Journal article
Elevation of Granulocyte Colony Stimulating Factor in Human AMD Donor RPE-Choroid
Investigative ophthalmology & visual science, Vol.65(14), 15
12/06/2024
DOI: 10.1167/iovs.65.14.15
PMCID: PMC11629913
PMID: 39641748
Abstract
Choroidal inflammation, complement deposition, and accumulation of C-reactive protein (CRP) are involved in age-related macular degeneration (AMD) pathology. The pro-inflammatory signals that regulate immune cell recruitment in the choroid of patients with AMD remain to be determined. We performed cytokine profiling of human AMD and age-matched control donor tissue to identify inflammatory molecules upregulated in AMD tissue.
Protein was isolated from 25 AMD and 21 control donor RPE/choroid macular punches. Total protein was quantified, and 50 µg assayed for expression of 40 cytokines using an inflammation array. We validated the elevated expression of granulocyte colony stimulating factor (G-CSF) protein by ELISA in a second cohort of 22 control and 26 AMD donors. To identify an AMD associated stressor responsible for upregulating G-CSF we assayed for changes in G-CSF protein secretion in RPE/choroid organ cultures treated with the monomeric (m)CRP, an inflammatory protein elevated in AMD.
Using a multiplex array, we identified elevated G-CSF protein in the choroid of AMD donors compared to age-matched non-AMD controls. Differential expression of G-CSF was confirmed via ELISA in an independent cohort of samples (P = 0.01). The mCRP, which is deposited in AMD choroids, increased G-CSF protein secretion in RPE/choroid organ cultures. Single nuclei RNA sequencing identified choroidal endothelial cells and fibroblasts as the primary cell types responsible for increased G-CSF secretion in response to mCRP. The G-CSF receptor is expressed primarily by choroidal macrophages and dendritic cells and anti-G-CSFR colocalizes with anti-CD45 and anti-CD68 in human donor choroid tissue.
Elevated G-CSF expression in AMD donor tissue as a result of increased levels of mCRP may be involved in immune cell recruitment in AMD contributing to inflammatory stress in the choroid.
Details
- Title: Subtitle
- Elevation of Granulocyte Colony Stimulating Factor in Human AMD Donor RPE-Choroid
- Creators
- Kelly Mulfaul - University of IowaAdnan H Khan - University of IowaSamantha G Schwarte - University of IowaAndrew P Voigt - University of IowaRachel F MooreLawrence A Potempa - Roosevelt UniversityKai Wang - University of IowaTodd E Scheetz - University of IowaEdwin M Stone - University of IowaBudd A Tucker - University of IowaRobert F Mullins - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Investigative ophthalmology & visual science, Vol.65(14), 15
- DOI
- 10.1167/iovs.65.14.15
- PMID
- 39641748
- PMCID
- PMC11629913
- NLM abbreviation
- Invest Ophthalmol Vis Sci
- ISSN
- 0146-0404
- eISSN
- 1552-5783
- Publisher
- ASSOC RESEARCH VISION OPHTHALMOLOGY INC
- Grant note
- National Institutes of health: EY024605, EY025580, EY033308 Elmer and Sylvia Sramek Charita-ble TrustBright Focus Foundation
Funded by the National Institutes of health grants EY024605, EY025580, and EY033308 the Elmer and Sylvia Sramek Charita-ble Trust, and the Bright Focus Foundation.
- Language
- English
- Date published
- 12/06/2024
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Neuroscience and Pharmacology; Ophthalmology and Visual Sciences
- Record Identifier
- 9984756245702771
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