Targeting circadian clock mechanisms in the intervertebral disc towards therapies for low back pain
Lead Research Organisation:
University of Manchester
Department Name: School of Biological Sciences
Abstract
Low back pain (LBP), caused by degeneration of the fibro-cartilaginous intervertebral disc in the spine, is amongst the most prevalent spinal diseases. It affects millions of individuals worldwide, causing severe pain and loss of mobility, with currently no long-term cure. Ageing is a major risk factor. However, we do not fully understand why age increases susceptibility to disc degeneration and LBP. Body clocks are responsible for generating 24 hour rhythms in behaviour and physiology. Rhythm disruptions by ageing or shift work can significantly increase disease risks. Crucially, our group, for the first time, has discovered a functional body clock in the intervertebral disc. This peripheral body clock weakens with age and its disruption leads to tissue damage characteristic of disc degeneration. This project will establish the disease relevance and explore the therapeutic potential of the body clocks in this tissue. Specifically, we will use our novel disc "clockless" mouse model, intervertebral disc explant culture and patient samples to test the following hypotheses: 1). Daily loading cycle associated with rest/activity can reset the IVD circadian rhythm; 2). Disruption to the IVD circadian rhythm compromises tissue homeostasis/integrity, and leads to increased susceptibility to degeneration; 3). Targeting IVD clock mechanisms provide a novel therapeutic approach for disc degeneration. Our research holds strong potential to identify novel therapeutic strategies for low back pain and therefore has an enormous potential for improving health and quality of life. Moreover, our findings emphasize the importance of maintaining robust body clocks through healthy life styles, which will have a long lasting impact on the health and well-being of the ageing population. This is particularly important because the greying of the population and the ever-increasing demands of our modern 24/7 society frequently disrupt our body clocks which can significantly increase risk of many diseases.
Technical Summary
Circadian clocks drive ~24 hour rhythms in nearly all aspects of physiology and behavior. Changes of circadian rhythms during ageing underpin many of the age-associated pathologies. Low back pain (LBP) is amongst the most prevalent spinal diseases associated with old age. It causes severe pain and loss of mobility, with currently no long-term cure. Progressive degeneration of the intervertebral disc (IVD) tissue is a major contributing factor. However, how susceptibility to IVD degeneration and LBP increases with age is poorly understood. Our preliminary data demonstrates cell autonomous circadian clocks in mouse intervertebral disc tissue and human disc cells. Significantly, our circadian transcriptome profiling data strongly implicate the circadian clock as a key regulatory mechanism for genes involved in disc tissue homeostasis. Therefore, it is timely to explicitly demonstrate the mechanistic links between the IVD clock and disc tissue homeostasis. Specifically, this research will combine our unique transgenic mouse, ex vivo disc explant and experiments utilising human IVDs to test the following hypotheses: 1) Daily osmotic changes associated with diurnal loading cycle can entrain the IVD circadian rhythm; 2) Disruption to the disc circadian rhythm compromises IVD homeostasis and leads to increased susceptibility to degeneration; 3) Targeting IVD clock mechanisms provides a novel approach to modulate the expression/activity of processes linked to disc degeneration. Key methodologies will include histology, RNAseq, qPCR and Western Blotting analysis to measure rhythmic patterns of gene expression, lentiviral gene delivery, real time bioluminescence monitoring of the dynamics of clock gene reporters, ChIP and functional luciferase assays. Importantly, this new avenue of research holds strong potential to identify new drugs (e.g. clock acting compounds) or improve current treatments (such as timing the delivery of drugs) for IVD degeneration and LBP.
Planned Impact
Low back pain (LBP), caused by degeneration of the intervertebral disc (IVD), is amongst the most prevalent spinal diseases. It affects millions of individuals, causing severe pain and loss of mobility, with currently no long-term cure. Ageing is a major risk factor. However, we do not fully understand why age increases susceptibility to disc degeneration and LBP. Our modern 24/7 society and ageing frequently disrupt circadian rhythms generated by our body clocks. We have discovered a functional clock in the IVD which weakens with age and its disruption leads to tissue damage. This project will establish the disease relevance and explore the therapeutic potential of the body clocks in this tissue. Our research may identify novel therapeutic strategies for spine diseases (including LBP), or improve the efficacy of current treatments and thus has enormous potential for improving health and quality of life.
