Optimization of Long-lasting Analgesia via Targeted Epigenetic Repression (LATER) for human use

National Center For Advancing Translational Sciences

Investigators

Abstract

NCATS and Navega have been working together to optimize transgene expression after transduction of iPSC-derived sensory neurons with the AAV zinc finger constructs provided by Navega. Optimization has included testing different AAV serotypes, promoter sequences, transduction times, and a variety of other protocol steps. These studies have revealed that a decrease in zinc finger expression is observed due to transitioning the zinc finger delivery system from a lentiviral to an AAV backbone. This year, to address this issue, AAV constructs that retain the zinc finger sequences were developed at Navega for repression of NaV1.7 gene expression in human DRG, these were validated at NCATS via RNAseq and functional analysis to demonstrate repression of NaV1.7 expression and its effect on the electrophysiological profile of human iPSC-derived nociceptor in vitro. Similarly, an additional series of AAV constructs were developed against NaV1.8, that were shown to repression expression in human iPSC-derived sensory neurons. As a next step, and to increase efficacy of this potential therapeutic, a dual NaV1.7-NaV1.8 AAV construct has been developed and is currently undergoing testing in human iPSC-derived sensory neurons for dual repression and its effects on electrophysiological activity.