cell leukemia (ATL), a fatal and aggressive CD4+ lymphocyte malignancy.[1] Despite current available treatment options, ATL mortality rate continues to remain high from organ complications, hypercalcemia and immunosuppression-caused infections supporting the need for new therapies.[2] Our study explores the histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA) also known as Vorinostat as a potential anti-cancer agent on four HTLV-I infected cell lines (MT2, C91, ATL-CR and ATL-TH cells) compared to one non HTLV-1 infected cell line (Jurkat cells).

All cell lines were incubated with 0 μM and 10 μM of SAHA. Post treatment, cells were stained and then analyzed by Flow Cytometry for apoptosis and necrosis.

We found that 10 μM of SAHA does not induce high levels of apoptosis amongst HTLV-I infected cells as compared to the control, Jurkat cells (non HTLV-I infected). Also, minimal levels of necrosis are induced in both HTLV-I and non HTLV-I infected cells with SAHA treatment.

In conclusion, we found that HTLV-I infected and ATL cells are more resistant to apoptosis compared to non-infected cells. Hence at 10 μM, SAHA does not represent an effective treatment option for ATL in vitro. Further experiments should be carried out to investigate the effect of SAHA on HTLV-I infected cells with higher concentrations and longer incubation periods or by synergizing SAHA with other anticancer agents.

References:

1. Matsuoka M, Jeang KT. Human T-cell leukaemia virus type 1 (HTLV-1) infectivity and cellular transformation. Nature reviews Cancer. 2007 Apr;7(4):270-80.

2. Tsukasaki K, Tobinai K. Clinical Trials and Treatment of ATL. Leukemia research and treatment. 2012;2012:101754.

  • Presenting Author: Mr Manraj-Singh Narulla
  • Supervisor: Dr Noreen Sheehy
  • Co-Supervisor: Dr Áine McCabe

45. High resolution genetic analysis in paediatric autism spectrum disorder.

Zulkifli ZF1, Regan R1,2,  Anney RJ3, Gallagher L3, Ennis S1

1UCD School of Medicine and Medical Science, University College of Dublin, Belfield, Dublin 4.

2National Children's Research Centre, Crumlin, Dublin.12

 3Department of Psychiatry, Trinity College Dublin, Dublin 1.

Autism Spectrum Disorder (ASD) is a highly heritable disorder of complex and heterogenous aetiology. It is characterised by impaired language and communication skills, poor social reciprocity and repetitive behaviour. The cause of the disorder is largely unknown but there is a strong genetic component. We investigated one type of genetic variation, Copy Number Variation (CNV) in a series of well defined ASD patients and assessed their contribution of the clinical phenotype.

DNA samples from 36 ASD patients were analysed by Array Comparative Genomic Hybridisation (CGH) using the Agilent SurePrint G3 4x180K microarrays protocol, with modifications, and CytoGenomics 2.5.8.11 software. Detected CNVs were filtered based on the following criteria; Hybridisation QA > 8, size >20kb and <2Mb and gene content. Assessment of the likelihood that the CNV was associated with ASD pathogenicity was based on the Database of Genomic Variants (DGV) and a review of published studies.

The array CGH protocol was optimised for 1000ng undigested DNA per sample.  Eight samples passed the quality assessment. 523 CNVs were detected, of which 375 were discounted based on size or genetic content, 148 were categorised as normal variants and 22 were considered rare and were investigated further.  In total, 14 CNVs were identified with genes either previously reported in ASD or were known to be involved in neurodevelopment e.g. PTEN, NPTX2 and GRIP2.   

The clinical heterogeneity of ASD may in part be attributed to the complexity of its genetic underpinnings. Our results highlighted a set of potentially medically relevant CNVs that following validation and further investigation, may help to unravel this complex disorder.

  • Presenting Author: Mr Zul Farhan Zulkifli
  • Supervisor: Dr Regina Regan
  • Co-Supervisor: Dr Sean Ennis

47. Maternal obesity and the outcome of labour

Rowan A, Fennessy AM, O’Higgins A1, Mullaney L, Turner MJ,1

1UCD Centre for Human Reproduction, Coombe Women and Infants University Hospital, Cork St, Dublin 8.

