Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome

Serena Colafrancesco; Cristiana Barbati; Roberta Priori; Erisa Putro; Federico Giardina; Angelica Gattamelata; Benedetta Monosi; Tania Colasanti; Alessandra Ida Celia; Bruna Cerbelli; Carla Giordano; Susanna Scarpa; Massimo Fusconi; Giulio Cavalli; Onorina Berardicurti; Saviana Gandolfo; Saba Nayar; Francesca Barone; Roberto Giacomelli; Salvatore De Vita; Cristiano Alessandri; Fabrizio Conti

doi:10.1002/art.42018

Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome

Serena Colafrancesco, Cristiana Barbati, Roberta Priori^*, Erisa Putro, Federico Giardina, Angelica Gattamelata, Benedetta Monosi, Tania Colasanti, Alessandra Ida Celia, Bruna Cerbelli, Carla Giordano, Susanna Scarpa, Massimo Fusconi, Giulio Cavalli, Onorina Berardicurti, Saviana Gandolfo, Saba Nayar, Francesca Barone, Roberto Giacomelli, Salvatore De VitaCristiano Alessandri, Fabrizio Conti

^*Corresponding author for this work

Inflammation and Ageing

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

Abstract

OBJECTIVE: Salivary gland epithelial cells (SGECs) are key cellular drivers in the pathogenesis of primary Sjögren's syndrome (SS); however, the mechanisms sustaining SGEC activation in primary SS remain unclear. We undertook this study to determine the role of autophagy in the survival and activation of SGECs in primary SS.

METHODS: Primary SGECs isolated from the minor SGs of patients with primary SS or sicca syndrome were evaluated by flow cytometry, immunoblotting, and immunofluorescence to assess autophagy (autophagic flux, light chain 3 IIB [LC3-IIB], p62, LC3-IIB+/lysosome-associated membrane protein 1 [LAMP-1] staining), apoptosis (annexin V/propidium iodide [PI], caspase 3), and activation (intercellular adhesion molecule, vascular cell adhesion molecule). Focus score and germinal center presence were assessed in the SGs from the same patients to assess correlation with histologic severity. Human SG (HSG) cells were stimulated in vitro with peripheral blood mononuclear cells (PBMCs) and serum from primary SS patients in the presence or absence of autophagy inhibitors to determine changes in autophagy and epithelial cell activation.

RESULTS: SGECs from primary SS patients (n = 24) exhibited increased autophagy (autophagic flux [P = 0.001]; LC3-IIB [P = 0.02]; p62 [P = 0.064]; and as indicated by LC3-IIB/LAMP-1+ staining), increased expression of antiapoptotic molecules (Bcl-2 [P = 0.006]), and reduced apoptosis (annexin V/PI [P = 0.002]; caspase 3 [P = 0.057]), compared to samples from patients with sicca syndrome (n = 16). Autophagy correlated with histologic disease severity. In vitro experiments on HSG cells stimulated with serum and PBMCs from primary SS patients confirmed activation of autophagy and expression of adhesion molecules, which was reverted upon pharmacologic inhibition of autophagy.

CONCLUSION: In primary SS SGECs, inflammation induces autophagy and prosurvival mechanisms, which promote SGEC activation and mirror histologic severity. These findings indicate that autophagy is a central contributor to the pathogenesis of primary SS and a new therapeutic target.

Original language	English
Pages (from-to)	654-664
Number of pages	11
Journal	Arthritis & Rheumatology (Hoboken)
Volume	74
Issue number	4
Early online date	8 Nov 2021
DOIs	https://doi.org/10.1002/art.42018
Publication status	Published - Apr 2022

Bibliographical note

Keywords

Annexin A5
Autophagy
Caspase 3/metabolism
Cell Adhesion Molecules/metabolism
Humans
Leukocytes, Mononuclear/metabolism
Sjogren's Syndrome
Transcription Factors/metabolism

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/art.42018

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Colafrancesco, S., Barbati, C., Priori, R., Putro, E., Giardina, F., Gattamelata, A., Monosi, B., Colasanti, T., Celia, A. I., Cerbelli, B., Giordano, C., Scarpa, S., Fusconi, M., Cavalli, G., Berardicurti, O., Gandolfo, S., Nayar, S., Barone, F., Giacomelli, R., ... Conti, F. (2022). Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome. Arthritis & Rheumatology (Hoboken), 74(4), 654-664. https://doi.org/10.1002/art.42018

