COMPLEMENT UK
Projects for PhD
Studentships
Table of Contents
PROJECT – 1: Investigating whether complement activation inside the eye is more important than
complement activation in the systemic circulation in the pathogenesis of age-related macular
degeneration, using a cell culture model ............................................................................................... 3
PROJECT – 2: Improved conjugate vaccines derived from a bacterial immunomodulatory protein ..... 4
PROJECT – 3: Genetic variations in complement and impact on eculizumab efficacy ........................... 5
PROJECT – 4: Activation and inactivation of complement anaphylastoxins and their receptors during
neutrophil dominated inflammation of the cystic fibrosis airway ......................................................... 6
PROJECT –5: Genetic variants of terminal pathway components in age-related macular degeneration
and atypical haemolytic uraemic syndrome ........................................................................................... 8
PROJECT –6: Factor H activation – a new strategy for therapeutic complement inhibition ................ 10
PROJECT –7: Molecular mechanisms for aHUS using a combined bioinformatics-molecular
interactions strategy for the disease-associated complement genes .................................................. 12
PROJECT –8: Unravelling the structural and functional defects in naturally occurring diseaseassociated variants of mannan-binding lectin ...................................................................................... 13
PROJECT –9: Optimising recombinant mouse factor H constructs for therapy in murine models of
kidney disease ....................................................................................................................................... 14
PROJECT – 1: Investigating whether complement activation inside the
eye is more important than complement activation in the systemic
circulation in the pathogenesis of age-related macular degeneration,
using a cell culture model
Project Description:
Age-related macular degeneration (AMD) is the leading cause of blindness in the developed
world. The complement system has an important role in the pathogenesis of AMD. Recent
studies indicate complement activation within the eye as opposed to complement activation
in the systemic circulation, may be more important in AMD pathogenesis. This project aims
to investigate the role of intraocular versus systemic complement in AMD at a cellular level.
The project will have both clinical and basic science components. Clinical component: An
ophthalmologist will collect vitreous samples from the eyes of patients with AMD, during
routine intravitreal injections / during intraocular surgery. Blood will be collected by
research nurses. The student will assist in recruitment of patients, processing and analysis of
vitreous/plasma samples. Samples will be analysed for complement protein activity using
novel multiple immune-assays at Cardiff University. The student will carry out DNA
extraction and genotyping for single nucleotide polymorphisms in complement genes
previously associated with AMD, to investigate the association of intraocular complement
activity and genotype. Basic science component: The student will set up a retinal pigment
epithelium (RPE) cell culture model and investigate the extent of complement deposition
and activation by adding vitreous / plasma from patients with AMD/controls. The benefits of
complement inhibition will be investigated in this in vitro model. This will help to
understand the role of intraocular versus systemic complement inhibition in the
management of AMD.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Professor Andrew Lotery
Research Institute & Dept: Clinical Neurosciences Research Group, Clinical and
Experimental Sciences, Faculty of Medicine, University of Southampton
Email: [email protected]
Supervisor profile url: http://www.southampton.ac.uk/medicine/about/staff/ajl.page?
2nd SUPERVISOR
Name of 2nd Supervisor: Professor B. Paul Morgan
Research Institute & Dept: Cardiff University; Infection and Immunity
Email: [email protected]
Supervisor profile url: http://medicine.cf.ac.uk/person/prof-paul-morgan/
PROJECT – 2: Improved conjugate vaccines derived from a bacterial
immunomodulatory protein
Project Description:
Our research aims to improve vaccination by developing a unique adjuvant protein with the
potential to provide easier delivery, increased safety and more accessible vaccination
programmes. An important aspect of our research is to improve the vaccine’s ability to be
given on a one hit basis and improve rates of protection, which would vastly reduce the
time needed to respond and reduce vaccination costs. We have already demonstrated that
our prototype vaccine conjugate is very effective in stimulating immune cells in vitro and we
now poised to evaluate this conjugate in vivo. With respect to Human vaccines there are a
number of significant unmet needs, particularly for vaccines against developing world
infectious diseases.



Influenza still presents a huge problem on a global basis, either seasonal or pandemic
Tuberculosis (TB) is a major concern as it infects approximately 9 million native
individuals, especially in the sib-Saharan African continent, Southeast Asia and in Eastern
Europe causing 1.7 million deaths every year
The technology could also address significant needs in the areas of autoimmune disease
and cancer
This project will focus on TB as we have access to standard vaccine antigens and can move
quickly to develop and test our novel vaccine conjugate. However, the scope of these
compounds is such that they can be applied to almost any antigen and the prototypic
conjugate has been shown to work in all species of animals tested. Thus, this vaccine
conjugate will have utility in animal vaccine programmes. This could be a very important
feature in tackling diseases that can jump between species (e.g. influenza).