Based on a Body Mass Index (BMI) > 29.9Kg/m2, maternal obesity has been associated with an increase in emergency caesarean section (CS).

Primigravids were recruited between July 2012 and July 2013 after sonographic confirmation of an on-going singleton pregnancy in the first trimester. Weight and height were measured and BMI calculated. Following individual chart reviews by a single researcher, data on 104 women was recorded.  Analysis was carried out using SPSS 20 (IBM Corp. 2012)

Table 1 contains patient characteristics. Of the patients 24% (n=25) delivered by CS; of those 24% (n=6) by elective CS and 76% (n=19) by emergency CS.  Overall 7.4% (n=4) of women in spontaneous labour required an emergency CS vs. 34.1% of all induced women (p=0.001). The rate of emergency CS were 17.3% (n=14) for non-obese women vs. 29.4% (n=5) in obese women (p=0.250). For obese women in spontaneous labour the emergency CS rate were 0% v 46% (n=5) in those with induced labour (p=0.049). Obese women were more likely to require induction 64.7% (n=11) vs. 40.7% (n=33) for non-obese women (p=0.071). Indications for induction were similar between obese and non-obese women.

The analysis shows that labour induction is the most significant factor for emergency CS in both obese and non-obese primigravids. Obese women are at a higher risk for induction of labour and subsequently emergency CS. As CS in obese women carries with it increased risks of morbidity and mortality, initiatives to decrease the CS rate must focus on reducing the incidence of pre-pregnancy obesity.

Characteristics of primigravid women (n=104)

Mean Age (years ± SD)

26.8  ± 5.8

Mean BMI (Kg/m² ± SD)

24.7 ± 5.2

% Obese (n=17)

16%

Gestation at delivery (weeks ± SD)

39.8 ± 1.6

Delivery <37 weeks (n=4)

3.8%

Birth weight( grams ± SD)

3390 ±  549

Spontaneous labour (n=54)

52%

Induced labour (n=44)

42.2%

Pre-labour cesarean (n=6)

5.8%

  • Presenting Author: Ms Ann Rowan
  • Supervisor: Dr Amy O’Higgins
  • Co-Supervisor: Prof Michael J Turner

48. Examination of the caesarean section rate in type i diabetes: use of the robson criteria to allow meaningful analysis of data

Carroll C1, Courtney W1, Higgins M2, Robson M2, McAuliffe F2, Foley M2

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2 UCD Obstetrics and Gynaecology, School of Medicine and Medical Science, University College Dublin, National Maternity Hospital, Holles Street, Dublin 2.

The aim of this study was to investigate the rate of caesarean section (CS) in women with type 1 diabetes (T1DM) attending a large tertiary level unit using the Robson Groups, and whether induction truly confers a higher CS delivery rate. The Robson Groups are internationally recognized as a method of meaningful analysis and comparison of CS rates.

Prospective study in a tertiary level unit with a large combined diabetic – obstetric service. Clinical data was recorded from multiple databases and cases assigned to their relevant Robson Group.

Between January 2005 and December 2012, 300 women with T1DM delivered in this institution. The CS delivery rate in T1DM was 58% (vs institutional CS rate of 19.6%). Nulliparous women undergoing induction of labour (Group 2a) had an increased CS delivery rate (48.5% in nulliparous vs 11.5% in multiparous women [Group 4a]). The vast majority of women with a previous CS delivered by repeat CS (97%). Those who presented in spontaneous labour with T1DM had a 5 to 25-fold increase in delivery by CS compared to the general population, though the absolute numbers are small.

This study analysed CS rates in T1DM women, confirming that high rates of induction of labour results in high rates of delivery by CS, especially in nulliparous women. Women with T1DM previously delivered by CS usually required a repeat CS delivery in subsequent pregnancies. With such high rates of delivery by CS, the case may be for planned elective CS in women with diabetes.

  • Presenting Author: Ms Ciara Carroll
  • Supervisor: Prof Michael Foley
  • Co-Supervisor: Dr Mary Higgins

49. The role of srf in epithelial to mesenchymal transition (emt) in advanced prostate cancer

Dilworth R1, Hanrahan K1, Watson RWG1,2 , Prencipe M1,2

1UCD School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin 4.

