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Cited by 53Retinal Diseases and TreatmentsTelomeres, Telomerase, and SenescenceRetinal Imaging and Analysis

Senescence Increases Choroidal Endothelial Stiffness and Susceptibility to Complement Injury: Implications for Choriocapillaris Loss in AMD

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Cabrera, Andrea P., Bhaskaran, Arun, Xu, Jun, Yang, Xiao, Scott, Harry A., Mohideen, Umar, Ghosh, Kaustabh·2016·Investigative Opthalmology & Visual Science
DOI 10.1167/iovs.16-19727
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Abstract

PURPOSE: Age-related macular degeneration (AMD) commonly causes blindness in the elderly. Yet, it is untreatable in the large fraction of all AMD patients that develop the early dry form. Dry AMD is marked by the deposition of membrane attack complex (MAC) on choriocapillaris (CC), which is implicated in CC degeneration and subsequent atrophy of overlying retinal pigment epithelium. Since MAC is also found on the CC of young eyes, here we investigated whether and how aging increases choroidal endothelial susceptibility to MAC injury. METHODS: Monkey chorioretinal endothelial cells (ECs, RF/6A) were cultured to high passages (>P60) to achieve replicative senescence. We treated ECs with complement-competent human serum to promote MAC deposition and injury, which were assessed by flow cytometry and trypan blue exclusion assay, respectively. Stiffness of EC was measured by atomic force microscopy indentation while Rho GTPase activity was quantified by Rho G-LISA assay. RESULTS: Our findings reveal that senescent ECs are significantly stiffer than their normal counterparts, which correlates with higher cytoskeletal Rho activity in these cells and their greater susceptibility to complement (MAC) injury. Importantly, inhibition of Rho activity in senescent ECs significantly reduced cell stiffness and MAC-induced lysis. CONCLUSIONS: By revealing an important role of senescence-associated choroidal EC stiffening in complement injury, these findings implicate CC stiffening as an important determinant of age-related CC atrophy seen in dry AMD. Future studies are needed to validate these findings in appropriate animal models so new therapeutic targets can be identified for treatment of dry AMD.

