Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes

Joshua Slysz; Arjun Sinha; Matthew DeBerge; Shalini Singh; Harris Avgousti; Inhyeok Lee; Kristofor Glinton; Reina Nagasaka; Prarthana Dalal; Shaina Alexandria; Ching Man Wai; Ricardo Tellez; Mariavittoria Vescovo; Ashwin Sunderraj; Xinkun Wang; Matthew Schipma; Ryan Sisk; Rishab Gulati; Jenifer Vallejo; Ryosuke Saigusa; Donald M. Lloyd-Jones; Jon Lomasney; Samuel Weinberg; Karen Ho; Klaus Ley; Chiara Giannarelli; Edward B. Thorp; Matthew J. Feinstein

doi:10.1172/jci.insight.171359

Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes

Joshua Slysz, Arjun Sinha, Matthew DeBerge, Shalini Singh, Harris Avgousti, Inhyeok Lee, Kristofor Glinton, Reina Nagasaka, Prarthana Dalal, Shaina Alexandria, Ching Man Wai, Ricardo Tellez, Mariavittoria Vescovo, Ashwin Sunderraj, Xinkun Wang, Matthew Schipma, Ryan Sisk, Rishab Gulati, Jenifer Vallejo, Ryosuke SaigusaDonald M. Lloyd-Jones, Jon Lomasney, Samuel Weinberg, Karen Ho, Klaus Ley, Chiara Giannarelli^*, Edward B. Thorp^*, Matthew J. Feinstein^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Femoral atherosclerotic plaques are less inflammatory than carotid plaques histologically, but limited cell-level data exist regarding comparative immune landscapes and polarization at these sites. We investigated intraplaque leukocyte phenotypes and transcriptional polarization in 49 patients undergoing femoral (n = 23) or carotid (n = 26) endarterectomy using single-cell RNA-Seq (scRNA-Seq; n = 13), flow cytometry (n = 24), and IHC (n = 12). Comparative scRNA-Seq of CD45⁺-selected leukocytes from femoral (n = 9; 35,265 cells) and carotid (n = 4; 30,655 cells) plaque revealed distinct transcriptional profiles. Inflammatory foam cell–like macrophages and monocytes comprised higher proportions of myeloid cells in carotid plaques, whereas noninflammatory foam cell–like macrophages and LYVE1-overexpressing macrophages comprised higher proportions of myeloid cells in femoral plaque (P < 0.001 for all). A significant comparative excess of CCR2⁺ macrophages in carotid versus plaque was observed by flow cytometry in a separate validation cohort. B cells were more prevalent and exhibited a comparatively antiinflammatory profile in femoral plaque, whereas cytotoxic CD8⁺ T cells were more prevalent in carotid plaque.

Original language	English (US)
Article number	e171359
Journal	JCI Insight
Volume	8
Issue number	17
DOIs	https://doi.org/10.1172/jci.insight.171359
State	Published - 2023

Funding

Acknowledgments We thank the Northwestern University IRB and Division of Cardiology (Department of Medicine), Division of Vascular Surgery (Department of Surgery), and Department of Pathology at the NUFSM. CG also acknowledges support by the NIH grants R01HL165258 and R01HL153712, by the American Heart Association (20SFRN35210252) and by the Chan Zuckerberg Initiative CZI NFL-2020-218415. CG is listed as an inventor on Tech 160808G PCT/US2022/017777 filed by the Icahn School of Medicine at Mount Sinai, which has no competing interest with this work. We thank the Northwestern University IRB and Division of Cardiology (Department of Medicine), Division of Vascular Surgery (Department of Surgery), and Department of Pathology at the NUFSM. CG also acknowledges support by the NIH grants R01HL165258 and R01HL153712, by the American Heart Association (20SFRN35210252) and by the Chan Zuckerberg Initiative CZI NFL-2020-218415. CG is listed as an inventor on Tech 160808G PCT/US2022/017777 filed by the Icahn School of Medicine at Mount Sinai, which has no competing interest with this work.

ASJC Scopus subject areas

General Medicine

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Slysz, J., Sinha, A., DeBerge, M., Singh, S., Avgousti, H., Lee, I., Glinton, K., Nagasaka, R., Dalal, P., Alexandria, S., Wai, C. M., Tellez, R., Vescovo, M., Sunderraj, A., Wang, X., Schipma, M., Sisk, R., Gulati, R., Vallejo, J., ... Feinstein, M. J. (2023). Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes. JCI Insight, 8(17), Article e171359. https://doi.org/10.1172/jci.insight.171359

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title = "Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes",

abstract = "Femoral atherosclerotic plaques are less inflammatory than carotid plaques histologically, but limited cell-level data exist regarding comparative immune landscapes and polarization at these sites. We investigated intraplaque leukocyte phenotypes and transcriptional polarization in 49 patients undergoing femoral (n = 23) or carotid (n = 26) endarterectomy using single-cell RNA-Seq (scRNA-Seq; n = 13), flow cytometry (n = 24), and IHC (n = 12). Comparative scRNA-Seq of CD45+-selected leukocytes from femoral (n = 9; 35,265 cells) and carotid (n = 4; 30,655 cells) plaque revealed distinct transcriptional profiles. Inflammatory foam cell–like macrophages and monocytes comprised higher proportions of myeloid cells in carotid plaques, whereas noninflammatory foam cell–like macrophages and LYVE1-overexpressing macrophages comprised higher proportions of myeloid cells in femoral plaque (P < 0.001 for all). A significant comparative excess of CCR2+ macrophages in carotid versus plaque was observed by flow cytometry in a separate validation cohort. B cells were more prevalent and exhibited a comparatively antiinflammatory profile in femoral plaque, whereas cytotoxic CD8+ T cells were more prevalent in carotid plaque.",

