The Proteomic Landscape of Extracellular Vesicles Derived from Human Intervertebral Disc Cells

Overview

Journal JOR Spine

Specialty Orthopedics

Date 2024 Nov 7

PMID 39507593

Authors

Li Li

Hadil Al-Jallad

Aiwei Sun

Miltiadis Georgiopoulos

Rakan Bokhari

Jean Ouellet

Peter Jarzem

Hosni Cherif

Lisbet Haglund

Affiliations

Soon will be listed here.

Abstract

Background: Extracellular vesicles (EVs) function as biomarkers and are crucial in cell communication and regulation, with therapeutic potential for intervertebral disc (IVD)-related low back pain (LBP). EV cargo is often affected by tissue health, which may affect the therapeutic potential. There is currently limited knowledge of how the cargo of IVD cell-derived EVs varies with tissue health and how differences in proteomic profile affect the predicted biological functions.

Methods: Our study purified EVs from human IVD cell conditioned media by size-exclusion chromatography. Nanoparticle tracking analysis was conducted to measure EV size and concentration. Transmission electron microscopy and Western blot were performed to examine EV structure and markers. Tandem mass tag-mass spectrometry was conducted to determine protein cargo.

Results: Most EVs were exosomes and intermediate microvesicles with an increasing amount linked to disease progression. Of the proteins detected, 88.6% were shared across the non-degenerate, mildly-degenerate, and degenerate samples. GO and KEGG analyses revealed that cargo from the mildly-degenerate samples was the most distinct, with the proteins in high abundance strongly associated with extracellular matrix (ECM) organization and structure. Shared proteins, highly expressed in the non-degenerate and degenerate samples, showed strong associations with cell adhesion, ECM-receptor interaction, and vesicle-mediated transport, respectively.

Conclusions: Our findings indicate that EVs from IVD cells from tissue with different degrees of degeneration share a majority of the cargo proteins. However, the level of expression differs with degeneration grade. Cargo from the mildly-degenerate samples exhibits the most differences. A better understanding of changes in EV cargo in the degenerative process may provide novel information related to molecular mechanisms underlying IVD degeneration and suggest new potential treatment modalities for IVD-related LBP.

Citing Articles

Alginate vs. Hyaluronic Acid as Carriers for Nucleus Pulposus Cells: A Study on Regenerative Outcomes in Disc Degeneration.

Ogasawara S, Schol J, Sakai D, Warita T, Susumu T, Nakamura Y Cells. 2024; 13(23).

PMID: 39682732 PMC: 11639827. DOI: 10.3390/cells13231984.

References

Matta A, Karim M, Isenman D, Erwin W . Molecular Therapy for Degenerative Disc Disease: Clues from Secretome Analysis of the Notochordal Cell-Rich Nucleus Pulposus. Sci Rep. 2017; 7:45623. PMC: 5372366. DOI: 10.1038/srep45623. View

Lischnig A, Bergqvist M, Ochiya T, Lasser C . Quantitative Proteomics Identifies Proteins Enriched in Large and Small Extracellular Vesicles. Mol Cell Proteomics. 2022; 21(9):100273. PMC: 9486130. DOI: 10.1016/j.mcpro.2022.100273. View

Cocozza F, Grisard E, Martin-Jaular L, Mathieu M, Thery C . SnapShot: Extracellular Vesicles. Cell. 2020; 182(1):262-262.e1. DOI: 10.1016/j.cell.2020.04.054. View

Yanez-Mo M, Siljander P, Andreu Z, Bedina Zavec A, Borras F, Buzas E . Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles. 2015; 4:27066. PMC: 4433489. DOI: 10.3402/jev.v4.27066. View

Millecamps M, Tajerian M, Sage E, S Stone L . Behavioral signs of chronic back pain in the SPARC-null mouse. Spine (Phila Pa 1976). 2010; 36(2):95-102. PMC: 3007098. DOI: 10.1097/BRS.0b013e3181cd9d75. View

Loreto C, Musumeci G, Castorina A, Loreto C, Martinez G . Degenerative disc disease of herniated intervertebral discs is associated with extracellular matrix remodeling, vimentin-positive cells and cell death. Ann Anat. 2011; 193(2):156-62. DOI: 10.1016/j.aanat.2010.12.001. View

