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Centre for Clinical Pharmacology and Therapeutics

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Research

Research Interests

The Centre for Clinical Pharmacology and Experimental Therapeutics is a collection of scientist and clinician/scientists working on topics ranging from imaging to cardiovascular pharmacology; from development of the cardiovascular system to inflammation research, cell biology and genetics. The central theme is excellence in research and teaching with a track record in fostering independent scientific careers and mentoring of doctoral training students. 
 

Professor Derek Gilroy

Research interests

We are interested in understanding the endogenous pathways that bring about the resolution of acute inflammation.

gilroy-fig1

Resolution of acute inflammation is no longer

considered a passive event where the response simply fizzles out, but an active process whose dysregulation may explain the complex aetiology of many chronic inflammatory diseases.

Our overall objective, therefore, is to identify pro-resolution factors that help switch inflammation off and to develop drugs based on their mode of action in order to drive on-going, chronic inflammatory diseases down a pro-resolution pathway.

We contend that this represents a novel approach to understanding and treating diseases driven by on-going inflammation. For detailed information on the Gilroy laboratory, its members, publications and research objectives go to derekgilroy.com
 

Technology in the Gilroy lab

Cantharidin skin blister

Following topical administration to the skin, cantharidin, a protein phosphatase inhibitor, induces atraumatic acantholysis and blister formation. This property is used to create defined skin windows which can be maintained for several days helping to profile the onset and resolving phases of inflammation.

Suction skin blister

In this model skin window is created by placing a suction cup on the fore-arm attached to negative pressure equipment. In addition to serving as a model of tissue injury, it can be modified to study in vivo cell function and effect of drugs on the onset and resolution phase.     

Systemic inflammation in humans

In order to investigate critical illness-induced immune dysfunction (CIIID) the group has designed, and is currently characterizing a robust clinically relevant 'two-hit' human pre-clinical model of systemic inflammation (mirroring sepsis, trauma, burn injury or major surgery) coupled with a biologically valid secondary infective challenge (mimicking hospital-acquired infection, HAI).

Intravenous endotoxaemia provides the primary insult, whilst the 'second hit' is induced by gram-positive bacterial challenge to suction-blister induced skin windows replicating central or peripheral venous access, surgical or traumatic wound infection, the second most common cause of HAI on intensive care (ICU).

Application of specially developed blister wells uniquely allows in-vivo analysis of the local inflammatory and immune response (cellular component [via flow cytometry], humoral mediators [via multiplex cytokine ELISA, with lipidomics via mass spectroscopy]) and its functionality at the infective site (fluorochrome-tagged bacterial phagocytosis).

Plasma and whole blood assays ex-vivo (stimulation and live bacterial killing) are complementary to the above, describing systemic disturbance in inflammo-immune homeostasis. Varying the time-point of the 'second hit' facilitates exploration of the time-course (and mediators) of immunosuppression, and sequential blister exudate aspiration the examination of different phases of inflammation.

In sum, the model provide a unique, controlled, adaptable translational platform upon which to dissect the molecular, cellular and genetic drivers of CIIID in humans, and a means of evaluating the efficacy of immunomodulatory therapies.

Experimental models of inflammation

The Gilroy lab has extensive experience and expertise in a number of in vivo inflammatory models, which is shown in the group's publication history.   These included the air pouch, zymosan elicited peritonitis, immunisation, extravasation, monoarthritis, polyarthritis, inflammation, pharmacokinetics, airway inflammation, and liver inflammation and fibrosis. 

We use a range of stimuli to induce inflammation, including zymosan, live bacteria, carrageenan, and methylated BSA. We also have expertise in cardiac punctures, dissection of a variety of tissues for subsequent analysis by FACS, qRT-PCR, primary cell culture and proliferation studies.

We maintain a good working relationship with the staff in our Biological Services Units to ensure our experimental systems are well cared for and monitored during the duration of our experiments, and always keep in mind the 3'R's during experimental design.

