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)

Failure in regression of hyaloid vasculature results in a

pathological condition called Persistant Fetal Vasculature

(PFV)

Arresten:α1 chain COLIV

Canstatin:α2 chain COLIV

Tumstatin: α3 chain COLIV

Endostatin: α1 chain COLXVIII

1Sottile J. Biochem BiophysActa. 20044;1654(1):13-22.

2Mundel TM, Kalluri R. Microvasc Res. 2007;74(2-3):85-9.

Arresten:α1 chain COLIV

Canstatin:α2 chain COLIV

Tumstatin: α3 chain COLIV

Endostatin: α1 chain COLXVIII

1Sottile J. Biochem BiophysActa. 20044;1654(1):13-22.

2Mundel TM, Kalluri R. Microvasc Res. 2007;74(2-3):85-9.

Regulation of Angiogenesis Collagen Derived Collagen Derived AAFsAAFs

Embryonic Stage : Lens covered by tunica vasculosa

lentis

However the exact mechanism of angiogenesis and regression is

not completely known

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

* 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

Amplification curves

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)

*

*P=0.01, # P=0.02,

Relative mRNA expression of AAFs and MMPs in PFV cases and ControlsIn 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

#

*

3Zhu M et al., Exp Eye Res. 2000;70(6):767-76 4Duh EJ et al., Ophthalmology. 2004; 111:1885-85Saint-Geniez M and D’Amore PA Int J Dev Biol 2004; 48:1045–1058 6Hackett SF et al., J Cell Physiol. 2002;192:182–187.7Reichel MB et al., Cell Death Differ. 1998;5:156 –162. 8Thornton JD et al., Invest Ophthalmol Vis Sci. 2007 ;48:491-99Rutland CS et al., Mol Vis.2007;19:47-56. 10Gehlbach 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

First study on human PHPV eyes

Future Direction Further studies on the protein levels, growth

factors and other inhibitors

Conclusion