Reducible PEG-POD/DNA Nanoparticles for Gene Transfer In Vitro and In Vivo: Application in a Mouse Model of Age-Related Macular Degeneration

Abstract

Non-viral gene delivery systems are being developed to address limitations of viral gene delivery. Many of these non-viral systems are modeled on the properties of viruses including cell surface binding, endocytosis, endosomal escape, and nuclear targeting. Most non-viral gene transfer systems exhibit little correlation between in vitro and in vivo efficiency, hampering a systematic approach to their development. Previously, we have described a 3.5 kDa peptide (peptide for ocular delivery [POD]) that targets cell surface sialic acid. When functionalized with polyethylene glycol (PEG) via a sulfhydryl group on the N-terminal cysteine of POD, PEG-POD could compact plasmid DNA, forming 120- to 180-nm homogeneous nanoparticles. PEG-POD enabled modest gene transfer and rescue of retinal degeneration in vivo. Systematic investigation of different stages of gene transfer by PEG-POD nanoparticles was hampered by their inability to deliver genes in vitro. Herein, we describe functionalization of POD with PEG using a reducible orthopyridyl disulfide bond. These reducible nanoparticles enabled gene transfer in vitro while retaining their in vivo gene transfer properties. These reducible PEG-POD nanoparticles were utilized to deliver human FLT1 to the retina in vivo, achieving a 50% reduction in choroidal neovascularization in a murine model of age-related macular degeneration.

Overcoming the cellular barriers and beyond: Recent progress on cell penetrating peptide modified nanomedicine in combating physiological and pathological barriers

“… Myocardial delivery PCM modified liposome WLSEAGPVVTVRALRGTGSW Prolong residence and accumulation in heart [ 91 , 92 ] Hph1–Hph1–dsRBD/siRNA complex Hph1: YARVRRRGPRR Enhance gene silencing effects [140] Lipoplexes modified with TAT and antimyosin antibody 2G4 RKKRRQRRR Increase accumulation in the ischemic rat myocardium [93] R7W-MP modified CaP NP DQRPDREAPRS Target the Cavb2 cytosolic subunit of the l -type calcium channel (LTCC), improve cardiac contractility in pathological conditions [94] TPSi NP RKKRRQRRR Increase both the cell survival rate and the delivery precision of stem cell transplantation [95] Ocular barrier POD-PEG NP GGGG(ARKKAAKA) 4 Target cell surface sialic acid. Reduce cell apoptosis and enhance the thickness of outer nuclear layer [141] POD-SS-PEG/DNA nanocomplex GGG[ARKKAAKA]4 Deliver FLT1 plasmid and achieve a 50% reduction in choroidal neovascularization in AMD model [142] POD conjugatedQD GGG[ARKKAAKA]4 Enter neural retina and localize to retinal pigment epithelium [143] Penetratin fused HDL mutants RQIKIWFQNRRMKWKK Enhance the delivery efficiency to the posterior segment of the eye [144] RFP plasmid /PAMAM/Penetratin RQIKIWFQNRRMKWKK Improve cellular uptake and transfection [102] cRGD/penetratin-PEG-PAMAM RQIKIWFQNRRMKWKK Deliver cargoes to the cornea and retina with a prolonged retention RGD/TAT-PEG-PLGA RKKRRQRRR Increase uptake in the choroid-retina [105] Mucosal barrier Penetratin- mesostructured silica NP ...…”

Smart Lipid–Polysaccharide Nanoparticles for Targeted Delivery of Doxorubicin to Breast Cancer Cells

“…For the reduction of disulfide bonds within Cys moieties, 5.0 mg PHYN was added to a 5.0 mL 10 mM 2-mercaptoethanol solution in PBS (pH 7.4, 0.01 M) and incubated at 25 • C for 2 h [51]. Then, after exhaustive dialysis (MWCO 12-14 kDa, Medicell International LTD, London, UK) against PBS (pH 7.4, 0.01 M), Ellman's assay was performed as follows [35]: 250 µL of dialyzed solution was mixed with 250 µL NaH 2 PO 4 /Na 2 HPO 4 phosphate buffer (pH 8.0, 0.5 M) and 500 µL Ellman's reagent consisting of 0.3 mg/mL DTNB in NaH 2 PO 4 /Na 2 HPO 4 phosphate buffer (pH 8.0, 0.05 M).…”

<p>Nanoparticle-Mediated Drug Delivery for the Treatment of Cardiovascular Diseases</p>

“…It ultimately leads to the decline (50%) in the choroidal neovascularization when tested in vivo. 134 Mannosylated chitosan nanoparticles are excellent carriers for DNA and siRNA delivery and, instead, are readily taken up by the macrophages. Thus, the application and future perspectives of mannosylated chitosan nanoparticles for subduing inflammation and reducing myocardial infarction await further studies.…”