Lentiviral Vector-Mediated Gene Transfer and RNA Silencing Technology in Neuronal Dysfunctions

Abstract

Lentiviral-mediated gene transfer in vivo or in cultured mammalian neurons can be used to address a wide variety of biological questions, to design animals models for specific neurodegenerative pathologies, or to test potential therapeutic approaches in a variety of brain disorders. Lentiviruses can infect non-dividing cells, thereby allowing stable gene transfer in post-mitotic cells such as mature neurons. An important contribution has been the use of inducible vectors: the same animal can thus be used repeatedly in the doxycycline-on or -off state, providing a powerful mean for assessing the function of a gene candidate in a disorder within a specific neuronal circuit. Furthermore, lentivirus vectors provide a unique tool to integrate siRNA expression constructs with the aim to locally knockdown expression of a specific gene, enabling to assess the function of a gene in a very specific neuronal pathway. Lentiviral vector-mediated delivery of short hairpin RNA results in persistent knockdown of gene expression in the brain. Therefore, the use of lentiviruses for stable expression of siRNA in brain is a powerful aid to probe gene functions in vivo and for gene therapy of diseases of the central nervous system. In this chapter I review the applications of lentivirus-mediated gene transfer in the investigation of specific gene candidates involved in major brain disorders and neurodegenerative processes. Major applications have been in polyglutamine disorders, such as synucleinopathies and Parkinson's disease, or in investigating gene function in Huntington's disease, dystonia, or muscular dystrophy. Recently, lentivirus gene transfer has been an invaluable tool for evaluation of gene function in behavioral disorders such as drug addiction and attention-deficit hyperactivity disorder or in learning and cognition.

Antioxidant gene therapy against neuronal cell death

“…However, because they insert their genetic material into the host genome they present an oncogenic risk. Major advantages of LV vectors are their ability to transduce non-dividing cells including differentiated neurons, and that large genes can be inserted and permanently incorporated into the host cell (Dreyer, 2011). …”

Chitosan-Mangafodipir nanoparticles designed for intranasal delivery of siRNA and DNA to brain

“…A major obstacle to fulfilling the therapeutic promise of gene therapy is the requirement for an invasive delivery system (stereotaxic injection of viral vectors or infusion of ASO directly into brain or intrathecal space) [9–11]. For diseases like HD and PD that gradually encompass the entire central nervous system (CNS), repeated injections of the agent (vector encoding DNA, siRNA or ASO) into multiple brain regions over the course of the illness make it unfeasible and unacceptable.…”

Gene-based therapy of Parkinson’s Disease: Translation from animal model to human clinical trial employing convection enhanced delivery

“…Recently, LV gene transfer has been an invaluable tool for evaluation of gene function in behavioral disorders such as drug addiction and attention-deficit hyperactivity disorder or in learning and cognition. 39 …”