Localization of SH3PXD2B in human eyes and detection of rare variants in patients with anterior segment diseases and glaucoma

Purpose Analysis of mutant mouse strains and linkage analysis with human families have both demonstrated that mutations influencing the podosomal adaptor protein SH3 and PX domains 2B (SH3PXD2B) can result in a congenital form of glaucoma. Here, we use immunohistochemistry to describe localization of the SH3PXD2B protein throughout the adult human eye and test whether sequence variants in SH3PXD2B occur in multiple other forms of glaucoma. Methods In immunohistochemical experiments, cryosections of human donor eyes were evaluated for SH3PXD2B immunoreactivity with a polyclonal antibody. In genetic experiments, exon sequences of SH3PXD2B from patients with primary congenital glaucoma (n=21), Axenfeld-Rieger syndrome (n=30), and primary open angle glaucoma (n=127) were compared to control subjects (n=89). The frequency of non-synonymous SH3PXD2B coding sequence variants were compared between patient cohorts and controls using Fisher’s exact test. Results Varying intensities of SH3PXD2B immunoreactivity were detected in almost all ocular tissues. Among tissues important to glaucoma, immunoreactivity was detected in the drainage structures of the iridocorneal angle, ciliary body, and retinal ganglion cells. Intense immunoreactivity was present in photoreceptor inner segments. From DNA analysis, a total of 11 non-synonymous variants were detected. By Fisher’s Exact test, there was not a significant skew in the overall frequency of these changes in any patient cohort versus controls (p-value >0.05). Each cohort contained unique variants not detected in other cohorts or patients. Conclusions SH3PXD2B is widely distributed in the adult human eye, including several tissues important to glaucoma pathogenesis. Analysis of DNA variants in three forms of glaucoma detected multiple variants unique to each patient cohort. While statistical analysis failed to support a pathogenic role for these variants, some of them may be rare disease-causing variants whose biologic significance warrants investigation in follow up replication studies and functional assays.

[1]  U. Schlötzer-Schrehardt,et al.  Variants in ASB10 are associated with open-angle glaucoma. , 2012, Human molecular genetics.

[2]  Michael G. Anderson,et al.  Reduction of ER stress via a chemical chaperone prevents disease phenotypes in a mouse model of primary open angle glaucoma. , 2011, The Journal of clinical investigation.

[3]  Young H. Kwon,et al.  Copy number variations on chromosome 12q14 in patients with normal tension glaucoma. , 2011, Human molecular genetics.

[4]  M. Brown,et al.  Genome-wide association study identifies susceptibility loci for open angle glaucoma at TMCO1 and CDKN2B-AS1 , 2011, Nature Genetics.

[5]  J. Fingert,et al.  Primary open-angle glaucoma genes , 2011, Eye.

[6]  Michael G. Anderson,et al.  Anterior segment dysgenesis and early-onset glaucoma in nee mice with mutation of Sh3pxd2b. , 2011, Investigative ophthalmology & visual science.

[7]  H. Stefánsson,et al.  Common variants near CAV1 and CAV2 are associated with primary open-angle glaucoma , 2010, Nature Genetics.

[8]  J. Wiggs,et al.  Glaucoma: genes, phenotypes, and new directions for therapy. , 2010, The Journal of clinical investigation.

[9]  J. Veltman,et al.  Disruption of the podosome adaptor protein TKS4 (SH3PXD2B) causes the skeletal dysplasia, eye, and cardiac abnormalities of Frank-Ter Haar Syndrome. , 2010, American journal of human genetics.

[10]  Michael G Anderson,et al.  Genetic dependence of central corneal thickness among inbred strains of mice. , 2010, Investigative ophthalmology & visual science.

[11]  T. Acott,et al.  Differential effects of ADAMTS-1, -4, and -5 in the trabecular meshwork. , 2009, Investigative ophthalmology & visual science.

[12]  Kathleen A. Marshall,et al.  Age-dependent effects of RPE65 gene therapy for Leber's congenital amaurosis: a phase 1 dose-escalation trial , 2009, The Lancet.

[13]  J. Vingerling,et al.  Heterozygous NTF4 mutations impairing neurotrophin-4 signaling in patients with primary open-angle glaucoma. , 2009, American journal of human genetics.

[14]  Michael G. Anderson,et al.  The podosomal-adaptor protein SH3PXD2B is essential for normal postnatal development , 2009, Mammalian Genome.

[15]  A. Caglayan,et al.  Frank-ter Haar syndrome with unusual clinical features. , 2009, European journal of medical genetics.

[16]  P. Russell,et al.  Gene expression profiling of TGFbeta2- and/or BMP7-treated trabecular meshwork cells: Identification of Smad7 as a critical inhibitor of TGF-beta2 signaling. , 2009, Experimental eye research.

[17]  D. Mackey,et al.  Null mutations in LTBP2 cause primary congenital glaucoma. , 2009, American journal of human genetics.

