TY - JOUR
T1 - Novel anti-angiogenic PEDF-derived small peptides mitigate choroidal neovascularization
AU - Sheibani, Nader
AU - Wang, Shoujian
AU - Darjatmoko, Soesiawati R.
AU - Fisk, Debra L.
AU - Shahi, Pawan K.
AU - Pattnaik, Bikash R.
AU - Sorenson, Christine M.
AU - Bhowmick, Reshma
AU - Volpert, Olga V.
AU - Albert, Daniel M.
AU - Melgar-Asensio, Ignacio
AU - Henkin, Jack
N1 - Funding Information:
This work was supported by an unrestricted award from Research to Prevent Blindness to the Department of Ophthalmology and Visual Sciences, Retina Research Foundation , P30 EY016665 , P30 CA014520 , EPA 83573701 , R24 EY022883 , and R01 EY026078 . CMS is supported by the RRF /Daniel M. Albert Chair. NS is a recipient of RPB Stein Innovation Award. DMA was supported by an Unrestricted Grant from Research to Prevent Blindness, New York , NY and P30 EY010572 NIH NEI . Zeta potential (ζ) was measured by the Analytical BioNanoTechnology Equipment Core (ANTEC) of Simpson Querrey Institute (SQI) at Northwestern University (NU), Chicago, IL, whose Peptide Synthesis Core produced peptides by solid state methods, supported by SHyNE Resource ( NSF ECCS-1542205 ). Rabbit eye drop delivery, also rabbit IVT injections and eye resections were performed by the NU Developmental Therapeutics Core (DTC), Evanston, IL.
Funding Information:
This work was supported by an unrestricted award from Research to Prevent Blindness to the Department of Ophthalmology and Visual Sciences, Retina Research Foundation, P30 EY016665, P30 CA014520, EPA 83573701, R24 EY022883, and R01 EY026078. CMS is supported by the RRF/Daniel M. Albert Chair. NS is a recipient of RPB Stein Innovation Award. DMA was supported by an Unrestricted Grant from Research to Prevent Blindness, New York, NY and P30 EY010572 NIH NEI. Zeta potential (?) was measured by the Analytical BioNanoTechnology Equipment Core (ANTEC) of Simpson Querrey Institute (SQI) at Northwestern University (NU), Chicago, IL, whose Peptide Synthesis Core produced peptides by solid state methods, supported by SHyNE Resource (NSF ECCS-1542205). Rabbit eye drop delivery, also rabbit IVT injections and eye resections were performed by the NU Developmental Therapeutics Core (DTC), Evanston, IL.
Publisher Copyright:
© 2019 Elsevier Ltd
PY - 2019/11
Y1 - 2019/11
N2 - Abnormal migration and proliferation of endothelial cells (EC) drive neovascular retinopathies. While anti-VEGF treatment slows progression, pathology is often supported by decrease in intraocular pigment epithelium-derived factor (PEDF), an endogenous inhibitor of angiogenesis. A surface helical 34-mer peptide of PEDF, comprising this activity, is efficacious in animal models of neovascular retina disease but remains impractically large for therapeutic use. We sought smaller fragments within this sequence that mitigate choroidal neovascularization (CNV). Expecting rapid intravitreal (IVT) clearance, we also developed a method to reversibly attach peptides to nano-carriers for extended delivery. Synthetic fragments of 34-mer yielded smaller anti-angiogenic peptides, and N-terminal capping with dicarboxylic acids did not diminish activity. Charge restoration via substitution of an internal aspartate by asparagine improved potency, achieving low nM apoptotic response in VEGF-activated EC. Two optimized peptides (PEDF 335, 8-mer and PEDF 336, 9-mer) were tested in a mouse model of laser-induced CNV. IVT injection of either peptide, 2–5 days before laser treatment, gave significant CNV decrease at day +14 post laser treatment. The 8-mer also decreased CNV, when administered as eye drops. Also examined was a nanoparticle-conjugate (NPC) prodrug of the 9-mer, having positive zeta potential, expected to display longer intraocular residence. This NPC showed extended efficacy, even when injected 14 days before laser treatment. Neither inflammatory cells nor other histopathologic abnormalities were seen in rabbit eyes harvested 14 days following IVT injection of PEDF 336 (>200 μg). No rabbit or mouse eye irritation was observed over 12–17 days of PEDF 335 eye drops (10 mM). Viability was unaffected in 3 retinal and 2 choroidal cell types by PEDF 335 up to 100 μM, PEDF 336 (100 μM) gave slight growth inhibition only in choroidal EC. A small anti-angiogenic PEDF epitope (G-Y-D-L-Y-R-V) was identified, variants (adipic-Sar-Y-N-L-Y-R-V) mitigate CNV, with clinical potential in treating neovascular retinopathy. Their shared active motif, Y - - - R, is found in laminin (Ln) peptide YIGSR, which binds Ln receptor 67LR, a known high-affinity ligand of PEDF 34-mer.
