Spatial Cholesterol Metabolism: A Mass Spectrometer for Better Diagnosis and Understanding of Disease
Lead Research Organisation:
Swansea University
Department Name: Institute of Life Science Medical School
Abstract
Disturbed cholesterol metabolism (including its synthesis and transport) is linked to many disease states including neurodegenerative, neurodevelopmental and metal health disorders, metabolic disease and microbial infections. In addition, many rare diseases show features of disturbed cholesterol metabolism. Research in many groups in the UK embraces cholesterol metabolism, but there are few laboratories which specifically focus on the analysis of the myriad of intermediates involved in pathways transforming cholesterol to bile acids and steroids. The Biomedical Mass Spectrometry Laboratory in Swansea University Medical School is one such laboratory and with new mass spectrometry (MS) equipment from the MRC Mid-Range Equipment Fund will provide a resource for researcher in the UK to access specialist MS equipment, expert knowledge in sample preparation, data analysis and its interpretation.
MS represents the primary technology in the study of cholesterol metabolism (cholesterolomics) and when combined with liquid chromatography (LC) is suitable for the unbiased identification and quantification of the greatest number of metabolites in a complex mixture. Security of metabolite identification is provided by orthogonal data including retention time, accurate mass measurement, and fragmentation patterns, preferably, in the case of cholesterol metabolites, multistage fragmentation (MSn). A recent addition to MS-tool box is MS-imaging (MSI) adding further to the available bioimaging technologies.
The current proposal is for a high-resolution MS instrument with MSn and UVPD capabilities which can be combined with existing interfaces for LC-MS, high spatial-resolution AP-MALDI-MSI and high sensitivity LESAplus-LC-MSI already in Swansea. The AP-MALDI source offers <10 micrometer spatial resolution while LESAplus-LC-MSI provides isomer separation and with pg/mg sensitivity. The instrument will be a resource for MRC-remit research in the UK. The new instrument will underpin scientific research in several key strategic priority areas for MRC which involve cholesterol metabolism, including (1) neuroscience and mental health; (2) infection and immunity; (3) molecular medicine; (4) clinical and translational research; and (5) global health.
MS represents the primary technology in the study of cholesterol metabolism (cholesterolomics) and when combined with liquid chromatography (LC) is suitable for the unbiased identification and quantification of the greatest number of metabolites in a complex mixture. Security of metabolite identification is provided by orthogonal data including retention time, accurate mass measurement, and fragmentation patterns, preferably, in the case of cholesterol metabolites, multistage fragmentation (MSn). A recent addition to MS-tool box is MS-imaging (MSI) adding further to the available bioimaging technologies.
The current proposal is for a high-resolution MS instrument with MSn and UVPD capabilities which can be combined with existing interfaces for LC-MS, high spatial-resolution AP-MALDI-MSI and high sensitivity LESAplus-LC-MSI already in Swansea. The AP-MALDI source offers <10 micrometer spatial resolution while LESAplus-LC-MSI provides isomer separation and with pg/mg sensitivity. The instrument will be a resource for MRC-remit research in the UK. The new instrument will underpin scientific research in several key strategic priority areas for MRC which involve cholesterol metabolism, including (1) neuroscience and mental health; (2) infection and immunity; (3) molecular medicine; (4) clinical and translational research; and (5) global health.
Technical Summary
There is considerable research interest in the involvement of cholesterol metabolism (including its synthesis and transport) in differing disease states. One way to learn more about such diseases is to monitor how cholesterol metabolism differs between patients and healthy people (and during disease progression) or between animal/cell models and wild type. This can be achieved by mass spectrometry analysis of fluids and tissue sample. By using mass spectrometry imaging (MSI) defined metabolites can be quantitatively visualised from tissue slices, revealing the metabolic differences between e.g. lesions and healthy tissue. Provision of a new mass spectrometer at Swansea, which is an acknowledged centre for the analysis of the cholesterol metabolic pathway, having developed EADSA technology (US patent US9851368B2), will provide a UK resource for biomedical scientists whose research requires an investigation of cholesterol metabolism.