In our pursuit of understanding B lymphocyte transdifferentiation, we have crossed transgenic B6.mb-1.iCre mouse harbouring a Cre recombinase gene integrated in the mb-1 locus (encodes the Ig-a signalling subunit of the B cell antigen receptor) with Rosa26R-eYFP (enhanced Yellow Fluorescent Protein) reporter mouse to obtain a B6.mb-1.iCre-Rosa26R-eYFP mouse. Hence, in Rosa26R-eYFP expression of florescent protein is dependent upon Cre recombinase, whose expression in B6.mb-1.iCre is restricted to B cell lineage enabling permanent labelling of these cells.

Therefore B6.mb-1.iCre-Rosa26R-eYFP mice are a valuable tool for tracing of the B-1 cell-derived MΦs.

 

Professor Mark Lythgoe

lythgoe-fig1

Research interests

Professor Mark Lythgoe is the Founder and Director of the UCL Centre for Advanced Biomedical Imaging (CABI), which is a new multidisciplinary research centre for experimental imaging.

Mark has established CABI as a world-class preclinical research centre, which offers researchers across the UK a range of platform imaging technologies to test their hypothesis in vivo.

The Centre now hosts ten state-of-the-art imaging modalities: MRI, photoacoustic imaging, ultrasound, bioluminescence and fluorescence imaging, PET and SPECT/CT as well as confocal endoscopy, optical projection tomography and light sheet imaging.

Mark also leads Biomedical Imaging at the new Francis Crick Institute, which has a strategic partnership with CABI to develop a joint, world-class imaging facility (opening 2015).

Mark has been awarded £43 million for his programme of imaging research and leads a group of 48 researchers.

Mark has published over 200 papers including publications in Nature, Nature Medicine and The Lancet. He is co-Director of the UCL EPSRC Centre for Doctoral Training in Medical Imaging.

In 2013 Mark was elected an Honorary Fellow of the British Science Association for his work in science and society and he has received the Davies Medal for a significant contribution to imaging science.

During Mark's tenure as Director of the Cheltenham Science Festival, it has become one of the largest science festivals in the world.

front cover Nature Photonics

front cover neuroimage

Keywords

Magnetic resonance imaging; nuclear imaging; bioluminescence; fluorescence imaging; photoacoustic imaging

 

Dr. Alastair O'Brien

Alastair O'Brien

Research interests

My research is directed towards combating infection in acute decompensation liver disease patients (AD). Having documented the extremely poor outcome for AD in intensive care (ICU), I believe the greatest gain will be achieved by tackling the major cause of ICU admission from the ward, infection, which is due to immune suppression. I have combined molecular science, clinical studies and large scale data analysis in order to pursue this. My research group consists of a post-doctoral research fellow, 3 PhD students, a data manager, clinical research nurse and clinical trial team at the UCL Comprehensive Clinical Trials Unit. This is funded by The Wellcome Trust, Department of Health,  Medical Research Council and Biotechnology and Biological Sciences Research Council.
My work identified the first molecular mechanism (elevated Prostaglandin E2, (PGE2) to explain immune suppression in AD patients (Nature Medicine). Importantly I identified that albumin could antagonise PGE2 which led to my randomised controlled trial (ATTIRE) to test whether albumin can prevent infection and improve mortality (www.attiretrial.com). This is the largest interventional study currently recruiting worldwide in patients with advanced liver disease at 30 hospitals (866 patients) throughout the UK. This will complete November 2018.

