Shifting expression of membrane Vascular Endothelial Growth Factor Receptor-2 (mVEGFR2) to soluble Vascular Endothelial Growth Factor Receptor-2 (sVEGFR2) reduces CNV volume and tumor growth

CNV Model: • CNV was induced in C57 mice using laser photocoagulation. • On day 1 and day 4 after photocoagulation, either control morpholino, sVEGFR2-inducing morpholino, or DBPS was injected intravitreously. • On day 7, CNV volumes were measured by confocal microscopy. Xenograft Tumor Model: • NMRI nu/nu mice received subcutaneous injections of HCT 116 colon cancer cells. • One week after the injection of cells, treatment began with control morpholino, sVEGFR2inducing morpholino, or HBSS. • Tumors were injected with treatment twice weekly for a period of three weeks. • Tumor size was measured twice weekly. • Fluorescent staining was performed on tumor sections. Isolectin conjugated Alexa488 (Invitrogen, # #I21411) and anti-LYVE-1 antibody (Abcam, #14917) were used for blood vessel and lymph vessel staining, respectively. Purpose • Angiogenesis plays a key role in both agerelated macular degeneration (AMD) and cancer growth. • VEGFR2 is present in both membrane-bound (mVEGFR2) and soluble (sVEGFR2) forms due to alternative splicing. • mVEGFR2 is pro-angiogenic, while sVEGFR2 is anti-angiogenic. • We have developed a morpholino-based gene therapy technique that shifts expression from mVEGFR2 to sVEGFR2. • We present the success of this technique in treating a murine model of choroidal neovascularization (CNV) and a xenograft tumor model. Shifting expression of membrane Vascular Endothelial Growth Factor Receptor-2 (mVEGFR2) to soluble Vascular Endothelial Growth Factor Receptor-2 (sVEGFR2) reduces CNV volume and tumor growth. Brian C. Stagg, Hironori Uehara, Bryce Radmall, Taylor Bates, Christina Mamalis, Leah Owen, Yang Kyung Cho, Tadashi Miya, Balamurali K. Ambati University of Utah School of Medicine, Salt Lake City, UT John A. Moran Eye Center, Salt Lake City, UT, St. Vincent Hospital, Catholic University or Korea, Seoul, Republic of Korea