CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages

Jonathan L Gillan; Mithil Chokshi; Gareth R Hardisty; Sara Clohisey Hendry; Daniel Prasca-Chamorro; Nicola J Robinson; Benjamin Lasota; Richard Clark; Lee Murphy; Moira K B Whyte; J Kenneth Baillie; Donald J Davidson; Gang Bao; Robert D Gray

doi:10.1126/sciadv.adg5128

CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages

Jonathan L Gillan, Mithil Chokshi, Gareth R Hardisty, Sara Clohisey Hendry, Daniel Prasca-Chamorro, Nicola J Robinson, Benjamin Lasota, Richard Clark, Lee Murphy, Moira K B Whyte, J Kenneth Baillie, Donald J Davidson, Gang Bao, Robert D Gray^*

^*Corresponding author for this work

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

Abstract / Description of output

An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5' end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.

Original language	English
Article number	eadg5128
Pages (from-to)	1-17
Number of pages	17
Journal	Science Advances
Volume	9
Issue number	21
Early online date	26 May 2023
DOIs	https://doi.org/10.1126/sciadv.adg5128
Publication status	Published - 26 May 2023

Keywords / Materials (for Non-textual outputs)

Humans
Cystic Fibrosis/genetics
Cystic Fibrosis Transmembrane Conductance Regulator/genetics
Macrophages/metabolism
Signal Transduction
Mutation
Pseudomonas aeruginosa

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10.1126/sciadv.adg5128

sciadv.adg5128Final published version, 1.92 MBLicence: Creative Commons: Attribution (CC-BY)

Genetics Core, Edinburgh Clinical Research Facility
Lee Murphy (Manager), Angie Fawkes (Other), Katarzyna Hafezi (Other), Richard Clark (Other), Amanda MacCallum (Other), Emma Aitken (Other) & Louise Macgillivray (Other)
Deanery of Clinical Sciences
Facility/equipment: Facility
Institute for Regeneration and Repair Flow Cytometry Facility
Shonna Johnston (Manager), Fiona Rossi (Manager), Claire Cryer (Other) & Ailsa Laird (Other)
Institute of Regeneration and Repair
Facility/equipment: Facility

Cite this

Gillan, J. L., Chokshi, M., Hardisty, G. R., Clohisey Hendry, S., Prasca-Chamorro, D., Robinson, N. J., Lasota, B., Clark, R., Murphy, L., Whyte, M. K. B., Baillie, J. K., Davidson, D. J., Bao, G., & Gray, R. D. (2023). CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages. Science Advances, 9(21), 1-17. Article eadg5128. Advance online publication. https://doi.org/10.1126/sciadv.adg5128

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title = "CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages",

abstract = "An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5' end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.",

keywords = "Humans, Cystic Fibrosis/genetics, Cystic Fibrosis Transmembrane Conductance Regulator/genetics, Macrophages/metabolism, Signal Transduction, Mutation, Pseudomonas aeruginosa",

author = "Gillan, {Jonathan L} and Mithil Chokshi and Hardisty, {Gareth R} and {Clohisey Hendry}, Sara and Daniel Prasca-Chamorro and Robinson, {Nicola J} and Benjamin Lasota and Richard Clark and Lee Murphy and Whyte, {Moira K B} and Baillie, {J Kenneth} and Davidson, {Donald J} and Gang Bao and Gray, {Robert D}",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors.",

year = "2023",

month = may,

day = "26",

doi = "10.1126/sciadv.adg5128",

language = "English",

volume = "9",

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publisher = "American Association for the Advancement of Science",

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Gillan, JL, Chokshi, M, Hardisty, GR , Clohisey Hendry, S, Prasca-Chamorro, D, Robinson, NJ, Lasota, B, Clark, R, Murphy, L, Whyte, MKB, Baillie, JK , Davidson, DJ, Bao, G & Gray, RD 2023, 'CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages', Science Advances, vol. 9, no. 21, eadg5128, pp. 1-17. https://doi.org/10.1126/sciadv.adg5128

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T1 - CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages

AU - Gillan, Jonathan L

AU - Chokshi, Mithil

AU - Hardisty, Gareth R

AU - Clohisey Hendry, Sara

AU - Prasca-Chamorro, Daniel

AU - Robinson, Nicola J

AU - Lasota, Benjamin

AU - Clark, Richard

AU - Murphy, Lee

AU - Whyte, Moira K B

AU - Baillie, J Kenneth

AU - Davidson, Donald J

AU - Bao, Gang

AU - Gray, Robert D

PY - 2023/5/26

Y1 - 2023/5/26

N2 - An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5' end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.

AB - An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5' end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.

KW - Humans

KW - Cystic Fibrosis/genetics

KW - Cystic Fibrosis Transmembrane Conductance Regulator/genetics

KW - Macrophages/metabolism

KW - Signal Transduction

KW - Mutation

KW - Pseudomonas aeruginosa

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DO - 10.1126/sciadv.adg5128

M3 - Article

C2 - 37235648

SN - 2375-2548

VL - 9

SP - 1

EP - 17

JO - Science Advances

JF - Science Advances

IS - 21

M1 - eadg5128

ER -

CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

Access to Document

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Equipment

Genetics Core, Edinburgh Clinical Research Facility

Institute for Regeneration and Repair Flow Cytometry Facility

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