Optimizing Enhanced Hammerhead Ribozymes for Retinal Nucleic Acid Therapeutics

NIH RePORTER · EY · R01 · $544,353 · view on reporter.nih.gov ↗

Abstract

Many mutations in the human rod opsin (hRHO) and peripherin genes cause autosomal dominant retinitis pigmentosa (adRP) and macular degenerations. The folded mRNAs are targets for mutation-independent hammerhead ribozyme (hhRz) gene therapy. Our long-range goal is to translate effective hhRz therapeutics for hRHO adRP and other genes into human clinical trials. We have discovered a potent form of hhRz, which we call Enhanced-hhRz (EhhRz) therapeutics. All or most known mutations in a given dominant disease gene could be treated with a mutation-independent EhhRz strategy that pairs knockdown (KD) of mutant (and WT) mRNA with reconstitution (RECON) of WT protein with expression of a cleavage-resistant mRNA. The current stage of development of EhhRzs (against hRHO) lead to kinetic turnover rates greater than 2-log orders improved over historical minimal hhRzs (mhhRz); one mhhRz partially rescued mutant RHO retinal degeneration in rodent models. EhhRzs function under substrate (target) excess conditions and physiological levels of cofactor Mg2+, which are optimum for intracellular KD therapeutics. If this exciting kinetic potential can be harnessed for the photoreceptor, retinal, or ocular cells then there is potential for nucleic acid drugs (injectable EhhRzs) as therapeutics without vector-based gene therapy, for diverse ocular diseases and inherited retinal degenerations. Recent clinical success with intravitreal injection of antisense agents incentivizes this development. The objective is to optimize EhhRzs for intracellular performance in cultured human cells and mouse photoreceptor cells that are “humanized” for RHO mRNAs (same targets as in clinical trials). The central hypothesis is that optimization of EhhRz kinetic performance and delivery will maximize target mRNA/protein KD; optimization requires quantitation of biophysical variables that underlie target: EhhRz interaction, rational and/or evolutionary engineering of RNA structure-related function, and identi

Key facts

NIH application ID
11332629
Project number
5R01EY035007-04
Recipient
STATE UNIVERSITY OF NEW YORK AT BUFFALO
Principal Investigator
JOHN M. SULLIVAN
Activity code
R01
Funding institute
EY
Fiscal year
2026
Award amount
$544,353
Award type
5
Project period
2023-08-01T00:00:00 → 2027-04-30T00:00:00