HDL Regulates TGFß-receptor Lipid Raft Partitioning, Restoring Contractile Features of Cholesterol-loaded Vascular Smooth Muscle Cells

Overview

Journal bioRxiv

Date 2023 Oct 31

PMID 37905061

Authors

Prashanth Thevkar Nagesh

Hitoo Nishi

Shruti Rawal

Tarik Zahr

Joseph M Miano

Mary Sorci-Thomas

Hao Xu

Naveed Akbar

Robin P Choudhury

Ashish Misra

Edward A Fisher

Affiliations

Soon will be listed here.

Abstract

Background: Cholesterol-loading of mouse aortic vascular smooth muscle cells (mVSMCs) downregulates , a master regulator of the contractile state downstream of TGFβ signaling. this results in transitioning from a contractile mVSMC to a macrophage-like state. This process likely occurs based on studies in mouse and human atherosclerotic plaques.

Objectives: To test whether cholesterol-loading reduces VSMC TGFβ signaling and if cholesterol efflux will restore signaling and the contractile state and .

Methods: Human coronary artery (h)VSMCs were cholesterol-loaded, then treated with HDL (to promote cholesterol efflux). For studies, partial conditional deletion of in lineage-traced VSMC mice was induced. Mice wild-type for VSMC or partially deficient (+/-) were made hypercholesterolemic to establish atherosclerosis. Mice were then treated with apoA1 (which forms HDL).

Results: Cholesterol-loading of hVSMCs downregulated TGFβ signaling and contractile gene expression; macrophage markers were induced. TGFβ signaling positively regulated expression, increasing expression and suppressing KLF4. Cholesterol-loading localized TGFβ receptors into lipid rafts, with consequent TGFβ signaling downregulation. Notably, in cholesterol-loaded hVSMCs HDL particles displaced receptors from lipid rafts and increased TGFβ signaling, resulting in enhanced expression and decreased KLF4-dependent macrophage features. ApoA1 infusion into +/- mice restored expression and decreased macrophage-marker expression in plaque VSMCs, with evidence of increased TGFβ signaling.

Conclusions: Cholesterol suppresses TGFβ signaling and the contractile state in hVSMC through partitioning of TGFβ receptors into lipid rafts. These changes can be reversed by promotion of cholesterol efflux, consistent with evidence .

References

Tabas I, Garcia-Cardena G, Owens G . Recent insights into the cellular biology of atherosclerosis. J Cell Biol. 2015; 209(1):13-22. PMC: 4395483. DOI: 10.1083/jcb.201412052. View

Feig J, Rong J, Shamir R, Sanson M, Vengrenyuk Y, Liu J . HDL promotes rapid atherosclerosis regression in mice and alters inflammatory properties of plaque monocyte-derived cells. Proc Natl Acad Sci U S A. 2011; 108(17):7166-71. PMC: 3084076. DOI: 10.1073/pnas.1016086108. View

Rong J, Li J, Reis E, Choudhury R, Dansky H, Elmalem V . Elevating high-density lipoprotein cholesterol in apolipoprotein E-deficient mice remodels advanced atherosclerotic lesions by decreasing macrophage and increasing smooth muscle cell content. Circulation. 2001; 104(20):2447-52. DOI: 10.1161/hc4501.098952. View

Robbins C, Hilgendorf I, Weber G, Theurl I, Iwamoto Y, Figueiredo J . Local proliferation dominates lesional macrophage accumulation in atherosclerosis. Nat Med. 2013; 19(9):1166-72. PMC: 3769444. DOI: 10.1038/nm.3258. View

Akbar N, Braithwaite A, Corr E, Koelwyn G, van Solingen C, Cochain C . Rapid neutrophil mobilization by VCAM-1+ endothelial cell-derived extracellular vesicles. Cardiovasc Res. 2022; 119(1):236-251. PMC: 10022859. DOI: 10.1093/cvr/cvac012. View

Iqbal A, Barrett T, Taylor L, McNeill E, Manmadhan A, Recio C . Acute exposure to apolipoprotein A1 inhibits macrophage chemotaxis in vitro and monocyte recruitment in vivo. Elife. 2016; 5. PMC: 5030090. DOI: 10.7554/eLife.15190. View

