Compacted DNA Nanoparticle Gene Transfer of GDNF to the Rat Striatum Enhances the Survival of Grafted Fetal Dopamine Neurons

Abstract

Previously it was established that infusion of glial cell line-derived neurotrophic factor (GDNF) protein into grafts of embryonic dopamine cells has a neurotrophic effect on the grafted cells. In this study we used a nonviral technique to transfer the gene encoding for GDNF to striatal cells. Plasmid DNA encoding for GDNF was compacted into DNA nanoparticles (DNPs) by 10 kDa polyethylene glycol (PEG)-substituted lysine 30-mers (CK 30 PEG10k) and then injected into the denervated striatum of rats with unilateral 6-hydroxydopamine lesions. Sham controls were injected with saline. One week later, experimental animals received either a ventral mesencephalic (VM) tissue chunk graft or a cell suspension VM graft implanted into the denervated striatum. Grafts were allowed to integrate for 4-6 weeks and during this period we monitored spontaneous and drug-induced motor activity. Using stereological cell counting we observed a 16-fold increase in the number of surviving TH + cells within tissue chunk grafts placed into the striatum pretreated with pGDNF DNPs (14,923 ± 4,326) when compared to grafts placed into striatum pretreated with saline (955 ± 343). Similarly, we observed a sevenfold increase in the number of TH + cells within cell suspension grafts placed into the striatum treated with pGDNF DNPs when compared to cell suspension grafts placed into the saline dosed striatum. Behaviorally, we observed significant improvement in rotational scores and in spontaneous forepaw usage of the affected forelimb in grafted animals receiving prior treatment with compacted pGDNF DNPs when compared to grafted animals receiving saline control pretreatment. Data analysis for protein, morphological, and behavioral measures suggests that compacted pGDNF DNPs injected into the striatum can result in transfected cells overexpressing GDNF protein at levels that provide neurotrophic support for grafted embryonic dopamine neurons.

Drug development in Parkinson's disease: From emerging molecules to innovative drug delivery systems

“…NP were then injected into the denervated striatum of rats with unilateral 6-OHDA lesion in order to enhance grafted DA neuron survival. Data suggest that compacted pGDNF could transfect cells to overexpress GDNF protein at levels that provide support for grafted stem cells [42].…”

DNA nanoparticle-mediated ABCA4 delivery rescues Stargardt dystrophy in mice

“…In contrast with many other nonviral approaches, these NPs drive long-term gene expression (1,2) after subretinal delivery to the mouse eye, making them an excellent choice for chronic retinal degenerations such as Stargardt. They have recently been used to mediate phenotypic rescue in rodent models of retinitis pigmentosa, Parkinson disease, and cystic fibrosis, and were safely employed in a phase I/II clinical trial for cystic fibrosis (2,8,9). One distinguishing feature of these NPs is their large capacity.…”

Diagnostic and Therapeutic Uses of Nanomaterials in the Brain

“…Regarding nanotechnology strategies for local administration of GDNF, Yurek et al published several studies using GDNF plasmid DNA compacted into NP using a polycation-like 10 kDa polyethylene glycol (PEG)-substituted lysine 30-mers (see Table 3). In the first paper the authors combine this nonviral gene therapy with neural grafts in order to improve the survival of the grafted cells and the recovery of parkinsonian rats [238]. Compacted DNA NP locally implanted into the striatum overexpressed GDNF in the lesioned striatum to levels able to provide support to grafted cells.…”