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UK Cilia Network

Projects menu

Research Projects

The various groups within the UK Cilia Network are engaged in numerous research projects investigating diverse aspects of cilia structure and function in development, ageing, health and disease. Many of these projects are collaborations between members of the group. The following are examples of some of the current funded research projects and PhD Studentship Projects

   

The Primary cilium in adult tissue homeostasis, inflammatory signalling and arthritis 

ARUK/KTRR Fellowship PI: Dr Angus Wann (Oxford) 

The human ciliopathies and associated in vivo model systems indicate the profound influence the ciliome exerts in development. This includes at the level of extracellular matrix regulation. In vitro evidence highlights that proteins associated with the cilium are important in how cells respond to signals important to physiological homeostasis and relevant in many disease contexts. This includes the response of cells to inflammatory cytokines. The aims of this project are to identify the influence the ciliome exerts in adult tissues, define how the ciliome tunes signalling downstream to pathophysiological challenge and identify exploitable mechanisms relevant to clinical disease by also screening disease for a 'cilia signature'.

Tissue-specific and inducible IFT creER models are being developed. A range of read-outs including uCT, histology and pain studies are being used to explore at a number of musculoskeletal focussed phenotypes.

In vitro ciliome targetting is being combined with biochemical, molecular assays and high resolution microscopy to assess how ciliary proteins regulate a number of cellular processes including endocytosis and NFkB signalling.

Finally, we are exploring links between GWAS hits associated with ciliary function.       

Osteoarthritis may be treated as an environmental ciliopathy

MRC Project Grant. PI: Prof Martin Knight (QMUL), CoI: Prof Phil Beales (UCL), Dr Hannah Mittchinson (UCL), Prof Paul Chapple (QMUL), Mr Manoj Ramachandran (Barts and the London NHS Trust)

Increasing evidence suggests that primary cilia and the associated signalling pathways are critical for the health of articular cartilage and that dysregulation is involved in the pathogenesis of osteoarthritis (OA). Recently we have shown that environmental/pathological stimuli regulate primary cilia length and that this modulates cilia function. We therefore hypothesise that pathological alterations in the cartilage microenvironment regulate chondrocyte primary cilia structure leading to fundamental changes in cilia signalling which drive cartilage degradation. Human articular chondrocytes will be subjected to mechanical trauma associated with development of OA and linked to changes in primary cilia length. Cilia will be imaged using confocal/super resolution microscopy and downstream changes in cilia signalling and matrix catabolism investigated using a combination of biochemical assays, molecular biology, imaging and biomechanical evaluation. We will determine whether modulation of primary cilia structure and proteome reduces catabolic signalling and cartilage degradation. Thus by the end of the project we will have identified the role of mechanical regulation of primary cilia length in cartilage degradation. We envisage that this will lead to the development of totally novel treatments for OA in the form of small molecule regulation of primary cilia structure-function.

 

Understanding centrosomes in cancer invasion

MRC Project Grant. PI: Dr Susana Godinho (QMUL)

In cancer, the tumour cells' DNA can become more and more disorganised as mutations build up and the cell loses the ability to fix errors during cell division. This disorganisation can be caused by centrosome amplification. Centrosomes are organising "stations" that help construct and manage protein "cable" structures inside the cell, called microtubules (collectively known as the spindle), that pull apart pairs of chromosomes during mitosis (cell division). They are essential in making sure the cell's copied DNA is separated evenly into the two daughter cells, to avoid the build-up of genetic abnormalities. Normally there are two centrosomes per dividing cell - each organising a "terminal" for the chromosomes to be pulled towards in cell division. When cells are not dividing, the centrosome contributes to cell organisation andmigration – a characteristic of invasive cancer.

Human tumours often carry abnormal numbers of centrosomes. Although the role of extra centrosomes in cancer development is still unclear, recent evidence suggests they are not only bystanders in cancer progression, but active in its promotion. This new work will contribute to our understanding of the effect of centrosome amplification in tumour progression.

​Mapping mechanotransduction mediated by primary cilia at nanoscale

Wellcome Trust Seed Funding. PI: Dr Pavel Novak

This exciting grant will combine the latest technology in scanning probe microscopy and nanopipette techniques with confocal microscopy, electrophysiology, and mechano-biology to break the existing resolution barrier in direct stimulation and recording of channel and receptor activity along the fine structure of primary cilia. The technique will be used to gather data on the distribution of mechano-chemical receptors in primary cilia to support future research proposals aimed at understanding the reorganisation of mechanotransduction in ciliated cells in disease.

