Defining the role and mechanisms of miRs in cartilage ageing and disease
Lead Research Organisation:
University of Liverpool
Department Name: Institute of Ageing and Chronic Disease
Abstract
The musculoskeletal system is severely affected by the ageing process, with many tissues undergoing changes that lead to loss of function and frailty. Articular cartilage is particularly susceptible to the age-related disease, osteoarthritis (OA). OA is the most common degenerative joint disorder worldwide, affecting 8.75 million people in the UK and presents with degradation of articular cartilage, leading to loss of joint mobility, function, and chronic pain. However the molecular mechanisms associated with age-specific and OA-specific changes in cartilage are poorly understood.
MicroRNAs (miRs) are novel gene expression regulators, which control tissue homeostasis in response to environment and intracellular changes. Their expression has been shown to be dysregulated during human and animal ageing in a number of different systems/tissues. miRNAs are differentially expressed in cartilage during ageing and in OA, and are thought to play a role in the regulation of important transcription factors involved in the development, maintenance, and repair of cartilage. Moreover, our small RNA-sequencing data from the femorotibial joint of C56BL6/J male mice, confirmed the differential expression of specific miRNAs with age and an in vivo model of OA (destabilisation of the medial meniscus). In addition we have preliminary data which has identified biologically relevant mRNAs and their targets in human knee OA and ageing cartilage.
We postulate that alterations in miRNA expression in the ageing cartilage result in a dysregulation of gene expression and hence altered chondrocyte phenotype contributing to the age-related disease OA. This will be tested by investigating the following overarching objectives:
1. Establish differentially expressed miRs in normal and OA chondrocytes during ageing using small RNAsequencing. This data will be produced as a by-product of a study being undertaken by the primary applicant to generate findings for small nucleolar RNAs and is funded by a Wellcome Trust Clinical Intermediate Fellowship.
2. Characterise selected target genes based on known or predicted function microRNAs validated from objective 1 through qPCR analysis and 3'UTR reporter assays, as well as microRNA gain- and loss-of-function approaches.
3. Investigating the role of physiologically relevant microRNA:target interactions predicted to regulate extracellular maintenance and cartilage homeostasis in chondrocytes. These studies will be undertaken in vitro and in vivo by analysing the phenotypes resulting from microRNA gain- and loss-of-function approaches.
MicroRNAs (miRs) are novel gene expression regulators, which control tissue homeostasis in response to environment and intracellular changes. Their expression has been shown to be dysregulated during human and animal ageing in a number of different systems/tissues. miRNAs are differentially expressed in cartilage during ageing and in OA, and are thought to play a role in the regulation of important transcription factors involved in the development, maintenance, and repair of cartilage. Moreover, our small RNA-sequencing data from the femorotibial joint of C56BL6/J male mice, confirmed the differential expression of specific miRNAs with age and an in vivo model of OA (destabilisation of the medial meniscus). In addition we have preliminary data which has identified biologically relevant mRNAs and their targets in human knee OA and ageing cartilage.
We postulate that alterations in miRNA expression in the ageing cartilage result in a dysregulation of gene expression and hence altered chondrocyte phenotype contributing to the age-related disease OA. This will be tested by investigating the following overarching objectives:
1. Establish differentially expressed miRs in normal and OA chondrocytes during ageing using small RNAsequencing. This data will be produced as a by-product of a study being undertaken by the primary applicant to generate findings for small nucleolar RNAs and is funded by a Wellcome Trust Clinical Intermediate Fellowship.
2. Characterise selected target genes based on known or predicted function microRNAs validated from objective 1 through qPCR analysis and 3'UTR reporter assays, as well as microRNA gain- and loss-of-function approaches.
3. Investigating the role of physiologically relevant microRNA:target interactions predicted to regulate extracellular maintenance and cartilage homeostasis in chondrocytes. These studies will be undertaken in vitro and in vivo by analysing the phenotypes resulting from microRNA gain- and loss-of-function approaches.
Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| MR/N013840/1 | 01/10/2016 | 30/09/2025 | |||
| 1794649 | Studentship | MR/N013840/1 | 05/09/2016 | 28/02/2020 | Panagiotis Balaskas |
| Description | Research Support Budget |
| Amount | £1,500 (GBP) |
| Organisation | University of Liverpool |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 02/2019 |
| End | 02/2019 |
| Description | Hope Scout Vlog |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | Who and how many people were involved - include both UoL staff/students and members of the public Mandy Peffers took part in both events James Anderson (PhD student), Aibek Smagul (PhD student), Panos Balaskas (PhD student), and Phil Dyer (research Technician) took part in the first event. What you did The project consisted of two parts. 1. On the initial visit to the Scout Group there will a number of activities. There was an oral presentation on the skeleton and joint ageing in man (15min) and small-groups of four-five scouts interacted with scientists using skeletons and joint models (30min). Two copies of a fact sheet were given to the scouts; one for them and one for the elderly participant. This was followed with a presentation on tips on how to make a 'video blog'. The video was to consist of the scout making a recording using their phone/I-Pad of an elderly participant on the elderly person's perception and experience of healthy (or unhealthy) joint and bone ageing. In a final small-group session scouts broke into twos and practiced using their phones or I-Pads (30min) to record their partner in a' mock' interview scenario. A handout containing a set of ideas for questions, and the format of the interview to be undertaken was given out (see below). The Scouts were asked to identify an elderly relative or colleague to make a two minute video with. The following week the participants were asked to email or put on pen-drives their videos for us to assess and make into a presentation for visit 2; the Showcase Event. 2. Visit two will be undertaken two weeks later at the Scout Hut. The scouts were invited to a 'Showcase Event'. The event included a short presentation consolidating the initial presentation (visit 1; skeleton and joint ageing and research we conduct at UoL in arthritis) and fact sheet given on visit one. A quiz was undertaken on bone which was marked by the scout leaders and prizes given for the top three results. As some of the vlogs were only given to me on the night I could not pre judge them. Therefore in order to engage the scouts more I gave each of them a piece of paper to put their name on and they were asked to vote for their favourite vlog. Out of 20 scouts from the first visit seven submitted vlogs (according to the scout leaders this was actually a good engagement rate!). Following this refreshments were served. Another quiz was given this time I read out some facts about arthritis and asked them to answer the multi choice questions straight away. A presentation followed with prizes for all those who submitted a vlog and further prizes for the top three. What were the outcomes, and you may want to think about this in terms of whether your project had an impact on any of the following (please include any supporting evidence): General I feel the event was successful. The quality of the vlogs was really good and each scout put their own spin on it. For instance one scout interviewed himself using a facemask to represent his older interviewee. Another took the role of a TV presenter. Talking to the scouts with a quiz immediately following was a great success as it made the scouts listen and think more. Your research The scouts were made aware that arthritis was an age-related disease and also learnt about other factors important in the disease. This was demonstrated by the second quiz in which all scouts got >50% of the questions correct. On the first visit using bones and joints from animals we were able to demonstrate anatomy but also that animals also get arthritis. The staff/students involved All my group were involved from the preplanning, making the interview script to running the first event. They all enjoyed it and some, who had not undertaken PE before were enthused! The public involved One of the outcome was that we were able to find out public opinion in an ageing group of what their joint problems were, what they thought about funding and if they had any tips for healthy musculoskeletal ageing. The University The scouts and interviewees were able to find out what research we do at Liverpool. I hope to put the vlogs on IACD website for further benefit of our work. |
| Year(s) Of Engagement Activity | 2018 |
| Description | INSPIRE Summer School |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Undergraduate students |
| Results and Impact | Mandy Peffers organised a 3 day INSPIRE funded summer school for veterinary undergraduates and UoL medical students. Panos assisted with some of the activities during the summer school. |
| Year(s) Of Engagement Activity | 2019,2020 |