Sphingosine-1-Phosphate receptor 1 signalling in bacterial-macrophage interactions: exploring novel anti-bacterial strategies using immunomodulatory t
Lead Research Organisation:
University of Exeter
Department Name: Biosciences
Abstract
In the quest for novel antimicrobial strategies, one area that remains under-explored is the development of agents that enhance immune system activity ("immunomodulators"). Such agents may be used as an alternative or adjunct to conventional antibiotics. This studentship focuses on the human sphingosine-1-phosphate (S1P) pathway, a key regulator of immune function and a viable target for such a therapeutic strategy. Modulation of the S1P pathway can be achieved using either selective S1P receptor (S1PR) agonists or inhibitors of S1P lyase (S1PL), an enzyme that controls S1P levels by catalysing its irreversible degradation.
Uniquely, S1P-targeted therapies have the potential to achieve beneficial immunomodulation whilst simultaneously directly inhibiting the pathogen. For example, certain S1PR agonists combine immunomodulatory activity with direct antimicrobial activity. Furthermore, we have recently characterized bacterial-encoded S1PLs that are required for the pathogenesis of Burkholderia pseudomallei, the causative agent of melioidosis. S1PL orthologues also exist in other bacterial species, including mycobacteria. Inhibitors of such bacterial S1PLs would be expected to impair virulence, thus combining immunomodulation with anti-virulence activity.
Through a multidisciplinary programme of research that sits at the interface between microbiology and cell biology, the student will aim to validate the S1P pathway as a viable therapeutic target for the treatment of infectious diseases. Specifically, the student will:
1) Undertake biochemical characterization of bacterial S1PLs and perform the first inhibitor studies against bacterial enzymes using a recently described S1PL inhibitor.
2) Define the impact of bacterial S1PLs and S1P pathway modulation during intracellular infection of macrophages. In particular, the student will investigate the linkage between sphingosine homeostasis, intracellular calcium mobilisation and lysosomal function (building on observations from the Lloyd-Evans group that changes in sphingosine/S1P levels alter lysosomal function).
3) Assess the activity of the latest generation of S1P pathway modulators (selective S1PR agonists and S1PL inhibitor) within relevant in vivo (larval) and in vitro infection models, with and without co-administration of conventional antibiotics.
The project offers outstanding training opportunities at the forefront of microbiology and cell biology and will benefit from complementary expertise of the supervisory team encompassing molecular bacteriology & host-pathogen interactions (Brown; Exeter), sphingolipid-lysosome biology & lysosomal calcium imaging (Lloyd-Evans; Cardiff), and enzymatic characterization of S1PLs (Campopiano; Edinburgh). The project also benefits from access to the latest generation of S1P pathway modulators from Novartis, including the first documented specific inhibitor of S1PL. Access to these compounds will fast-track clinical translation.
Uniquely, S1P-targeted therapies have the potential to achieve beneficial immunomodulation whilst simultaneously directly inhibiting the pathogen. For example, certain S1PR agonists combine immunomodulatory activity with direct antimicrobial activity. Furthermore, we have recently characterized bacterial-encoded S1PLs that are required for the pathogenesis of Burkholderia pseudomallei, the causative agent of melioidosis. S1PL orthologues also exist in other bacterial species, including mycobacteria. Inhibitors of such bacterial S1PLs would be expected to impair virulence, thus combining immunomodulation with anti-virulence activity.
Through a multidisciplinary programme of research that sits at the interface between microbiology and cell biology, the student will aim to validate the S1P pathway as a viable therapeutic target for the treatment of infectious diseases. Specifically, the student will:
1) Undertake biochemical characterization of bacterial S1PLs and perform the first inhibitor studies against bacterial enzymes using a recently described S1PL inhibitor.
2) Define the impact of bacterial S1PLs and S1P pathway modulation during intracellular infection of macrophages. In particular, the student will investigate the linkage between sphingosine homeostasis, intracellular calcium mobilisation and lysosomal function (building on observations from the Lloyd-Evans group that changes in sphingosine/S1P levels alter lysosomal function).
3) Assess the activity of the latest generation of S1P pathway modulators (selective S1PR agonists and S1PL inhibitor) within relevant in vivo (larval) and in vitro infection models, with and without co-administration of conventional antibiotics.
The project offers outstanding training opportunities at the forefront of microbiology and cell biology and will benefit from complementary expertise of the supervisory team encompassing molecular bacteriology & host-pathogen interactions (Brown; Exeter), sphingolipid-lysosome biology & lysosomal calcium imaging (Lloyd-Evans; Cardiff), and enzymatic characterization of S1PLs (Campopiano; Edinburgh). The project also benefits from access to the latest generation of S1P pathway modulators from Novartis, including the first documented specific inhibitor of S1PL. Access to these compounds will fast-track clinical translation.