The new instrument will be capable of fluid analysis and with already existing equipment in Swansea, mass spectrometry imaging (MSI). To maximize the security of metabolite identification the instrument will be able to record high-resolution (500,000) accurate mass (<1ppm) spectra and will be able to generate MSn fragmentation trees using different modes of fragmentation, i.e. quadrupole-like, ion-trap-like and UVPD. We have found MSn to be particularly important for the structural characterisation of isomeric oxysterols and bile acid precursors, while UVPD has been shown to be able to locate double bond positions in sterol esters. The instrument will support research in the MRC strategic priority areas including neuroscience and mental health; infection and immunity; molecular medicine; clinical and translational research; and global health. The results generated will help define biomarker panels for the definition and monitoring of the progression of disease and assist in the development of therapy.
The new instrument will be capable of fluid analysis and with already existing equipment in Swansea, mass spectrometry imaging (MSI). To maximize the security of metabolite identification the instrument will be able to record high-resolution (500,000) accurate mass (<1ppm) spectra and will be able to generate MSn fragmentation trees using different modes of fragmentation, i.e. quadrupole-like, ion-trap-like and UVPD. We have found MSn to be particularly important for the structural characterisation of isomeric oxysterols and bile acid precursors, while UVPD has been shown to be able to locate double bond positions in sterol esters. The instrument will support research in the MRC strategic priority areas including neuroscience and mental health; infection and immunity; molecular medicine; clinical and translational research; and global health. The results generated will help define biomarker panels for the definition and monitoring of the progression of disease and assist in the development of therapy.
Organisations
- Swansea University (Lead Research Organisation)
- UNIVERSITY OF OXFORD (Collaboration)
- Eberhard Karls University of Tübingen (Collaboration)
- University of Manchester (Collaboration)
- University Hospital of Wales (Collaboration)
- Karolinska Institute (Collaboration)
- National University of Singapore (Collaboration)
- Sheffield Children's NHS Foundation Trust (Collaboration)
Publications
![publication icon](/resources/img/placeholder-60x60.png)
Griffiths W
(2023)
Role of Bile Acid Pathway Intermediates in Pathology of CTX
![publication icon](/resources/img/placeholder-60x60.png)
Griffiths W
(2023)
Role of Bile Acid Pathway Intermediates in Pathology of CTX
![publication icon](/resources/img/placeholder-60x60.png)
Griffiths WJ
(2023)
Role of Bile Acid Pathway Intermediates in Pathology of CTX
Description | This award was for mass spectrometry equipment delivered to Swansea less than 12 months ago. The award has enabled us to optimize our methodology to allow the detection of almost all cholesterol metabolites in a single analysis. This is being utilized in new collaborations with Sheffield Children' Hospital and the University Hospital of Wales and existing collaborations with the St Mary's Hospital in Manchester, the Wellcome Centre for Human Genetics at the University of Oxford, Oxford Centre for Diabetes Endocrinology and Metabolism, the Center of Neurology and Hertie-Institute for Clinical Brain Research in Tubingen and Karolinska Institutet in Sweden. The award has also allowed us to develop closer links with the pharmaceutical industry and a small startup company. |
Exploitation Route | Cholesterol metabolism is an area of great interest with regard to health and disease. The methods we are exploiting using the instrument funded by this award are opening new avenues for diagnosis in the NHS and providing new directions for exploitation by the pharmaceutical industry. |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
Description | Department of Health workshop on non-genetic conditions |
Geographic Reach | National |
Policy Influence Type | Contribution to a national consultation/review |
Description | NHS Wales Rare Diseases Implementation Group |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | Bile Acid Ring Trial |
Organisation | National University of Singapore |
Country | Singapore |
Sector | Academic/University |
PI Contribution | We are one of a number of groups world wide providing bile acid measurement on a standard reference material. |
Collaborator Contribution | We are one of a number of groups world wide providing bile acid measurement on a standard reference material. Commercial partners are providing authentic standards free of charge. |
Impact | An expert protocol has been developed for bile acid analysis in human plasma. |
Start Year | 2020 |
Description | Cardiff University & UHW |
Organisation | University Hospital of Wales |
Country | United Kingdom |
Sector | Hospitals |
