March 2012
Volume 53, Issue 14
ARVO Annual Meeting Abstract  |   March 2012
Arresten Regulates Choroidal Angiogenesis By Fasl, Caspase-3 And Parp Mediated Apoptosis In-vitro And In-vivo
Author Affiliations & Notes
  • Raj K. Verma
    Cell Signalling Retinal angiogenesis, Boys Town National Research Hospital, Omaha, Nebraska
  • Venugopal Gunda
    Genetics, Boys Town Nt'l Research Hosp, Omaha, Nebraska
  • Chandra S. Boosani
    Genetics, Boys Town Nt'l Research Hosp, Omaha, Nebraska
  • Sudhakar A. Yakkanti
    Genetics/ Retinal Cell Signaling, Boys Town Natl Res Hospital, Omaha, Nebraska
  • Footnotes
    Commercial Relationships  Raj K. Verma, None; Venugopal Gunda, None; Chandra S. Boosani, None; Sudhakar A. Yakkanti, None
  • Footnotes
    Support  5R01CA143128-02
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 2949. doi:
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      Raj K. Verma, Venugopal Gunda, Chandra S. Boosani, Sudhakar A. Yakkanti; Arresten Regulates Choroidal Angiogenesis By Fasl, Caspase-3 And Parp Mediated Apoptosis In-vitro And In-vivo. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2949.

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

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Purpose: : Type IV collagen plays a crucial role in regulation of angiogenesis. Proteolytic degradation of type IV collagen in the vascular basement membrane (VBM) generates antiangiogenic molecules. We previously reported that the antiangiogenic non-collagenous (NC1) domain derived from α1 chain of type IV collagen, α1(IV)NC1 (arresten) induces apoptosis in endothelial cells. However, active site of arresten responsible for its antiangiogenic properties is not yet identified. In the present study, the N- and C-terminal regions of arresten were cloned as two individual subunits and studied their anti-angiogenic and pro-apoptotic properties.

Methods: : Both the N- and C-terminal subunits of arresten were cloned in the pET22b (+) expression vector and expressed in Escherichia coli. Expressed subunits were purified using affinity and size exclusion chromatographies with simultaneous renaturation.

Results: : Our preliminary in-vitro results with mouse choroidal endothelial cells (MCECs) have shown inhibition of VEGF and bFGF induced endothelial cells proliferation, migration and tube formation by both the subunits. In addition, arresten promoted FasL mediated apoptosis through caspase-3 and PARP both in-vitro and in-vivo.

Conclusions: : Both N- and C-terminal subunits of arresten exhibited antiangiogenic activity in MCECs by showing inhibitory effects on VEGF and bFGF induced proliferation, migration and tube formation. The FasL mediated apoptosis activation by arresten is also identified.

Keywords: apoptosis/cell death • choroid: neovascularization • extracellular matrix 

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