Organisations
People |
ORCID iD |
Alan Brown (Primary Supervisor) | |
Joshua Dyer (Student) |
Studentship Projects
Project Reference | Relationship | Related To | Start | End | Student Name |
---|---|---|---|---|---|
MR/N013794/1 | 01/10/2016 | 30/09/2025 | |||
1773789 | Studentship | MR/N013794/1 | 01/10/2016 | 31/03/2020 | Joshua Dyer |
Description | UK National Productivity Investment Fund (NPIF) Innovation Placement via the GW4 BioMed MRC DTP |
Amount | £9,984 (GBP) |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 04/2020 |
End | 08/2020 |
Title | S. aureus Gentamicin Protection Assay |
Description | The gentamicin protection assay has been used since the 1980's to assess the survival of bacteria within macrophages. I adapted the gentamicin protection assay incrementally over the course of a year to allow for an optimal assessment of S. aureus's survival within macrophages over long-periods of time. Whilst this wouldn't constitute an entirely new method, the methodology of my approach differs from others in the field in the following ways. - Amount of bacteria added to macrophage cells - Concentrations of gentamicin to kill extracellular bacteria - Care of cells by replacing media frequently to allow for long-term observation of intracellular survival - Taking measures of cell health, extracellular bacterial survival and intracellular bacterial survival in the same assay to provide a more rounded view of the intracellular survival dynamics. |
Type Of Material | Model of mechanisms or symptoms - in vitro |
Year Produced | 2018 |
Provided To Others? | No |
Impact | Advances in the understanding of S. aureus' intracelluar survival in macrophages and the effect of sphingosine-1-phosphate receptor 1 agonists on bacterial intracellular survival in macrophages. |
Title | 3D Fluoresence Image Distance Analysis |
Description | A simple python script that works with 3D confocal image sets. It identifies objects in channel 1 (e.g. a fluorescent bacteria) and the objects in channel 2 (e.g. the nucleus). It then calculates the distance of the object in channel 1 from the nearest object in channel 2. This allows the relative localization of two objects in a cell to be assessed in 3D. I |
Type Of Material | Data analysis technique |
Year Produced | 2019 |
Provided To Others? | No |
Impact | Insights into the intracellular lifestyle of S. aureus in macrophages. Insights into the effects of S1PR1 agonists on macrophage antibacterial activity. |
Title | S1PR1 agonist treated macrophages - dynamics of intracellular abcteria |
Description | Confocal images taken over long timescales looking at the intracellular survival of S. aureus in macrophages treated with Sphingosine-1-phosphate receptor agonists. |
Type Of Material | Database/Collection of data |
Year Produced | 2019 |
Provided To Others? | No |
Impact | Insights into the effects of sphingosine-1-phosphate receptor agonists on macrophage antimicrobial activity and how this activity is mediated. |
Title | PyDist & PD measurements |
Description | A python module (soon to be) released as an open-source tool was generated. It provides a new way of quantifying immunofluoresence microscopy images using a set of measurements we have defined as Perimeter Distance (PD) measurements In essence, it allows the distance between biomarkers (e.g. fluoresently labelled cellular compartments) to be quantified, generating large amounts of data from microscopy images. |
Type Of Technology | Software |
Year Produced | 2020 |
Impact | Interesting findings were generated by the use of this analysis approach from my research that has formed a large portion of my thesis. This leads us to believe that PyDist and PD measurements could be highly useful for the large community of researchers that use immunofluoresence microscopy in their studies. |
Description | Organization of CLESCon; a multi-disciplinary post-graduate student led conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Postgraduate students |
Results and Impact | I led an organizational committee of 10 post-graduate students in the development of CLESCon; a multi-disciplinary conference with data presented from researchers within the following departments: Biosciences, Geography, Sports & Health Science and Psychology. We had 60 scientific posters in the poster sessions comprised consisting mostly of post-graduate students alongside a small number of post-docs. We also had 16 post-graduate talks throughout the day alongside 4 'keynote' academic speakers from each of the four departments. Key outcomes from this exercise were as follows: - provided an opportunity for post-graduate students to practice presentations skills and receive feedback - provided an opportunity for post-graduate students and academics from different drastically different fields to meet, possibly stimulating new ideas and collaborations. - Strong indications that this conference would be invited to be put on again in 2019 |
Year(s) Of Engagement Activity | 2018 |
URL | http://www.exeter.ac.uk/research/events/clescon/ |