Proteomic Analyses of Human Trabecular Meshwork

Summary

Principal Investigator: Sanjoy Bhattacharya
Abstract: [unreadable] DESCRIPTION (provided by applicant): Glaucoma refers collectively to a group of eye diseases whose molecular basis is poorly understood. Except in "normal-pressure" glaucoma, the diseases are associated with increased intraocular pressure. Increased resistance to aqueous outflow through the trabecular meshwork (TM) appears to play a key role in the onset and progression of primary open angle glaucoma (POAG). The hypothesis of the proposal is that POAG associated proteins and protein modifications contribute to blockage of aqueous outflow in the TM. Proteomic analyses of TM tissue from in vivo trabeculectomy and left over rim-tissue from cornea transplant will be pursued to determine protein differences between normal and glaucomatous donors. Liquid chromatography tandem mass spectrometry and bioinformatic methods will be used to identify POAG associated protein and protein modifications. In preliminary studies, 4-hydorxynonenal (HNE) and Argpyrimidine oxidative protein modifications appear to be more prevalent in glaucomatous tissue. Furthermore, cochlin, a protein associated with inheritable deafness, has been found only in glaucomatous tissue. Western analysis and immunohistochemistry will be used to probe for differences in protein modifications between normal and glaucomatous TM tissue and verify the prevalence of HNE modification in glaucomatous TM tissue. Western analysis and immunohistochemistry will also be used to test the hypothesis that cochlin is associated with glaucomatous trabecular meshwork plaques/mucopolysaccharide deposits. The long-term goal of this proposal is to better understand mechanisms of glaucoma pathogenesis and facilitate the development of effective therapies for limiting disease progression. [unreadable] [unreadable]
Funding Period: 2005-09-01 - 2008-08-31
more information: NIH RePORT

Top Publications

  1. pmc Aberrant glycosylation in the human trabecular meshwork
    Adam E Sienkiewicz
    Bascom Palmer Eye Institute, University of Miami, Miami, FL, USA
    Proteomics Clin Appl 8:130-42. 2014
  2. pmc COCH transgene expression in cultured human trabecular meshwork cells and its effect on outflow facility in monkey organ cultured anterior segments
    Eun Suk Lee
    Departments of Ophthalmology and Visual Sciences, Sodaemungu Shinchondong, Seoul 120 752, Korea
    Invest Ophthalmol Vis Sci 51:2060-6. 2010
  3. pmc Decreased carbohydrate metabolism enzyme activities in the glaucomatous trabecular meshwork
    Anna K Junk
    Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA
    Mol Vis 16:1286-91. 2010
  4. pmc Cochlin induced TREK-1 co-expression and annexin A2 secretion: role in trabecular meshwork cell elongation and motility
    Manik Goel
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida, United States of America
    PLoS ONE 6:e23070. 2011
  5. pmc Cochlin, intraocular pressure regulation and mechanosensing
    Manik Goel
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida, United States of America
    PLoS ONE 7:e34309. 2012
  6. pmc Increased isolevuglandin-modified proteins in glaucomatous astrocytes
    Bharathi Govindarajan
    Department of Chemistry, Case Western Reserve University, Cleveland, OH, USA
    Mol Vis 15:1079-91. 2009
  7. pmc Laser trabeculoplasty induces changes in the trabecular meshwork glycoproteome: a pilot study
    Adriana Amelinckx
    Bascom Palmer Eye Institute, Ophthalmic Biophysics Center, University of Miami Miller School of Medicine, Miami, Forida 33136, USA
    J Proteome Res 8:3727-36. 2009
  8. pmc Cochlin expression in anterior segment organ culture models after TGFbeta2 treatment
    Sanjoy K Bhattacharya
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida 33136, USA
    Invest Ophthalmol Vis Sci 50:551-9. 2009
  9. pmc Neuroprotection in glaucoma using calpain-1 inhibitors: regional differences in calpain-1 activity in the trabecular meshwork, optic nerve and implications for therapeutics
    Bharathi Govindarajan
    Bascom Palmer Eye Institute, University of Miami, Miami, FL 33136, USA
    CNS Neurol Disord Drug Targets 7:295-304. 2008
  10. pmc Technical Brief: a novel strategy for enrichment of trabecular meshwork protease proteome
    Renata Picciani
    Bascom Palmer Eye Institute, University of Miami, Miami, FL 33136, USA
    Mol Vis 14:871-7. 2008