 

Aberrant chondrocyte primary cilia signalling drives cartilage degradation in response to IL-1

ARUK Project Grant. PI: Prof Martin Knight (QMUL). CoI: Prof Paul Chapple (QMUL)

Primary cilia are poorly understood cytoskeletal organelles that projects into the extracellular milieu. The chondrocytes cilium acts as a centre for Hedgehog (Hh) signalling which has recently been shown to be up-regulated in osteoarthritis where it drives cartilage degradation [31]. However the mechanisms leading to aberrant Hh signalling have not been identified. Preliminary data show that the inflammatory cytokine, IL-1β, which is up-regulated in osteoarthritis, triggers a significant increase in primary cilia length. We hypothesise that this cytokine-mediated ciliogenesis activates changes in cilia function, including Hh signalling. We will therefore identify the underlying mechanisms through which cytokines modulate cilia length, including the involvement of adenyly cyclise, calcium signalling and the actin remodelling. We will then determine the consequences for cilia-mediated Hh signalling, and downstream cartilage matrix synthesis and catabolism. Studies will utilise isolated primary chondrocytes and cartilage explants as well as transgenic chondrocytes that lack primary cilia. We will ultimately determine whether manipulation of ciliogenesis can down-regulate cytokine-induced changes in primary cilia function and thus identify novel cilia-related therapeutic targets to attenuate cartilage degradation.

Preventing tumour progression in Von Hippel–Lindau disease by remodulating primary cilia function

Barts Charity Clinical Research Training Fellowship: Dr Sam O’Toole. Supervisor: Prof Paul Chapple (QMUL)

Disruption of primary cilia is an increasingly recognised feature of a number of cancers. Both dysregulation of cilia-mediated signalling pathways and loss of cell cycle checkpoints are potentially important mechanisms in tumour development.

Von Hippel-Lindau disease is an inherited cancer syndrome which features kidney cancer, phaeochromocytomas (tumours of the adrenal gland), cysts of the pancreas and kidneys and haemangioblastomas of the central nervous system and is due to a mutation in the VHL gene.  One function of VHL is in maintenance of the primary cilium function. If the VHL gene is faulty, the structure and therefore function of the primary cilium may be disrupted, which could partly explain why these tumours develop. This is the case in kidney cancer in VHL, but it is unknown whether it is important in the other tumours that occur in VHL.

The objectives of this research are to determine whether primary cilium structure and function is disrupted in phaeochromocytomas (both when they occur in patients with VHL and when they don't) and whether this is important in tumour development.

The overall aim is to modulate primary cilia function as a strategy to prevent tumour progression.

CANBUILD - Engineering the tumour microenvironment

ERC Senior Fellowship
CANBUILD Team: Professor Frances Balkwill (PI), Dr Michelle Lockley, Dr John Connelly, Professor Ian MacKenzie and Professor Martin Knight. 

This project aims to revolutionise the field of cancer cell research by using bioengineering techniques to grow the first complex 3-dimensional human "tumour microenvironment" in the laboratory. In the CANBUILD project the multi-disciplinary team of scientists are using the latest advances in tissue engineering, biomechanics, imaging and stem cell biology which they believe will make it possible to engineer, for the first time, a complex 3-dimensional human tumour in which the different cell types of the tumour microenvironment will communicate, evolve and grow in vitro. The CANBUILD goal is to recreate the tumour microenvironment of human high-grade serious ovarian cancer, the subtype that leads to 70 per cent of all ovarian cancer deaths, but the research may have implications for several other cancers as well. The vision is that this project will replace inadequate techniques where human cancer cells are grown in isolation on plastic surfaces. Success in the CANBUILD project may also provide better ways of testing new drugs that target the human tumour microenvironment. The five-year research plan involves: - "Deconstruction" of the human ovarian cancer tumour microenvironment - "Construction" of the tissue engineered tumour microenvironment incorporating the artificial scaffold, different cell types,- "Testing" new treatments that target the tumour microenvironment. Part of the research involves examining the role of primary cilia in sensing the tumour microenvironment and regulating tumourogenesis.

 


PhD Studentship Projects:

The ciliome in adult cartilage homeostasis and pathogenesis 

Kennedy Trust prize DPhil studentship (Clarissa Coveney) Supervisors Dr Angus Wann and Prof Tonia Vincent (All Oxford)

Joint development depends upon multiple cellular responses and signalling that are, in part, regulated by the proteins associated with the primary cilium. This manifests in ciliopathy models that demonstrate roles for the cilium in cell differentiation and matrix turnover. Little is known about what influence the ciliome has in adult cartilage and bone homeostasis and pathogenesis. This project will use a cartilage specific transgenic model developed for inducible knock-downs in mature cartilage and a complementary pan-tissue model. Importantly, the ciliome, by means of targeting IFT, will be perterbed  at and after skeletal maturity. Both naive and pathologically challenged systems are being investigated. The project will also investigate a link between IFT and endocytotic regulation of matrix turnover and GWAS data that indicates a potential role for a cilia-linked protein in cartilage homeostasis and pathogenesis.  

The project has so far identified that the ciliome remains influential beyond development and is unpicking how the ciliome apparently regulates extracellular aggrecanase activity  in a post-translational manner.   By using a range of in vivo and in vitro techniques from the level of the whole joint to the single cell the project aims to define adult and arthritis roles for the ciliome with a view towards translational exploitation.

