The role of small non-coding RNA extracellular vesicle cargo in age-related anterior cruciate ligament degeneration

Background: The anterior cruciate ligment (ACL) is the most frequently damaged ligament of the human knee joint following injuries and age-related ligament breakdown, leading to the development of osteoarthritis (OA). The incidence of age-related ACL breakdown is increasing in our ageing population and currently the mechanisms by which these changes leads to knee joint OA are poorly understood. Early intervention in ACL disease may be able to decelerate OA development/progression with significant benefits for patients and OA-related health care costs.

We have identified changes in small molecules (small non-coding RNAs) in diseased ACLs from OA knee joints. These small molecules can be transferred between tissues in small cell-derived particles (extracellular vesicles) and may have an important regulatory role in the development of OA. However, identification of these small molecules within the ageing ACL as well as the cargo within the associated extracellular vesicles and their influence on other ageing knee joint tissues is unknown.

This project will identify functional differences in small non-coding RNAs (sncRNAs) in ACL and ACL extracellular vesicles (EVs) that could potentially play an important role in the development of knee joint OA.

Objectives:

1)     Identify and characterise the sncRNAs within healthy young and healthy aged ACLs.

2)     Identify the sncRNAs within extracellular vesicles from healthy young and healthy aged ACLs.

3)     Determine the biological role of age-related sncRNAs on other knee joint tissues. 

Novelty: This project is extremely novel in examining the role of the ageing ACL degeneration on other joint tissues within the knee joint organ. ACL degeneration as an initiator of OA has long been overlooked. Our approach determining sncRNAs within ACL and ACL-derived EVs using an in vitro knee model system is innovative and feasible given the multidisciplinary structure of the research team.

Timeliness : Small non-coding RNAs (sncRNAs) have proven highly reliable as biomarkers and preliminary data from our group has identified altered sncRNA expression in cartilage and ACLs from OA joints exposing underlying age-related mechanisms. However alterations of sncRNA expression with age in the ACL and ACL-derived EVs has not been yet investigated and therefore this project is timely in its further expansion of other knee joint tissues’ contribution to joint ageing.

Experimental Approaches 

·        Small RNA sequencing of ACLs and ACL derived EVs sncRNAs

·        EV isolation and characterisation

·        Incubation of knee joint cells lines with identified age-relevant sncRNAs

·        Determining cellular phenotypic shift and matrix changes with qPCR, glycosaminoglycans and hydroxyproline assays

Our group is multidisciplinary with expertise in both wet laboratory work and computational biology skills. All supervisors are unique in our ‘One Health’ approach to musculoskeletal disease, ageing and RNA as we are fundamental, biomedical and veterinary scientists. These supervisors will apply their skills to both human and animal datasets and tissues and therefore would be able to provide students with an interdisciplinary approach to research, preparing them to meet the challenges inherent in the renewed momentum for ‘One Health’.

Other Training to be given:

As part of their degree registration, the student will have access to the Liverpool Doctoral College and its training resources including lectures on time management, team building, career development as well as thesis writing support and viva practice. Other modules such as study design and statistical training are available through the Department of Biostatistics at Liverpool. The student will be a member of the Comparative Musculoskeletal disease group at the Institute of Life Course and Medical Sciences. This research environment will provide a great opportunity for the PhD student to learn new skills in study design, participate in clinical related research, bioinformatics, data analysis and preparing work for publication.

An Undergraduate Degree in Biological Sciences or related; 2:1 or equivalent is required.

Informal enquiries or expressions of interest to Professor Eithne Comerford on [Email Address Removed] or Professor Mandy Peffers on [Email Address Removed].