2UCD Conway Institute of Biomedical and Biomolecular Research, University College Dublin, Belfield, Dublin 4.

Prostate cancer is the most frequently diagnosed cancer (excluding non-melanoma skin cancers) in Irish males, with incidence rates in Ireland among the highest in Europe.1 Epithelial to Mesenchymal Transition (EMT), a process through which cells lose their epithelial characteristics and acquire a mesenchymal phenotype, has been associated with cancer cell migration, invasion, and therefore metastasis. Recently, Serum Responsive Factor (SRF) was identified as a key transcription factor involved in the development of castrate resistant prostate cancer.2 Given the involvement of SRF in cellular differentiation and cytoskeleton organisation, it was hypothesized that SRF plays a role in the induction of EMT in prostate cancer cells.

Two isogenic prostate cancer cell lines were used to investigate this hypothesis: the LNCaP parental cell line and their castration-resistant LNCaP Abl sub-cell line.  Both lines were transfected with SRF-targeting siRNA and scramble (control) siRNA to knock down SRF. The effectiveness of SRF knockdown and the expression of epithelial and mesenchymal protein markers were assessed by Western blotting. Cell migration was assessed by the scratch assay, and cell invasion was assessed using Matrigel™-coated inserts.

SRF siRNA significantly knocked down SRF protein expression. Invasion assays showed significantly decreased cellular invasion in the parental cell line after SRF knockdown.  Western blotting showed upregulation of E-cadherin, an important epithelial marker, after SRF knockdown in both parental and Abl cells.

Our findings demonstrate that SRF plays a role in inducing EMT-like processes in prostate cancer cells. SRF represents a promising therapeutic target for reducing the metastatic potential of advancing prostate cancer.

References:

1. National Cancer Registry Ireland. Cancer in Ireland 2013: Annual Report of the National Cancer Registry. Cork: National Cancer Registry; 2013. Available from: http://www.ncri.ie/publications/statistical-reports/cancer-ireland-2013-annual-report-national-cancer-registry

2. Prencipe M, Madden SF, O’Neill A, O’Hurley G, Culhane A, O’Connor D, Klocker H, Kay EW, Gallagher WM,  Watson RW (2013) Identification of transcription factors associated with castration-resistance: is the serum responsive factor a potential therapeutic target? Prostate 73(7):743-753

  • Presenting Author: Ms Rachel Dilworth
  • Supervisor: Dr Maria Prencipe
  • Co-Supervisor: Prof R William G Watson

50. Whole blood gene expression and alternative splicing in sickle cell anemia adults with high and low ldh sub-phenotypes

Murphy C1, Wang Z2, Diaw L2, Darbari D2, Quinn M2, Barb J2, Munson P2,  Raghavachari N2, Klings E2, Taylor VI JG2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Genomic Medicine Section, Hematology Branch, NHLBI, National Institute of Health, Building 10, Room 5N101/103, 10 Center Drive, MSC-1476, Bethesda, MD 20892-1476.

Sickle cell anemia (SCA) is a common monogenic disease characterised by a wide spectrum of phenotypic variation.  High serum LDH is associated with vascular complications in SCA including leg ulceration, tricuspid regurgitation, less avascular necrosis and possibly early mortality.  These observations support the hypothesis that steady state LDH may be a biomarker for SCA sub-phenotypes.

We utilized whole blood gene expression profiling to further assess the LDH’s validity as a phenotypic biomarker.  SCA subjects with steady state LDH measurements had RNA isolated from peripheral blood PAX gene tubes.  Ten males and 10 females with high LDH were age and sex matched to 20 subjects with low steady state LDH and 20 normal volunteers.  Genome-wide expression profiling used Affymetrix Exon Arrays. Paired expression analyses were performed. 

Genes selected for validation had statistically significant differences in alternative splicing between groups.  Reverse transcribed mRNA was PCR amplified to validate alternative splicing for eight genes from high LDH, low LDH and control groups. Sanger sequencing confirmed alternative splicing.

