Periodic Reporting for period 1 - Pept-AGE (Elucidating the molecular mechanisms of age-related muscle loss to inform Artificial Intelligence-based discovery of novel peptide therapeutics for sarcopenia.)
Reporting period: 2020-06-02 to 2022-06-01
Ageing is often viewed as an inevitability – it has been with us for as long as life has existed. With age, there is a gradual decline in physical function. One of the most impactful aspects of this age-related physical decline is sarcopenia. Sarcopenia, or age-related muscle loss, is a primary cause of loss of physical independence in the elderly. Through increased rates of serious falls, loss of physical capability and by promoting a wide range of other age-associated diseases, sarcopenia has a huge impact on society, particularly given that it is considered an accepted side-effect of growing old. The global population is growing older, and with that comes a significant rise in age-associated disease, including sarcopenia – which is expected to explode in incidence in the coming decades. The need for effective treatment strategies that can slow, or reverse age-related muscle loss is essential if a future crisis is to be avoided. Despite the obvious problems posed by sarcopenia, there are no effective therapies currently on the market.
One possible reason for this is that the scientific community has no reliable model of sarcopenia “in a dish”. Often the knowledge used to design drugs relies heavily on basic research, which usually provides information about diseases and illnesses at a molecular level to identify what particular cellular process is no longer working as intended in a particular disease. These discoveries are often based on research carried out using in vitro models – or cells in a dish. Without an easy way to test hypotheses, screen potential therapies before moving into animal models, or even to help identify the molecular pathways responsible for the disease, sarcopenia research is at a massive disadvantage.
To try to address these problems, Pept-AGE had several aims.
1. The primary goal of Pept-AGE was to develop a new model of age-related muscle loss in a dish by pushing skin cells from young and old donors down a different developmental pathway to make young and old muscle cells
2. Add to the current knowledge of the processes that cause sarcopenia using the muscle cells developed in Aim 1
3. Using the model developed in Aim 1 and the newfound mechanisms identified in Aim 2, Nuritas’ AI to identify a peptide with the potential to slow age-related muscle loss down at the cellular level
4. A major goal of all MSCA Individual Fellowships is to expand the skills, training and career prospects of the researcher and so one of the main aims of Pept-AGE in addition to the aims above is development of the fellow as an independent scientist
Importantly, Pept-AGE has succeeded in developing a novel in vitro model of age-related muscle loss and used that model to identify a potentially important pathway that may play a role in age related muscle loss. As such, Pept-AGE has provided an important new tool that will hopefully allow speed up progress in the field of sarcopenia research and is currently in use at Nuritas to test muscle health peptides prior to clinical trials.
One possible reason for this is that the scientific community has no reliable model of sarcopenia “in a dish”. Often the knowledge used to design drugs relies heavily on basic research, which usually provides information about diseases and illnesses at a molecular level to identify what particular cellular process is no longer working as intended in a particular disease. These discoveries are often based on research carried out using in vitro models – or cells in a dish. Without an easy way to test hypotheses, screen potential therapies before moving into animal models, or even to help identify the molecular pathways responsible for the disease, sarcopenia research is at a massive disadvantage.
To try to address these problems, Pept-AGE had several aims.
1. The primary goal of Pept-AGE was to develop a new model of age-related muscle loss in a dish by pushing skin cells from young and old donors down a different developmental pathway to make young and old muscle cells
2. Add to the current knowledge of the processes that cause sarcopenia using the muscle cells developed in Aim 1
3. Using the model developed in Aim 1 and the newfound mechanisms identified in Aim 2, Nuritas’ AI to identify a peptide with the potential to slow age-related muscle loss down at the cellular level
4. A major goal of all MSCA Individual Fellowships is to expand the skills, training and career prospects of the researcher and so one of the main aims of Pept-AGE in addition to the aims above is development of the fellow as an independent scientist
Importantly, Pept-AGE has succeeded in developing a novel in vitro model of age-related muscle loss and used that model to identify a potentially important pathway that may play a role in age related muscle loss. As such, Pept-AGE has provided an important new tool that will hopefully allow speed up progress in the field of sarcopenia research and is currently in use at Nuritas to test muscle health peptides prior to clinical trials.
The work carried out during the fellowship has made important strides in progressing the development of an in vitro model of muscle loss, especially in the context of advancing age. In addition, the work carried out during this fellowship was particularly beneficial in developing the skills and training of the fellow. A brief summary of the work carried out, as well as key outcomes can be found below.
