Dataset Information

Sox9 accelerates vascular ageing by regulating extracellular matrix composition and stiffness

ABSTRACT: Vascular calcification and increased extracellular matrix (ECM) stiffness are hallmarks of vascular ageing. Sox9 (SRY-Box Transcription Factor 9) is a master regulator of chondrogenesis, also expressed in the vasculature, that has been implicated in vascular smooth muscle cell (VSMC) osteo-chondrogenic conversion. Here, we investigated the relationship between vascular ageing, calcification and Sox9-driven ECM regulation in VSMCs. Immunohistochemistry in human aortic samples showed that Sox9 was not spatially associated with vascular calcification but correlated with the senescence marker p16. Analysis of Sox9 expression in vitro showed it was mechanosensitive with increased expression and nuclear translocation in senescent cells and on stiff matrices. Manipulation of Sox9 via overexpression and depletion, combined with atomic force microscopy (AFM) and proteomics, revealed that Sox9 regulates ECM stiffness and organisation by orchestrating changes in collagen expression and reducing VSMC contractility, leading to the formation of an ECM that mirrored that of senescent cells. These ECM changes promoted phenotypic modulation of VSMCs whereby senescent cells plated onto ECM synthesized from cells depleted of Sox9 returned to a proliferative state, while proliferating cells on a matrix produced by Sox9 expressing cells showed reduced proliferation and increased DNA damage, reiterating features of senescent cells. Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 (LH3) was identified as a Sox9 target, and key regulator of ECM stiffness. LH3 is packaged into extracellular vesicles (EVs) and Sox9 promoted EV secretion, leading to increased LH3 deposition within the ECM. These findings identify cellular senescence and Sox9 as a key regulators of ECM stiffness during VSMC ageing and highlight a crucial role for ECM structure and composition in regulating VSMC phenotype. We identify a positive feedback cycle whereby cellular senescence and increased ECM stiffening promote Sox9 expression which drives further ECM modifications that act to accelerate vascular stiffening and cellular senescence.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Steven Lynham

LAB HEAD: Prof Cathy Shanahan

PROVIDER: PXD046001 | Pride | 2024-06-16

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	PR667_MF1_1_35F_P10_YoungCtrl_rep1.mzid.gz	Mzid
	PR667_MF1_1_35F_P10_YoungCtrl_rep1.mzid_PR667_MF1_1_35F_P10_YoungCtrl_rep1.MGF	Mzid
	PR667_MF1_1_35F_P10_YoungCtrl_rep1.raw	Raw
	PR667_MF1_1_35F_P10_YoungCtrl_rep2.mzid.gz	Mzid
	PR667_MF1_1_35F_P10_YoungCtrl_rep2.mzid_PR667_MF1_1_35F_P10_YoungCtrl_rep2.MGF	Mzid

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Publications

Sox9 Accelerates Vascular Aging by Regulating Extracellular Matrix Composition and Stiffness.

Faleeva Maria M Ahmad Sadia S Theofilatos Konstantinos K Lynham Steven S Watson Gabriel G Whitehead Meredith M Marhuenda Emilie E Iskratsch Thomas T Cox Susan S Shanahan Catherine M CM

Circulation research 20240105 3

<h4>Background</h4>Vascular calcification and increased extracellular matrix (ECM) stiffness are hallmarks of vascular aging. Sox9 (SRY-box transcription factor 9) has been implicated in vascular smooth muscle cell (VSMC) osteo/chondrogenic conversion; however, its relationship with aging and calcification has not been studied.<h4>Methods</h4>Immunohistochemistry was performed on human aortic samples from young and aged patients. Young and senescent primary human VSMCs were induced to produce EC ...[more]

PMID: 38179698

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