Scientific Experts

  • G Hoppe
  • Sanjoy Bhattacharya
  • Eun Suk Lee
  • Anna K Junk
  • Renata Picciani
  • Bharathi Govindarajan
  • Adam E Sienkiewicz
  • Manik Goel
  • Richard K Lee
  • Robert G Salomon
  • Adriana Amelinckx
  • James Laird
  • Teresia A Carreon
  • Genea Edwards
  • Brandon N Rosenberg
  • Jianhua Wang
  • Mabel Algeciras
  • Jean Marie A Parel
  • Eleut Hernandez
  • TinThu Lee
  • Nelson Salas
  • Esdras Arrieta-Quintero
  • Maria Castello
  • Ronald Sherman
  • Tiziana Cogliati
  • Jasenka Guduric-Fuchs
  • Cynthia C Morton
  • Kavita Desai

Detail Information

Publications20

  1. pmc Aberrant glycosylation in the human trabecular meshwork
    Adam E Sienkiewicz
    Bascom Palmer Eye Institute, University of Miami, Miami, FL, USA
    Proteomics Clin Appl 8:130-42. 2014
    ..To determine the difference in protein glycosylation and glycosylation enzyme levels between glaucomatous and control trabecular meshwork (TM)...
  2. pmc COCH transgene expression in cultured human trabecular meshwork cells and its effect on outflow facility in monkey organ cultured anterior segments
    Eun Suk Lee
    Departments of Ophthalmology and Visual Sciences, Sodaemungu Shinchondong, Seoul 120 752, Korea
    Invest Ophthalmol Vis Sci 51:2060-6. 2010
    ..Cochlin expression was effective in decreasing OF and increasing ASP in MOCAS, suggesting possible involvement of cochlin in IOP elevation in vivo. COCH gene delivery has potential for use in developing a glaucoma model...
  3. pmc Decreased carbohydrate metabolism enzyme activities in the glaucomatous trabecular meshwork
    Anna K Junk
    Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA
    Mol Vis 16:1286-91. 2010
    ..To determine whether activity of carbohydrate metabolism enzymes (aldolase, pyruvate kinase, isocitrate dehydrogenase, and malate dehydrogenase) are altered in the glaucomatous trabecular meshwork (TM) compared to controls...
  4. pmc Cochlin induced TREK-1 co-expression and annexin A2 secretion: role in trabecular meshwork cell elongation and motility
    Manik Goel
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida, United States of America
    PLoS ONE 6:e23070. 2011
    ..Annexin A2 expression and secretion follows cochlin-TREK-1 syn-expression and correlates with cell elongation. Thus cytoskeleton changes in response to fluid shear sensed by cochlin are further mediated by TREK-1 and annexin A2...
  5. pmc Cochlin, intraocular pressure regulation and mechanosensing
    Manik Goel
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida, United States of America
    PLoS ONE 7:e34309. 2012
    ..Taken together with previous studies, we show cochlin is involved in regulation of intraocular pressure in DBA/2J potentially through mechanosensing of the shear stress...
  6. pmc Increased isolevuglandin-modified proteins in glaucomatous astrocytes
    Bharathi Govindarajan
    Department of Chemistry, Case Western Reserve University, Cleveland, OH, USA
    Mol Vis 15:1079-91. 2009
    ..In addition, we examined whether the iso[4]LGE(2) protein adducts are altered following exposure of astrocytes to elevated pressure...
  7. pmc Laser trabeculoplasty induces changes in the trabecular meshwork glycoproteome: a pilot study
    Adriana Amelinckx
    Bascom Palmer Eye Institute, Ophthalmic Biophysics Center, University of Miami Miller School of Medicine, Miami, Forida 33136, USA
    J Proteome Res 8:3727-36. 2009
    ..Further investigation is anticipated to provide insight into how glycosylation changes affect TM proteins and TM regulation of aqueous outflow in response to laser trabeculoplasty...
  8. pmc Cochlin expression in anterior segment organ culture models after TGFbeta2 treatment
    Sanjoy K Bhattacharya
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida 33136, USA
    Invest Ophthalmol Vis Sci 50:551-9. 2009
    ..To determine the effect of transforming growth factor (TGF)-beta2 treatment on intraocular pressure (IOP), outflow facility, and cochlin expression in vitro in monkey and pig organ-cultured anterior segments (MOCAS and POCAS)...