There was no alternative splicing in ABCG2 (array p=1.58x10-2) or UBXN10 (array p=1.62x10-2). Alternative splicing was validated for ARL4A (array p=1.21x10-2), a GTP binding protein family member(1).

The absence of alternative splicing in 2 genes highlights the importance of validation of findings using hybridization based microarrays.  Alternative splicing in ARL4A is associated with high and low LDH groups in adults with SCA.  Further work is needed to determine if alternative ARL4A transcripts explain wide phenotypic variation in SCA.

Reference:

Li CC, Wu TS, Huang CF, Jang LT, Liu YT, You ST, Liou GG, Lee FJ.. GTP-binding-

defective ARL4D alters mitochondrial morphology and membrane potential.. PLoSOne 2012;8(e43552):.http://www.ncbi.nlm.nih.gov/pubmed/22927989 (accessed ).

  • Presenting Author: Ms Catherine Murphy
  • Supervisor: Prof James Taylor
  • Co-Supervisor: Dr Zhengyuan Wang

51. Investigation of the localisation of cysteine rich transmembrane bmp regulator 1 (crim-1) in the hypoxic lung

Boyle N, Ali N, McLoughlin P1,2, Costello CM. 1,2

1UCD School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin 4.

2UCD Conway Institute of Biomedical and Biomolecular Research, University College Dublin, Belfield, Dublin 4.

Sustained pulmonary hypoxia is a common complication of chronic lung diseases leading to the development of pulmonary hypertension (PH). The molecular pathogenesis of this process remains poorly understood, however reduced BMP signalling, mediated by the upregulation of BMP antagonists such as CRIM-1, may be important. The aim of this study was to examine the cellular localisation of CRIM-1 in the hypoxic lung.

Western Blotting was used to validate the CRIM-1 antibody (ab135326; Abcam). CRIM-1 localisation was then examined by performing immunohistochemistry on n=7 hypoxic mice (2 days at 10%O2) using a purified rabbit IgG (ab27478; Abcam) as a negative control.

A single band at the expected molecular weight (114kDa) was observed by Western Blot analysis. An initial dilution series indicated that a working concentration of 50µg/ml of antibody was optimal. Immunohistochemistry identified CRIM-1 expression in the endothelial cells of small and large blood vessels and the smooth muscle layer of larger arteries. CRIM-1 was also expressed in pulmonary epithelium and Type 2 pneumocytes. CRIM-1 was present in cells extravasating from blood vessels into alveolar spaces notably granular leukocytes (eosinophils, basophils) however expression was minimal in non-granular leukocytes or erythrocytes.

We report, for the first time, that CRIM-1 is expressed in a model of hypoxic lung disease. Protein expression was observed in blood vessels and infiltrating cells. This is striking given that vascular remodelling and inflammation are hallmarks of the disease condition. Further studies are required to establish if CRIM1 may have a role to play in the pathogenesis of PH.

  • Presenting Author: Ms Niamh Boyle
  • Supervisor: Dr Christine Costello
  • Co-Supervisor: Prof Paul McLoughlin

56. Il6 targeting in triple negative breast cancer (tnbc) enhances the efficacy of the anti-proliferative chemotherapeutic paclitaxel (taxol®).

O’ Malley C1, Gorzel K2, O’Reilly E2, McCann A1,2

1UCD School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin 4.

2UCD Conway Institute of Biomedical and Biomolecular Research, University College Dublin, Belfield, Dublin 4.

Triple Negative Breast Cancer (TNBC) is a subset of breast cancer immunohistochemically negative for the expression of the oestrogen receptor, progesterone receptor and the tyrosine kinase receptor HER-2. TNBC is associated with poor prognosis, early relapse and a significantly shorter survival rate following disease recurrence when compared to non-TNBC cancers [1]. This aggressive subset of breast cancer relies critically on the cytokine IL-6 to drive tumour growth and suppress apoptosis [2].

The aim of this research was to (a) investigate what impact targeting the IL-6 pathway in triple negative breast cancer cells has on IL6- mediated growth and (b) investigate how this affects the resultant cellular response to the anti-mitotic chemotherapeutic drug Paclitaxel (Taxol®).