obrien-fig

Selected Publications

    1. China L, Skene S, Shabir Z, Maini A, Sylvestre Y, Bennett K, Bevan S, O'Beirne J, Forrest E, Portal J, Ryder S, Wright G, Gilroy DW & O'Brien AJ. Administration of Albumin Solution Increases Serum Levels of Albumin in Patients with Chronic Liver Failure in a Single-arm Feasibility Trial. Clin Gastroenterol Hepatol. 2017 Sep 12. pii: S1542-3565(17)31098-4. [Epub ahead of print]
    2. China L, Maini A, Skene S, Shabir Z, Sylvestre, Colas R, Ly L, Becares Salles N, Belloti V, Dalli J, Gilroy DW & O'Brien AJ. Albumin Counteracts Immune-suppressive Effects of Lipid Mediators in Patients With Advanced Liver Disease. Clin Gastroenterol Hepatol. 2017 Aug 28. pii: S1542-3565(17)30993-X. doi: 10.1016/j.cgh.2017.08.027. [Epub ahead of print]
    3. ATTIRE: Albumin To prevenT Infection in chronic liveR failurE: study protocol for a single-arm feasibility trial. China L, Muirhead N, Skene SS, Shabir Z, De Maeyer RP, Maini A, Gilroy DW, O'Brien AJ. BMJ Open. 2016 Jan 25;6(1):e010132.
    4. Shallcross L, O'Brien A. Antimicrobial resistance in liver disease: better diagnostics are needed. Lancet Gastroenterol Hepatol. 2017 Mar;2(3):151-153.
    5. Immunosuppression in acutely decompensated cirrhosis is mediated by prostaglandin E2. O'Brien AJ, Fullerton JN, Massey KA, Auld G, Sewell G, James S, Newson J, Karra E, Winstanley A, Alazawi W, Garcia-Martinez R, Cordoba J, Nicolaou A, Gilroy DW. Nature Medicine. 2014;20(5):518-23.
    6. O'Brien AJ, Welch CA, Singer, M, Harrison DA.  Prevalence and outcome of cirrhosis patients admitted to UK intensive care. A comparison against dialysis-dependent chronic renal failure patients. Intensive Care Med. 2012; 38:991-1000.

    Keywords

    Liver cirrhosis, Immune suppression, Eicosanoids, Albumin
     

    Dr. Nephtali Marina-Gonzalez

    Biography

    Dr Marina-Gonzalez is a cardiovascular neuroscientist based in the UCL Department of Clinical Pharmacology.

    He studied Medicine in Mexico where he did a Masters and a PhD in Neuroscience.

    He completed postdoctoral training in UCL and Cambridge before obtaining a British Heart Foundation Intermediate Basic Science Research Fellowship to study the interaction between circulating adipokines and peripheral and central chemoreceptors in the development of obesity-related hypertension.

    He is organiser of the Heart and Circulation module for 3rd year undergraduate medical students.

    Research Interests

    In my lab we study the role of the autonomic nervous system in the pathogenesis of hypertension and heart failure.

    We employ a wide array of cutting-edge techniques, including optogenetics and pharmacogenetics for selective activation or inhibition of astrocytic and neuronal networks that control cardio-respiratory activity.

    This involves active collaboration with Prof Alexander Gourine (UCL),  Profs Sergey Kasparov and Prof Julian Paton (University of Bristol), Prof Andyrew Tinker (William Harvey Heart Centre), Prof Michael Spyer (UCL), Dr Vidya Mohamed-Ali (ADL, Qatar) Dr Stefan Trapp (Imperial College London) and Prof Gregory Funk (University of Alberta).

    In 2011, I was awarded a British Heart Foundation Intermediate Research Science Fellowship to investigate the role of peripheral and central adipokine sensors in the development of obesity-related hypertension.

    Teaching Summary

    In 2013 I became course organiser of the Heart & Circulation module
    Lecturing in the following courses: Circulation & Breathing, Heart & Circulation, Respiration in health and disease.

    Other academic activities: Workshops, practicals and tutorials for 1st and 3rd years MBBS students.

    In 2010 I was awarded a Postgraduate certificate in learning and teaching in higher education by the Centre for Advancement of Learning and Teaching, UCL. 