WP 1 successfully developed and refined the method to make young and old muscle cells from young and old fibroblasts
• A differentiation protocol to generate DIMs was developed and refined - resulting in generation of age-appropriate myoblasts from fibroblasts
• IMR90s were shown to be an excellent source of fibroblasts from which to differentiate DIMs at scale
• DIMs were extensively characterised and found to compare favourably to primary human skeletal muscle cells (HSkMCs) in many regards, and even outperform primary HSkMCs in others
Building on the success of the model developed in WP1, WP2 looked at the expression of thousands of proteins simultaneously to identify changes in aged DIMs compared to young
• Extensive proteomic characterisation of the new DIMs model was carried out identifying important differences between young and old DIMs relevant to age related muscle loss (further validating the model)
• Mitochondrial function was identified as one of the most highly dysregulated processes in old DIMs in agreement with current literature - this preliminary data formed the hypothesis for a follow-on grant which was recently awarded (€2.44 million from Enterprise Ireland)
• A potentially unreported mode of age-related muscle loss was identified in down-regulation of both subunits of LAT1 in old DIMs
• The DIMs developed and characterised in WP1 and WP2 are currently being used to help repurpose an existing Nuritas muscle health product and provide data in an aging context before moving into clinical trials
WP4, 5 & 6 dealt with transfer of knowledge, outreach and project management respectively
• The fellow received extensive training and experience in the business facing aspects of R&D in industry – something which the fellow had no previous experience with
• The fellow significantly expanded the experimental toolkit at Nuritas
• Pept-AGE hosted secondary school students for a week of hands-on experiments, saw the fellow lecture to Secondary school students on STEM career paths, and hold a career Q&A with MSCA early-stage researchers
• The findings of Pept-AGE have been presented at the key sarcopenia meeting (ICFSR, Boston)
WP 1 successfully developed and refined the method to make young and old muscle cells from young and old fibroblasts
• A differentiation protocol to generate DIMs was developed and refined - resulting in generation of age-appropriate myoblasts from fibroblasts
• IMR90s were shown to be an excellent source of fibroblasts from which to differentiate DIMs at scale
• DIMs were extensively characterised and found to compare favourably to primary human skeletal muscle cells (HSkMCs) in many regards, and even outperform primary HSkMCs in others
Building on the success of the model developed in WP1, WP2 looked at the expression of thousands of proteins simultaneously to identify changes in aged DIMs compared to young
• Extensive proteomic characterisation of the new DIMs model was carried out identifying important differences between young and old DIMs relevant to age related muscle loss (further validating the model)
• Mitochondrial function was identified as one of the most highly dysregulated processes in old DIMs in agreement with current literature - this preliminary data formed the hypothesis for a follow-on grant which was recently awarded (€2.44 million from Enterprise Ireland)
• A potentially unreported mode of age-related muscle loss was identified in down-regulation of both subunits of LAT1 in old DIMs
• The DIMs developed and characterised in WP1 and WP2 are currently being used to help repurpose an existing Nuritas muscle health product and provide data in an aging context before moving into clinical trials
WP4, 5 & 6 dealt with transfer of knowledge, outreach and project management respectively
• The fellow received extensive training and experience in the business facing aspects of R&D in industry – something which the fellow had no previous experience with
• The fellow significantly expanded the experimental toolkit at Nuritas
• Pept-AGE hosted secondary school students for a week of hands-on experiments, saw the fellow lecture to Secondary school students on STEM career paths, and hold a career Q&A with MSCA early-stage researchers
• The findings of Pept-AGE have been presented at the key sarcopenia meeting (ICFSR, Boston)
This MSCA has expanded the state of the art in several ways. Firstly, the DIMs model developed in PeptAge has the potential to form an important, and desperately lacking, test platform for basic research in sarcopenia. This was highlighted by the fact that the model generated significant attention when presented at the ICFSR conference. The approach can be modified and used to model genetic forms of muscle loss too and so is particularly flexible as a research tool. While Pept-AGE has not yet managed to identify peptide therapeutics to treat age related muscle loss, the data generated in this MSCA have identified a novel and possibly important mechanism that could play a significant role in promoting age-related muscle loss.
The anticipated impacts of the research carried out in this project are potentially very significant. We are on the cusp of a very significant health-care crisis resulting from an ageing population. If the data generated as part of Pept-AGE can contribute in some way or provide a model for future researchers to make progress in sarcopenia research, then the project has been absolutely worthwhile.
The anticipated impacts of the research carried out in this project are potentially very significant. We are on the cusp of a very significant health-care crisis resulting from an ageing population. If the data generated as part of Pept-AGE can contribute in some way or provide a model for future researchers to make progress in sarcopenia research, then the project has been absolutely worthwhile.