Type I IFN–related NETosis in ataxia telangiectasia and Artemis deficiency - 20/12/17

Doi : 10.1016/j.jaci.2017.10.030

Ersin Gul, MSc ^a,^{^{∗
These 3 authors contributed equally to this work.}}, Esra Hazar Sayar, MD ^b,^{^{∗
These 3 authors contributed equally to this work.}}, Bilgi Gungor, PhD ^a,^{^{∗
These 3 authors contributed equally to this work.}}, Fehime Kara Eroglu, MD ^c, Naz Surucu, MSc ^a, Sevgi Keles, MD ^b, Sukru Nail Guner, MD ^b, Siddika Findik, MD ^d, Esin Alpdundar, MSc ^a, Ihsan Cihan Ayanoglu, MSc ^a, Basak Kayaoglu, MSc ^a, Busra Nur Geckin, BSc ^a, Hatice Asena Sanli, BSc ^a, Tamer Kahraman, PhD ^c, Cengiz Yakicier, MD ^e, Meltem Muftuoglu, PhD ^e, Berna Oguz, MD ^f, Deniz Nazire Cagdas Ayvaz, MD, PhD ^g, Ihsan Gursel, PhD ^c, Seza Ozen, MD ^h, Ismail Reisli, MD ^b, Mayda Gursel, PhD ^a,^⁎
^a Department of Biological Sciences, Middle East Technical University, Ankara, Turkey
^b Department of Immunology and Allergy, Meram Medical Faculty, Necmettin Erbakan University, Konya, Turkey
^c Thorlab, Therapeutic Oligodeoxynucleotide Research Laboratory, Department of Molecular Biology and Genetics, Ihsan Dogramaci Bilkent University, Ankara, Turkey
^d Department of Pathology, Meram Medical Faculty, Necmettin Erbakan University, Konya, Turkey
^e Department of Molecular Biology and Genetics, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey
^f Department of Radiology, Hacettepe University Medical Faculty, Ankara, Turkey
^g Department of Pediatric Immunology, Hacettepe University Medical Faculty, Ankara, Turkey
^h Department of Pediatric Rheumatology, Hacettepe University Medical Faculty, Ankara, Turkey

^∗Corresponding author: Mayda Gursel, PhD, Department of Biological Sciences, Middle East Technical University, 06800 Ankara, Turkey.Department of Biological SciencesMiddle East Technical UniversityAnkara06800Turkey

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Abstract

Background

Pathological inflammatory syndromes of unknown etiology are commonly observed in ataxia telangiectasia (AT) and Artemis deficiency. Similar inflammatory manifestations also exist in patients with STING-associated vasculopathy in infancy (SAVI).

Objective

We sought to test the hypothesis that the inflammation-associated manifestations observed in patients with AT and Artemis deficiency stem from increased type I IFN signature leading to neutrophil-mediated pathological damage.

Methods

Cytokine/protein signatures were determined by ELISA, cytometric bead array, or quantitative PCR. Stat1 phosphorylation levels were determined by flow cytometry. DNA species accumulating in the cytosol of patients' cells were quantified microscopically and flow cytometrically. Propensity of isolated polymorhonuclear granulocytes to form neutrophil extracellular traps (NETs) was determined using fluorescence microscopy and picogreen assay. Neutrophil reactive oxygen species levels and mitochondrial stress were assayed using fluorogenic probes, microscopy, and flow cytometry.

Results

Type I and III IFN signatures were elevated in plasma and peripheral blood cells of patients with AT, Artemis deficiency, and SAVI. Chronic IFN production stemmed from the accumulation of DNA in the cytoplasm of AT and Artemis-deficient cells. Neutrophils isolated from patients spontaneously produced NETs and displayed indicators of oxidative and mitochondrial stress, supportive of their NETotic tendencies. A similar phenomenon was also observed in neutrophils from healthy controls exposed to patient plasma samples or exogeneous IFN-α.

Conclusions

Type I IFN–mediated neutrophil activation and NET formation may contribute to inflammatory manifestations observed in patients with AT, Artemis deficiency, and SAVI. Thus, neutrophils represent a promising target to manage inflammatory syndromes in diseases with active type I IFN signature.

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Graphical abstract

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Key words : Primary immunodeficiencies, autoinflammation, ataxia telangiectasia, Artemis deficiency, type I IFN, interferonopathy, neutrophil extracellular traps, NETosis

Abbreviations used : AT, ATM, dsDNA, recIFN-α, SAVI, ssDNA, NET, SLE, IP-10, ISGs, ROS

Plan

Methods

Participants

Cytokine measurements and functional studies

Results

Autoimmune/autoinflammatory manifestations in patients with AT, Artemis deficiency, and SAVI in relation to elevated type I IFN activity

DNA spontaneously accumulates in the cytosol of AT and Artemis-deficient patient cells

AT, Artemis-deficient and SAVI patient neutrophils spontaneously produce NETs

AT, Artemis-deficient and SAVI patient plasma samples and recombinant IFN-α trigger NET release from healthy neutrophils

Type I IFNs mediate oxidative and mitochondrial stress in neutrophils

Discussion

	This project was partially supported by the Scientific and Technological Research Council of Turkey (TUBITAK) (grant nos. 315S125 and 115S430).
	Disclosure of potential conflict of interest: I. Gursel has received a grant from TUBITAK (grant no. 315S125). S. Ozen has consultant arrangements with Novartis and Pfizer. M. Gursel has received a grant from TUBITAK (grant no. 115S430). The rest of the authors declare that they have no relevant conflicts of interest.