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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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Improved Synthesis and In Vitro Evaluation of an Aptamer Ribosomal Toxin Conjugate

Linsley Kelly1, Christina Kratschmer1, Keith E Maier1

  • 1Department of Biochemistry, Albert Einstein College of Medicine , Bronx, New York City, New York.

Nucleic Acid Therapeutics
|May 27, 2016
PubMed

Insights

Researchers developed a streamlined method for creating aptamer-toxin conjugates for cancer therapy. This new approach uses a minimized aptamer for efficient delivery of toxins like gelonin to prostate cancer cells.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Therapeutics

Background:

  • Antibody-toxin conjugates (immunotoxins) show promise in cancer treatment but face challenges like bulkiness and immunogenicity.
  • Previous work utilized a full-length aptamer (A9) to deliver recombinant gelonin (rGel) to prostate-specific membrane antigen (PSMA)-expressing cells.
  • A need exists for more streamlined and scalable methods for generating aptamer-toxin conjugates.

Purpose of the Study:

  • To develop a simplified and scalable method for generating aptamer-rGel conjugates.
  • To evaluate the binding specificity and toxicity of the novel aptamer-toxin conjugate.
  • To assess the in vitro stability of the conjugate for potential in vivo applications.

Main Methods:

  • Chemically synthesized a minimized A9 aptamer with a 5' terminal thiol.
  • Generated aptamer-toxin conjugates using a reducible disulfide linkage.
  • Assessed conjugate binding specificity and cytotoxicity on PSMA(+) LNCaP prostate cancer cells.
  • Performed serum stability analysis of the conjugate in mouse serum.

Main Results:

  • The A9.min-rGel conjugate demonstrated specific binding and toxicity against PSMA(+) LNCaP cells with an IC50 of approximately 60 nM.
  • The conjugate exhibited a half-life (t1/2) of approximately 4 hours in mouse serum.
  • The streamlined chemical synthesis facilitated large-scale production of the aptamer-toxin conjugate.

Conclusions:

  • A minimized aptamer approach offers an efficient and scalable method for producing aptamer-toxin conjugates.
  • The A9.min-rGel conjugate shows potential for targeted cancer therapy due to its specificity and stability.
  • This strategy paves the way for further in vivo studies and development of novel targeted cancer treatments.

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