Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Journal of Neuroinflammation
Published in: Journal of Neuroinflammation 1/2016

Open Access 01-12-2016 | Research

Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)

Authors: Felix Grassmann, Stuart Cantsilieris, Anja-Sabrina Schulz-Kuhnt, Stefan J. White, Andrea J Richardson, Alex W Hewitt, Brendan J. Vote, Denise Schmied, Robyn H Guymer, Bernhard H.F. Weber, Paul N. Baird

Published in: Journal of Neuroinflammation | Issue 1/2016

Login to get access

Abstract

Background

Age-related macular degeneration (AMD) is the leading cause of vision loss in Western societies with a strong genetic component. Candidate gene studies as well as genome-wide association studies strongly implicated genetic variations in complement genes to be involved in disease risk. So far, no association of AMD with complement component 4 (C4) was reported probably due to the complex nature of the C4 locus on chromosome 6.

Methods

We used multiplex ligation-dependent probe amplification (MLPA) to determine the copy number of the C4 gene as well as of both relevant isoforms, C4A and C4B, and assessed their association with AMD using logistic regression models.

Results

Here, we report on the analysis of 2645 individuals (1536 probands and 1109 unaffected controls), across three different centers, for multiallelic copy number variation (CNV) at the C4 locus. We find strong statistical significance for association of increased copy number of C4A (OR 0.81 (0.73; 0.89);P = 4.4 × 10−5), with the effect most pronounced in individuals over 78 years (OR 0.67 (0.55; 0.81)) and females (OR 0.77 (0.68; 0.87)). Furthermore, this association is independent of known AMD-associated risk variants in the nearby CFB/C2 locus, particularly in females and in individuals over 78 years.