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Dr Kevin Marchbank
Research Institute & Dept: Newcastle University, Institute of Cellular Medicine
Email: [email protected]
Supervisor profile url:
http://www.ncl.ac.uk/biomedicine/research/groups/profile/kevin.marchbank
2nd SUPERVISOR
Name of 2nd Supervisor: Dr Jean van den Elsen
Research Institute & Dept: University of Bath, Dept of Biology and Biochemistry
Email: [email protected]
Supervisor profile url: http://www.bath.ac.uk/biosci/contacts/academics/jean_van_den_elsen//
PROJECT – 3: Genetic variations in complement and impact on eculizumab
efficacy
Project Description:
Paraoxysmal Nocturnal Haemoglobinuria (PNH) is a clonal haematopoietic disorder resulting
in the production of red blood cells lacking surface receptors required to protect them from
complement-mediated lysis. The absence of these complement regulatory proteins allows
for the activation of the complement cascade, leading ultimately to the generation of a
membrane attack complex and a high level of the intravascular haemolysis.
Unlike PNH, aHUS is a genetic disorder of complement over-activation. Loss of function
mutations in complement regulatory proteins, factor I, factor H and membrane cofactor
protein (CD46) and gain of function mutations in the complement components C3 and
factor B all predispose to disease. Haemolysis in aHUS is secondary to a microangiopathic
haemolytic anaemia, although the role of direct complement lysis of erythrocytes has not
been extensively examined.
Despite the complement regulatory difference between aHUS and PNH, eculizumab has
proven effective in both conditions. Thus comparison of erythrocytes in aHUS and PNH
patients treated with eculizumab will provide a unique insight into differing pathological
processes in these conditions.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Professor Peter Hillmen
Research Institute & Dept: University of Leeds, Section of Experimental Haematology
Email: [email protected]
Supervisor profile url:
http://medhealth.leeds.ac.uk/info/940/experimental_haematology/1066/translational_hae
matology_group
2nd SUPERVISOR
Name of 2nd Supervisor: Dr David Kavanagh
Research Institute & Dept: Newcastle University, Institute of Genetics Medicine
Email: [email protected]
Supervisor profile url: http://www.ncl.ac.uk/igm/staff/profile/david.kavanagh
PROJECT – 4: Activation and inactivation of complement anaphylastoxins and
their receptors during neutrophil dominated inflammation of the cystic
fibrosis airway
Project Description:
Cystic fibrosis (CF) is the most commonly inherited fatal disorder in the UK. Chronic
colonisation of the airway by microbial pathogens leads to the recruitment of large numbers
of neutrophils. However, the normal biology of the neutrophil is hindered in the CF airway,
leading to the accumulation of large numbers of dead and dying cells. Consequently many
neutrophil derived anti-microbial molecules including proteases and reactive oxygen
intermediates are released into the airway and in the absence of appropriate counterregulation; they attack the delicate lung tissue. Furthermore, many of these normally antimicrobial molecules also inactivate elements of the immune system itself including IL-8
receptors, CR1 and IL-6 receptor, further exacerbating the inflammatory response.
The complement system is critical part of the innate immune host defence which initiates
inflammatory and anti-microbial responses to infection. Amongst many consequences of
complement activation, the anaphylatoxins C3a and C5a stand out as important regulators
of neutrophil recruitment and activation may be important contributors to CF airway
disease. Little is known about the expression of behaviour of C3a and C5a in CF or how
these molecule and their receptors are affected by the hostile environment of the CF
airway. We will test the hypothesis that poorly regulated neutrophil activity in the CF lung
exacerbates inflammation and hinders anti-microbial responses by targeting C3a, C5a and
their receptors.
The study will use a variety of molecular and cellular approaches including ELISA, flow
cytometry, primary cell isolation, cell culture, SDS-PAGE, mass-spectrometry and specific
enzyme assays to assess the expression and function of C3a, C5a and their receptors in lung
secretions from patients with CF. This project follows on from recent work by this group
which has identified a number of defects in the innate immune system associated with the
inflamed CF airway.