The cytotoxic effects of the anti IL-6 monoclonal antibody MAB206 and Paclitaxel on MDA-MB-231 TNBC cells was assessed using the MTT assay.  MDA-MB-231 cells were seeded (5000 cells/well) in 96-well plates in culture medium containing the Paclitaxel IC50 concentration in one treatment and then Paclitaxel and MAB206 in combination. Levels of the cytokine IL6 were analysed using an IL-6 ELISA (Immuntools ®). Cell viability decreased when MDA-MB-231 cells were treated their Paclitaxel IC50 values in combination with MAB206 compared to Paclitaxel treatment alone. This was concomitant with clinical data that shows low IL-6 expression is associated with a better disease free survival in breast cancer patients (DFS).

The results demonstrate that the combination of an IL-6 monoclonal antibody and Paclitaxel attenuated cellular proliferation in a synergistic manner and enhanced the cytotoxic effects of the chemotherapeutic Paclitaxel.

References:

1. Rodler E, Korde L, Gralow J, et al. Current treatment options in triple negative breast cancer. Breast Disease. 2010, vol. 32(1-2):99-122.

2. Hartman Z, Poage G, Hollander P, et al. Growth of Triple Negative Breast Cancer Relies upon Coordinate Autocrine Expression of the Proinflammatory Cytokine IL-6 and IL-8. Cancer Res. 2013; vol 73(11); 1-11. 

  • Presenting Author: Mr Cian O’Malley
  • Supervisor: Dr Amanda McCann
  • Co-Supervisor: Ms Karolina Weiner Gorzel

 

57. An investigation of cellular hypoxia and its influence on circadian gene expression

Jones-O’Connor M1, Altman BJ2, Hsieh A2, Dang CV2

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, PA 19104, USA

The heterodimeric transcription factor HIF-1 has numerous known target genes, with wide-ranging cellular effects (1). However, little is known about the effects of hypoxia and HIF-1 on circadian genes. In addition, the strict regulation of its α-subunit means experimental protein detection can be erroneous. Our aims were: (a) to find a method of detection of HIF-1α that provided consistent results and (b) to examine the influence of hypoxia on circadian and metabolic genes.

HIF-1 expression was induced in U2OS osteosarcoma cells with both native hypoxia and a hypoxia mimetic, 1mM dimethyloxalylglycine (DMOG). Western blots were used for HIF-1α detection. Cells were lysed indirectly, according to the Qiagen protocol, or directly on-plate. Antibodies were sourced from BD Biosciences. We used real-time quantitative polymerase chain reaction (qPCR) to analyse the levels of gene expression.  Copy threshold values from comparative quantitation (ΔΔ-Ct) were analysed in comparison with a housekeeping gene, B2M.

DMOG treatment with direct cell lysis yielded the clearest immunoblots. Indirect protein extraction yielded lower HIF-1α levels. Negligible levels of HIF-1α were visible from native hypoxia treatment. The experimental data from qPCR suggests that DMOG treatment affects the cellular expression of named circadian and metabolic genes.

Our work suggests DMOG paired with indirect cell lysis as a model for inducing and detecting HIF-1α. The findings relating to gene expression need further investigation to elucidate a mechanism for these effects. Similar investigations using native hypoxia should be completed to confirm that these are in vivo effects and not solely effects of DMOG treatment.

Reference:

Liu W, Shen SM, Zhao X, Chen GQ. Targeted genes and interacting proteins of hypoxia inducible factor-1. Int J Biochem Mol Biol. 2012; 3(2): 165–178

  • Presenting Author: Ms Maeve Jones-O’Connor
  • Supervisor: Dr Chi Van Dang
  • Co-Supervisor: Dr Brian Altman

58. The role of epithelial mpges1 in her2/neu driven mammary tumours

Smith N1, Markosyan N2, Chen E2, Ndong V2, Smyth E2, Fitzgerald G2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Institute of Translational Medicine, University of Pennsylvania, Philadelphia.