Conclusions

Our data strengthen the notion that complement dysregulation plays a crucial role in AMD etiology, an important finding for early intervention strategies and future therapeutics. In addition, for the first time, we provide evidence that multiallelic CNVs are associated with AMD pathology.
Appendix
Available only for authorised users
Literature
1.
Grassmann F, Fauser S, Weber BHF: The genetics of age-related macular degeneration (AMD)—novel targets for designing treatment options? Eur J Pharm Biopharm. 2015;95(Pt B):194–202.
2.
Grassmann F, Ach T, Brandl C, Heid IM, Weber BHF. What does genetics tell us about age-related macular degeneration? Annu Rev Vis Sci. 2015;1:73–96.CrossRef
3.
Fritsche LG, Fariss RN, Stambolian D, Abecasis GR, Curcio CA, Swaroop A. Age-related macular degeneration: genetics and biology coming together. Annu Rev Genomics Hum Genet. 2014;15:151–71.CrossRefPubMedPubMedCentral
4.
Swaroop A, Branham KE, Chen W, Abecasis G: Genetic susceptibility to age-related macular degeneration: a paradigm for dissecting complex disease traits. Hum Mol Genet 2007, 16 Spec No:R174–82.
5.
Gold B, Merriam JE, Zernant J, Hancox LS, Taiber AJ, Gehrs K, et al. Variation in factor B (BF) and complement component 2 (C2) genes is associated with age-related macular degeneration. Nat Genet. 2006;38:458–62.CrossRefPubMedPubMedCentral
6.
Hageman GS, Anderson DH, Johnson LV, Hancox LS, Taiber AJ, Hardisty LI, et al. A common haplotype in the complement regulatory gene factor H (HF1/CFH) predisposes individuals to age-related macular degeneration. Proc Natl Acad Sci U S A. 2005;102:7227–32.CrossRefPubMedPubMedCentral
7.
Seddon JM, Yu Y, Miller EC, Reynolds R, Tan PL, Gowrisankar S, et al. Rare variants in CFI, C3 and C9 are associated with high risk of advanced age-related macular degeneration. Nat Genet. 2013;45:1366–70.CrossRefPubMedPubMedCentral
8.
Zhan X, Larson DE, Wang C, Koboldt DC, Sergeev YV, Fulton RS, et al. Identification of a rare coding variant in complement 3 associated with age-related macular degeneration. Nat Genet. 2013;45:1375–9.CrossRefPubMedPubMedCentral
9.
Fritsche LG, Igl W, Bailey JNC, Grassmann F, Sengupta S, Bragg-Gresham JL, et al.: A large genome-wide association study of age-related macular degeneration highlights contributions of rare and common variants. Nat Genet 2015, in press.
10.
Cantsilieris S, White SJ. Correlating multiallelic copy number polymorphisms with disease susceptibility. Hum Mutat. 2013;34:1–13.CrossRefPubMed
11.
Hollox E. The challenges of studying complex and dynamic regions of the human genome, Genomic Structural Variants. New York: Springer; 2012.CrossRef
12.
Pinto D, Darvishi K, Shi X, Rajan D, Rigler D, Fitzgerald T, et al. Comprehensive assessment of array-based platforms and calling algorithms for detection of copy number variants. Nat Biotechnol. 2011;29:512–20.CrossRefPubMedPubMedCentral
13.
Aldhous MC, Abu Bakar S, Prescott NJ, Palla R, Soo K, Mansfield JC, et al. Measurement methods and accuracy in copy number variation: failure to replicate associations of beta-defensin copy number with Crohn’s disease. Hum Mol Genet. 2010;19:4930–8.CrossRefPubMedPubMedCentral
14.
Cantsilieris S, Western PS, Baird PN, White SJ. Technical considerations for genotyping multi-allelic copy number variation (CNV), in regions of segmental duplication. BMC Genomics. 2014;15:329.CrossRefPubMedPubMedCentral
15.
Hollox EJ, Huffmeier U, Zeeuwen PLJM, Palla R, Lascorz J, Rodijk-Olthuis D, et al. Psoriasis is associated with increased beta-defensin genomic copy number. Nat Genet. 2008;40:23–5.CrossRefPubMedPubMedCentral
16.
Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G. Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res. 2002;30:e57.CrossRefPubMedPubMedCentral
17.
Grassmann F, Friedrich U, Fauser S, Schick T, Milenkovic A, Schulz HL, et al. A candidate gene association study identifies DAPL1 as a female-specific susceptibility locus for age-related macular degeneration (AMD). NeuroMolecular Med. 2015;17:111–20.CrossRefPubMedPubMedCentral
