Down-regulation of KLF2 in lung fibroblasts is linked with COVID-19 immunofibrosis and restored by combined inhibition of NETs, JAK-1/2 and IL-6 signaling

Chrysanthopoulou, Akrivi; Antoniadou, Christina; Natsi, Anastasia-Maria; Gavriilidis, Efstratios; Papadopoulos, Vasileios; Xingi, Evangelia; Didaskalou, Stylianos; Mikroulis, Dimitrios; Tsironidou, Victoria; Kambas, Konstantinos; Koffa, Maria; Skendros, Panagiotis; Ritis, Konstantinos

doi:10.1016/j.clim.2023.109240

Cited by 13 publications

(28 citation statements)

References 68 publications

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“…Our functional studies indicated that NET-enriched extracellular mediators (eNETs) ex-vivo isolated from peripheral neutrophils of CD patients, and not the CD serum directly, were able to transform healthy intestinal fibroblasts toward the distinct CD phenotype characterized by negative KLF2 and high CCN2 expression, leading to collagen production. Similar immunofibrotic phenotype of lung fibroblasts has been also described in severe COVID-19 (13) . However, eNETs from peripheral neutrophils of UC patients did not have a similar potential, suggesting differential neutrophil/NETs plasticity on fibroblasts between the two IBDs.…”

Section: Discussionsupporting

confidence: 76%

“…To study the intracellular protein levels of CCN2 or KLF2, PIFs were cultured in 96-well high-binding microplates (655061; Thermo Fisher Scientific) in the presence of distinct stimuli for 4 h, as previously described (13, 37) . Briefly, goat anti-CCN2 (2 μg/mL; sc14939; Santa Cruz Biotechnology Inc) or mouse anti-KLF2 (6 μg/mL; MA5-24300; Thermo Fisher Scientific) were used as primary monoclonal antibodies.…”

Section: Supplementary Materials and Methodsmentioning

confidence: 99%

“…PIFs were cultured in 8-well-chambered slides (80806; Ibidi), and treated with various stimuli for 4 h, as previously described (13, 33) . In brief, a rabbit anti-CCN2 polyclonal antibody (1:600 dilution; ab5097; Αbcam), a mouse anti-KLF2 monoclonal antibody (6 μg/mL; MA5-24300; Thermo Fisher Scientific), a mouse anti-IL-8 monoclonal antibody (15 μg/mL; MAB208-100; R&D systems), a rabbit anti-desmin polyclonal antibody (1:200; ab15200; Abcam), and/or a rabbit anti-Vimentin monoclonal antibody (1:250; ab92547; Abcam), were used as primary antibodies.…”

Section: Supplementary Materials and Methodsmentioning

confidence: 99%

“…References (13,33,37,45,(51)(52)(53)(54)(55)(56) used as primary monoclonal antibodies. Horseradish peroxidase-conjugated donkey anti-goat IgG (1:1000 dilution, HAF109; R&D Systems) or donkey anti-mouse IgG (1:1000 dilution, HAF018; R&D Systems) were used as secondary antibodies.…”

Section: Fig S1 To S7mentioning

confidence: 99%

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Neutrophil-fibroblast crosstalk drives immunofibrosis in Crohn’s disease through IFNα pathway

