Abstract
The ability to precisely upregulate genes in inflamed brain holds great therapeutic promise. Here we report a novel class of vectors, genetically modified macrophages that carry reporter and therapeutic genes to neural cells. Systemic administration of macrophages transfected ex vivo with a plasmid DNA (pDNA) encoding a potent antioxidant enzyme, catalase, produced month-long expression levels of catalase in the brain resulting in three-fold reductions in inflammation and complete neuroprotection in mouse models of Parkinson's disease (PD). This resulted in significant improvements in motor functions in PD mice. Mechanistic studies revealed that transfected macrophages secreted extracellular vesicles, exosomes, packed with catalase genetic material, pDNA and mRNA, active catalase, and NF-κb, a transcription factor involved in the encoded gene expression. Exosomes efficiently transfer their contents to contiguous neurons resulting in de novo protein synthesis in target cells. Thus, genetically modified macrophages serve as a highly efficient system for reproduction, packaging, and targeted gene and drug delivery to treat inflammatory and neurodegenerative disorders.
Highlights
Development of new delivery systems for gene and drug transport for neurodegenerative disorders, including Parkinson’s and Alzheimer’s diseases (PD and AD), is greatly needed
The best results were obtained, when macrophages were transfected using GenePORTER 3000 transfection agent incubated with 2 mg/ml plasmid DNA (pDNA) encoded green fluorescent protein (GFP), or catalase for four hours, and cultured in complete media with 20% fetal bovine serum (FBS)
A sustained protein expression and prolonged release into the media for at least 21 days was detected by GFP fluorescence, or catalase enzyme activity (Fig. 1 B), and confirmed by confocal microscopy in catalase-transfected macrophages (Fig. 1 D)
Summary
Development of new delivery systems for gene and drug transport for neurodegenerative disorders, including Parkinson’s and Alzheimer’s diseases (PD and AD), is greatly needed. The pathobiology of PD and AD is linked to microglial activation and subsequent secretion of neurotoxic factors. These include reactive oxygen and nitrogen species (ROS and RNS) leading to oxidative stress [1,2,3,4], which affects neuronal, astrocyte, and microglia function by inducing ion transport and calcium mobilization, and activating apoptotic programs. The mitochondrial respiratory chain affects oxidative phosphorylation and is responsible for ROS production. Such pathways lead to neuronal demise and underlie the pathobiology of PD and AD [6]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
