Project ID CM-HD2024_08


Co Supervisor 1A Faculty of Life Sciences & Medicine, School of Cardiovascular and Metabolic Medicine & Sciences, Vascular themeWebsite

Co Supervisor 1B Faculty of Life Sciences & Medicine, School of Cardiovascular and Metabolic Medicine & Sciences, Cardiac themeWebsite

Additional Supervisor Mona Bafadhel

Epigenetic regulation of arterial calcification in vascular ageing and disease.

Vascular calcification causes hardening of the arteries and leads to cardiovascular death and currently there are no treatments. Our work has shown that a key driver of vascular calcification is a change in the function of vascular smooth muscle cells (VSMCs) of the vessel wall from a contractile to an osteogenic-like cell. This project will focus on understanding the role of epigenetic factors in mediating this phenotype change. We have shown that the SWI/SNF (SWItch/Sucrose Non-Fermentable), subfamily of ATP-dependent chromatin complexes are deregulated in the vessel wall during calcification and ageing and their manipulation in vitro can block osteogenic change in VSMCs. The aims of this project will be to understand what the key components of the complex are that regulate calcification and to understand the targets of the complex in response to stimuli that induce calcification. We will verify these pathways in human tissue samples and animal models of calcification where the complex has been disrupted.

Techniques and Skills:
Primary human VSMC culture and calcification assays. ‘Cut and Run’ ChIP-seq experiments, ATAC and RNA-seq and downstream bioinformatics analysis to identify transcriptional targets and pathways. ChIP-PCR, RT-PCR, IP (immunoprecipitation) and Western blotting to verify targets. Proteomics Mass-Spec to identify complex components under control and calcifying conditions. Immunohistochemistry (IHC) in human tissue sections from healthy and calcified patients. Analysis of animal models of calcification where the SWI/SNF complex has been disrupted genetically or with drugs
Year 1-2 – Identification of timecourse of expression of candidate SWI/SNF components during calcification and characterisation of complex changes using IP and Mass Spec.
Year 2 – ChIP-seq and RNA-seq experiments and bioinformatics to identify transcriptional targets. Target verification and effects on calcification using siRNA/inhibitors assays.
Year 3 – Selected targets are verified in vivo using IHC and human and animal models.

Representative Publications

Runx2 (Runt-Related Transcription Factor 2) Links the DNA Damage Response to Osteogenic Reprogramming and Apoptosis of Vascular Smooth Muscle Cells. Cobb AM, Yusoff S, Hayward R, Ahmad S, Sun M, Verhulst A, D’Haese PC, Shanahan CM. Arterioscler Thromb Vasc Biol. 2021 Apr;41(4):1339-1357. doi: 10.1161/ATVBAHA.120.315206. Poly(ADP-Ribose) Links the DNA Damage Response and Biomineralization. Müller KH, Hayward R, Rajan R, Whitehead M, Cobb AM, Ahmad S, Sun M, Goldberga I, Li R, Bashtanova U, Puszkarska AM, Reid DG, Brooks RA, Skepper JN, Bordoloi J, Chow WY, Oschkinat H, Groombridge A, Scherman OA, Harrison JA, Verhulst A, D’Haese PC, Neven E, Needham LM, Lee SF, Shanahan CM, Duer MJ.Cell Rep. 2019 Jun 11;27(11):3124-3138.e13. doi: 10.1016/j.celrep.2019.05.038 Arterial “inflammaging” drives vascular calcification in children on dialysis. Sanchis P, Ho CY, Liu Y, Beltran LE, Ahmad S, Jacob AP, Furmanik M, Laycock J, Long DA, Shroff R, Shanahan CM.Kidney Int. 2019 Apr;95(4):958-972. doi: 10.1016/j.kint.2018.12.014.
Novel nesprin-1 mutations associated with dilated cardiomyopathy cause nuclear envelope disruption and defects in myogenesis. Zhou C, Li C, Zhou B, Sun H, Koullourou V, Holt I, Puckelwartz MJ, Warren DT, Hayward R, Lin Z, Zhang L, Morris GE, McNally EM, Shackleton S, Rao L, Shanahan CM, Zhang Q. Hum Mol Genet. 2017 Jun 15;26(12):2258-2276. doi: 10.1093/hmg/ddx116