[18]  D. Epstein,et al.  Alterations in gene expression induced by cyclic mechanical stress in trabecular meshwork cells , 2009, Molecular vision.

[19]  I. Pass,et al.  The novel adaptor protein Tks4 (SH3PXD2B) is required for functional podosome formation. , 2009, Molecular biology of the cell.

[20]  Xiangyin Kong,et al.  Removal of Hsf4 leads to cataract development in mice through down-regulation of γS-crystallin and Bfsp expression , 2009, BMC Molecular Biology.

[21]  T. Acott,et al.  Specialized podosome- or invadopodia-like structures (PILS) for focal trabecular meshwork extracellular matrix turnover. , 2008, Investigative ophthalmology & visual science.

[22]  Edwin M Stone,et al.  Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics , 2008, Proceedings of the National Academy of Sciences.

[23]  Andrew J. Grimm,et al.  Interpreting missense variants: comparing computational methods in human disease genes CDKN2A, MLH1, MSH2, MECP2, and tyrosinase (TYR) , 2007, Human mutation.

[24]  R. Lavker,et al.  Transcriptional Profiling of Enriched Populations of Stem Cells Versus Transient Amplifying Cells , 2006, Journal of Biological Chemistry.

[25]  Shunsuke Kato,et al.  Computational approaches for predicting the biological effect of p53 missense mutations: a comparison of three sequence analysis based methods , 2006, Nucleic acids research.

[26]  David J. Calkins,et al.  Microarray analysis of retinal gene expression in the DBA/2J model of glaucoma. , 2006, Investigative ophthalmology & visual science.

[27]  H. Quigley,et al.  The number of people with glaucoma worldwide in 2010 and 2020 , 2006, British Journal of Ophthalmology.

[28]  R. Ritch,et al.  Identification of a novel adult-onset primary open-angle glaucoma (POAG) gene on 5q22.1. , 2005, Human molecular genetics.

[29]  R. Lewis,et al.  Cytochrome P4501B1 mutations cause only part of primary congenital glaucoma in Ecuador , 2004, Ophthalmic genetics.

[30]  Steven Henikoff,et al.  SIFT: predicting amino acid changes that affect protein function , 2003, Nucleic Acids Res..

[31]  D. Stephan,et al.  Effects of prostaglandin analogues on human ciliary muscle and trabecular meshwork cells. , 2003, Investigative ophthalmology & visual science.

[32]  P. Bork,et al.  Human non-synonymous SNPs: server and survey. , 2002, Nucleic acids research.

[33]  V. P. Costa,et al.  Molecular genetics of primary congenital glaucoma in Brazil. , 2002, Investigative ophthalmology & visual science.

[34]  I. Kimura,et al.  Novel cytochrome P4501B1 (CYP1B1) gene mutations in Japanese patients with primary congenital glaucoma. , 2001, Investigative ophthalmology & visual science.

[35]  Jean Bennett,et al.  Gene therapy restores vision in a canine model of childhood blindness , 2001, Nature Genetics.

[36]  W. Alward,et al.  Axenfeld-Rieger syndrome in the age of molecular genetics. , 2000, American journal of ophthalmology.

[37]  Stefan M. Larson,et al.  The identification of conserved interactions within the SH3 domain by alignment of sequences and structures , 2000, Protein science : a publication of the Protein Society.

[38]  M. Cargill.  Characterization of single-nucleotide polymorphisms in coding regions of human genes , 1999, Nature Genetics.

[39]  J. Morissette,et al.  Mutations of the forkhead/winged-helix gene, FKHL7, in patients with Axenfeld-Rieger anomaly. , 1998, American journal of human genetics.

[40]  V. Sheffield,et al.  The forkhead transcription factor gene FKHL7 is responsible for glaucoma phenotypes which map to 6p25 , 1998, Nature Genetics.

[41]  R. Ferrell,et al.  A novel homeobox gene PITX3 is mutated in families with autosomal-dominant cataracts and ASMD , 1998, Nature Genetics.

[42]  M. Sarfarazi,et al.  Identification of three different truncating mutations in cytochrome P4501B1 (CYP1B1) as the principal cause of primary congenital glaucoma (Buphthalmos) in families linked to the GLC3A locus on chromosome 2p21. , 1997, Human molecular genetics.

[43]  H. Quigley.  Number of people with glaucoma worldwide. , 1996, The British journal of ophthalmology.

[44]  J. Thornton,et al.  A revised set of potentials for β‐turn formation in proteins , 1994 .

[45]  J. Thornton,et al.  A revised set of potentials for beta-turn formation in proteins. , 1994, Protein science : a publication of the Protein Society.

[46]  S. Henikoff,et al.  Amino acid substitution matrices from protein blocks. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[47]  P. Raymond,et al.  Improved method for obtaining 3-microns cryosections for immunocytochemistry. , 1990, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[48]  B. Cohen,et al.  Megalocornea associated with multiple skeletal anomalies: a new genetic syndrome? , 1973, Journal de genetique humaine.