Retinal angiogenesis suppression through small molecule activation of p53
Sai H. Chavala, Younghee Kim, Laura Tudisco, Valeria Cicatiello, Till Milde, Nagaraj Kerur, Nidia Claros, Susan Yanni, Victor H. Guaiquil, William W. Hauswirth, John S. Penn, Shahin Rafii, Sandro De Falco, Thomas C. Lee, Jayakrishna Ambati
Sai H. Chavala, Younghee Kim, Laura Tudisco, Valeria Cicatiello, Till Milde, Nagaraj Kerur, Nidia Claros, Susan Yanni, Victor H. Guaiquil, William W. Hauswirth, John S. Penn, Shahin Rafii, Sandro De Falco, Thomas C. Lee, Jayakrishna Ambati
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Research Article Vascular biology

Retinal angiogenesis suppression through small molecule activation of p53

Abstract

Neovascular age-related macular degeneration is a leading cause of irreversible vision loss in the Western world. Cytokine-targeted therapies (such as anti-vascular endothelial growth factor) are effective in treating pathologic ocular angiogenesis, but have not led to a durable effect and often require indefinite treatment. Here, we show that Nutlin-3, a small molecule antagonist of the E3 ubiquitin protein ligase MDM2, inhibited angiogenesis in several model systems. We found that a functional p53 pathway was essential for Nutlin-3–mediated retinal antiangiogenesis and disruption of the p53 transcriptional network abolished the antiangiogenic activity of Nutlin-3. Nutlin-3 did not inhibit established, mature blood vessels in the adult mouse retina, suggesting that only proliferating retinal vessels are sensitive to Nutlin-3. Furthermore, Nutlin-3 inhibited angiogenesis in nonretinal models such as the hind limb ischemia model. Our work demonstrates that Nutlin-3 functions through an antiproliferative pathway with conceivable advantages over existing cytokine-targeted antiangiogenesis therapies.

Authors

Sai H. Chavala, Younghee Kim, Laura Tudisco, Valeria Cicatiello, Till Milde, Nagaraj Kerur, Nidia Claros, Susan Yanni, Victor H. Guaiquil, William W. Hauswirth, John S. Penn, Shahin Rafii, Sandro De Falco, Thomas C. Lee, Jayakrishna Ambati

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Figure 5

p53 is necessary for Nutlin-3–mediated cell death.

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p53 is necessary for Nutlin-3–mediated cell death. (A and B) HUVECs were...
(A and B) HUVECs were infected with a lentiviral vector expressing p53 or control shRNA for 48 hours and then underwent puromycin selection for 72 hours. (A) Cell lysates for Western blot analysis were used to confirm knockdown of p53 in shRNA-infected HUVECs. (B) p53 shRNA– and control shRNA–infected HUVECs were seeded at 1 × 106 cells in 6-well plates and incubated with FGF-2 and either Nutlin-3A (7.5 μM) or vehicle (DMSO) for 48 hours. Cell proliferation was measured at 48 hours by manual counting using trypan blue exclusion. Data represent mean ± SD. *P < 0.05.