Project ID CM-HD2024_57


Co Supervisor 1A Faculty of Dentistry, Oral & Craniofacial Sciences, Centre for Craniofacial & Regenerative BiologyWebsite

Co Supervisor 1B Faculty of Life Sciences & Medicine, School of Basic & Medical Biosciences, Centre for Human & Applied Physiological SciencesWebsite

Molecular dissection of ageing associated changes to muscle stem cell function

This project aims to test whether candidate ageing-related factors affect muscle stem cell (muSC) function in vivo (using a zebrafish model) and in vitro (using a human primary cell culture model). It is becoming clear that although ageing is genetically programmed there are a number of factors that impact muSC function including DNA damage and chromatin packaging. Our long-term goal is to identify factors that promote an older unhealthy phenotype that can be targeted by available biopharmaceuticals to enhance muscle function in old age. Using zebrafish to visualise tissue and cell behaviour in vivo together with cells isolated from human muscle we will test whether factors identified in ageing muscle affect muscle function and repair.

We have previously identified several factors with age-associated changes in muscle (Lam et al, 2021; Agley et al. 2017). We will test whether candidate ageing factors alter muSC responses to injury in a genetic zebrafish ageing model using live cell imaging. Gene function will be modified using drugs, gene over-expression and knockdown. Factors that affect muSCs in zebrafish will then be functionally tested on human muSCs by pharmacological and transient gene over-expression to determine how they affect cell proliferation, differentiation and metabolism.

Aims for the project are:
Year 1: test whether putative ageing-associated factors affect muSC function in zebrafish and humans
Year 2: determine the molecular consequences of manipulating putative ageing-associated factors
Year 3: test specific ageing-associated factors in zebrafish and human ageing models

Techniques and skills:
Confocal and multiphoton microscopy, transgenesis and CRISPR/Cas9 gene manipulation, zebrafish genetics, human muscle cell culture, qRT-PCR, Western blotting, histology

Representative Publications

Lam S, Hartmann N, Benfeitas R, Zhang C, Arif M, Turkez H, Uhlén M, Englert C, Knight R, Mardinoglu A. Systems Analysis Reveals Ageing-Related Perturbations in Retinoids and Sex Hormones in Alzheimer’s and Parkinson’s Diseases. Biomedicines. 2021 Sep 24;9(10):1310. doi: 10.3390/biomedicines9101310. Sultan, S.H.A., C. Dyer, and Knight, R. D. Notch Signaling Regulates Muscle Stem Cell Homeostasis and Regeneration in a Teleost Fish. Frontiers in Cell and Developmental Biology, 2021. 9(2501). Haroon M, Klein-Nulend J, Bakker A. D., Jin J, Seddiqi H, Offringa C, de Wit G. M. J., Le Grand F, Giordani L, Liu K. J., Knight R. D., Jaspers R. T. Myofiber stretch induces tensile and shear deformation of muscle stem cells in their native niche. Biophys J. 2021 Jul 6;120(13):2665-2678. doi: 10.1016/j.bpj.2021.05.021.
Francis TG, Jaka O, Ellison-Hughes G, Lazarus NR, Harridge SDR (2022) Induced senescence in human primary skeletal muscle-derived myoblasts and fibroblasts reveals different phenotypes. The Journal of Cachexia, Sarcopenia and Muscle: Rapid Communications 5: 226– 238. Agley CC, Lewis FC, Jaka O, Lazarus NR, Velloso C, Francis-West P, Ellison-Hughes GM, Harridge SDR (2017) Active GSK3β and an intact β-catenin TCF complex are essential for the differentiation of human myogenic progenitor cells. Scientific Reports 2017 Oct 13;7(1):13189. doi: 10.1038/s41598-017-10731-1. Battey E, Hoang A, Ross JA, Pollock RD, Kalakoutis M, Wilson DGS, Pugh JN, Close GL, Lazarus NR, Iskratsch T, Harridge SDR, Ochala J, & Stroud MJ (2023) Exercise mitigates myonuclear dysfunction in young and aged humans and mice. Journal of Physiology DOI: 10.1113/JP284128