SUPERVISOR DETAILS
st
1 SUPERVISOR
Name of 1st Supervisor: Dr Eamon McGreal
Research Institute & Dept: Institute of Molecular and Experimental Medicine, Cardiff
University School of Medicine
Email: [email protected]
Supervisor profile url:
http://medicine.cf.ac.uk/person/eamon-mcgreal
2nd SUPERVISOR
Name of 2nd Supervisor: Dr Carmon W van den Berg
Research Institute & Dept: Dept of Pharmacology, Institute of Molecular and Experimental
Medicine, Cardiff University
Email: [email protected]
Supervisor profile url: http://medicine.cf.ac.uk/person/carmen-vandenber/
3rd SUPERVISOR
Name of 3rd Supervisor: Dr Peter Monk
Research Institute & Dept: Dept of Infection and Immunity, Sheffield University Medical
School
Email: [email protected]
Supervisor profile url:
http://www.sheffield.ac.uk/infectionandimmunity/staffprofiles/monk
PROJECT –5: Genetic variants of terminal pathway components in age-related
macular degeneration and atypical haemolytic uraemic syndrome
Project Description:
Complement is an ancient defence system which coats both foreign cells (such as bacteria
and viruses) and our own cells (including those of the kidney and eye) with a substance
called C9. This substance will eventually lead to the destruction of bacteria and viruses by
damaging the cell wall. Our own cells are protected against C9 by expression of regulatory
proteins that protect them suffering a similar fate.
Age-related macular degeneration (AMD) and atypical haemolytic uraemic syndrome (aHUS)
are both disorders of complement over-activation. AMD is the commonest cause of
irreversible blindness in the elderly in the developed World while aHUS results in
irreversible kidney failure. We have recently demonstrated that both these conditions are
associated with a rare genetic variant in C9. We hypothesise that this variant results in a
hyperactive complement system which damages host kidney and eye tissues.
To assess the functional consequences of this rare genetic variant, human and recombinant
wild type and risk variant protein will be purified and made as recombinant proteins.
Haemolytic assays with reconstituted C9 depleted serum will initially be performed. Surface
plasmon resonance will be used to compare binding of the risk and wild type variants to
other complement proteins and to the complement regulator CD59. Monoclonal antibodies
specific for the variant forms of C9 will be generated and used to develop a simple, robust
clinical assay. Genetic analysis of large numbers of aHUS individuals will be carried out to
assess the frequency of this C9 genetic variant and whether other changes in related
complement proteins occur in aHUS patients.
The student will receive training in a broad range of techniques including: protein chemistry,
molecular biology, recombinant protein production, cell culture, hybridoma generation,
surface plasmon resonance, next generation sequencing and Multiplex ligation-dependent
probe amplification.
References:
1. De Cordoba SR, et al. (2012). Immunobiology; 217(11): 1034-46.
2. Heurich M, et al. (2011). Proc Natl Acad Sci USA; 108(21): 8761-6.
3. Martínez-Barricarte R, et al. (2010). J Clin Invest; 120(10):3702-12.
4. Seddon JM, et al. (2013). Nat Genet 2013;in press.
5. Hakobyan S, et al. (2010). Kidney Int; 78(8):782-8.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Professor Paul Morgan
Research Institute & Dept: Cardiff University, Institute of Infection and Immunity
Email: [email protected]
Supervisor profile url: http://me icine cf ac u infect-immun
2nd SUPERVISOR
Name of 2nd Supervisor: Dr David Kavanagh
Research Institute & Dept: Newcastle University, Institute of Genetics Medicine
Email: [email protected]
Supervisor profile url: http://www.ncl.ac.uk/igm/staff/profile/david.kavanagh
PROJECT –6: Factor H activation – a new strategy for therapeutic complement
inhibition
Project Description:
The studentship involves testing a potential treatment, inspired by bacteria, for some
evastating iseases that occur when the immune system ‘self harms’
The first onslaught a bacterium must survive when it invades the blood stream is launched
by the complement system. This is a team of proteins that can rapidly coat a bacterium with
the protein C3b that acts as an ‘eat me’ signal to the immune cells The complement system
can also punch thousands of holes in a cell membrane. Finally, complement acts in a similar
but gentler way to eradicate damaged or senescent human cells.