 


The role of primary cilia in tendinopathy

Institute of Bioengineering PhD Studentship (Dan Rowsen) Supervisors: Prof Martin Knight and Dr Hazel Screen

Tendon is mechanosensitive, maintaining tissue health in response to applied loads. Overload is a key contributor to the development of tendon pathologies, know as tendinopathies; a range of highly debilitating and increasingly prevalent conditions2-3. However, the mechanisms associated with tendinopathy development remain unclear. Current evidence supports a combined mechanical and cellular pathway, with early structural damage initiating a catabolic cell response. Primary cilia consist of a single microtubule axoneme, 2-4µm in length, which projects into the extracellular environment and regulates fundamental signalling pathways including mechanotransduction, hedgehog (Hh) and Wnt signalling which are important in tissue development, health and disease. Although primary cilia have been identified in tendon, their precise function is unknown. Recent studies of cilia in cartilage indicate that they are essential for tissue development and mechanotransduction mediated matrix synthesis. Furthermore cilia may play a major role in the aetiology of osteoarthritis (OA) through upregulation of Hh signalling8 and alterations in hypoxia inducible factor (HIF) expression. Work from Knight’s group also suggests that physicochemical changes in the OA microenvironment, including mechanical loading and inflammatory cytokines, disrupt primary cilia structure and associated signalling pathways. As a tissue damaged through injurious overload, it is reasonable to hypothesise that similar cilia mediated mechanotransduction pathways may be present in tendon. This studentship tests the overall hypothesis that injurious mechanical loading influences primary cilia structure & function, creating a catabolic cell response & leading to the development of tendinopathy.

 


Primary cilia structure and function in mesenchymal stem cells

Institute of Bioengineering PhD Studentship (Melis Dalbay) Supervisors: Prof Martin Knight, Dr Paul Chapple and Dr John Connelly

I am investigating roles of primary cilia in hMSC differentiation. Primary cilia are single non-motile organelles that protrude out of the cell and provide a specialised compartment for sensing extracellular mechanical, chemical stimuli and coupling them into cellular responses with signalling proteins enriched in them. They are crucial for cell development and metabolism, defects in them leading to various pathological conditions. My project focuses on investigating if changes in primary cilia structure in response to chemical stimuli or surface topography regulate hMSC differentiation. We have shown that primary cilia length and prevalence change during hMSC differentiation in a lineage specific manner in response to chemical stimuli. Primary cilia elongation was observed during adipogenic differentiation of hMSCs which was associated with increased IGFR-1β trafficking into the cilium, which is involved in adipogenic differentiation induction. Inhibition of this elongation by siRNAs that target ciliary proteins involved in cilia formation and trafficking resulted in inhibition of adipogenic differentiation through decreased ciliary translocation of IGFR-1β. We are currently investigating structural changes in primary cilia of hMSCs in response to surface topography. hMSCs grown on tri-calcium phosphate ceramic discs with different surface structure were shown to respond differently, cells growing on nano scale surfaces resulting in osteogenic differentiation of hMSC in vitro and in-vivo in the absence of osteogenic factors. We have shown that primary cilia structure changes in response to these topographical cues and are currently investigating their role in regulating osteogenic differentiation on these materials. Our recent findings implicate the importance of primary cilium in controlling hMSC differentiation which could prove to be a new target to be used in tissue regeneration studies as well as cilia targeted therapies. (Melis Dalbay)

 


Collagen nanomechanics and the role of primary cilia in articular cartilage

Institute of Bioengineering PhD Studentship (Sheetal Inamdar) Supervisors: Dr Himadri Gupta and Prof Martin Knight

Determining the depth-dependent relationship between the mechanical behaviour and the composition and structure of articular cartilage is crucial in understanding the changes that develop during osteoarthritic degradation. Currently, little is known as to how the networks of collagen fibrils contribute to the tissue’s mechanics, with the fibrils acting as the structural framework of the tissue. Furthermore, there is evidence that has been developed in Knight’s group that would suggest there may be a link between primary cilia dysfunction, an organelle found on eukaryotic cells that regulate many complex signalling pathways, and mechanical changes in the microenvironment.

By using high brilliance synchrotron x-ray diffraction, in situ experiments allow the fibrillar network to be probed for various different parameters such as fibril strain, fibril orientation and fibrillar disorganisation to potentially build a multi-scale model. This information will allow us to understand how changes to primary cilia in terms of structure and function may be influencing the nanoscale mechanics of the tissue, and therefore allow us to predict and understand the mechanisms that lead to changes in tissue mechanics in osteoarthritis.

This studentship is focussed on characterising the nanoscale deformations in healthy cartilage and measuring the changes that occur with manipulation of primary cilia as well as tissue composition, by using small angle x-ray scattering (SAXS) combined with macro-scale mechanics.

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