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Cell-compatible, integrin-targeted cryptophane-129Xe NMR biosensors.

Garry K Seward1, Yubin Bai1, Najat S Khan1

  • 1University of Pennsylvania, Department of Chemistry, 231 South 34 St, Philadelphia, PA 19104-6323, USA.

Chemical Science
|November 4, 2014
PubMed
Summary

Peptide-modified cryptophane sensors show enhanced targeting to cancer cells expressing αvβ3 integrin receptors. This advancement enables sensitive detection of protein analytes using hyperpolarized 129Xe NMR spectroscopy for improved cancer diagnostics.

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Area of Science:

  • Biomedical Engineering
  • Chemical Biology
  • Molecular Imaging

Background:

  • αvβ3 integrin receptor is overexpressed in many human cancers, making it a key target for cancer diagnostics.
  • Hyperpolarized 129Xe NMR spectroscopy offers high sensitivity for detecting biomolecular interactions.
  • Cryptophane molecules can be functionalized for targeted delivery and detection of specific analytes.

Purpose of the Study:

  • To develop and evaluate a peptide-modified cryptophane biosensor for targeted delivery to αvβ3 integrin-expressing cancer cells.
  • To assess the cellular internalization, specificity, and biosensing capabilities of the modified cryptophane using hyperpolarized 129Xe NMR.
  • To determine the non-toxicity and efficacy of the cryptophane conjugate for potential in vivo applications.

Main Methods:

  • Functionalization of cryptophane with cyclic RGDyK peptide for αvβ3 integrin targeting and propionic acid groups for water solubility.
  • Confocal laser scanning microscopy and flow cytometry to monitor cellular internalization and specificity.
  • MTT assay to evaluate cytotoxicity in normal human lung fibroblasts.
  • Hyperpolarized 129Xe NMR spectroscopy to detect cryptophane binding to integrin receptors.

Main Results:

  • Peptide-cryptophane conjugate demonstrated specific and enhanced cellular internalization (4-fold higher) in cancer cells overexpressing αvβ3 integrin.
  • Competitive blocking assays confirmed receptor-mediated endocytosis.
  • The biosensor exhibited nanomolar inhibitory concentrations (IC50 = 20-30 nM) for integrin binding.
  • Hyperpolarized 129Xe NMR showed distinct spectral changes upon binding to αIIbβ3 integrin, enabling sensitive detection.

Conclusions:

  • Peptide-functionalized cryptophane serves as a potent and specific biosensor for αvβ3 integrin, enabling sensitive detection via hyperpolarized 129Xe NMR.
  • The developed cryptophane conjugate is non-toxic and shows improved targeting to cancer cells, highlighting its potential for cancer imaging and diagnostics.
  • This approach advances the application of hyperpolarized 129Xe NMR spectroscopy for sensitive and targeted molecular imaging in oncology.