In self- efence many bacteria have evolve to exploit a wea lin in complement’s chain of
command. This is the protein factor H (FH) that prevents our own cells becoming coated
with C3b and ensuring injury. Importantly, several diseases – notably those involving
damage to the eye or kidney - are caused by dysfunctional FH. Because FH circulates in the
blood, it is readily hijacked by a bacterium and used to protect the bacterial cell.
This student will receive supervision from protein scientists in Edinburgh and immunologists
and clinicians in Newcastle. A highly marketable, possibly unique, cross-disciplinary skill-set
will be acquired spanning molecular sciences and studies of animal models of diseases. The
project will involve producing recombinant polypeptides derived from a bacterial protein
that has the remarkable ability to both capture FH on the bacterial surface and to activate it
tenfold. The polypeptides are soluble and can be used to turbo-charge FH in plasma and
thus enhance its ability to suppress the complement system. Having fully characterised their
recombinant polypeptides using structural and functional techniques, the student will
investigate their efficacy in two mouse models of disease. We predict that these studies will
prove that FH activation is an exciting novel strategy for treating human diseases that are
caused by complement dysregulation.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Professor Paul Barlow
Research Institute & Dept: Edinburgh University, School of Chemistry
Email: [email protected]
Supervisor profile url: http://www.chem.ed.ac.uk/staff/academic/barlow.html
2nd SUPERVISOR
Name of 2nd Supervisor: Dr Kevin Marchbank
Research Institute & Dept: Newcastle University, Institute of Cellular Medicine
Email: [email protected]
Supervisor profile url:
http://www.ncl.ac.uk/biomedicine/research/groups/profile/kevin.marchbank
3rd SUPERVISOR
Name of 3rd Supervisor: Dr David Kavanagh
Research Institute & Dept: Newcastle University, Institute of Genetics Medicine
Email: [email protected]
Supervisor profile url: http://www.ncl.ac.uk/igm/staff/profile/david.kavanagh
PROJECT –7: Molecular mechanisms for aHUS using a combined
bioinformatics-molecular interactions strategy for the disease-associated
complement genes
Project Description:
The cost of DNA sequencing has fallen to the extent that it is now inexpensive to sequence
an in ivi ual’s genome This “next generation sequencing” technology is being applie to
patients with both rare and common diseases associated with defects of the ancient
immune pathway called complement. Atypical haemolytic uraemic syndrome is a rare
disease characterised by irreversible renal failure in children and young adults. Age-related
macular degeneration is the commonest cause of blindness in the UK. Sequencing the
complement genes of patients and healthy individuals has provided a wealth of information
about how genetic variants may cause these diseases. By constructing an interactive
database of these mutations, alongside the use of predictive and protein structural tools to
evaluate their mechanistic roles, the most crucial disease-associated residues will be
rationally predicted in order to plan mutagenesis experiments. Next, these mutagenesis
predictions will be used to plan molecular interaction experiments with recombinant small
fragments of complement Factor H and C3b and small heparin fragments of sizes dp6-dp36.
Methods will include state-of-the-art surface plasmon resonance, isothermal calorimetry,
analytical ultracentrifugation and X-ray scattering. The combination of these methods will
provide reliable quantitative measures of the strength of the interaction (dissociation
constants) and how these are affected by mutations. The outcome of the project will
provide key clarifications of the molecular mechanism of these diseases, the clinical
importance of the observed mutations, and the relative importance of the Factor H
interactions with the C3d fragment of C3b vs. those with heparin fragments. The PhD
project thus offers a broad multidisciplinary training in computational biology (database
development, programming, statistics and web design) and biomedical aspects of
complement (sequencing, structural analysis, molecular interactions technology). There will
be opportunities to meet complement researchers and clinicians in Europe and the USA,
including visits to their laboratories.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Professor Stephen Perkins
Research Institute & Dept: Dept of Structural and Molecular Biology, University College
London
Email: [email protected]
Supervisor profile url: http://www.ucl.ac.uk/smb/perkins
2nd SUPERVISOR
Name of 2nd Supervisor: Professor Timothy Goodship
Research Institute & Dept: Institute of Human Genetics, Newcastle University
Email: [email protected]
Supervisor profile url:
http://www.ncl.ac.uk/biomedicine/research/groups/profile/tim.goodship
PROJECT –8: Unravelling the structural and functional defects in naturally
occurring disease-associated variants of mannan-binding lectin
Project Description:
Blood proteins called "complement" act to recognise and destroy invading foreign bacteria.
