September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
The expression of complement genes by retinal pigment epithelial cells from donors with and without AMD
Author Affiliations & Notes
  • Selina McHarg
    Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester, Manchester, England, United Kingdom
  • Nicole Brace
    Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester, Manchester, England, United Kingdom
  • Rahat Perveen
    Institute of Human Development, Centre for Genomic Medicine, University of Manchester, Manchester, United Kingdom
  • Graeme C.M. Black
    Institute of Human Development, Centre for Genomic Medicine, University of Manchester, Manchester, United Kingdom
    Centre for Genomic Medicine, Saint Mary's Hospital, Central Manchester University Hospitals, Manchester, United Kingdom
  • Richard Unwin
    Centre for Advanced Discovery and Experimental Therapeutics (CADET), Central Manchester University Hospitals NHS Foundation Trust, Institute of Human Development, University of Manchester, Manchester, United Kingdom
  • Gregory S Hageman
    Moran Center for Translational Medicine, Department of Ophthalmology & Visual Sciences, University of Utah, Salt Lake City, Utah, United States
  • Anthony John Day
    Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom
  • Simon John Clark
    Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester, Manchester, England, United Kingdom
  • Paul N Bishop
    Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester, Manchester, England, United Kingdom
    Centre for Advanced Discovery and Experimental Therapeutics (CADET), Central Manchester University Hospitals NHS Foundation Trust, Institute of Human Development, University of Manchester, Manchester, United Kingdom
  • Footnotes
    Commercial Relationships   Selina McHarg, None; Nicole Brace, None; Rahat Perveen, None; Graeme Black, None; Richard Unwin, None; Gregory Hageman, Voyant Biotherapeutics LLC. (I); Anthony Day, None; Simon Clark, None; Paul Bishop, None
  • Footnotes
    Support  Fight for Sight grant R117520
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 2638. doi:
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      Selina McHarg, Nicole Brace, Rahat Perveen, Graeme C.M. Black, Richard Unwin, Gregory S Hageman, Anthony John Day, Simon John Clark, Paul N Bishop; The expression of complement genes by retinal pigment epithelial cells from donors with and without AMD. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2638.

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      © ARVO (1962-2015); The Authors (2016-present)

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  • Supplements
Abstract

Purpose : Aberrant activation of the complement cascade is implicated in the pathogenesis of age-related macular degeneration (AMD), with genetic variation in complement genes known to be a major risk determinant. However, exactly how common variants in complement genes modify AMD risk remains poorly understood. The purpose of this study was to investigate the expression of complement genes in primary cultures of human retinal pigment epithelial (RPE) cells.

Methods : RPE cells isolated from adult human eye donor tissue were cultured for 1-2 weeks, and the RNA extracted. Quantitative polymerase chain reaction was used to determine the expression of the genes encoding factor H (FH), factor H-like protein 1 (FHL-1) and the factor H related proteins 1-5 (FHR1-5). Their relative expression was compared in RPE cells derived from donors with and without AMD.

Results : The genes encoding FH and FHL-1 were expressed by cultured human RPE cells. The expression levels in RPE cells derived from unaffected donors were relatively tightly clustered, but those in RPE cells derived from donors with AMD were much more variable and overall expression levels of FH and FHL-1 were both significantly increased, by over 2-fold. The gene encoding FHR3 was weakly expressed in RPE cells derived from subjects both with and without AMD, whilst weak gene expression of FHR1 and 5 was detected only in RPE cells derived from donors with AMD. Expression of the genes encoding FHR2 and FHR4 was not detected in any of the cultured RPE cells.

Conclusions : We demonstrate that primary cultures of human RPE cells express genes encoding FH, FHL-1, FHR1, FHR3 and FHR5; therefore locally derived complement proteins expressed by the RPE are likely to contribute to regulation of the complement system in the macula. The increased variability in expression of genes encoding FH and FHL-1 in RPE cells derived from donors with AMD, compared to those from unaffected donors, could be explained by the underlying genotypes and we are currently genotyping the donor tissue to explore this possibility.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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