New MiniPromoter Ple345 (NEFL) drives strong and specific expression in retinal ganglion cells of mouse and primate retina

Elizabeth M. Simpson, Andrea J. Korecki, Oriol Fornes, Trevor McGill, Jorge Luis Cueva-Vargas, Jessica Agostinone, Rachelle A. Farkas, Jack W. Hickmott, Siu Ling Lam, Anthony Mathelier, Lauren M. Renner, Jonathan Stoddard, Michelle Zhou, Adriana Di Polo, Martha Neuringer, Wyeth W. Wasserman

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Retinal gene therapy is leading the neurological gene therapy field, with 32 ongoing clinical trials of recombinant adeno-associated virus (rAAV)-based therapies. Importantly, over 50% of those trials are using restricted promoters from human genes. Promoters that restrict expression have demonstrated increased efficacy and can limit the therapeutic to the target cells thereby reducing unwanted off-target effects. Retinal ganglion cells are a critical target in ocular gene therapy; they are involved in common diseases such as glaucoma, rare diseases such as Leber's hereditary optic neuropathy, and in revolutionary optogenetic treatments. Here, we used computational biology and mined the human genome for the best genes from which to develop a novel minimal promoter element(s) designed for expression in restricted cell types (MiniPromoter) to improve the safety and efficacy of retinal ganglion cell gene therapy. Gene selection included the use of the first available droplet-based single-cell RNA sequencing (Drop-seq) dataset, and promoter design was bioinformatically driven and informed by a wide range of genomics datasets. We tested seven promoter designs from four genes in rAAV for specificity and quantified expression strength in retinal ganglion cells in mouse, and then the single best in nonhuman primate retina. Thus, we developed a new human-DNA MiniPromoter, Ple345 (NEFL), which in combination with intravitreal delivery in rAAV9 showed specific and robust expression in the retinal ganglion cells of the nonhuman-primate rhesus macaque retina. In mouse, we also developed MiniPromoters expressing in retinal ganglion cells, the hippocampus of the brain, a pan neuronal pattern in the brain, and peripheral nerves. As single-cell transcriptomics such as Drop-seq become available for other cell types, many new opportunities for additional novel restricted MiniPromoters will present.

Original languageEnglish (US)
Pages (from-to)257-272
Number of pages16
JournalHuman Gene Therapy
Volume30
Issue number3
DOIs
StatePublished - Mar 1 2019

Fingerprint

Retinal Ganglion Cells
Primates
Retina
Genetic Therapy
Dependovirus
Genes
Optogenetics
Leber's Hereditary Optic Atrophy
RNA Sequence Analysis
Brain
Human Genome
Cell- and Tissue-Based Therapy
Rare Diseases
Genomics
Computational Biology
Macaca mulatta
Peripheral Nerves
Glaucoma
Hippocampus
Therapeutics

Keywords

  • brain
  • enhancer
  • gene therapy
  • intravitreal injection
  • promoter
  • rAAV9

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Simpson, E. M., Korecki, A. J., Fornes, O., McGill, T., Cueva-Vargas, J. L., Agostinone, J., ... Wasserman, W. W. (2019). New MiniPromoter Ple345 (NEFL) drives strong and specific expression in retinal ganglion cells of mouse and primate retina. Human Gene Therapy, 30(3), 257-272. https://doi.org/10.1089/hum.2018.118

New MiniPromoter Ple345 (NEFL) drives strong and specific expression in retinal ganglion cells of mouse and primate retina. / Simpson, Elizabeth M.; Korecki, Andrea J.; Fornes, Oriol; McGill, Trevor; Cueva-Vargas, Jorge Luis; Agostinone, Jessica; Farkas, Rachelle A.; Hickmott, Jack W.; Lam, Siu Ling; Mathelier, Anthony; Renner, Lauren M.; Stoddard, Jonathan; Zhou, Michelle; Di Polo, Adriana; Neuringer, Martha; Wasserman, Wyeth W.

In: Human Gene Therapy, Vol. 30, No. 3, 01.03.2019, p. 257-272.

Research output: Contribution to journalArticle

Simpson, EM, Korecki, AJ, Fornes, O, McGill, T, Cueva-Vargas, JL, Agostinone, J, Farkas, RA, Hickmott, JW, Lam, SL, Mathelier, A, Renner, LM, Stoddard, J, Zhou, M, Di Polo, A, Neuringer, M & Wasserman, WW 2019, 'New MiniPromoter Ple345 (NEFL) drives strong and specific expression in retinal ganglion cells of mouse and primate retina', Human Gene Therapy, vol. 30, no. 3, pp. 257-272. https://doi.org/10.1089/hum.2018.118
Simpson, Elizabeth M. ; Korecki, Andrea J. ; Fornes, Oriol ; McGill, Trevor ; Cueva-Vargas, Jorge Luis ; Agostinone, Jessica ; Farkas, Rachelle A. ; Hickmott, Jack W. ; Lam, Siu Ling ; Mathelier, Anthony ; Renner, Lauren M. ; Stoddard, Jonathan ; Zhou, Michelle ; Di Polo, Adriana ; Neuringer, Martha ; Wasserman, Wyeth W. / New MiniPromoter Ple345 (NEFL) drives strong and specific expression in retinal ganglion cells of mouse and primate retina. In: Human Gene Therapy. 2019 ; Vol. 30, No. 3. pp. 257-272.
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