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