    Selected publications

    1. Nephtali Marina, Natalie D. Bull & Keith R Martin. A semi-automated targeted sampling method to assess optic nerve axonal loss in a rat model of glaucoma. Nature Protocols 2010 Sep;5(10):1642-51. Citations: 12. Impact factor: 11.74
    2. Alexander V. Gourine, Vitaliy Kasymov, Nephtali Marina, Feige Tang, Melina F. Figueiredo, Samantha Lane, Anja G. Teschemacher, K. Michael Spyer, Karl Deisseroth, and Sergey Kasparov. Astrocytes Control Breathing Through pH-Dependent Release of ATP. Science. 2010; 329(5991): 571-575. Citations: 144. Impact factor: 31
    3. Nephtali Marina, Ana PL Abdala, Alla Korsak, Annabel E Simms,  Andrew M Allen, Julian FR Paton and Alexander V Gourine. Control of sympathetic vasomotor tone by catecholaminergic C1 neurones of the rostral ventrolateral medulla oblongata. Cardiovascular Research 2011; 91(4):703-710. Citations: 12. Impact factor: 5.94
    4. Nephtali Marina, Melina Figueiredo, Vitaliy Kasymov, Feige Tang,  Vidya Mohamed-Ali, Anja G Teschemacher, Sergey Kasparov and Alexander V Gourine. Purinergic signalling in the rostral ventro-lateral medulla controls sympathetic drive and contributes to the progression of heart failure following myocardial infarction in rats. Basic Research in Cardiology. 2013: 108(1) 317 Citations: 2. Impact factor: 5.9
    5. Nephtali Marina, Ana P Abdala, Stefan Trapp, Aihua Li, Eugene E. Nattie, Jeffrey C. Smith, Julian FR Paton, Alexander V Gourine. Essential Role of Phox2b-Expressing Ventrolateral Brainstem Neurons in the Chemosensory Control of Inspiration and Expiration. Journal of Neuroscience 2010 Sep 15;30(37):12466-73. Citations: 27. Impact factor: 7.12
    Dr. Reecha Sofat

    Reecha Sofat is a Clinical Pharmacologist practising general internal medicine, cardiovascular risk and stroke as a sub-speciality.  She is deputy Clinical Divisional Director of Medical Specialities at ULCH, the largest Division spanning all medical specialities. She chairs the ULCH Formulary Committee and is Vice Chair of the Regional Joint Formulary Committee of North Central London.

    Research

    Reecha's group is interested in harnessing routinely collected health care data to improve quality and safety of care but also to be able to generate knowledge and hypotheses by linking this information to next generation -omics technologies. The aim is that this will improve outcomes not only for individuals but also will be of public health benefit.  

    She is the PI of a group of studies, CORUM that aims to embed research into routine health care. By nature clinical care phenotypes patients and the sub-types of disease in great detail, be it through the written clinical record, routine blood tests and imaging that are carried out. Linked to next generation technologies including genomics, transcriptomics, proteomics and metabolomics these data could allow understanding aetiology of disease and specifically sub-types of disease. Moreover, because this research is embedded within the health care system, linking these data to the NHS number allows long term follow up and establishment of prognostic cohorts. The scale allows generation of knowledge that is of public health value and the focus on disease sub-types linked to next generation technologies allows precision medicine to become a reality. This framework will facilitate the next generation of randomised controlled trials for disease sub-types, but importantly by feeding this information back into the health care system it aims to improve the quality and safety of care of patients now and in the future. The first of the studies, SIGNUM, focuses on stroke. This is currently the highest recruiting study nationally. Other studies will focus on heart failure (CARDIUM), atrial fibrillation (ATRIUM), venous thromboembolism (COAGULUM) and conditions that may be related to vascular outcomes (VASCULARUM). These will open in early 2018.

    By nature the work is multi-disciplinary and includes collaborations with clinical services and scientist, health care delivery organisations, epidemiologists, computational biology and engineering amongst others.

    Dr. Ines Pineda-Torra

    Our group aims to better understand how genes are modulated by a subclass of lipid activated nuclear receptors (the LXRs) in the context of atherosclerosis to help develop improved LXR-based therapies for the treatment of this and other vascular and inflammatory diseases.

    Summary of Research

    Atherosclerosis is the hardening of major blood vessels resulting from the accumulation of cholesterol. This lesion eventually causes the majority of cardiovascular diseases, the leading cause of mortality worldwide. Abnormally high levels of cholesterol are a risk factor for atherosclerosis and in recent years inflammatory pathways have also been shown to extensively contribute to the development of atherosclerosis. Thus, molecules providing a tight control of cholesterol levels with additional anti-inflammatory properties represent ideal anti-atherosclerotic drugs. The Liver X receptors (LXRs) behave like cholesterol sensors: they are lipid-activated transcription factors that control the expression of key genes to tightly maintain cholesterol levels. Moreover, LXR activation inhibits inflammation in diverse settings. These actions contribute to the reduced atherosclerosis observed in several experimental models treated with LXR ligands which is why LXRs are considered promising targets to develop therapies against atherosclerosis.

