The role of intermediary metabolism on skin ageing and interplay between inflammation and senescence
Lead Research Organisation:
Queen Mary University of London
Department Name: Blizard Institute of Cell and Molecular
Abstract
Objectives
We propose (a) intrinsic and extrinsic ageing of fibroblasts promote a switch from glycolytic to OXPHOS metabolism in keratinocytes (b) resulting in increased ROS promoting signs of ageing including changes in skin thickness, metalloproteinase and extracellular matrix (ECM) production and impaired hair growth. (c) Changes in skin metabolism due to intrinsic and extrinsic ageing impact on its ability to respond to UV and oxidative stress promoting further ageing.
The overarching aim is to provide a rationale for modulating keratinocyte metabolism as a strategy for promoting healthy ageing. The combination of diverse strengths from each party involved provides a chance for a novel way of investigating ageing.
Work plan
We will utilise 2D cell cultures combined with 3D living skin equivalents (LSE) to investigate the effect of dermal fibroblasts (DF) & DP on keratinocyte metabolism (glycolysis and OXPHOS) and investigate the effects of UV and oxidative stress on these metabolic pathways and correlate our findings in ex vivo human skin and hair.
Key aims
1a Establish rates of glycolysis and OXPHOS in neonatal, intrinsically aged and senescent DF and balding/non-balding DP using transcriptomics, Seahorse extracellular flux analysis, ATP and NADP/NADPH, mitochondrial uptake of fluorescently labelled deoxy-glucose and mitochondrial activity by imaging rhodamine dye accumulation (TMRM).
1b Investigate impact of UV and Oxidative stress on glycolysis and OXPHOS, ATP and NADP/NADPH, glucose uptake and TMRN in neonatal, intrinsically aged and senescent DF and DP.
2a Use transcriptomics to validate metabolic profile in keratinocytes incorporated into LSE containing neonatal, intrinsically aged and senescent DF and prioritise metabolic pathways for further follow-up using LC-MS/MS.
2b Investigate glycolysis, OXPHOS, ATP and NADP/NADPH, glucose uptake and TMRN in LSE incorporating neonatal, intrinsically aged and senescent DF and keratinocyte DP co-cultures and impact of UV and oxidative stress on these metabolic parameters.
3a Characterise the mitochondrial metabolomic changes which occur during the ageing processes and UV/Oxidative stress using LC-MS/MS under the supervision of Dr Katiuscia Bianchi. Although 13C analysis is technically challenging and considered higher risk, Dr Bianchi is currently establishing this technique in her laboratory in collaboration with Dr Christian Frezza, MRC Cancer Unit, Cambridge. 13C techniques will provide more detailed metabolic profile as multiple metabolites can be measured. Additional support will be available with access to state-of-the-art equipment at Unilever's partners at The University of Liverpool.
We propose (a) intrinsic and extrinsic ageing of fibroblasts promote a switch from glycolytic to OXPHOS metabolism in keratinocytes (b) resulting in increased ROS promoting signs of ageing including changes in skin thickness, metalloproteinase and extracellular matrix (ECM) production and impaired hair growth. (c) Changes in skin metabolism due to intrinsic and extrinsic ageing impact on its ability to respond to UV and oxidative stress promoting further ageing.
The overarching aim is to provide a rationale for modulating keratinocyte metabolism as a strategy for promoting healthy ageing. The combination of diverse strengths from each party involved provides a chance for a novel way of investigating ageing.
Work plan
We will utilise 2D cell cultures combined with 3D living skin equivalents (LSE) to investigate the effect of dermal fibroblasts (DF) & DP on keratinocyte metabolism (glycolysis and OXPHOS) and investigate the effects of UV and oxidative stress on these metabolic pathways and correlate our findings in ex vivo human skin and hair.
Key aims
1a Establish rates of glycolysis and OXPHOS in neonatal, intrinsically aged and senescent DF and balding/non-balding DP using transcriptomics, Seahorse extracellular flux analysis, ATP and NADP/NADPH, mitochondrial uptake of fluorescently labelled deoxy-glucose and mitochondrial activity by imaging rhodamine dye accumulation (TMRM).
1b Investigate impact of UV and Oxidative stress on glycolysis and OXPHOS, ATP and NADP/NADPH, glucose uptake and TMRN in neonatal, intrinsically aged and senescent DF and DP.
2a Use transcriptomics to validate metabolic profile in keratinocytes incorporated into LSE containing neonatal, intrinsically aged and senescent DF and prioritise metabolic pathways for further follow-up using LC-MS/MS.
2b Investigate glycolysis, OXPHOS, ATP and NADP/NADPH, glucose uptake and TMRN in LSE incorporating neonatal, intrinsically aged and senescent DF and keratinocyte DP co-cultures and impact of UV and oxidative stress on these metabolic parameters.
3a Characterise the mitochondrial metabolomic changes which occur during the ageing processes and UV/Oxidative stress using LC-MS/MS under the supervision of Dr Katiuscia Bianchi. Although 13C analysis is technically challenging and considered higher risk, Dr Bianchi is currently establishing this technique in her laboratory in collaboration with Dr Christian Frezza, MRC Cancer Unit, Cambridge. 13C techniques will provide more detailed metabolic profile as multiple metabolites can be measured. Additional support will be available with access to state-of-the-art equipment at Unilever's partners at The University of Liverpool.
People |
ORCID iD |
| Michael Philpott (Primary Supervisor) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/R505420/1 | 01/10/2017 | 31/12/2021 | |||
| 1962581 | Studentship | BB/R505420/1 | 01/10/2017 | 30/09/2021 |
| Description | This project aims to study biological mechanisms of the process of ageing in human skin. To understand the process better the project has allowed to develop two physiologically relevant study models of skin ageing. These models induce cell damage, cell cycle arrest and proliferation loss (which is termed as 'senescence') via: 1) treating human cells with ultraviolet radiation, or 2) through continous cell culture until cell proliferation capacity is exhausted. Therefore, this project has allowed to: 1) implement new culture models to study senescence, 2) study the differences and biological relevance of these models 3) collaborate with various academic leads and bring their expertise and views to the project 4) attract interest from industrial collaborator, providing experimental guidance as well as the financial support to equip the project 5) improved my research skills and knowledge in the field through providing additional training and attendance of specialist courses (e.g SysMIC, Cellular Bioenergetics Course on metabic fluxes) |
| Exploitation Route | Research methods and the knowledge obtained from the project will greatly benefit to the field of senescence, ageing and skin biology. More specifically, identification of specific genes, proteins and metabolites associated with ageing will enhance the development of therapeutics and novel active agents to promote human longevity and wellbeing. |
| Sectors | Chemicals,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| URL | https://gtr.ukri.org/projects?ref=studentship-1962581 |