Identification of Aptamers That Bind to Sickle Hemoglobin and Inhibit Its Polymerization

Nucleic Acid Ther. 2017 Dec;27(6):354-364. doi: 10.1089/nat.2016.0646. Epub 2017 Oct 17.

Abstract

The pathophysiology of sickle cell disease (SCD) is dependent on the polymerization of deoxygenated sickle hemoglobin (HbS), leading to erythrocyte deformation (sickling) and vaso-occlusion within the microvasculature. Following deoxygenation, there is a delay time before polymerization is initiated, during which nucleation of HbS monomers occurs. An agent with the ability to extend this delay time or slow polymerization would therefore hold a therapeutic, possibly curative, potential. We used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method to screen for HbS-binding RNA aptamers modified with nuclease-resistant 2'-fluoropyrimidines. Polymerization assays were employed to identify aptamers with polymerization-inhibitory properties. Two noncompeting aptamers, DE3A and OX3B, were found to bind hemoglobin, significantly increase the delay time, and reduce the rate of polymerization of HbS. These modifiable, nuclease-resistant aptamers are potential new therapeutic agents for SCD.

Keywords: RNA; aptamer; hemoglobin; polymerization; sickle cell disease.

MeSH terms

  • Anemia, Sickle Cell / drug therapy
  • Anemia, Sickle Cell / metabolism
  • Antisickling Agents / administration & dosage*
  • Antisickling Agents / chemical synthesis
  • Antisickling Agents / chemistry*
  • Aptamers, Nucleotide / administration & dosage
  • Aptamers, Nucleotide / chemical synthesis
  • Aptamers, Nucleotide / chemistry*
  • Cell-Free System
  • DNA, Complementary
  • Deoxycytosine Nucleotides / chemistry
  • Deoxyuracil Nucleotides / chemistry
  • Dose-Response Relationship, Drug
  • Drug Discovery / methods
  • Hemoglobin, Sickle / chemistry*
  • Humans
  • Polymerization / drug effects*
  • Pyrimidines / chemistry
  • SELEX Aptamer Technique / methods

Substances

  • Antisickling Agents
  • Aptamers, Nucleotide
  • DNA, Complementary
  • Deoxycytosine Nucleotides
  • Deoxyuracil Nucleotides
  • Hemoglobin, Sickle
  • Pyrimidines
  • deoxyuridine triphosphate
  • 2'-deoxycytidine 5'-triphosphate