Role of microRNAs in ageing at the blood-brain barrier: integrated studies in human and mouse models

Lead Research Organisation: University of Sheffield
Department Name: Neurosciences

Abstract

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Technical Summary

The aim of the proposal is to test the hypothesis that age-induced changes in gene expression in cerebral endothelium leading to cerebrovascular dysfunction are mediated, at least partially, by changes in microRNA levels. MicroRNAs (miRs) are endogenous non-coding small RNAs that mediate gene silencing by translation inhibition and/or by mRNA decay of target genes. Evidence suggests a role for miRs in regulating the life span of model organisms but the mechanisms involved in this process are largely unknown. One of the first signs of ageing is the loss of specialized functions by cells. The objective of the work is to elucidate general mechanisms by which miRs may modulate normal ageing in a specialized biological system that is amenable to functional, cellular and molecular investigations, the blood-brain barrier (BBB), across two model organisms with differing life spans, humans and mice. First, we will characterize progressive changes in BBB function in humans and mice during normal ageing. Second, using microarray technology, age-deregulated miRs in the cerebral endothelium and age-associated shifts in the global pattern of target gene expression will be analysed so that changes predicted to be directed by alterations in miR levels at the BBB are identified. Third, specific age-deregulated miRs will be selected using their ability to modulate specialized BBB function and/or known pathways of cellular senescence in vitro. Finally, we will validate actions of specific miRs on putative gene targets and confirm age-induced changes in levels of selected miR/target gene pairs at the BBB. BBB dysfunction is recently emerging as an important factor leading to age-associated loss of homeostasis in the central nervous system and we envisage that age-induced phenotypic changes at the BBB mediated by miRs will be identified that may well be not only specific to cerebrovascular function but also constitute general pathways of ageing across biological systems.

Planned Impact

The proposed research seeks to determine the roles of microRNAs in the endothelium of the blood-brain barrier (BBB) during normal brain ageing. It has two main aims: first to systematically characterise changes occurring in brain endothelial cells during ageing and, second, to identify the roles of microRNAs in these changes, and thus their possible involvement in age-induced BBB dysfunction. Comparison of microRNA changes with alterations in the general endothelial transcriptome will provide a powerful data set of altered endothelial gene expression in ageing. BBB dysfunction and endothelial pathology have been implicated in the development and progression of degenerative brain disease, such as Alzheimer's Disease. However, these processes, their interaction with brain parenchyma ageing and their roles in disease remain poorly defined. The results from the work will therefore have a major impact both on our knowledge base in the areas of microRNA functions, ageing and neuroscience and also in the future, for health and wellbeing, as the results should lead to knowledge that can be used in the development of strategies and therapies to prevent, delay or alleviate age-associated BBB dysfunction in neuropathological conditions. Such novel approaches are essential given the limitations of current strategies.
The immediate beneficiaries of the proposed research are academics, the public sector and third sectors. In the longer term, potential future beneficiaries include health professionals, the commercial private sector and general public.
The academic community will benefit from the proposed research because at present our knowledge of the roles of microRNAs is in its infancy. Evidence suggests that these molecules play a role in ageing and in endothelial cell biology, and the proposed work will provide new information in both these important areas.
Within the public and third sectors, the specific groups to whom this research is relevant are research organisations (including BBSRC, MRC), professional bodies (such as the British Society for Research on Ageing, the British Neuropathological Society and the British Neuroscience Association) and charities (e.g. Age UK, Alzheimer's Research Trust, Alzheimer's Society, Alzheimer's Association). These groups will benefit from the knowledge generated from the project, as it will inform decisions on the future research priorities and development activities in the areas of ageing and neuroscience, in particular neuropathology.
We do not anticipate any early developments that will warrant commercial exploitation, although there is potential for such exploitation in the future. We will ensure that any such developments are taken forward, with appropriate partners and IP procedures.
The health and wellbeing of the general public will benefit from this research in the longer term. For example, understanding how the process of normal ageing affects blood-brain barrier function, in particular that related to transporter activity which influences CNS availability of many commercial drugs, will impact on the development of therapeutic regimes targeted at the ageing population with neurological conditions. The public should also have the benefit of being informed about the work.