    Elucidating the signaling pathways that regulate its activity is critical to our understanding of LXR function. My group aims to better understand how genes are modulated by LXR in the context of atherosclerosis. We are particularly interested in pathways regulated by LXR in a non-canonical fashion, e.g. by modulating post-translational modifications of the receptor (such as phosphorylation) or through crosstalk with other transcriptional and/or signaling pathways. We are generating unique tools and experimental models to investigate whether changes observed in cultured cells are translated in vivo in a pathological context.

    Main Research Projects

    Impact of LXRalpha phosphorylation on vascular inflammation and atherosclerosis

    Previous work identified a phosphorylation site in LXRalpha that is important to selectively inhibit the transcription of specific LXR target genes and expands the range of LXRalpha-responsive genes in macrophages (Pineda-Torra I, Mol Cell Biol 2008). We are now investigating whether changes in the phosphorylation status of LXR affect macrophage inflammation levels and the development of atherosclerosis. To this end, we have generated a novel experimental model that expresses a phosphorylation-deficient mutant of the receptor (LXRa-S196A) in a cell/tissue-selective manner in an atherosclerotic background. This innovative approach is at the forefront of nuclear receptor and atherosclerosis research and may lead to the development of novel and more efficient LXR activators to reduce atherosclerosis.

    Funded by MRC New Investigator Award

    Role of LXRalpha phosphorylation in lung inflammation

    LXR agonists anti-inflammatory properties have also been shown in models of acute lung injury. LXRs are expressed in several alveolar cell types were they inhibit cytokine expression, diminish reactive oxygen species and reduce neutrophil recruitment.  We now aim to

    1. Investigate LXR?lpha phosphorylation in lung as a means to modulate its activity alternatively to ligand activation
    2. Examine its impact on severe lung inflammation and
    3. Uncover how changes in LXR?lpha phosphorylation impact lung target gene expression during persistent severe inflammation.

    We will employ our unique experimental model expressing a mutation that abolishes LXRa phosphorylation.

    Funded by Grand Challenges PhD Studentship

    Impact of interferon signaling on LXR anti-atherosclerotic actions

    Our group at UCL (Pourcet B., Circulation Res 2011) demonstrated that LXR induces the expression of an important anti-inflammatory protein (arginase 1) in a novel way that involves two other transcription factors (IRF8 and PU.1). We now have evidence that in addition, other novel LXR target genes affecting pathways relevant to atherosclerosis are dependent on IRF8 for their response to LXR ligands. Thus we are exploring how IRF8 impacts the anti-atherosclerotic actions of LXR ligands by generating a novel experimental model deficient in IRF8 on an atherosclerotic background and determine how IRF8-dependent LXR regulated genes are regulated by the LXR/IRF8 crosstalk. We additionally aim to better understand how LXR ligands induce IRF8 interactions with other proteins to regulate LXR target gene expression.

    Funded by a British Heart Foundation Project Grant

    Technology in the Pineda-Torra Laboratory

    Cell culture:

    • Isolation of bone marrow-preparation of murine (peritoneal and bone marrow derived) macrophages, isolation and culture of human macrophages
    • murine and human macrophage-like cell lines (RAW 267.4, THP-1 cells)
    • other cells lines commonly used to overexpress proteins and study nuclear receptor activity (Hek293, Cos)

    RNA expression:

    • Real time quantitative RT-PCR
    • RNA interference
    • Microarray analysis of whole genome gene expression

    Assays commonly used to study nuclear receptor/transcription factor  activity:

    • Luciferase reporter assays
    • Cloning of promoters /enhancers into reporter vectors
    • Promoter/enhancer mutagenesis
    • Chromatin immunoprecipitation assays to study transcription factor binding to chromatin

    Protein:protein interaction assays

    • Immunoprecipitation assays
    • GST pulldowns

    Protein expression:

    • Immunoblotting
    • ELISAs
    • FACs

    Phosphorylation assays:

    • Generation of phosphospecific antibodies
    • Detection of phosphorylation with phosphospecific antibodies (immunoblotting, immunocytochemistry and immunohistochemistry)
    • Kinase assays