@article{9f5cbded7df84e72b0a9a9331bbc5935,

title = "Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sj{\"o}gren's Syndrome",

abstract = "OBJECTIVE: Salivary gland epithelial cells (SGECs) are key cellular drivers in the pathogenesis of primary Sj{\"o}gren's syndrome (SS); however, the mechanisms sustaining SGEC activation in primary SS remain unclear. We undertook this study to determine the role of autophagy in the survival and activation of SGECs in primary SS.METHODS: Primary SGECs isolated from the minor SGs of patients with primary SS or sicca syndrome were evaluated by flow cytometry, immunoblotting, and immunofluorescence to assess autophagy (autophagic flux, light chain 3 IIB [LC3-IIB], p62, LC3-IIB+/lysosome-associated membrane protein 1 [LAMP-1] staining), apoptosis (annexin V/propidium iodide [PI], caspase 3), and activation (intercellular adhesion molecule, vascular cell adhesion molecule). Focus score and germinal center presence were assessed in the SGs from the same patients to assess correlation with histologic severity. Human SG (HSG) cells were stimulated in vitro with peripheral blood mononuclear cells (PBMCs) and serum from primary SS patients in the presence or absence of autophagy inhibitors to determine changes in autophagy and epithelial cell activation.RESULTS: SGECs from primary SS patients (n = 24) exhibited increased autophagy (autophagic flux [P = 0.001]; LC3-IIB [P = 0.02]; p62 [P = 0.064]; and as indicated by LC3-IIB/LAMP-1+ staining), increased expression of antiapoptotic molecules (Bcl-2 [P = 0.006]), and reduced apoptosis (annexin V/PI [P = 0.002]; caspase 3 [P = 0.057]), compared to samples from patients with sicca syndrome (n = 16). Autophagy correlated with histologic disease severity. In vitro experiments on HSG cells stimulated with serum and PBMCs from primary SS patients confirmed activation of autophagy and expression of adhesion molecules, which was reverted upon pharmacologic inhibition of autophagy.CONCLUSION: In primary SS SGECs, inflammation induces autophagy and prosurvival mechanisms, which promote SGEC activation and mirror histologic severity. These findings indicate that autophagy is a central contributor to the pathogenesis of primary SS and a new therapeutic target.",

keywords = "Annexin A5, Autophagy, Caspase 3/metabolism, Cell Adhesion Molecules/metabolism, Humans, Leukocytes, Mononuclear/metabolism, Sjogren's Syndrome, Transcription Factors/metabolism",

author = "Serena Colafrancesco and Cristiana Barbati and Roberta Priori and Erisa Putro and Federico Giardina and Angelica Gattamelata and Benedetta Monosi and Tania Colasanti and Celia, {Alessandra Ida} and Bruna Cerbelli and Carla Giordano and Susanna Scarpa and Massimo Fusconi and Giulio Cavalli and Onorina Berardicurti and Saviana Gandolfo and Saba Nayar and Francesca Barone and Roberto Giacomelli and {De Vita}, Salvatore and Cristiano Alessandri and Fabrizio Conti",

note = "{\textcopyright} 2021 American College of Rheumatology.",

year = "2022",

month = apr,

doi = "10.1002/art.42018",

language = "English",

volume = "74",

pages = "654--664",

journal = "Arthritis & Rheumatology (Hoboken)",

issn = "2326-5191",

publisher = "Wiley",

number = "4",

}

Colafrancesco, S, Barbati, C, Priori, R, Putro, E, Giardina, F, Gattamelata, A, Monosi, B, Colasanti, T, Celia, AI, Cerbelli, B, Giordano, C, Scarpa, S, Fusconi, M, Cavalli, G, Berardicurti, O, Gandolfo, S, Nayar, S, Barone, F, Giacomelli, R, De Vita, S, Alessandri, C & Conti, F 2022, 'Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome', Arthritis & Rheumatology (Hoboken), vol. 74, no. 4, pp. 654-664. https://doi.org/10.1002/art.42018

