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(99m)Tc-MORF oligomers specific for bacterial ribosomal RNA as potential specific infection imaging agents

Ling Chen1, Yi Wang, Dengfeng Cheng

  • 1Division of Nuclear Medicine, Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655, United States.

Bioorganic & Medicinal Chemistry
|September 24, 2013
PubMed
Abstract

Insights

Technetium-99m-labeled phosphorodiamidate morpholino (MORF) oligomers targeting bacterial 16S rRNA show high binding affinity and specific accumulation in bacteria. These MORF oligomers show promise as novel imaging agents for bacterial infections.

Area of Science:

  • Molecular Biology
  • Medical Imaging
  • Antimicrobial Research

Background:

  • Bacterial infections pose a significant global health challenge.
  • Accurate and early detection of bacterial infections is crucial for effective treatment.
  • Current diagnostic methods have limitations in sensitivity and specificity.

Purpose of the Study:

  • To investigate radiolabeled oligomers complementary to bacterial 16S rRNA as potential agents for bacterial infection imaging.
  • To evaluate the binding affinity and specificity of different oligomer backbones (MORF, PNA, PS-DNA) to bacterial RNA.
  • To assess the in vivo accumulation and imaging potential of the most promising oligomer in a murine model.

Main Methods:

  • Synthesis and (99m)Tc-labeling of phosphorodiamidate morpholino (MORF), peptide nucleic acid (PNA), and phosphorothioate DNA (PS-DNA) oligomers.
  • In vitro evaluation of RNA binding affinity using Escherichia coli strains.
  • In situ hybridization and flow cytometry to assess cellular uptake in fixed and live bacteria.
  • In vivo imaging studies in mice with induced bacterial infection.

Main Results:

  • MORF oligomers exhibited significantly higher binding affinity to bacterial RNA compared to PNA and PS-DNA.
  • MORF oligomers demonstrated specific accumulation in live bacteria, confirmed by fluorescence microscopy and flow cytometry.
  • (99m)Tc-labeled MORF showed targeted accumulation in infected tissues in a mouse model, with significantly higher signal in infected thighs compared to controls.
  • High binding of (99m)Tc-MORF to bacterial RNA from E. coli strains was quantified.

Conclusions:

  • A (99m)Tc-labeled MORF oligomer targeting bacterial 16S rRNA demonstrates effective binding to bacterial RNA in vitro.
  • Specific accumulation of MORF oligomers into live bacteria was observed, indicating potential for targeted delivery.
  • Radiolabeled MORF oligomers antisense to bacterial rRNA represent a promising new class of agents for imaging bacterial infections.
Keywords:
(99m)TcAF633Alexa Fluor 633 carboxylic acid succinimidyl esterBacterial infection imagingD-PBSDulbecco’s PBSE. coliEscherichia coliFISHK. pneumoniaKlebsiella pneumoniaMAG(3)MORFODPNAPS-DNAS-acetyl NHS-MAG(3)S. aureusSDSSPECT/CTStaphylococcus aureusfluorescence in situ hybridizationoptical densitypeptide nucleic acidphosphorodiamidate morpholinophosphorothioate DNArRNAribosomal RNAsodium dodecyl sulfatetechnetium-99m

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