The Convergence of Accelerated and Physiological Vascular Ageing: A Multi-Omic Analysis of 2D and 3D in vitro models of Progeria

Cellular and molecular hallmarks of ageing contribute heavily to onset of vascular disease with age. With the average age of global populations increasing, the associated rise in vascular diseases presents a huge challenge. A key obstacle in vascular gerontology research is a lack of suitable in vitro human models which allow the study of human ageing in a highly controlled experimental environment. Induced pluripotent stem cells derived from patients suffering from the accelerated ageing disease, Hutchinson Gilford Progeria Syndrome (HGPS), provide an indefinite source of human cells which recapitulate several hallmarks of ageing. Furthermore, the cardiovascular pathology of HGPS closely resembles that of elderly individuals, with patients showing severe vascular stiffening and thickening, leading to premature death in the second decade of life from heart attacks or strokes as a result of severe atherosclerosis. Despite the phenotypic similarities, the extent to which HGPS truly captures ageing signatures remains uncertain. In this study, we have used CRISPR-Cas9 gene editing to rescue molecular signatures of ageing in HGPS- vascular smooth muscle cells (VSMCs), a cell type highly important in vascular function, and heavily affected by HGPS. We performed transcriptomic analyses comparing HGPS-VSMCs to isogenic controls and wildtype VSMCs, before performing an unbiased comparison to transcriptomic data from primary aged SMCs, thus identifying core convergence of ageing signatures shared across both physiological and accelerated ageing. Furthermore, we have presented and validated a mid-throughput 3D engineered vascular tissue to provide a clinically-relevant yet scalable model for the functional analyses of these cells. Our multi-omic analyses of both 2D and 3D accelerated ageing models identified aquaporin 1 (AQP1) as a potential therapeutic target in the prevention and treatment of age-related vascular dysfunction. Here, we showed that AQP1 is upregulated in both accelerated and physiological ageing, and is differentially expressed in response to differences in ECM deposition or stiffness. Moreover, small molecule inhibition of AQP1 resulted in a significant reduction in reactive oxygen species: a key hallmark in the progression of ageing. Together, these results have provided an overview of the key converging pathways which are common to ageing phenotypes, identified a potential therapeutic target in HGPS and age-related vascular disease and has provided a platform for further investigation and testing of such pathways.

Name dc:type.qualificationname

Doctor of Philosophy (PhD)

Level dc:type.qualificationlevel

Doctoral

Grantor dc:publisher.institution

University of Cambridge

Year dc:date.issued

2026

Author dc:creator

• Pullinger, Anna

Advisor dc:contributor.advisor

• Sinha, Sanjay

• Aquaporin
• HGPS
• Multi-Omic
• Stem Cell
• Vascular Ageing
• VSMC

Licence

• https://www.repository.cam.ac.uk/bitstreams/ede8cf71-91d7-40e2-9232-7ca2b8ea2415/download
• http://purl.org/NET/rdflicense/allrightsreserved

Language dc:language

eng

DOI dc:identifier.doi

https://doi.org/10.17863/CAM.131856

OAI identifier oai:identifier

oai:www.repository.cam.ac.uk:1810/405725