Regular use of non steroidal anti-inflammatory drugs (NSAIDs), non selective cyclo-oxygenase (COX) inhibitors, causes a reduction in breast cancer incidence (1). COX-2 induces expression of microsomal prostaglandin E2 synthase-1 (mPGES1), which is involved in generation of prostaglandin (PG) E2. NSAIDs and current treatments targeting COX-2 specifically are unsatisfactory due to their side effect profiles (2). Our aim is to see if inhibition of mPGES1 specifically in mammary epithelial cells (MEC) affects tumour development, in which case this enzyme could be a target for research into novel breast cancer treatment.

mPGES1 wild type (WT) and knock out (KO) mice were crossed with mice expressing activated HER2/neu oncogene under mmtv promoter. Mice were genotyped using Polymerase Chain Reaction (PCR). Lentiviral shRNA delivery was used to generate knock down (KD) mPGES1 in HER2/neu transformed breast cancer cells and injected into WT mice to examine orthotopic tumour growth. Mammary glands were harvested from 22 week old mice to compare tumour development. Unpaired student t-test was used.

Our findings indicated that there was a significant decrease in tumour volume in mPGES1 KD mice each week post-injection. (p=<0.05, n=6). We also saw that tumour multiplicity was decreased significantly in mPGES1MEC KO mice (WT = 4.167 ± 0.9458 N=6, KO = 0.5000 ± 0.2887 n=4, p=0.028).

This work shows that removal of mPGES1 from MEC decreases tumour multiplicity and slows the growth of orthotopic tumours. Our results indicate that knocking out mPGES1 in MEC appears protective against mammary tumours in mice.

Acknowledgment:

Funded by UCD North American Medical Graduates Association and the University of Pennsylvania.

References:

1. Harris, R.E et al. (2003). Breast cancer and nonsteroidal anti-inflammatory drugs: prospective results from the Women's Health Initiative. Cancer Res. Sep 15;63(18):6096-101

2. Howe, L.R et al. (2002). Celecoxib, a selective cyclooxygenase 2 inhibitor, protects against human epidermal growth factor receptor 2 (HER-2)/neu-induced breast cancer. Cancer Res. Oct 1;62(19):5405-7

  • Presenting Author: Ms Naomi Smith
  • Supervisor: Dr Nune Markosyan

59. Analysis of caesarean section rates in gestational diabetes: use of the robson groups to allow meaningful examination of data

Courtney W, Carroll C, Courtney D, Higgins M, Robson M, McAuliffe F1, Foley Mq

1UCD Obstetrics and Gynaecology, UCD School of Medicine and Medical Science, University College Dublin, National Maternity Hospital, Holles Street, Dublin 2.

This was a prospective study examining patients who attended this large tertiary level obstetric unit from January 2005 to December 2012. It aimed to examine the rate of caesarean section (CS) in gestational diabetes (GDM) patients as classified by the Robson Criteria, an internationally recognised mode of analysis.

Over this time, 61,116 women delivered infants greater than 24 weeks gestation in this unit, of which, 1126(1.8%) were diagnosed with gestational diabetes.

During this time, the overall hospital CS rate was 19.6%, compared to 32.5% rate in the GDM population. Nulliparous women undergoing induction of labour (IOL)(Group 2a) had a 63% increase in CS delivery rate over the non-GDM population; multiparous women (Group 4a) had a 20% increase over the non GDM population. By comparison, those with GDM who went into spontaneous labour incurred a 19% (nulliparous) and 1.2% (multiparous) increase in CS rates over the non-GDM population. The majority of women with a previous CS delivered by repeat CS (71%).

Overall, the GDM cohort had a 65% increased rate of delivery by CS compared to the general population (p<.0001). The highest rate of CS was amongst women with a previous CS. Those undergoing IOL (groups 2a and 4a) were more likely to need CS than those who went into SL. This data supports a policy of thoughtful IOL in women with GDM and realistic counselling of GDM women, particularly those in the nulliparous category, regarding the possible outcomes of IOL. In this instance, elective CS could be meaningfully discussed. 

  • Presenting Author: Mr William Courtney        
  • Supervisor: Prof Michael Foley
  • Co-Supervisor: Dr Mary Higgins