Scientific Impact
This project will initially benefit a wide range of researchers (see Academic Beneficiaries). We will reveal for the first time the genome-wide rhythmic target genes in the IVD, and establish mechanistic links between IVD clock dysregulation and tissue damage. This knowledge will significantly advance our understanding of disc physiology and disease. Second, our research will reveal a completely new mechanism of clock entrainment by dynamic osmotic changes associated with the daily loading cycle. This new mechanism may also apply to other musculoskeletal tissues, and therefore will impact on researchers in other fields of skeletal biology or chronobiology. Finally, we will link circadian rhythms to human IVD degeneration for the first time, and explore the clock-targeting potential to modify disc pathways.
Novel Technology
To address the functional significance of the IVD peripheral clock, we have generated a unique conditional IVD-"clockless" mouse model. This novel mouse line is essential because current available models of circadian rhythm disruptions (e.g. whole body clock mutant mice, or mice kept under weekly reversal of light/dark cycle) lead to global disruptions to circadian rhythms, which will severely confound the interpretation of results. In addition, the use of the clock gene reporter mouse (PER2::Luciferase) and human IVD clock reporter cell line models will greatly facilitate the accurate evaluation of the molecular circadian clocks.
Translational impact
The pathways identified by this project that link circadian clocks to intervertebral disc biology (such as REV-ERBa and RORa the possibilities of targeting NHRs (PPARs and LXR) for skeletal diseases. Over the last few years, QJM has established collaborative links with GSK and Pfizer, which will ensure rapid translation of the research findings.
Societal and health impact
Our studies will have profound impacts on public understanding of rhythm disruptions and disease risks. We expect that the outcomes will be media worthy and be of great interest to the sufferers of LBP, healthcare professionals, the general public and the elderly population. The rhythmic nature of the clock targets identified in this study calls for novel treatment strategies for spine diseases, which take into accounts the time-of-day effect, i.e., chronopharmacology. Furthermore, the quest for prognostic biomarkers also has to consider their potential daily variations. Our research will also reveal detrimental effects of rotating shift work to spine health and LBP, and therefore could influence the policy-makers regarding how to best set up schedules for shift workers. Finally, understanding the time-of-day exacerbation of their symptoms may help LBP sufferers better manage their pain and mobility.
Scientific Impact
This project will initially benefit a wide range of researchers (see Academic Beneficiaries). We will reveal for the first time the genome-wide rhythmic target genes in the IVD, and establish mechanistic links between IVD clock dysregulation and tissue damage. This knowledge will significantly advance our understanding of disc physiology and disease. Second, our research will reveal a completely new mechanism of clock entrainment by dynamic osmotic changes associated with the daily loading cycle. This new mechanism may also apply to other musculoskeletal tissues, and therefore will impact on researchers in other fields of skeletal biology or chronobiology. Finally, we will link circadian rhythms to human IVD degeneration for the first time, and explore the clock-targeting potential to modify disc pathways.
Novel Technology
To address the functional significance of the IVD peripheral clock, we have generated a unique conditional IVD-"clockless" mouse model. This novel mouse line is essential because current available models of circadian rhythm disruptions (e.g. whole body clock mutant mice, or mice kept under weekly reversal of light/dark cycle) lead to global disruptions to circadian rhythms, which will severely confound the interpretation of results. In addition, the use of the clock gene reporter mouse (PER2::Luciferase) and human IVD clock reporter cell line models will greatly facilitate the accurate evaluation of the molecular circadian clocks.
Translational impact
The pathways identified by this project that link circadian clocks to intervertebral disc biology (such as REV-ERBa and RORa the possibilities of targeting NHRs (PPARs and LXR) for skeletal diseases. Over the last few years, QJM has established collaborative links with GSK and Pfizer, which will ensure rapid translation of the research findings.
Societal and health impact
Our studies will have profound impacts on public understanding of rhythm disruptions and disease risks. We expect that the outcomes will be media worthy and be of great interest to the sufferers of LBP, healthcare professionals, the general public and the elderly population. The rhythmic nature of the clock targets identified in this study calls for novel treatment strategies for spine diseases, which take into accounts the time-of-day effect, i.e., chronopharmacology. Furthermore, the quest for prognostic biomarkers also has to consider their potential daily variations. Our research will also reveal detrimental effects of rotating shift work to spine health and LBP, and therefore could influence the policy-makers regarding how to best set up schedules for shift workers. Finally, understanding the time-of-day exacerbation of their symptoms may help LBP sufferers better manage their pain and mobility.