Publications

10 25 50
 
Description We have now published a paper showing changes in blood brain barrier across age in both humans and mice. We have now completed and published a second paper detailing changes in gene expression in endothelium across the ageing spectrum in humans and comparing this to mice.
Exploitation Route It would be good to take forward key pathway targets into functional studies.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description One data paper was published showing that blood brain barrier is increasingly leaky with age in mouse and human. We also published a second paper on gene expression changes. To complete the work for this, we provided a BMedSci student project. This was successfully completed and the student presented at a national conference.
First Year Of Impact 2019
Sector Healthcare
 
Description Interdisciplinary research grant
Amount £237,984 (GBP)
Funding ID ARUK-IRG2014-10 
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2014 
End 12/2016
 
Description Occludin as a Mediator of Neuroglial RNA Dysmetabolism in Dementia
Amount £47,703 (GBP)
Funding ID ARUK-PPG2019B-005 
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2021 
End 10/2021
 
Description PhD Scholarship
Amount £84,450 (GBP)
Funding ID ARUK-PhD2016-5 
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 08/2016 
End 08/2019
 
Description Preparatory clinical research fellowship
Amount £69,998 (GBP)
Funding ID ARUK-PCRF2016A-1 
Organisation Alzheimer's Research UK 
Department Alzheimers Research UK, Cambridge
Sector Charity/Non Profit
Country United Kingdom
Start 08/2016 
End 09/2017
 
Description Project grant. Basis of post stroke dementia
Amount £378,751 (GBP)
Funding ID 386 
Organisation Alzheimer's Society 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2018 
End 01/2021
 
Title Development of immuno-directed laser capture microdissection. 
Description Protocols for cell-type specific laser capture in the CNS. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? Yes  
Impact Several papers over the years, most recently brought together in a methods chapter Simpson JE, Wharton SB, Heath PR. Immuno-laser-capture microdissection for the isolation of enriched glial populations from frozen post-mortem human brain. In: Murray G (eds) Laser Capture Microdissection. Methods in Molecular Biology vol 1723; pp273-284, Humana Press, New York, NY 
 
Title Age associated endothelial gene expression changes in mouse and human 
Description Changes in mRNA in endothelium across age span in human brain and mice. Freely available at Gene Expression Omnibus (human array data accession number GSE127710 and mouse array data accession number GSE127709). 
Type Of Material Database/Collection of data 
Year Produced 2019 
Provided To Others? Yes  
Impact Availability of data to bioinformatics researchers. 
 
Description A Cellular Pathology and Gene Expression Approach to Understand the Basis of Progressive Dementia Occurring After Stroke 
Organisation Newcastle University
Country United Kingdom 
Sector Academic/University 
PI Contribution Joint study of post stroke dementia. We will provide laser capture and gene expression analysis.
Collaborator Contribution Collaboration has just begun.
Impact Just started
Start Year 2018
 
Description Defective nutrient signalling and dementia: an epidemiological neuropathology approach 
Organisation Newcastle University
Country United Kingdom 
Sector Academic/University 
PI Contribution Pathological analysis of CFAS tissue studying effects of diabetes on the brain.
Collaborator Contribution Tissue analysis.
Impact Conference presentations.
Start Year 2015
 
Description Defective nutrient signalling and dementia: an epidemiological neuropathology approach 
Organisation University of Cambridge
Department Cambridge Institute of Public Health
Country United Kingdom 
Sector Academic/University 
PI Contribution Pathological analysis of CFAS tissue studying effects of diabetes on the brain.
Collaborator Contribution Tissue analysis.
Impact Conference presentations.
Start Year 2015
 
Description Neurovascular dysfunction in Alzheimer's disease: the search for early biomarkers 
Organisation University of Sheffield
Department Department of Psychology
Country United Kingdom 
Sector Academic/University 
PI Contribution Collaboration with Psychology bringing together our tissue expertise and linking to physiological analysis.
Collaborator Contribution Tissue analysis, laser capture and gene expression microarrays.
Impact Review paper detailed in pulications.
Start Year 2014
 
Description ARUK lab tour for lay group. 
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 Public/other audiences
Results and Impact Lab tour and talks informing members of public about our research
Year(s) Of Engagement Activity 2018
 
Description Alzheimer Society Local Group Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Member of out group spoke about our dementia research predominantly to dementia support workers.

