Optimization of a 40-mer Antimyelin DNA Aptamer Identifies a 20-mer with Enhanced Properties for Potential Multiple Sclerosis Therapy

Brandon Wilbanks, John Smestad, Robin M. Heider, Arthur E. Warrington, Moses Rodriguez, L James Maher III

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We previously reported the in vitro selection and characterization of a DNA aptamer capable of stimulating remyelination in a mouse model of multiple sclerosis. This aptamer was selected for its ability to bind to suspensions of crude murine myelin in vitro. Our initial studies in vitro and in vivo involved a 40-nucleotide derivative (LJM-3064) of the original 100-nucleotide aptamer. LJM-3064 retained robust myelin-binding properties. Structural characterization of LJM-3064 revealed that the guanosine-rich 5′ half of the sequence forms different G-quadruplex-type structures that are variably stable in the presence of physiologically relevant ions. We hypothesized that this structured domain is sufficient for myelin binding. In this study, we confirm that a 20-nucleotide DNA, corresponding to the 5′ half of LJM-3064, retains myelin-binding properties. We then optimize this minimal myelin-binding aptamer via systematic evolution of ligands by exponential enrichment after sparse rerandomization. We report a sequence variant (LJM-5708) of the 20-nucleotide myelin-binding aptamer with enhanced myelin-binding properties and the ability to bind cultured human oligodendroglioma cells in vitro, providing the first evidence of cross-species reactivity of this myelin-binding aptamer. As our formulation of DNA aptamers for in vivo remyelination therapy involves conjugation to streptavidin, we verified that the myelin-binding properties of LJM-5708 were retained in conjugates to avidin, streptavidin, and neutravidin. DNA aptamer LJM-5708 is a lead for further preclinical development of remyelinating aptamer technologies.

Original languageEnglish (US)
Pages (from-to)126-135
Number of pages10
JournalNucleic Acid Therapeutics
Volume29
Issue number3
DOIs
StatePublished - Jun 1 2019

Fingerprint

Nucleotide Aptamers
Myelin Sheath
Multiple Sclerosis
Streptavidin
Nucleotides
Avidin
Guanosine
Therapeutics
Suspensions
SELEX Aptamer Technique
Ions
Ligands
Derivatives
G-Quadruplexes
Oligodendroglioma
LJM-3064
DNA
Technology

Keywords

  • aptamer
  • conjugate
  • G-quadruplex
  • multiple sclerosis
  • myelin

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery

Cite this

Optimization of a 40-mer Antimyelin DNA Aptamer Identifies a 20-mer with Enhanced Properties for Potential Multiple Sclerosis Therapy. / Wilbanks, Brandon; Smestad, John; Heider, Robin M.; Warrington, Arthur E.; Rodriguez, Moses; Maher III, L James.

In: Nucleic Acid Therapeutics, Vol. 29, No. 3, 01.06.2019, p. 126-135.

Research output: Contribution to journalArticle

Wilbanks, Brandon ; Smestad, John ; Heider, Robin M. ; Warrington, Arthur E. ; Rodriguez, Moses ; Maher III, L James. / Optimization of a 40-mer Antimyelin DNA Aptamer Identifies a 20-mer with Enhanced Properties for Potential Multiple Sclerosis Therapy. In: Nucleic Acid Therapeutics. 2019 ; Vol. 29, No. 3. pp. 126-135.
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