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  1. [1]1. Cook HL, Patel PJ, Tufail A. Age-related macular degeneration: diagnosis and management. Br Med Bull . 2008;85:127149.
  2. [2]2. Friedman DS, O'Colmain BJ, Munoz B, et al. Prevalence of agerelated macular degeneration in the United States. Arch Ophthalmol . 2004;122:564-572.
  3. [3]3. Ambati J, Fowler BJ. Mechanisms of age-related macular degeneration. Neuron . 2012;75:26-39.
  4. [4]4. Zhang K, Zhang L, Weinreb RN. Ophthalmic drug discovery: novel targets and mechanisms for retinal diseases and glaucoma. Nat Rev Drug Discov . 2012;11:541-559.
  5. [5]5. Whitmore SS, Sohn EH, Chirco KR, et al. Complement activation and choriocapillaris loss in early AMD: implications for pathophysiology and therapy. Prog Retin Eye Res . 2015;45: 1-29.
  6. [6]6. Mullins RF, Johnson MN, Faidley EA, Skeie JM, Huang J. Choriocapillaris vascular dropout related to density of drusen in human eyes with early age-related macular degeneration. Invest Ophthalmol Vis Sci . 2011;52:1606-1612.
  7. [7]7. McLeod DS, Grebe R, Bhutto I, Merges C, Baba T, Lutty GA. Relationship between RPE and choriocapillaris in age-related macular degeneration. Invest Ophthalmol Vis Sci . 2009;50: 4982-4991.
  8. [8]8. McLeod DS, Taomoto M, Otsuji T, Green WR, Sunness JS, Lutty GA. Quantifying changes in RPE and choroidal vasculature in eyes with age-related macular degeneration. Invest Ophthalmol Vis Sci . 2002;43:1986-1993.
  9. [9]9. Parmeggiani F , Romano MR, Costagliola C, et al. Mechanism of inflammation in age-related macular degeneration. Mediators Inflamm . 2012;2012:546786.
  10. [10]10. Anderson DH, Mullins RF , Hageman GS, Johnson LV . A role for local inflammation in the formation of drusen in the aging eye. Am J Ophthalmol . 2002;134:411-431.
  11. [11]11. Mullins RF , Schoo DP , Sohn EH, et al. The membrane attack complex in aging human choriocapillaris: relationship to macular degeneration and choroidal thinning. Am J Pathol . 2014;184:3142-3153.
  12. [12]12. Friedman E, Ivry M, Ebert E, Glynn R, Gragoudas E, Seddon J. Increased scleral rigidity and age-related macular degeneration. Ophthalmology . 1989;96:104-108.
  13. [13]13. Kotliar KE, Baumann M, Vilser W, Lanzl IM. Pulse wave velocity in retinal arteries of healthy volunteers. Br J Ophthalmol . 2011;95:675-679.
  14. [14]14. Wang M, Monticone RE, Lakatta EG. Proinflammation of aging central arteries: a mini-review. Gerontology . 2014;60:519-529.
  15. [16]16. Kothapalli D, Liu SL, Bae YH, et al. Cardiovascular protection by ApoE and ApoE-HDL linked to suppression of ECM gene expression and arterial stiffening. Cell Rep . 2012;2:1259-1271.
  16. [17]17. Ghosh K, Thodeti CK, Dudley AC, Mammoto A, Klagsbrun M, Ingber DE. Tumor-derived endothelial cells exhibit aberrant Rho-mediated mechanosensing and abnormal angiogenesis in vitro. Proc Natl Acad Sci U S A . 2008;105:11305-11310.
  17. [18]18. Mammoto A, Mammoto T, Kanapathipillai M, et al. Control of lung vascular permeability and endotoxin-induced pulmonary oedema by changes in extracellular matrix mechanics. Nat Commun . 2013;4:1759.
  18. [19]19. Mammoto A, Mammoto T, Ingber DE. Rho signaling and mechanical control of vascular development. Curr Opin Hematol . 2008;15:228-234.
  19. [20]20. Huynh J, Nishimura N, Rana K, et al. Age-related intimal stiffening enhances endothelial permeability and leukocyte transmigration. Sci Transl Med . 2011;3:112ra122.
  20. [21]21. Yang X, Scott HA, Ardekani S, Williams M, Talbot P , Ghosh K. Aberrant cell and basement membrane architecture contribute to sidestream smoke-induced choroidal endothelial dysfunction. Invest Ophthalmol Vis Sci . 2014;55:3140-3147.
  21. [22]22. Yang X, Scott HA, Monickaraj F , et al. Basement membrane stiffening promotes retinal endothelial activation associated with diabetes. FASEB J . 2016;30:601-611.
  22. [23]23. Scott HA, Quach B, Yang X, et al. Matrix stiffness exerts biphasic control over monocyte-endothelial adhesion via Rhomediated ICAM-1 clustering. Integr Biol (Camb) . 2016;8:869878.
  23. [24]24. Debacq-Chainiaux F , Erusalimsky JD, Campisi J, Toussaint O. Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc . 2009;4:1798-1806.
  24. [25]25. Campisi J, Robert L. Cell senescence: role in aging and agerelated diseases. Interdiscip Top Gerontol . 2014;39:45-61.
  25. [27]27. Campisi J, d'Adda di Fagagna F . Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol . 2007;8: 729-740.
  26. [28]28. Johnson LV , Forest DL, Banna CD, et al. Cell culture model that mimics drusen formation and triggers complement activation associated with age-related macular degeneration. Proc Natl Acad Sci U S A . 2011;108:18277-18282.
  27. [29]29. Gorham RD Jr, Forest DL, Tamamis P , et al. Novel compstatin family peptides inhibit complement activation by drusen-like deposits in human retinal pigmented epithelial cell cultures. Exp Eye Res . 2013;116:96-108.
  28. [30]30. Chen CS, Mrksich M, Huang S, Whitesides GM, Ingber DE. Geometric control of cell life and death. Science . 1997;276: 1425-1428.
  29. [31]31. Ghosh K, Ingber DE. Micromechanical control of cell and tissue development: implications for tissue engineering. Adv Drug Deliv Rev . 2007;59:1306-1318.
  30. [32]32. Kovacic JC, Moreno P , Nabel EG, Hachinski V , Fuster V . Cellular senescence vascular disease, and aging: part 2 of a 2-part review: clinical vascular disease in the elderly. Circulation . 2011;123:1900-1910.
  31. [33]33. Kovacic JC, Moreno P , Hachinski V , Nabel EG, Fuster V . Cellular senescence, vascular disease, and aging: part 1 of a 2-part review. Circulation . 2011;123:1650-1660.
  32. [34]34. Huveneers S, Daemen MJ, Hordijk PL. Between Rho(k) and a hard place: the relation between vessel wall stiffness endothelial contractility, and cardiovascular disease. Circ Res . 2015;116:895-908.
  33. [35]35. Cuerrier CM, Gagner A, Lebel R, Gobeil F Jr, Grandbois M. Effect of thrombin and bradykinin on endothelial cell mechanical properties monitored through membrane deformation. J Mol Recognit . 2009;22:389-396.
  34. [36]36. Birukova AA, Smurova K, Birukov KG, Kaibuchi K, Garcia JG, Verin AD. Role of Rho GTPases in thrombin-induced lung vascular endothelial cells barrier dysfunction. Microvasc Res . 2004;67:64-77.
  35. [37]37. Birukova AA, Tian X, Cokic I, Beckham Y, Gardel ML, Birukov KG. Endothelial barrier disruption and recovery is controlled by substrate stiffness. Microvasc Res . 2013;87:50-57.
  36. [38]38. Krisinger MJ, Goebeler V, Lu Z, et al. Thrombin generates previously unidentified C5 products that support the terminal complement activation pathway. Blood . 2012;120:17171725.
  37. [39]39. Sennlaub F , Auvynet C, Calippe B, et al. CCR2( þ ) monocytes infiltrate atrophic lesions in age-related macular disease and mediate photoreceptor degeneration in experimental subretinal inflammation in Cx3cr1 deficient mice. EMBO Mol Med . 2013;5:1775-1793.
  38. [40]40. Skeie JM, Mullins RF . Macrophages in neovascular age-related macular degeneration: friends or foes? Eye (Lond) . 2009;23: 747-755.
  39. [42]42. Lou D, Hu F. Co-distribution of von Willebrand factor and fibronectin in cultured rhesus endothelial cells. Histochem J . 1987;19:431-438.
  40. [43]43. Ottino P, Finley J, Rojo E, et al. Hypoxia activates matrix metalloproteinase expression and the VEGF system in monkey choroid-retinal endothelial cells: involvement of cytosolic phospholipase A2 activity. Mol Vis . 2004;10:341-350.
  41. [44]44. Amrite AC, Kompella UB. Celecoxib inhibits proliferation of retinal pigment epithelial and choroid-retinal endothelial cells by a cyclooxygenase-2-independent mechanism. J Pharmacol Exp Ther . 2008;324:749-758.
  42. [45]45. Kim NW, Piatyszek MA, Prowse KR, et al. Specific association of human telomerase activity with immortal cells and cancer. Science . 1994;266:2011-2015.
  43. [46]46. Camous L, Roumenina L, Bigot S, et al. Complement alternative pathway acts as a positive feedback amplification of neutrophil activation. Blood . 2011;117:1340-1349.
  44. [47]47. Rutar M, Valter K, Natoli R, Provis JM. Synthesis and propagation of complement C3 by microglia/monocytes in the aging retina. PLoS One . 2014;9:e93343.
  45. [48]48. Luo C, Zhao J, Madden A, Chen M, Xu H. Complement expression in retinal pigment epithelial cells is modulated by activated macrophages. Exp Eye Res . 2013;112:93-101.
  46. [49]49. Cheung TM, Yan JB, Fu JJ, Huang J, Yuan F , Truskey GA. Endothelial cell senescence increases traction forces due to age-associated changes in the glycocalyx and SIRT1. Cell Mol Bioeng . 2014;8:63-75.
  47. [50]50. Lehenkari PP , Charras GT, Nykanen A, Horton MA. Adapting atomic force microscopy for cell biology. Ultramicroscopy . 2000;82:289-295.
  48. [51]51. Adams WJ, O'Grady ML, Ghosh K, Gibbs AD, Geisse NA, Ingber DE, Parker KK. Viscoelastic indentation of extremely soft biological samples. Biophysical J . 2009;96:398a.
  49. [54]54. Francis PJ, Appukuttan B, Simmons E, et al. Rhesus monkeys and humans share common susceptibility genes for age-related macular disease. Hum Mol Genet . 2008;17:2673-2680.
  50. [55]55. Acosta J, Hettinga J, Fluckiger R, et al. Molecular basis for a link between complement and the vascular complications of diabetes. Proc Natl Acad Sci U S A . 2000;97:5450-5455.
  51. [56]56. Lee MS, Jones T, Song DY, Jang JH, Jung JU, Gao SJ. Exploitation of the complement system by oncogenic Kaposi's sarcoma-associated herpesvirus for cell survival and persistent infection. PLoS Pathog . 2014;10:e1004412.
  52. [58]58. Barker FM II, Snodderly DM, Johnson EJ, et al. Nutritional manipulation of primate retinas, V: effects of lutein, zeaxanthin, and n-3 fatty acids on retinal sensitivity to blue-lightinduced damage. Invest Ophthalmol Vis Sci . 2011;52:39343942.

