Quantitatively Visualizing Tumor-Related Protease Activity in Vivo Using a Ratiometric Photoacoustic Probe

J Am Chem Soc. 2019 Feb 20;141(7):3265-3273. doi: 10.1021/jacs.8b13628. Epub 2019 Feb 8.


The abnormal expression of tumor-related proteases plays a critical role in cancer invasion, progression, and metastasis. Therefore, it is considerably meaningful to non-invasively assess the proteases' activity in vivo for both tumor diagnosis and therapeutic evaluation. Herein, we report an activatable probe constructed with a near-infrared dye (Cy5.5) and a quencher (QSY21) covalently linked through a peptide substrate of matrix metalloproteinases-2 (MMP-2) that was chosen as a model for tumor-associated proteases. Upon cleavage with activated MMP-2, this probe emitted an MMP-2-concentration-dependent fluorescence. Quite unexpectedly, owing to the variation in the aggregation state of both the dye and its quencher as a consequence of the cleavage, the responsive probe presented a dramatic MMP-2-concentration-dependent absorption at around 680 nm, while that at around 730 nm was MMP-2 concentration independent. These features allowed detection of MMP-2 activity via both fluorescence and photoacoustic (PA) imaging in vitro, respectively. Moreover, taking the PA signal at 730 nm as an internal reference, the PA signal at 680 nm allowed quantitative detection of MMP-2 expression in breast cancer in vivo. We thus envision that our current approach would offer a useful tool for studying the malignant impacts of versatile tumor-associated proteases in vivo.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Carbocyanines / chemistry
  • Cell Line, Tumor
  • Female
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Mammary Neoplasms, Animal / diagnostic imaging
  • Matrix Metalloproteinase 2 / analysis*
  • Mice, Inbred BALB C
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / methods
  • Nucleotides / chemistry
  • Photoacoustic Techniques / methods
  • Rhodamines / chemistry


  • CY5.5 cyanine dye
  • Carbocyanines
  • Fluorescent Dyes
  • Nucleotides
  • QSY 21 compound
  • Rhodamines
  • Matrix Metalloproteinase 2
  • Mmp2 protein, mouse