18.
Cantsilieris S, White SJ, Richardson AJ, Guymer RH, Baird PN. Comprehensive analysis of copy number variation of genes at chromosome 1 and 10 loci associated with late age related macular degeneration. PLoS One. 2012;7:e35255.CrossRefPubMedPubMedCentral
19.
White SJ, Vissers LE, Geurts van Kessel A, de Menezes RX, Kalay E, Lehesjoki AE, et al. Variation of CNV distribution in five different ethnic populations. Cytogenet Genome Res. 2007;118:19–30.CrossRefPubMed
20.
White SJ, Vink GR, Kriek M, Wuyts W, Schouten J, Bakker B, et al. Two-color multiplex ligation-dependent probe amplification: detecting genomic rearrangements in hereditary multiple exostoses. Hum Mutat. 2004;24:86–92.CrossRefPubMed
21.
Barba GM, Braun-Heimer L, Rittner C, Schneider PM. A new PCR-based typing of the Rodgers and Chido antigenic determinants of the fourth component of human complement. Eur J Immunogenet. 1994;21:325–39.CrossRefPubMed
22.
O’Connell J, Gurdasani D, Delaneau O, Pirastu N, Ulivi S, Cocca M, et al. A general approach for haplotype phasing across the full spectrum of relatedness. PLoS Genet. 2014;10:e1004234.CrossRefPubMedPubMedCentral
23.
Higgins JPT, Thompson SG. Quantifying heterogeneity in a meta-analysis. Stat Med. 2002;21:1539–58.CrossRefPubMed
24.
Wouters D, van Schouwenburg P, van der Horst A, de Boer M, Schooneman D, Kuijpers TW, et al. High-throughput analysis of the C4 polymorphism by a combination of MLPA and isotype-specific ELISA’s. Mol Immunol. 2009;46:592–600.CrossRefPubMed
25.
Yu CY, Belt KT, Giles CM, Campbell RD, Porter RR. Structural basis of the polymorphism of human complement components C4A and C4B: gene size, reactivity and antigenicity. EMBO J. 1986;5:2873–81.PubMedPubMedCentral
26.
Dodds AW, Ren XD, Willis AC, Law SK. The reaction mechanism of the internal thioester in the human complement component C4. Nature. 1996;379:177–9.CrossRefPubMed
27.
Boteva L, Morris DL, Cortés-Hernández J, Martin J, Vyse TJ, Fernando MMA. Genetically determined partial complement C4 deficiency states are not independent risk factors for SLE in UK and Spanish populations. Am J Hum Genet. 2012;90:445–56.CrossRefPubMedPubMedCentral
28.
Yang Y, Chung EK, Wu YL, Savelli SL, Nagaraja HN, Zhou B, et al. Gene copy-number variation and associated polymorphisms of complement component C4 in human systemic lupus erythematosus (SLE): low copy number is a risk factor for and high copy number is a protective factor against SLE susceptibility in European America. Am J Hum Genet. 2007;80:1037–54.CrossRefPubMedPubMedCentral
29.
Fernando MMA, Boteva L, Morris DL, Zhou B, Wu YL, Lokki M-L, et al. Assessment of complement C4 gene copy number using the paralog ratio test. Hum Mutat. 2010;31:866–74.CrossRefPubMedPubMedCentral
30.
31.
Ersoy L, Ristau T, Hahn M, Karlstetter M, Langmann T, Dröge K, et al. Genetic and environmental risk factors for age-related macular degeneration in persons 90 years and older. Investig Opthalmology Vis Sci. 2014;55:1842.CrossRef
32.
33.
34.
35.
Johnson LV, Ozaki S, Staples MK, Erickson PA, Anderson DH. A potential role for immune complex pathogenesis in drusen formation. Exp Eye Res. 2000;70:441–9.CrossRefPubMed
36.
Yang Y, Chung EK, Zhou B, Blanchong CA, Yu CY, Füst G, et al. Diversity in intrinsic strengths of the human complement system: serum C4 protein concentrations correlate with C4 gene size and polygenic variations, hemolytic activities, and body mass index. J Immunol. 2003;171:2734–45.CrossRefPubMed
37.
Candore G, Modica MA, Lio D, Colonna-Romano G, Listì F, Grimaldi MP, et al. Pathogenesis of autoimmune diseases associated with 8.1 ancestral haplotype: a genetically determined defect of C4 influences immunological parameters of healthy carriers of the haplotype. Biomed Pharmacother. 2003;57:274–7.CrossRefPubMed
38.
