Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata

IntroductionAlopecia areata (AA) is an autoimmune disorder characterized by non-scarring hair loss ranging from mild, self-limiting episodes to severe and chronic forms. While prior research has primarily focused on lesional skin, the contribution of systemic immune cells remains underexplored.Metho...

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Main Authors: Jesus Gay-Mimbrera, Pedro Jesús Gómez-Arias, Pablo Álvarez-Heredia, Alexander Batista-Duharte, Irene Rivera-Ruiz, Macarena Aguilar-Luque, Miguel Juan-Cencerrado, Carmen Mochón-Jiménez, Álvaro Cebrián-García, Eloísa Andújar-Pulido, Mónica Pérez-Alegre, Alejandra Pera, Juan Ruano
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Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1565241/full
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author Jesus Gay-Mimbrera
Pedro Jesús Gómez-Arias
Pedro Jesús Gómez-Arias
Pablo Álvarez-Heredia
Alexander Batista-Duharte
Irene Rivera-Ruiz
Macarena Aguilar-Luque
Miguel Juan-Cencerrado
Miguel Juan-Cencerrado
Carmen Mochón-Jiménez
Carmen Mochón-Jiménez
Álvaro Cebrián-García
Eloísa Andújar-Pulido
Mónica Pérez-Alegre
Alejandra Pera
Alejandra Pera
Juan Ruano
Juan Ruano
Juan Ruano
author_facet Jesus Gay-Mimbrera
Pedro Jesús Gómez-Arias
Pedro Jesús Gómez-Arias
Pablo Álvarez-Heredia
Alexander Batista-Duharte
Irene Rivera-Ruiz
Macarena Aguilar-Luque
Miguel Juan-Cencerrado
Miguel Juan-Cencerrado
Carmen Mochón-Jiménez
Carmen Mochón-Jiménez
Álvaro Cebrián-García
Eloísa Andújar-Pulido
Mónica Pérez-Alegre
Alejandra Pera
Alejandra Pera
Juan Ruano
Juan Ruano
Juan Ruano
author_sort Jesus Gay-Mimbrera
collection DOAJ
description IntroductionAlopecia areata (AA) is an autoimmune disorder characterized by non-scarring hair loss ranging from mild, self-limiting episodes to severe and chronic forms. While prior research has primarily focused on lesional skin, the contribution of systemic immune cells remains underexplored.MethodsWe performed integrated single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) on peripheral blood mononuclear cells (PBMCs) from patients with mild and severe AA, as well as healthy controls. A total of 32,453 high-quality cells were analyzed across 36 immune cell subtypes.ResultsIn AA patients, especially those with severe disease, we observed increased transcriptional heterogeneity, cytokine and chemokine pathway activation, and upregulation of antigen-presentation machinery enriched in TH1, TH2, and TH17 signatures. Chromatin accessibility profiling revealed 42,248 significant peaks with pronounced epigenetic remodeling in CD14+ monocytes, NK cells, and CD8+ T cells. Mild AA showed early immune regulatory failure, with elevated exhaustion markers in double-negative T cells and increased apoptosis in myeloid populations. Pseudotime and transcription factor analyses indicated altered differentiation trajectories, and inferred cell-cell communication networks highlighted monocytes, NK cells, and memory T cells as key signaling hubs.DiscussionOur results provide the first integrated single-cell chromatin and transcriptomic map of peripheral immune dysregulation in AA. These findings uncover systemic alterations associated with disease severity and identify candidate pathways for immune modulation and therapeutic targeting.