TY - JOUR

T1 - Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome

AU - Colafrancesco, Serena

AU - Barbati, Cristiana

AU - Priori, Roberta

AU - Putro, Erisa

AU - Giardina, Federico

AU - Gattamelata, Angelica

AU - Monosi, Benedetta

AU - Colasanti, Tania

AU - Celia, Alessandra Ida

AU - Cerbelli, Bruna

AU - Giordano, Carla

AU - Scarpa, Susanna

AU - Fusconi, Massimo

AU - Cavalli, Giulio

AU - Berardicurti, Onorina

AU - Gandolfo, Saviana

AU - Nayar, Saba

AU - Barone, Francesca

AU - Giacomelli, Roberto

AU - De Vita, Salvatore

AU - Alessandri, Cristiano

AU - Conti, Fabrizio

PY - 2022/4

Y1 - 2022/4

N2 - OBJECTIVE: Salivary gland epithelial cells (SGECs) are key cellular drivers in the pathogenesis of primary Sjögren's syndrome (SS); however, the mechanisms sustaining SGEC activation in primary SS remain unclear. We undertook this study to determine the role of autophagy in the survival and activation of SGECs in primary SS.METHODS: Primary SGECs isolated from the minor SGs of patients with primary SS or sicca syndrome were evaluated by flow cytometry, immunoblotting, and immunofluorescence to assess autophagy (autophagic flux, light chain 3 IIB [LC3-IIB], p62, LC3-IIB+/lysosome-associated membrane protein 1 [LAMP-1] staining), apoptosis (annexin V/propidium iodide [PI], caspase 3), and activation (intercellular adhesion molecule, vascular cell adhesion molecule). Focus score and germinal center presence were assessed in the SGs from the same patients to assess correlation with histologic severity. Human SG (HSG) cells were stimulated in vitro with peripheral blood mononuclear cells (PBMCs) and serum from primary SS patients in the presence or absence of autophagy inhibitors to determine changes in autophagy and epithelial cell activation.RESULTS: SGECs from primary SS patients (n = 24) exhibited increased autophagy (autophagic flux [P = 0.001]; LC3-IIB [P = 0.02]; p62 [P = 0.064]; and as indicated by LC3-IIB/LAMP-1+ staining), increased expression of antiapoptotic molecules (Bcl-2 [P = 0.006]), and reduced apoptosis (annexin V/PI [P = 0.002]; caspase 3 [P = 0.057]), compared to samples from patients with sicca syndrome (n = 16). Autophagy correlated with histologic disease severity. In vitro experiments on HSG cells stimulated with serum and PBMCs from primary SS patients confirmed activation of autophagy and expression of adhesion molecules, which was reverted upon pharmacologic inhibition of autophagy.CONCLUSION: In primary SS SGECs, inflammation induces autophagy and prosurvival mechanisms, which promote SGEC activation and mirror histologic severity. These findings indicate that autophagy is a central contributor to the pathogenesis of primary SS and a new therapeutic target.

AB - OBJECTIVE: Salivary gland epithelial cells (SGECs) are key cellular drivers in the pathogenesis of primary Sjögren's syndrome (SS); however, the mechanisms sustaining SGEC activation in primary SS remain unclear. We undertook this study to determine the role of autophagy in the survival and activation of SGECs in primary SS.METHODS: Primary SGECs isolated from the minor SGs of patients with primary SS or sicca syndrome were evaluated by flow cytometry, immunoblotting, and immunofluorescence to assess autophagy (autophagic flux, light chain 3 IIB [LC3-IIB], p62, LC3-IIB+/lysosome-associated membrane protein 1 [LAMP-1] staining), apoptosis (annexin V/propidium iodide [PI], caspase 3), and activation (intercellular adhesion molecule, vascular cell adhesion molecule). Focus score and germinal center presence were assessed in the SGs from the same patients to assess correlation with histologic severity. Human SG (HSG) cells were stimulated in vitro with peripheral blood mononuclear cells (PBMCs) and serum from primary SS patients in the presence or absence of autophagy inhibitors to determine changes in autophagy and epithelial cell activation.RESULTS: SGECs from primary SS patients (n = 24) exhibited increased autophagy (autophagic flux [P = 0.001]; LC3-IIB [P = 0.02]; p62 [P = 0.064]; and as indicated by LC3-IIB/LAMP-1+ staining), increased expression of antiapoptotic molecules (Bcl-2 [P = 0.006]), and reduced apoptosis (annexin V/PI [P = 0.002]; caspase 3 [P = 0.057]), compared to samples from patients with sicca syndrome (n = 16). Autophagy correlated with histologic disease severity. In vitro experiments on HSG cells stimulated with serum and PBMCs from primary SS patients confirmed activation of autophagy and expression of adhesion molecules, which was reverted upon pharmacologic inhibition of autophagy.CONCLUSION: In primary SS SGECs, inflammation induces autophagy and prosurvival mechanisms, which promote SGEC activation and mirror histologic severity. These findings indicate that autophagy is a central contributor to the pathogenesis of primary SS and a new therapeutic target.

KW - Annexin A5

KW - Autophagy

KW - Caspase 3/metabolism

KW - Cell Adhesion Molecules/metabolism

KW - Humans

KW - Leukocytes, Mononuclear/metabolism

KW - Sjogren's Syndrome

KW - Transcription Factors/metabolism

U2 - 10.1002/art.42018

DO - 10.1002/art.42018

M3 - Article

C2 - 34748286

SN - 2326-5191

VL - 74

SP - 654

EP - 664

JO - Arthritis & Rheumatology (Hoboken)

JF - Arthritis & Rheumatology (Hoboken)

IS - 4

ER -

Maladaptive Autophagy in the Pathogenesis of Autoimmune Epithelitis in Sjögren's Syndrome

Abstract

Bibliographical note

Keywords

UN SDGs

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