Better public information
Year(s) Of Engagement Activity 2014
 
Description Alzheimer's Society Network Volunteers 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 Patients, carers and/or patient groups
Results and Impact Visit by lay group. Talks given on the various projects we have underway.
Year(s) Of Engagement Activity 2016
 
Description Alzheimers Research Network Volunteers Visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Supporters
Results and Impact Attendance by a group of lay monitors. Presented talks in relation to our projects and lab visit. Opportunities for informal discussion and advice from the lay members.
Year(s) Of Engagement Activity 2017
 
Description Annual meeting of the ARUK Network Yorkshire Branch Open Public Meeting. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Series of talks from members of the Neuroscience Dept in Sheffield and premier of film Alive Inside - documentary about living with dementia. Prof Paul Ince from the Neuropathology group gave a talk about different pathological causes of dementia - to raise awareness of the non-AD types.

Public information
Year(s) Of Engagement Activity 2014
 
Description Dementia Futures Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Conference designed to present results on dementia to lay public. 3 of our group's PhD students presented.
Year(s) Of Engagement Activity 2018
 
Description Dementia futures conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Conference to present work of PhD Students to members of the public. Covered by BBC radio Sheffield. Three PhD students from our group gave talks. Also attended by representative of local city council.
Year(s) Of Engagement Activity 2019
URL http://tinyurl.com/DementiaFuturesConference2019
 
Description Dr Garwood presentation at Festival of Science and Engineering Your amazing brain 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Schools
Results and Impact Stimulated interest in children

Questions
Year(s) Of Engagement Activity 2015
 
Description Invited speaker British Neuroscience Association Symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Generate questions and discussion. Improved professional relationships that might lead to collaboration.

Communication both immediately after and by email
Year(s) Of Engagement Activity 2015
 
Description Memory Outreach Clinic 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Patients, carers and/or patient groups
Results and Impact Discussing our work on dementia with relatives at a local memory café.
Year(s) Of Engagement Activity 2020
 
Description Radio interview by Dr Claire Garwood on our group's work, BBC Radio Sheffield 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Media (as a channel to the public)
Results and Impact Informed public via radio

Not formally assessed.
Year(s) Of Engagement Activity 2015
 
Description Research Network volunteer visit (Alz Soc) Aug 2017 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Visit of Alz Soc research network volunteer. Presented the research work of our group.
Year(s) Of Engagement Activity 2017
 
Description SITraN Open Day 
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 Public/other audiences
Results and Impact Institute open day with range of talks
Year(s) Of Engagement Activity 2018
 
Description SITraN Open Day July 2014 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Open day at SITraN allowing lay persons to look at work on neurodegeneration, in including dementia in Sheffield.

Public information and interest in donations
Year(s) Of Engagement Activity 2014
 
Description Short film about dementia research in Sheffield 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Claire Garwood received funding from the Engaged Curriculum to produce a short film about the broad range of dementia research at the University of Sheffield.
Year(s) Of Engagement Activity 2015
 
Description Talk to Ignite Academy Event 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Talk given by member of our group "Astrocytes:Guardians of the Brain"

Better public information
Year(s) Of Engagement Activity 2014
 
Description Visit by Alzheimer's Society network volunteers 
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 Supporters
Results and Impact Stephen Wharton Organised visit by Alzheimer's Society network volunteers. Nov 10 2015. 3 visitors - meet the group, 2 talks and lab tour.
Year(s) Of Engagement Activity 2015
 
Description Visit by lay monitors 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact Visit by lay monitors to discuss our work on dementia
Year(s) Of Engagement Activity 2019
 
Description Visit to local primary school 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Visit to local primary school by team members, Key stages 1-2. Talk and workshop focused on the brain.
Year(s) Of Engagement Activity 2019
 
Description Workshops in primary schools 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Claire Garwood, Laura Ratcliffe and Irina Villasenor. Sheffield Festival of Science and Engineering. 3 Half day workshops in local primary schools "Your Amazing Brain".
Year(s) Of Engagement Activity 2015