1. The authors have no financial interest in this presentation Iladevi Cataract and IOL Research Centre, Ahmedabad, India Shail A. Vasavada, DO Devarshi Gajjar, PhD Kaid Johar SR, PhD Abhay Vasavada, MS, FRCS (England)

2. Failure in regression of hyaloid vasculature results in a pathological condition called Persistant Fetal Vasculature (PFV)  Arresten: α 1 chain COLIV  Canstatin: α 2 chain COLIV  Tumstatin: α 3 chain COLIV  Endostatin: α 1 chain COLXVIII 1Sottile J. Biochem BiophysActa. 20044;1654(1):13-22. 2Mundel TM, Kalluri R. Microvasc Res. 2007;74(2-3):85-9. Is by Proteolytic cleavage of collagen in the lens capsule Stimulated by release of growth factors from ECM Inhibited by release of Antiangiogenic factors (AAF) from ECM Matrix Metalloproteases(MMPs) regulate the proteolytic cleavage of ECM Is by Proteolytic cleavage of collagen in the lens capsule Stimulated by release of growth factors from ECM Inhibited by release of Antiangiogenic factors (AAF) from ECM Matrix Metalloproteases(MMPs) regulate the proteolytic cleavage of ECM Regulation of Angiogenesis Collagen Derived AAFs Embryonic Stage : Lens covered by tunica vasculosa lentis However the exact mechanism of angiogenesis and regression is not completely known

3. Study Design Case-Control Study Cases – Pediatric eyes with anterior PFV (n=6) Controls – Age-matched Congenital cataracts (n=6) Purpose To measure mRNA levels of Antiangiogenic factors (arresten, canstatin, tumstatin, endostatin) and Matrix Metalloproteinases (MMP-2 and MMP-9) in lens epithelial cells(LECs) of eyes with and without PFV

4. * All the anterior capsules collected from pediatric eyes during the surgery were in accordance with Declaration of Helsinki. Ethical approval for the study and use of human LECs was obtained from the Institutional Ethics Committee Material and Methods Representative PFV Cases: membranous cataract, traction on ciliary processes, fibrovascular plaque

5. Amplification curves mRNA levels were expressed as a ratio, using the delta-delta method for comparing the relative expression results between cases and controls Non-parametric Mann-Whitney U test performed Melting peaks Melting curves Amplification curves Standardization for qRT-PCR Quantitative real time RT-PCR performed to determine the mRNA levels of arresten, canstatin, tumstatin, endostatin, MMP-2, MMP-9, GAPDH and Beta actin (Housekeeping genes)

6. * *P=0.01, # P=0.02, Relative mRNA expression of AAFs and MMPs in PFV cases and Controls In eyes with PFV...  The mRNA level of MMP-2 was 4 times higher in PFV cases  No change was observed in MMP-9 mRNA levels (p = 0.82) Results AAFs Arresten 1.6 times Tumstatin 2.5 times Canstatin 2.3 times # *

7. Literature Review Apoptotic and necrotic cell death of endothelial cells and pericytes of the blood vessels play an important role in the regression of hyaloid vasculature 3 Potential role for endostatin in fetal vascular remodeling of the eye was shown in a patient with Knobloch syndrome and PFV 4 VEGF, FGF, PEGF, Factors implicated in developmental and pathological changes in vessel formation and regression during development 5 Mouse models for PFV have implicated the role of proteins, Angiopoietin-2(Ang-2) 6, P53 7, Arf 8, VEGF-A 188 9, NUC1 10 3 Zhu M et al., Exp Eye Res. 2000;70(6):767-76 4 Duh EJ et al., Ophthalmology. 2004; 111:1885-8 5 Saint-Geniez M and D’Amore PA Int J Dev Biol 2004; 48:1045–1058 6 Hackett SF et al., J Cell Physiol. 2002;192:182–187. 7 Reichel MB et al., Cell Death Differ. 1998;5:156 –162. 8 Thornton JD et al., Invest Ophthalmol Vis Sci. 2007 ;48:491-9 9 Rutland CS et al., Mol Vis.2007;19:47-56. 10 Gehlbach P et al., Neuroscience. 2006;137(2):447-61. Speculation In the present study we speculate that increased level of canstatin and tumstatin could be to cope up with increased angiogenesis in the pathological condition of PFV Decreased levels of Arresten could be because of the developmental shift in collagen IV of the lens capsule Speculation In the present study we speculate that increased level of canstatin and tumstatin could be to cope up with increased angiogenesis in the pathological condition of PFV Decreased levels of Arresten could be because of the developmental shift in collagen IV of the lens capsule

8. -The levels of Anti-Angiogenic Factors are altered in eyes with PHPV -AAFs appear to play a pivotal role in maintaining the lens avascularity, which gets altered during the pathogenesis of PFV First study on human PHPV eyes Future Direction Further studies on the protein levels, growth factors and other inhibitors Conclusion