  9. pmc Neuroprotection in glaucoma using calpain-1 inhibitors: regional differences in calpain-1 activity in the trabecular meshwork, optic nerve and implications for therapeutics
    Bharathi Govindarajan
    Bascom Palmer Eye Institute, University of Miami, Miami, FL 33136, USA
    CNS Neurol Disord Drug Targets 7:295-304. 2008
    ..Therapeutic neuroprotection strategies using calpain-1 inhibitors will require consideration of such anatomic differences in its activity and biosynthesis...
  10. pmc Technical Brief: a novel strategy for enrichment of trabecular meshwork protease proteome
    Renata Picciani
    Bascom Palmer Eye Institute, University of Miami, Miami, FL 33136, USA
    Mol Vis 14:871-7. 2008
    ..Both serine and cysteine proteases were captured using this strategy with improved coverage compared to our previous identification without affinity enrichment...
  11. ncbi Cochlin deposits in the trabecular meshwork of the glaucomatous DBA/2J mouse
    Sanjoy K Bhattacharya
    Exp Eye Res 80:741-4. 2005
    ..Studies of DBA/2J mice offer promise for understanding the role cochlin may play in glaucoma...
  12. pmc Focus on molecules: cochlin
    Sanjoy K Bhattacharya
    Evelyn F and William L McKnight Vision Research Center, Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, FL 33136, USA
    Exp Eye Res 82:355-6. 2006
  13. pmc Cochlin and glaucoma: a mini-review
    Sanjoy K Bhattacharya
    Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, OH 44195, USA
    Vis Neurosci 22:605-13. 2005
    ..Here we review these findings and describe how future studies in DBA/2J mice can help resolve whether cochlin plays a causal role in mechanisms of POAG and elevated IOP...
  14. ncbi Proteomics implicates peptidyl arginine deiminase 2 and optic nerve citrullination in glaucoma pathogenesis
    Sanjoy K Bhattacharya
    Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, Ohio, USA
    Invest Ophthalmol Vis Sci 47:2508-14. 2006
    ..Peptidyl arginine deiminase 2 (PAD2), an enzyme that converts protein arginine to citrulline, was found only in POAG optic nerve and was probed further for a mechanistic role in glaucoma...
  15. ncbi Molecular basis for the redox control of nuclear transport of the structural chromatin protein Hmgb1
    George Hoppe
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA
    Exp Cell Res 312:3526-38. 2006
    ..Our cysteine targeted mutational analysis suggests that Cys23 and 45 induce conformational changes in response to oxidative stress, whereas Cys106 appears to be critical for the nucleocytoplasmic shuttling of Hmgb1...
  16. ncbi Modulation of peptidyl arginine deiminase 2 and implication for neurodegeneration
    Sanjoy K Bhattacharya
    Bascom Palmer Eye Institute, University of Miami, Miami, Florida, USA
    Curr Eye Res 31:1063-71. 2006
    ..To demonstrate that elevated pressure increases the peptidyl arginine deiminase 2 (PAD2) expression in cultured astrocytes in vitro that can be modulated by pharmacological agents modulating intracellular calcium...
  17. pmc Cochlin in the eye: functional implications
    Renata Picciani
    Bascom Palmer Eye Institute, University of Miami, Miami, FL 33136, USA
    Prog Retin Eye Res 26:453-69. 2007
    ..In vitro, cochlin induces aggregation of primary TM cells suggesting a role in cell adhesion, possibly in mechanosensation, and in modulation of the TM filter...
  18. pmc Isolevuglandin-modified proteins, including elevated levels of inactive calpain-1, accumulate in glaucomatous trabecular meshwork
    Bharathi Govindarajan
    Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, USA
    Biochemistry 47:817-25. 2008
    ..These data suggest that interference with proteasomal activity, owing to protein modification by isoLGs, could contribute to glaucoma pathophysiology by decreasing the ability of the TM to modulate outflow resistance...
  19. pmc Proteomics reveal Cochlin deposits associated with glaucomatous trabecular meshwork
    Sanjoy K Bhattacharya
    Cole Eye Institute and Lerner Research Institute i31, Cleveland Clinic Foundation, Cleveland, OH 44195, USA
    J Biol Chem 280:6080-4. 2005
    ....