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Authors
Cabrera, A. P., Bhaskaran, A., Xu, J., Yang, X., Scott, H. A., Mohideen, U., & Ghosh, K.
Affiliations
University of California, Riverside (US)
Corresponding author
Ghosh, Kaustabh
ORCID
Venue
Investigative Opthalmology & Visual Science
Publisher
Association for Research in Vision and Ophthalmology (ARVO)
Published
2016-11-01
Volume
57
Issue
14
Pages
5910
ISSN
  • 0146-0404 (ISSN-L)
  • 1552-5783
DOI
10.1167/iovs.16-19727
OpenAlex
W2547543108
Cited by
53
References
4
Citations by year
2026: 2 · 2025: 2 · 2024: 10 · 2023: 5 · 2022: 7 · 2021: 4 · 2020: 6 · 2019: 7 · 2018: 7 · 2017: 3
FWCI
6.02
Type
journal article
Language
en
Open access
Licence
Locations
  • Investigative Ophthalmology & Visual Science (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, public-domain)
  • eScholarship (California Digital Library) (submittedVersion)
Topics
  • Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Retinal Imaging and Analysis
Keywords
age-related macular degeneration, dry age-related macular degeneration, choroidal endothelial cells, replicative senescence, cell stiffness, membrane attack complex, Rho GTPases, cytoskeleton, endothelial cell lysis, aging
MeSH terms
Aged, Animals, Haplorhini, Cells, Cultured, Choroid, Complement System Proteins, Humans, Macular Degeneration, Male, Retina, Complement Membrane Attack Complex, Cellular Senescence, Microscopy, Atomic Force, Endothelial Cells

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