    Atherosclerosis:

    • Foam cell formation in vitro
    • Dissection of aortic roots and arches, thoracic and abdominal aortas
    • Oil Red O staining of lipids in foam cells (in vitro, atherosclerotic plaques)
    • Immunohistochemistry to assess inflammatory markers and  proliferation within atherosclerotic plaques
    • Generation of experimental in vivo models

    Present and Past Lab members

    Ines Pineda-Torra (Group Leader)
    i.torra@ucl.ac.uk

    Oscar Pello (Senior Research Associate)
    o.pello@ucl.ac.uk

    Matthew Gage (Senior Research Associate)
    m.gage@ucl.ac.uk

    Natalia Becares Salles (PhD student)
    natalia.becares.13@ucl.ac.uk

    Benoit Pourcet
    (currently at the Institut Pasteur de Lille, France benoit.pourcet@inserm.fr)

    Main National and International Collaborations
    Michael Garabedian, Prof in Urology and Microbiology, NYU Langone Medical Center, USA
    Edward Fisher, Leon H Charney Prof of Cardiovascular Medicine, NYU Langone Medical Center, USA
    Sidney Morris, Prof Emeritus in Microbiology and Molecular Genetics, University of Pittsburgh
    Keyko Ozato, Prof in Universityof Maryland College Park
    Xiaoyu Hu, Assistant Professor of Immunology, Department of Medicine, Weill Cornell
    Antonio Castrillo, Group Leader in the Department of Metabolism and signaling, Instituto de Investigaciones Biomedics "Alberto Sols"
    Ian Zachary, Prof in Cardiovascular Biology
    Liz Jury, Principle Research Associate, Centre for Rheumatology,  UCL




     

    Dr. Simon Yona

     

    yona-fig1

    Research interest

     Our lab is interested in the inflammatory response. Inflammation is an organism's defensive response to foreign bodies and injury. While this is beneficial, it can go into disarray resulting in tissue damage and the development of chronic inflammatory conditions - including arthritis, fibrosis, multiple sclerosis etc. We are interested in cells of the mononuclear phagocyte system, are a group of white blood cells that play an important role in the body's immune response. The Mononuclear Phagocyte System consists of three types of cell: monocytes, macrophages and dendritic cells (DC), each with distinct functions during both steady state and disease.

    In the lab our research focuses on monocyte, DC and macrophage dynamics and function under steady state and during disease to gain insights into how these cells are regulated and regulate physiological and pathological processes. This will potentially enable the development of new treatments, based on a better understanding of how the body responds to inflammation.

    Keywords

    Monocyte, macrophage, inflammation.

    Standard techniques: Flow cytometry, cell sorting, adoptive transfer.

    Selected Recent Publication

    • Yona S & Gordon S (2015) From the Reticuloendothelial to Mononuclear Phagocyte System - The Unaccounted Years. Front Immunol 1;6:328.
    • Aychek T, Mildner A, Yona S, Kim KW, Lampl N, Reich-Zeliger S, Boon L, Yogev N, Waisman A, Cua DJ, Jung S (2015) IL-23-mediated mononuclear phagocyte crosstalk protects mice from Citrobacter rodentium-induced colon immunopathology. (2015) Nature Commun 12;6:6525
    • Guilliams M, Ginhoux F, Jakubzick C, Naik SH, Onai N, Schraml BU, Segura E, Tussiwand R, Yona S (2014) Dendritic cells, monocytes and macrophages: a unified nomenclature based on ontogeny. Nature Rev Immunol 14(8):571-8
    • Yona S, Kim K, Wolf Y, Mildner A, Breker M, Ayali D, Viukov S, Guilliams M, Misharin A, Hume D, Perlman A, Malissen B, Zelzer E & Jung S (2013) Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis. Immunity 38 79-91
    • Avraham-Davidi I, Yona S, Grunewald M, Landsman L, Cochain C, Silvestre JS, Mizrahi H, Faroja M, Strauss-Ayali D, Mack M, Jung S, Keshet E (2013) On-site education of VEGF-recruited monocytes improves their performance as angiogenic and arteriogenic accessory cells. J Exp Med 18;210(12):2611-25  (Joint First author)