CLT-009

CLT-009 is a peptide that is a signalling receptor antagonist.  CLT-009 prevents the induction of signalling pathways in the retina and RPE that lead to angiogenesis.

 

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Inhibition of VEGF expression by signalling blockade with CLT-009

Inhibition of the receptor-signaling and VEGF expression by CLT-009.  Vldlr-/- mice received a subretinal injection of 5 µg purified mouse CLT-009 into the right eyes and the same amount BSA into the left eyes.  The eyecups were dissected 24 h after the injection for the following analyses. A, phosphorylated -marker P1 (a marker for signaling blockade) and VEGF levels were measured by Western blot analysis of a representative mouse eyecup using respective antibodies.  B, VEGF protein levels in Vldlr-/- eyecups were measured using ELISA.  C, VEGF mRNA levels in the Vldlr-/- eyecups were measured using quantitative real-time RT-PCR.  Values are the mean ± S.D. (n = 4).  D-G, Vldlr-/- mice were injected with 5 µg/eye BSA into the subretinal space of the left eye (D and F) and the same amount of CLT-009 into the right eye (E and G).  The ocular sections were immunostained with an anti-VEGF antibody (red) (D and E) and an anti-marker P1 (green) (F and G) and counter-stained with 4',6-diamidino-2-phenylindole (blue), 400x. PRL, photoreceptor layer. H and I, HUVEC were transfected with the VLDLR siRNA. The transfected cells were treated with 10 µg/ml human CLT-009 or BSA.  VEGF protein was measured in the culture medium by ELISA (H), and the VEGF mRNA was measured in the cells by real-time RT-PCR, normalized by 18 S RNA, and expressed as arbitrary unit (mean ± S.D., n = 3) (I) 24 h after the addition of CLT-009.