Gavriilidis,

Divolis,

Natsi

et al. 2023

Preprint

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Crohn’s disease (CD) is characterized by chronic inflammation and intestinal fibrosis leading to lifelong complications. However, disease pathogenesis is elusive and the therapeutic options are limited. We investigated the interaction between neutrophils and intestinal fibroblasts in the development of CD immunofibrosis. Peripheral neutrophils, enriched neutrophil extracellular traps (eNETs), serum, primary intestinal fibroblasts (PIFs) and intestinal biopsies from CD and ulcerative colitis (UC) patients, and healthy individuals (HI), were studied. Neutrophil-targeted transcriptomics, multi-cytokine profiling and cell-based functional assays at mRNA/protein level were performed. Compared to UC, PIFs from CD patients displayed a distinct fibrotic phenotype characterized by negative Krüppel-like Factor-2 (KLF2) and increased cellular communication network factor-2 (CCN2) expression leading to collagen production. In UC and CD, PIFs-derived IL-8 appears as a culprit chemoattractant of neutrophils in the intestine, where CD neutrophils were accumulated close to fibrotic lesions. Functionally, only CD neutrophilsviaeNETs can induce a CD-like phenotype in HI PIFs, suggesting their fibrotic plasticity. High IFNα in serum and IFΝ-responsive signature in peripheral neutrophils were observed in CD, distinguishing it from UC. Moreover, CD serum can stimulate the release of fibrogenic eNETs from neutrophils in an IFNα-dependent manner, suggesting the priming role of IFNα in circulating neutrophils. Inhibition of eNETs or JAK signaling in neutrophils or PIFs prevented the neutrophil-mediated fibrotic effect on PIFs. Furthermore, serum IFNα and transcripts of key IFN-signaling components in neutrophils were well-correlated with CD severity. This study reveals the important role of IFNα/neutrophil/fibroblast axis in CD immunofibrosis, suggesting candidate biomarkers and putative therapeutic targets.

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Section: Discussionsupporting

confidence: 76%

Section: Supplementary Materials and Methodsmentioning

confidence: 99%

Section: Supplementary Materials and Methodsmentioning

confidence: 99%

Section: Fig S1 To S7mentioning

confidence: 99%

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Neutrophil-fibroblast crosstalk drives immunofibrosis in Crohn’s disease through IFNα pathway

Gavriilidis,

Divolis,

Natsi

et al. 2023

Preprint

Self Cite

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“…As one modulating factor of COVID-19 disease clinical outcomes, control of air pollution and short- and long-term exposure to toxic air pollutants are essential for improving public health measures, especially to protect the vulnerable population in urban dense areas ( Clerbaux et al, 2022 ). In addition is well known that coinfection with the influenza A virus and other viral infections enhances SARS-CoV-2 infectivity in lower immunity human systems ( Bai et al, 2021 ; Chrysanthopoulou et al, 2023 ). However, outdoor and indoor environmental exposures ( Yang et al, 2023 ) are interlinked with social, demographic, and evolutionary factors ( Destoumieux-Garzon et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Peculiar weather patterns effects on air pollution and COVID-19 spread in Tokyo metropolis

Zoran

Savastr

Săvăstru

et al. 2023

Environmental Research

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Comprehensive transcription factor perturbations recapitulate fibroblast transcriptional states

Southard,

Ardy,

Tang

et al. 2024

Preprint

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SummaryCell atlas projects have nominated recurrent transcriptional states as drivers of biological processes and disease, but their origins, regulation, and properties remain unclear. To enable complementary functional studies, we developed a scalable approach for recapitulating cell statesin vitrousing CRISPR activation (CRISPRa) Perturb-seq. Aided by a novel multiplexing method, we activated 1,836 transcription factors in two cell types. Measuring 21,958 perturbations showed that CRISPRa activated targets within physiological ranges, that epigenetic features predicted activatable genes, and that the protospacer seed region drove an off-target effect. Perturbations recapitulatedin vivofibroblast states, including universal and inflammatory states, and identifiedKLF4andKLF5as key regulators of the universal state. Inducing the universal state suppressed disease-associated states, highlighting its therapeutic potential. Our findings cement CRISPRa as a tool for perturbing differentiated cells and indicate thatin vivostates can be elicited via perturbation, enabling studies of clinically relevant statesex vivo.

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Down-regulation of KLF2 in lung fibroblasts is linked with COVID-19 immunofibrosis and restored by combined inhibition of NETs, JAK-1/2 and IL-6 signaling

Cited by 13 publications

References 68 publications

Neutrophil-fibroblast crosstalk drives immunofibrosis in Crohn’s disease through IFNα pathway

Neutrophil-fibroblast crosstalk drives immunofibrosis in Crohn’s disease through IFNα pathway

Peculiar weather patterns effects on air pollution and COVID-19 spread in Tokyo metropolis

Comprehensive transcription factor perturbations recapitulate fibroblast transcriptional states

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