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spelling doaj-art-cca6efea7f214d829238a6c1968d525e2025-08-20T02:37:38ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15652411565241Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areataJesus Gay-Mimbrera0Pedro Jesús Gómez-Arias1Pedro Jesús Gómez-Arias2Pablo Álvarez-Heredia3Alexander Batista-Duharte4Irene Rivera-Ruiz5Macarena Aguilar-Luque6Miguel Juan-Cencerrado7Miguel Juan-Cencerrado8Carmen Mochón-Jiménez9Carmen Mochón-Jiménez10Álvaro Cebrián-García11Eloísa Andújar-Pulido12Mónica Pérez-Alegre13Alejandra Pera14Alejandra Pera15Juan Ruano16Juan Ruano17Juan Ruano18Inflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainDepartment of Dermatology, Reina Sofía University Hospital, Córdoba, SpainImmunology and Allergy Group (GC01), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Cordoba, SpainImmunology and Allergy Group (GC01), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Cordoba, SpainDepartment of Dermatology, Reina Sofía University Hospital, Córdoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainDepartment of Dermatology, Reina Sofía University Hospital, Córdoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainDepartment of Dermatology, Reina Sofía University Hospital, Córdoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainGenomic Unit, Andalusian Molecular Biology and Regenerative Medicine Center (CABIMER), CSIC-University of Seville-University Pablo de Olavide, Seville, SpainGenomic Unit, Andalusian Molecular Biology and Regenerative Medicine Center (CABIMER), CSIC-University of Seville-University Pablo de Olavide, Seville, SpainImmunology and Allergy Group (GC01), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Cordoba, SpainDepartment of Cell Biology, Physiology and Immunology, University of Cordoba, Cordoba, SpainInflammatory Immune-Mediated Chronic Skin Diseases Laboratory (GC26), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Córdoba, SpainImmunology and Allergy Group (GC01), Maimonides Biomedical Research Institute of Cordoba (IMIBIC)/University of Cordoba/Reina Sofia University Hospital, Cordoba, SpainDepartment of Medicine, University of Córdoba, Córdoba, SpainIntroductionAlopecia areata (AA) is an autoimmune disorder characterized by non-scarring hair loss ranging from mild, self-limiting episodes to severe and chronic forms. While prior research has primarily focused on lesional skin, the contribution of systemic immune cells remains underexplored.MethodsWe performed integrated single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) on peripheral blood mononuclear cells (PBMCs) from patients with mild and severe AA, as well as healthy controls. A total of 32,453 high-quality cells were analyzed across 36 immune cell subtypes.ResultsIn AA patients, especially those with severe disease, we observed increased transcriptional heterogeneity, cytokine and chemokine pathway activation, and upregulation of antigen-presentation machinery enriched in TH1, TH2, and TH17 signatures. Chromatin accessibility profiling revealed 42,248 significant peaks with pronounced epigenetic remodeling in CD14+ monocytes, NK cells, and CD8+ T cells. Mild AA showed early immune regulatory failure, with elevated exhaustion markers in double-negative T cells and increased apoptosis in myeloid populations. Pseudotime and transcription factor analyses indicated altered differentiation trajectories, and inferred cell-cell communication networks highlighted monocytes, NK cells, and memory T cells as key signaling hubs.DiscussionOur results provide the first integrated single-cell chromatin and transcriptomic map of peripheral immune dysregulation in AA. These findings uncover systemic alterations associated with disease severity and identify candidate pathways for immune modulation and therapeutic targeting.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1565241/fullAlopecia areataseverityperipheral bloodimmune cellssingle-cell RNA sequencingchromatin accessibility
spellingShingle Jesus Gay-Mimbrera
Pedro Jesús Gómez-Arias
Pedro Jesús Gómez-Arias
Pablo Álvarez-Heredia
Alexander Batista-Duharte
Irene Rivera-Ruiz
Macarena Aguilar-Luque
Miguel Juan-Cencerrado
Miguel Juan-Cencerrado
Carmen Mochón-Jiménez
Carmen Mochón-Jiménez
Álvaro Cebrián-García
Eloísa Andújar-Pulido
Mónica Pérez-Alegre
Alejandra Pera
Alejandra Pera
Juan Ruano
Juan Ruano
Juan Ruano
Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
Frontiers in Immunology
Alopecia areata
severity
peripheral blood
immune cells
single-cell RNA sequencing
chromatin accessibility
title Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
title_full Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
title_fullStr Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
title_full_unstemmed Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
title_short Integrated single-cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
title_sort integrated single cell chromatin and transcriptomic analyses of peripheral immune cells in patients with alopecia areata
topic Alopecia areata
severity
peripheral blood
immune cells
single-cell RNA sequencing
chromatin accessibility
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1565241/full
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