author = "Joshua Slysz and Arjun Sinha and Matthew DeBerge and Shalini Singh and Harris Avgousti and Inhyeok Lee and Kristofor Glinton and Reina Nagasaka and Prarthana Dalal and Shaina Alexandria and Wai, {Ching Man} and Ricardo Tellez and Mariavittoria Vescovo and Ashwin Sunderraj and Xinkun Wang and Matthew Schipma and Ryan Sisk and Rishab Gulati and Jenifer Vallejo and Ryosuke Saigusa and Lloyd-Jones, {Donald M.} and Jon Lomasney and Samuel Weinberg and Karen Ho and Klaus Ley and Chiara Giannarelli and Thorp, {Edward B.} and Feinstein, {Matthew J.}",

note = "Publisher Copyright: Copyright: {\textcopyright} 2023, Slysz et al.",

year = "2023",

doi = "10.1172/jci.insight.171359",

language = "English (US)",

volume = "8",

journal = "JCI Insight",

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Slysz, J, Sinha, A, DeBerge, M, Singh, S, Avgousti, H, Lee, I, Glinton, K, Nagasaka, R, Dalal, P, Alexandria, S, Wai, CM, Tellez, R, Vescovo, M, Sunderraj, A, Wang, X , Schipma, M, Sisk, R, Gulati, R, Vallejo, J, Saigusa, R, Lloyd-Jones, DM, Lomasney, J , Weinberg, S, Ho, K, Ley, K, Giannarelli, C, Thorp, EB & Feinstein, MJ 2023, 'Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes', JCI Insight, vol. 8, no. 17, e171359. https://doi.org/10.1172/jci.insight.171359

TY - JOUR

T1 - Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes

AU - Slysz, Joshua

AU - Sinha, Arjun

AU - DeBerge, Matthew

AU - Singh, Shalini

AU - Avgousti, Harris

AU - Lee, Inhyeok

AU - Glinton, Kristofor

AU - Nagasaka, Reina

AU - Dalal, Prarthana

AU - Alexandria, Shaina

AU - Wai, Ching Man

AU - Tellez, Ricardo

AU - Vescovo, Mariavittoria

AU - Sunderraj, Ashwin

AU - Wang, Xinkun

AU - Schipma, Matthew

AU - Sisk, Ryan

AU - Gulati, Rishab

AU - Vallejo, Jenifer

AU - Saigusa, Ryosuke

AU - Lloyd-Jones, Donald M.

AU - Lomasney, Jon

AU - Weinberg, Samuel

AU - Ho, Karen

AU - Ley, Klaus

AU - Giannarelli, Chiara

AU - Thorp, Edward B.

AU - Feinstein, Matthew J.

PY - 2023

Y1 - 2023

N2 - Femoral atherosclerotic plaques are less inflammatory than carotid plaques histologically, but limited cell-level data exist regarding comparative immune landscapes and polarization at these sites. We investigated intraplaque leukocyte phenotypes and transcriptional polarization in 49 patients undergoing femoral (n = 23) or carotid (n = 26) endarterectomy using single-cell RNA-Seq (scRNA-Seq; n = 13), flow cytometry (n = 24), and IHC (n = 12). Comparative scRNA-Seq of CD45+-selected leukocytes from femoral (n = 9; 35,265 cells) and carotid (n = 4; 30,655 cells) plaque revealed distinct transcriptional profiles. Inflammatory foam cell–like macrophages and monocytes comprised higher proportions of myeloid cells in carotid plaques, whereas noninflammatory foam cell–like macrophages and LYVE1-overexpressing macrophages comprised higher proportions of myeloid cells in femoral plaque (P < 0.001 for all). A significant comparative excess of CCR2+ macrophages in carotid versus plaque was observed by flow cytometry in a separate validation cohort. B cells were more prevalent and exhibited a comparatively antiinflammatory profile in femoral plaque, whereas cytotoxic CD8+ T cells were more prevalent in carotid plaque.

AB - Femoral atherosclerotic plaques are less inflammatory than carotid plaques histologically, but limited cell-level data exist regarding comparative immune landscapes and polarization at these sites. We investigated intraplaque leukocyte phenotypes and transcriptional polarization in 49 patients undergoing femoral (n = 23) or carotid (n = 26) endarterectomy using single-cell RNA-Seq (scRNA-Seq; n = 13), flow cytometry (n = 24), and IHC (n = 12). Comparative scRNA-Seq of CD45+-selected leukocytes from femoral (n = 9; 35,265 cells) and carotid (n = 4; 30,655 cells) plaque revealed distinct transcriptional profiles. Inflammatory foam cell–like macrophages and monocytes comprised higher proportions of myeloid cells in carotid plaques, whereas noninflammatory foam cell–like macrophages and LYVE1-overexpressing macrophages comprised higher proportions of myeloid cells in femoral plaque (P < 0.001 for all). A significant comparative excess of CCR2+ macrophages in carotid versus plaque was observed by flow cytometry in a separate validation cohort. B cells were more prevalent and exhibited a comparatively antiinflammatory profile in femoral plaque, whereas cytotoxic CD8+ T cells were more prevalent in carotid plaque.

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ER -

Single-cell profiling reveals inflammatory polarization of human carotid versus femoral plaque leukocytes

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