Fritsche LG, Chen W, Schu M, Yaspan BL, Yu Y, Thorleifsson G, et al. Seven new loci associated with age-related macular degeneration. Nat Genet. 2013;45:433–9. 439e1–2.CrossRefPubMed
39.
Kopplin LJ, Igo RP, Wang Y, Sivakumaran TA, Hagstrom SA, Peachey NS, et al. Genome-wide association identifies SKIV2L and MYRIP as protective factors for age-related macular degeneration. Genes Immun. 2010;11:609–21.CrossRefPubMedPubMedCentral
40.
Cipriani V, Leung H-T, Plagnol V, Bunce C, Khan JC, Shahid H, et al. Genome-wide association study of age-related macular degeneration identifies associated variants in the TNXB-FKBPL-NOTCH4 region of chromosome 6p21.3. Hum Mol Genet. 2012;21:4138–50.CrossRefPubMedPubMedCentral
41.
Anderson DH, Radeke MJ, Gallo NB, Chapin EA, Johnson PT, Curletti CR, et al. The pivotal role of the complement system in aging and age-related macular degeneration: hypothesis re-visited. Prog Retin Eye Res. 2010;29:95–112.CrossRefPubMedPubMedCentral
42.
Feng Y, Wang Y, Li L, Wu L, Hoffmann S, Gretz N, et al. Gene expression profiling of vasoregression in the retina—involvement of microglial cells. PLoS One. 2011;6:e16865.CrossRefPubMedPubMedCentral
43.
Newsome DA, Hewitt AT, Huh W, Robey PG, Hassell JR. Detection of specific extracellular matrix molecules in drusen, Bruch’s membrane, and ciliary body. Am J Ophthalmol. 1987;104:373–81.CrossRefPubMed
44.
Flachsbart F, Franke A, Kleindorp R, Caliebe A, Blanché H, Schreiber S, et al. Investigation of genetic susceptibility factors for human longevity—a targeted nonsynonymous SNP study. Mutat Res. 2010;694:13–9.CrossRefPubMed
45.
Kramer J, Fülöp T, Rajczy K, Nguyen AT, Füst G. A marked drop in the incidence of the null allele of the B gene of the fourth component of complement (C4B*Q0) in elderly subjects: C4B*Q0 as a probable negative selection factor for survival. Hum Genet. 1991;86:595–8.CrossRefPubMed
46.
Hughes AE, Orr N, Esfandiary H, Diaz-torres M, Goodship T, Chakravarthy U. A common CFH haplotype, with deletion of CFHR1 and CFHR3, is associated with lower risk of age-related macular degeneration. Nat Genet. 2007;38:1173–8.CrossRef
47.
Fritsche LG, Loenhardt T, Janssen A, Fisher SA, Rivera A, Keilhauer CN, et al. Age-related macular degeneration is associated with an unstable ARMS2 (LOC387715) mRNA. Nat Genet. 2008;40:892–6.CrossRefPubMed
48.
Stuart PE, Hüffmeier U, Nair RP, Palla R, Tejasvi T, Schalkwijk J, et al. Association of β-defensin copy number and psoriasis in three cohorts of European origin. J Invest Dermatol. 2012;132:2407–13.CrossRefPubMedPubMedCentral
49.
Bay JT, Schejbel L, Madsen HO, Sørensen SS, Hansen JM, Garred P. Low C4 gene copy numbers are associated with superior graft survival in patients transplanted with a deceased donor kidney. Kidney Int. 2013;84:562–9.CrossRefPubMed
Metadata
Title
Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)
Authors
Felix Grassmann
Stuart Cantsilieris
Anja-Sabrina Schulz-Kuhnt
Stefan J. White
Andrea J Richardson
Alex W Hewitt
Brendan J. Vote
Denise Schmied
Robyn H Guymer
Bernhard H.F. Weber
Paul N. Baird
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Journal of Neuroinflammation / Issue 1/2016
Electronic ISSN: 1742-2094
DOI
https://doi.org/10.1186/s12974-016-0548-0

Other articles of this Issue 1/2016

Journal of Neuroinflammation 1/2016 Go to the issue

Research

1-Oleyl-lysophosphatidic acid (LPA) promotes polarization of BV-2 and primary murine microglia towards an M1-like phenotype

Research

Spatio-temporal profile, phenotypic diversity, and fate of recruited monocytes into the post-ischemic brain

Research

Intra-amniotic LPS causes acute neuroinflammation in preterm rhesus macaques

Research

Minocycline attenuates interferon-α-induced impairments in rat fear extinction

Research

N-Docosahexaenoylethanolamine ameliorates LPS-induced neuroinflammation via cAMP/PKA-dependent signaling

Research

Interferon-γ blocks signalling through PDGFRβ in human brain pericytes