These proteins serves as a first line of immune defence by both destroying invading
pathogens directly and helping the adaptive immune system to distinguish between what is
foreign (a bacterial cell) and what is self (host cell). For this PhD project, we will study a
protein called mannan-binding lectin (MBL) which specifically recognises arrays of sugars
found on bacterial pathogens but not present on our own human cells. MBL binding then
triggers a change in a second protein called MASP, causing it to switch from an inactive to an
active form. These two events then lead to activation of other complement proteins, which
ultimately trigger the destruction of the bacterium, as well as stimulating other immune
processes and protective functions. We are already studying how MBL and MASP bind to
each other and how MASP activation takes place. The PhD project will investigate what
happens to the structure of MBL when it is modified in genetic diseases. A range of common
genetic alterations in MBL lead to a broad spectrum of disorders. These include increased
susceptibility to infections in early childhood, to instances in adults where adaptive
immunity becomes ineffective such as during AIDS or cancer chemotherapy. We will
determine new protein crystal structures and use new X-ray scattering and analytical
ultracentrifugation experiments combined with advanced molecular graphics modelling to
study mutant MBL structures. In particular we aim to explain why some mutant MBLs overactivates MASP while others result in no MASP activation.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Dr Russell Wallis
Research Institute & Dept: Dept of Infection, Immunity and Inflammation, University of
Leicester
Email: [email protected]
Supervisor profile url: http://www2.le.ac.uk/departments/biochemistry/staff/wallis
2nd SUPERVISOR
Name of 2nd Supervisor: Professor Stephen Perkins
Research Institute & Dept: Dept of Structural and Molecular Biology, University College
London
Email: [email protected]
Supervisor profile url: http://www.ucl.ac.uk/smb/perkins/
PROJECT –9: Optimising recombinant mouse factor H constructs for therapy
in murine models of kidney disease
Project Description:
Our complement system provides a first line of defence against invading microbes but can
attack and destroy our own tissues. The kidney is highly sensitive to inadequate regulation
of the complement system. Exemplified by two rare but devastating kidney diseases,
atypical haemolytic uremic syndrome (aHUS) and dense deposit disease (DDD).
Many academic groups and commercial companies are now racing to design and develop
therapeutic approaches to bring the complement system back into balance. Several labs are
pursuing a strategy of engineering a natural protein found in blood – called factor H (FH) –
that normally regulates the complement system very effectively. Inherited defects of FH are
linked to aHUS and DDD. Administration of human FH (purified from blood) was shown to
be sufficient to restore normal complement function in animal models. This suggested that
replacement if defective FH and/or boosting of protective FH to super-physiological levels,
would present a valid approach to control complement mediated disease.
Demonstrating which engineered version of FH is most appropriate for clinical purposes is
now essential. In this project, the successful candidate will use the recombinant DNA
technology to generate both full-length mouse FH and a panel of minimal versions of FH
lacking apparently non-essential regions. Training in this phase will be provided in the
Barlow laboratory in Edinburgh. Subsequently, the various recombinant versions of FH will
be tested in Newcastle, using mouse models of disease. Training for this work will be
provided by the Kavanagh and Marchbank groups in Newcastle. The project therefore
provides a possibly unique opportunity to acquire a wide range of valuable skills that span
the disciplines of biochemistry and immunology, including DNA manipulation, culture of
Pichia pastoris, protein purification and biophysical characterisation, Surface Plasmon
Resonance, ELISA, immunohistrochemistry and flow cytometry. The data produced by the
student will have a clear impact on the eventual use of inhibitors based on human FH for
therapy in aHUS and DDD.
SUPERVISOR DETAILS
1st SUPERVISOR
Name of 1st Supervisor: Dr David Kavanagh
Research Institute & Dept: Newcastle University, Institute of Genetics Medicine
Email: [email protected]
Supervisor profile url: http://www.ncl.ac.uk/igm/staff/profile/david.kavanagh
2nd SUPERVISOR
Name of 2nd Supervisor: Professor Paul Barlow
Research Institute & Dept: Edinburgh University, School of Chemistry
Email: [email protected]
Supervisor profile url: http://www.chem.ed.ac.uk/staff/academic/barlow.html
3rd SUPERVISOR
Name of 3rd Supervisor: Dr Kevin Marchbank
Research Institute & Dept: Newcastle University, Institute of Cellular Medicine
Email: [email protected]
Supervisor profile url:
http://www.ncl.ac.uk/biomedicine/research/groups/profile/kevin.marchbank