Intrathecal Delivery of Therapeutic Oligonucleotides for Potent Modulation of Gene Expression in the Central Nervous System.

Zachary Kennedy,Bruno M D C Godinho,James W Gilbert

doi:10.1007/978-1-0716-2010-6_24

Zachary Kennedy, Bruno M D C Godinho + Show 1 more

Open Access

https://doi.org/10.1007/978-1-0716-2010-6_24

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Abstract

Therapeutic oligonucleotides hold tremendous potential for treating central nervous system (CNS) disorders. The route of administration of oligonucleotides significantly impacts both distribution and silencing efficiency. Here, we describe a technically simple, clinically relevant method to administer oligonucleotide compounds into the CNS via direct intrathecal injections. This method achieves distribution throughout the CNS rapidly and permits high-throughput testing of oligonucleotide efficacy and potency in mammals.

Highlights

Therapeutic oligonucleotides hold tremendous potential for treating disorders of the central nervous system (CNS), but numerous factors can hinder the delivery of these compounds
Among the major determinants affecting delivery of drugs to the CNS is the route of administration
Drug delivery to the CNS can be performed through two general approaches: (a) systemic, where oligonucleotides are initially delivered outside of the CNS, or (b) direct, where oligonucleotides are delivered directly within the CNS

Summary

Introduction

Therapeutic oligonucleotides hold tremendous potential for treating disorders of the central nervous system (CNS), but numerous factors can hinder the delivery of these compounds. Systemic delivery strategies (e.g., intravenous or subcutaneous) are technically simple and easy to perform, but require significantly higher doses and result in very low efficacy in the CNS, mostly due to the inability of oligonucleotides to pass the blood–brain barrier [1–3]. IT administration is favored from a clinical perspective because it is less invasive than ICV injections; IT injections do not require brain surgery and can be performed as an outpatient procedure. IT injections are the only clinically approved route of administration for therapeutic oligonucleotide treatment of CNS disorders [8]. Given its ability to deliver to the CNS, the comparatively quick operation time, and the clinical relevance of the route of administration, IT injections are an attractive delivery method for which to screen oligonucleotide compounds for CNS applications. Special attention is given to the preparation of the oligonucleotide compounds, prepping the mouse for the injections, and ensuring successful delivery

Direct Intrathecal Injections

Preparation of Test Oligonucleotides

Direct Intrathecal Injection

Suggestions for Tissue Collection and Processing