Tsamchoe M, Petrillo S, Lazaris A, Metrakos P . Isolation of extracellular vesicles from human plasma samples: The importance of controls. Biotechnol J. 2023; 18(6):e2200575. DOI: 10.1002/biot.202200575. View

Kim H, Choi D, Yun S, Choi S, Kang J, Jung J . Proteomic analysis of microvesicles derived from human mesenchymal stem cells. J Proteome Res. 2011; 11(2):839-49. DOI: 10.1021/pr200682z. View

Palviainen M, Saari H, Karkkainen O, Pekkinen J, Auriola S, Yliperttula M . Metabolic signature of extracellular vesicles depends on the cell culture conditions. J Extracell Vesicles. 2019; 8(1):1596669. PMC: 6461113. DOI: 10.1080/20013078.2019.1596669. View

10.

Reiss K, Saftig P . The "a disintegrin and metalloprotease" (ADAM) family of sheddases: physiological and cellular functions. Semin Cell Dev Biol. 2008; 20(2):126-37. DOI: 10.1016/j.semcdb.2008.11.002. View

11.

Yuan X, Sun L, Jeske R, Nkosi D, York S, Liu Y . Engineering extracellular vesicles by three-dimensional dynamic culture of human mesenchymal stem cells. J Extracell Vesicles. 2022; 11(6):e12235. PMC: 9206229. DOI: 10.1002/jev2.12235. View

12.

Venturella M, Criscuoli M, Carraro F, Naldini A, Zocco D . Interplay between Hypoxia and Extracellular Vesicles in Cancer and Inflammation. Biology (Basel). 2021; 10(7). PMC: 8301089. DOI: 10.3390/biology10070606. View

13.

Hao Y, Zhu G, Yu L, Ren Z, Zhang P, Zhu J . Extracellular vesicles derived from mesenchymal stem cells confer protection against intervertebral disc degeneration through a microRNA-217-dependent mechanism. Osteoarthritis Cartilage. 2022; 30(11):1455-1467. DOI: 10.1016/j.joca.2022.08.009. View

14.

Scheiber A, Clark C, Kaito T, Iwamoto M, Horwitz E, Kawasawa Y . Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles. Int J Mol Sci. 2022; 23(3). PMC: 8834965. DOI: 10.3390/ijms23031017. View

15.

Liu D, Upton F, Rees E, Limb C, Jiao L, Krell J . Size-Exclusion Chromatography as a Technique for the Investigation of Novel Extracellular Vesicles in Cancer. Cancers (Basel). 2020; 12(11). PMC: 7693800. DOI: 10.3390/cancers12113156. View

16.

Gruber H, Ingram J, Leslie K, Hanley Jr E . Cellular, but not matrix, immunolocalization of SPARC in the human intervertebral disc: decreasing localization with aging and disc degeneration. Spine (Phila Pa 1976). 2004; 29(20):2223-8. DOI: 10.1097/01.brs.0000142225.07927.29. View

17.

Gruber H, Hanley Jr E . Human disc cells in monolayer vs 3D culture: cell shape, division and matrix formation. BMC Musculoskelet Disord. 2001; 1:1. PMC: 32203. DOI: 10.1186/1471-2474-1-1. View

18.

Yang Y, Morand E, Leech M . Annexin A1: potential for glucocorticoid sparing in RA. Nat Rev Rheumatol. 2013; 9(10):595-603. DOI: 10.1038/nrrheum.2013.126. View

19.

Chen J, Jing L, Gilchrist C, Richardson W, Fitch R, Setton L . Expression of laminin isoforms, receptors, and binding proteins unique to nucleus pulposus cells of immature intervertebral disc. Connect Tissue Res. 2009; 50(5):294-306. PMC: 2929695. View

20.

Monguio-Tortajada M, Moron-Font M, Gamez-Valero A, Carreras-Planella L, Borras F, Franquesa M . Extracellular-Vesicle Isolation from Different Biological Fluids by Size-Exclusion Chromatography. Curr Protoc Stem Cell Biol. 2019; 49(1):e82. DOI: 10.1002/cpsc.82. View