Cellular and molecular signature of cardioPICs.
It is well known that different cell types can in part influence either negatively or positively muscle homeostasis, including the resident fibro/adipogenic progenitors (FAPs) expressing platelet-derived growth-factor receptor (PDGFR)-α (Joe et al., 2010; Uezumi et al., 2011) and the PW1/Peg3 positive interstitial cells (myoPiCS) (Lewis et al., 2014; Pannerec et al., 2013). Despite the presence in the heart of a population of (cardioPICs) (Ellison et al., 2013; Lewis et al., 2014; Vicinanza et al., 2017) with myogenic potential, there is no evidence at present about their role in cardiac homeostasis or repair.
ESR9 will isolate the cardioPIC from transgenic mice expressing identifiable reporter genes (Pannerec et al., 2013) and use protocols proven to be successful for myoPICs (Lewis et al., 2014) to test their differentiation phenotype in vitro. In depth single cell RNA-Seq will be performed to determine the cardioPiCs heterogeneity and transcriptional profile.
ESR9 will determine the aged phenotype of cardioPICs. Single-cell transcriptome sequencing will be performed on cardioPICs isolated from aged mice and human hearts. Both p16pos-senescent and p16neg non-senescent cardioPICs will be characterised for growth, senescence and regenerative potential both in vitro and in the myocardial-infarction regeneration model in vivo.
Finally, ESR9 will consider variation in single progenitor cells (single cell RNA-seq) isolated from old and young human hearts (young refer to adult patients less than 50 years old) in terms of gene expression, somatic mutations and mitochondria RNA phenotypes and then consider systematic changes in gene expression as hearts age.
ESR9 will also study the potential causal mechanisms of these changes by looking at the accumulation of somatic mutations in nuclear genome across single cells of both young and old tissue, and also survey mutations, post-transcriptional modification events and gene expression in the mitochondria genome. These findings will then be correlated to patient characteristics (age, heart failure class, genetic hereditary factors). Besides gold-standard RNA sequencing analysis pipelines, work on mitochondrial gene expression and detection of mitochondrial RNA modifications will rely on novel computational techniques developed at KCL
To isolate and characterize cardioPICs from young and aged mouse and human hearts
To assess differentiation potential of cardioPICs in vitro and in vivo
To test and develop computational methods/pipelines for the analysis of single cell RNA sequencing data
To identify heterogeneity in gene expression profiles at the single cell level in cardioPICs
To identify molecular signatures that define the ageing process in human heart tissue
Enrolment in Doctoral degree
PhD programme in Basic and Medical Biosciences King’s College London
Ellison, G.M., Vicinanza, C., Smith, A.J., Aquila, I., Leone, A., Waring, C.D., Henning, B.J., Stirparo, G.G., Papait, R., Scarfo, M., et al. (2013). Adult c-kit(pos) cardiac stem cells are necessary and sufficient for functional cardiac regeneration and repair. Cell 154, 827-842.
Joe, A.W.B., Yi, L., Natarajan, A., Le Grand, F., So, L., Wang, J., Rudnicki, M.A., and Rossi, F.M.V. (2010). Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis. Nature Cell Biology 12, 153-163.
Lewis, F.C., Henning, B.J., Marazzi, G., Sassoon, D., Ellison, G.M., and Nadal-Ginard, B. (2014). Porcine skeletal muscle-derived multipotent PW1pos/Pax7neg interstitial cells: isolation, characterization, and long-term culture. Stem cells translational medicine 3, 702-712.
Pannerec, A., Formicola, L., Besson, V., Marazzi, G., and Sassoon, D.A. (2013). Defining skeletal muscle resident progenitors and their cell fate potentials. Development 140, 2879-2891.
Uezumi, A., Ito, T., Morikawa, D., Shimizu, N., Yoneda, T., Segawa, M., Yamaguchi, M., Ogawa, R., Matev, M.M., Miyagoe-Suzuki, Y., et al. (2011). Fibrosis and adipogenesis originate from a common mesenchymal progenitor in skeletal muscle. Journal of Cell Science 124, 3654-3664.
Vicinanza, C., Aquila, I., Scalise, M., Cristiano, F., Marino, F., Cianflone, E., Mancuso, T., Marotta, P., Sacco, W., Lewis, F.C., et al. (2017). Adult cardiac stem cells are multipotent and robustly myogenic: c-kit expression is necessary but not sufficient for their identification. Cell Death Differ 24, 2101-2116.