Feil S, Fehrenbacher B, Lukowski R, Essmann F, Schulze-Osthoff K, Schaller M . Transdifferentiation of vascular smooth muscle cells to macrophage-like cells during atherogenesis. Circ Res. 2014; 115(7):662-7. DOI: 10.1161/CIRCRESAHA.115.304634. View

Feinberg M, Wara A, Cao Z, Lebedeva M, Rosenbauer F, Iwasaki H . The Kruppel-like factor KLF4 is a critical regulator of monocyte differentiation. EMBO J. 2007; 26(18):4138-48. PMC: 2230668. DOI: 10.1038/sj.emboj.7601824. View

Shankman L, Gomez D, Cherepanova O, Salmon M, Alencar G, Haskins R . KLF4-dependent phenotypic modulation of smooth muscle cells has a key role in atherosclerotic plaque pathogenesis. Nat Med. 2015; 21(6):628-37. PMC: 4552085. DOI: 10.1038/nm.3866. View

10.

Allahverdian S, Chaabane C, Boukais K, Francis G, Bochaton-Piallat M . Smooth muscle cell fate and plasticity in atherosclerosis. Cardiovasc Res. 2018; 114(4):540-550. PMC: 5852505. DOI: 10.1093/cvr/cvy022. View

11.

MacDonald J, Pike L . A simplified method for the preparation of detergent-free lipid rafts. J Lipid Res. 2005; 46(5):1061-7. DOI: 10.1194/jlr.D400041-JLR200. View

12.

Alencar G, Owsiany K, Karnewar S, Sukhavasi K, Mocci G, Nguyen A . Stem Cell Pluripotency Genes Klf4 and Oct4 Regulate Complex SMC Phenotypic Changes Critical in Late-Stage Atherosclerotic Lesion Pathogenesis. Circulation. 2020; 142(21):2045-2059. PMC: 7682794. DOI: 10.1161/CIRCULATIONAHA.120.046672. View

13.

Boscher C, Nabi I . Caveolin-1: role in cell signaling. Adv Exp Med Biol. 2012; 729:29-50. DOI: 10.1007/978-1-4614-1222-9_3. View

14.

Wirka R, Wagh D, Paik D, Pjanic M, Nguyen T, Miller C . Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis. Nat Med. 2019; 25(8):1280-1289. PMC: 7274198. DOI: 10.1038/s41591-019-0512-5. View

15.

Chattopadhyay A, Kwartler C, Kaw K, Li Y, Kaw A, Chen J . Cholesterol-Induced Phenotypic Modulation of Smooth Muscle Cells to Macrophage/Fibroblast-like Cells Is Driven by an Unfolded Protein Response. Arterioscler Thromb Vasc Biol. 2020; 41(1):302-316. PMC: 7752246. DOI: 10.1161/ATVBAHA.120.315164. View

16.

Castiglioni S, Monti M, Arnaboldi L, Canavesi M, Buscherini G, Calabresi L . ABCA1 and HDL are required to modulate smooth muscle cells phenotypic switch after cholesterol loading. Atherosclerosis. 2017; 266:8-15. DOI: 10.1016/j.atherosclerosis.2017.09.012. View

17.

Murphy A, Woollard K, Hoang A, Mukhamedova N, Stirzaker R, McCormick S . High-density lipoprotein reduces the human monocyte inflammatory response. Arterioscler Thromb Vasc Biol. 2008; 28(11):2071-7. DOI: 10.1161/ATVBAHA.108.168690. View

18.

Khera A, Cuchel M, de la Llera-Moya M, Rodrigues A, Burke M, Jafri K . Cholesterol efflux capacity, high-density lipoprotein function, and atherosclerosis. N Engl J Med. 2011; 364(2):127-35. PMC: 3030449. DOI: 10.1056/NEJMoa1001689. View

19.

Chin D, Poon C, Wang J, Joo J, Ong V, Jiang Z . miR-145 micelles mitigate atherosclerosis by modulating vascular smooth muscle cell phenotype. Biomaterials. 2021; 273:120810. PMC: 8152375. DOI: 10.1016/j.biomaterials.2021.120810. View

20.

Long X, Miano J . Transforming growth factor-beta1 (TGF-beta1) utilizes distinct pathways for the transcriptional activation of microRNA 143/145 in human coronary artery smooth muscle cells. J Biol Chem. 2011; 286(34):30119-29. PMC: 3191051. DOI: 10.1074/jbc.M111.258814. View