PI Contribution | We have provided expertise, intellectual input and access to equipment to assist in the diagnosis of rare diseases. |
Collaborator Contribution | Our collaborators have provided access to appropriate samples and clinical expertise. |
Impact | We have assisted in patient diagnosis where genomic data was equivocal. |
Start Year | 2023 |
Description | Hereditary Spastic Paraplegia |
Organisation | Eberhard Karls University of Tübingen |
Department | Centre of Neurology and Hertie-Institute for Clinical Brain Research |
Country | Germany |
Sector | Academic/University |
PI Contribution | Expertise in oxysterol and sterol analysis. |
Collaborator Contribution | Hepatocyte and cortical neuron differentiation from iPS cells. Clinical samples from patients with in born errors of metabolism |
Impact | doi: 10.1172/JCI68506 doi.org/10.1016/j.aca.2021.338259 Analytical science and medicine |
Start Year | 2014 |
Description | Karolinska |
Organisation | Karolinska Institute |
Department | Department of Medical Biochemistry and Biophysics |
Country | Sweden |
Sector | Academic/University |
PI Contribution | We have brought essential data and insight to the collaboration. We are bringing new technology |
Collaborator Contribution | They have brought essential data and insight to the collaboration. Our partners are bringing transgenic animal material and expertise to the collaboration. |
Impact | This is a multidisciplinary collaboration between neuroscientists and clinical chemists at Karolinska Institute and analytical scientists in Swansea. doi: 10.3390/biom9040149 doi: 10.1074/jbc.RA118.005639 doi: 10.1016/j.bbalip.2018.11.006 doi: 10.1172/JCI68506 doi: 10.1038/nchembio.1156 WO2014132052A2 EP3044192B1 |
Start Year | 2006 |
Description | Manchester: Collaboration on inborn errors of metabolism |
Organisation | University of Manchester |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Analysis of plasma and urine samples in the study of inborn errors of cholesterol biosynthesis and metabolism. |
Collaborator Contribution | Provision of samples for the study of inborn errors of cholesterol biosynthesis and metabolism. |
Impact | doi: 10.1016/j.freeradbiomed.2019.04.020 doi: 10.1194/jlr.D083246 doi: 10.1016/j.jsbmb.2020.105794 doi: 10.1016/j.jsbmb.2016.03.018 doi: 10.1373/clinchem.2014.231332 doi: 10.1172/JCI68506 doi: 10.1016/j.freeradbiomed.2012.07.027 Multidisciplinary, medicine and analytical science. |
Start Year | 2012 |
Description | Oxford Centre for Diabetes, Endocrinology & Metabolism |
Organisation | University of Oxford |
Department | Oxford Hub |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise in the analysis of oxysterols |
Collaborator Contribution | Provision of valuable biological material |
Impact | Joint grant application made. |
Start Year | 2023 |
Description | SHEFFIELD CHILDREN'S NHS FOUNDATION TRUST |
Organisation | Sheffield Children's NHS Foundation Trust |
Country | United Kingdom |
Sector | Public |
PI Contribution | Collaboration to facilitate the diagnosis of rare disease |
Collaborator Contribution | Facilitate the collection of relevant samples, provide background information and clinical information. |
Impact | The collaboration has assisted in clinical diagnosis where genetics were equivocal. |
Start Year | 2023 |
Description | Wellcome Centre for Human Genetics |
Organisation | University of Oxford |
Department | Oxford Hub |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We are performing sterol analysis to clarify the biochemistry behind rare mutations in human and in a mouse model. |
Collaborator Contribution | Oxford are providing biological material. |
Impact | Grant application. |
Start Year | 2022 |
Description | Cardiff Huntington's Disease Centre Public Engagement Event |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | Presentation to the Cardiff Huntington's Disease Centre Public Engagement Event. Great interest from the patients and carers about the work we are doing to support future clinical trials. |
Year(s) Of Engagement Activity | 2023 |
Description | DHSC |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Presentation made to DHSC workshop on non-genetic conditions and how to address some of the barriers to improved outcomes and experience. |
Year(s) Of Engagement Activity | 2023 |
Description | NHS WALES RDID |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Invited presentation to NHS Wales Rare Diseases Implementation Group. The presentation made the group aware of the potential of metabolomics/lipidomics for rare disease diagnosis. |
Year(s) Of Engagement Activity | 2023 |
Description | Welsh Government Visit |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Policymakers/politicians |
Results and Impact | Eluned Morgan AS/MS, Minister for Health and Social Services Welsh Government visited our laboratories to learn more about how metabolomics/lipidomics can be used in the diagnosis of rare diseases. |
Year(s) Of Engagement Activity | 2023 |
URL | https://www.swansea.ac.uk/press-office/news-events/news/2023/12/swansea-university-opens-specialist-... |