AB - Abnormal migration and proliferation of endothelial cells (EC) drive neovascular retinopathies. While anti-VEGF treatment slows progression, pathology is often supported by decrease in intraocular pigment epithelium-derived factor (PEDF), an endogenous inhibitor of angiogenesis. A surface helical 34-mer peptide of PEDF, comprising this activity, is efficacious in animal models of neovascular retina disease but remains impractically large for therapeutic use. We sought smaller fragments within this sequence that mitigate choroidal neovascularization (CNV). Expecting rapid intravitreal (IVT) clearance, we also developed a method to reversibly attach peptides to nano-carriers for extended delivery. Synthetic fragments of 34-mer yielded smaller anti-angiogenic peptides, and N-terminal capping with dicarboxylic acids did not diminish activity. Charge restoration via substitution of an internal aspartate by asparagine improved potency, achieving low nM apoptotic response in VEGF-activated EC. Two optimized peptides (PEDF 335, 8-mer and PEDF 336, 9-mer) were tested in a mouse model of laser-induced CNV. IVT injection of either peptide, 2–5 days before laser treatment, gave significant CNV decrease at day +14 post laser treatment. The 8-mer also decreased CNV, when administered as eye drops. Also examined was a nanoparticle-conjugate (NPC) prodrug of the 9-mer, having positive zeta potential, expected to display longer intraocular residence. This NPC showed extended efficacy, even when injected 14 days before laser treatment. Neither inflammatory cells nor other histopathologic abnormalities were seen in rabbit eyes harvested 14 days following IVT injection of PEDF 336 (>200 μg). No rabbit or mouse eye irritation was observed over 12–17 days of PEDF 335 eye drops (10 mM). Viability was unaffected in 3 retinal and 2 choroidal cell types by PEDF 335 up to 100 μM, PEDF 336 (100 μM) gave slight growth inhibition only in choroidal EC. A small anti-angiogenic PEDF epitope (G-Y-D-L-Y-R-V) was identified, variants (adipic-Sar-Y-N-L-Y-R-V) mitigate CNV, with clinical potential in treating neovascular retinopathy. Their shared active motif, Y - - - R, is found in laminin (Ln) peptide YIGSR, which binds Ln receptor 67LR, a known high-affinity ligand of PEDF 34-mer.
KW - Angiogenesis
KW - Choroidal neovascularization
KW - Laminin receptor
KW - Macular degeneration
KW - PEDF
KW - Peptides
KW - Retinopathy
KW - YIGSR
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U2 - 10.1016/j.exer.2019.107798
DO - 10.1016/j.exer.2019.107798
M3 - Article
C2 - 31520600
AN - SCOPUS:85072194310
SN - 0014-4835
VL - 188
JO - Experimental Eye Research
JF - Experimental Eye Research
M1 - 107798
ER -