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Rationally Engineering a pH-Low Insertion Peptide Variant Probe for High-Sensitivity Imaging of Early Tumors.

Xiaoxuan Yu1, Jiantong Dong1, Hengwei Bian1

  • 1State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Molecular Recognition and Biosensing, Frontiers Science Center for New Organic Matter, College of Chemistry, Nankai University, Tianjin 300071, China.

ACS Applied Materials & Interfaces
|June 16, 2025
PubMed
Summary

A novel fluorescent probe, 4 M pHLIP-Cy5.5, effectively detects early-stage tumors by targeting their acidic microenvironment. This advancement offers highly sensitive in vivo imaging for improved cancer diagnosis.

Keywords:
In vivocancer diagnosisfluorescent probesmall tumortumor microenvironment

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Area of Science:

  • Biomedical Imaging
  • Oncology
  • Molecular Probes

Background:

  • Early-stage tumor detection is crucial but challenging in oncology.
  • The acidic tumor microenvironment presents a target for diagnostic tools.

Purpose of the Study:

  • To develop a highly sensitive fluorescent probe for early-stage tumor imaging.
  • To evaluate the efficacy of a novel pH-sensitive peptide probe in vivo.

Main Methods:

  • Development of a pH-low insertion peptide (pHLIP) variant, 4 M pHLIP-Cy5.5, conjugated with a fluorescent dye.
  • In vitro characterization of pH-dependent conformational changes and binding affinity.
  • In vivo imaging studies in a mouse model of breast cancer.

Main Results:

  • 4 M pHLIP-Cy5.5 demonstrated ultrasensitive pH-dependent fluorescence changes and enhanced cancer cell binding.
  • The probe detected small tumors (<20 mm³) with improved signal-to-background ratio compared to wild-type.
  • Prolonged tumor retention was observed, with fluorescence significantly higher than in healthy tissue at 72 hours post-injection.

Conclusions:

  • 4 M pHLIP-Cy5.5 enables high-sensitivity imaging of early-stage tumors.
  • The probe's pH sensitivity, rapid targeting, and retention offer a promising strategy for cancer diagnosis.
  • This fluorescent probe advances early tumor detection and monitoring capabilities.