Publications
Yeung CC
(2023)
Mmp14 is required for matrisome homeostasis and circadian rhythm in fibroblasts.
in Matrix biology : journal of the International Society for Matrix Biology
Yang N
(2020)
Quantitative live imaging of Venus::BMAL1 in a mouse model reveals complex dynamics of the master circadian clock regulator.
in PLoS genetics
Streuli CH
(2019)
Influence of the extracellular matrix on cell-intrinsic circadian clocks.
in Journal of cell science
Sherratt MJ
(2019)
Circadian rhythms in skin and other elastic tissues.
in Matrix biology : journal of the International Society for Matrix Biology
Preston R
(2022)
The dynamic kidney matrisome - is the circadian clock in control?
in Matrix biology : journal of the International Society for Matrix Biology
Naven MA
(2022)
Development of human cartilage circadian rhythm in a stem cell-chondrogenesis model.
in Theranostics
Morris H
(2021)
Tissue physiology revolving around the clock: circadian rhythms as exemplified by the intervertebral disc.
in Annals of the rheumatic diseases
Gonçalves C
(2019)
Timing metabolism in cartilage and bone: links between circadian clocks and tissue homeostasis
in Journal of Endocrinology
Dudek M
(2023)
The clock transcription factor BMAL1 is a key regulator of extracellular matrix homeostasis and cell fate in the intervertebral disc.
in Matrix biology : journal of the International Society for Matrix Biology
Dudek M
(2017)
The intervertebral disc contains intrinsic circadian clocks that are regulated by age and cytokines and linked to degeneration
in Annals of the Rheumatic Diseases
Dudek M
(2023)
The circadian clock and extracellular matrix homeostasis in aging and age-related diseases.
in American journal of physiology. Cell physiology
Dudek M
(2023)
Mechanical loading and hyperosmolarity as a daily resetting cue for skeletal circadian clocks
in Nature Communications
Dudek M
(2021)
Circadian time series proteomics reveals daily dynamics in cartilage physiology.
in Osteoarthritis and cartilage
Chang J
(2020)
Circadian control of the secretory pathway maintains collagen homeostasis.
in Nature cell biology
Title | One of our research images was chosen as the cover image of Nature Reviews Rheumatology 2018 (all 12 issues) |
Description | An experimental image of degenerative articular cartilage in the clockless mouse model was chosen as the cover image of Nature Reviews Rheumatology 2018 (all 12 issues) |
Type Of Art | Image |
Year Produced | 2018 |
Impact | All readers of the NRR journal will be able to see this image and the research story associated with it. |
URL | http://www.nature.com/nrrheum/journal/v14/n2/covers/index.html |
Title | https://www.asmb.net/image-contest-winners |
Description | In 2019, Honor Morris' research image was one of the winners of the ASMB (American Society for Matrix Biology) image contest. |
Type Of Art | Image |
Year Produced | 2019 |
Impact | The image showcases our cutting edge techniques in imaging collagen matrix. |
URL | https://www.asmb.net/image-contest-winners |
Description | MRC DTP PhD studentship |
Amount | £100,011 (GBP) |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 08/2017 |
End | 08/2021 |
Description | MRC DTP studentship |
Amount | £98,000 (GBP) |
Organisation | MRC Doctoral Training Program |
Sector | Academic/University |
Country | United Kingdom |
Start | 09/2021 |
End | 09/2025 |
Description | MRC project grant |
Amount | £546,000 (GBP) |
Funding ID | MR/T016744/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2020 |
End | 12/2022 |
Description | Clocks in musculoskeletal system |
Organisation | University of Hong Kong |
Country | Hong Kong |
Sector | Academic/University |
PI Contribution | My lab has started working with Prof. Danny Chan (Hong Kong University) on circadian rhythms in the musculoskeletal system. |
Collaborator Contribution | The Chan lab has strong expertise in cartilage/intervertebral disc biology, development and genetics. We have started by exchange of visiting scientists. |
Impact | New collaboration. |
Start Year | 2018 |
Description | Smart cells chondrocytes |
Organisation | Washington University in St Louis |
Country | United States |
Sector | Academic/University |
PI Contribution | We have a joint publication currently under review at Science Advances. |
Collaborator Contribution | The Guilak lab generated smart cells chondrocytes which can produce anti-inflammatory agents upon inflammation. |
Impact | Multi-disciplinary collaboration as it spans genetic engineering, immunology and circadian biology. |
Start Year | 2020 |
Description | BBC Breakfast |
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 | I was interviewed by BBC Breakfast to discuss my body clock research and its implications in chronotherapy and shift work |
Year(s) Of Engagement Activity | 2018 |
Description | BBC Breakfast |
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 | I was invited to BBC Breakfast to comment on a BBC Documentary on Body Clocks |
Year(s) Of Engagement Activity | 2018 |