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Updated: May 21, 2025

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Detecting microsatellite instability in cancer via multiplexed orthogonal gap-enhanced Raman tags.

Guowei Fu1,2, Jin Li2, Qian Zhang3

  • 1Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Pharmacy and Food Engineering, Wuyi University 529020 Jiangmen China wyuchemwrh@126.com.

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Summary

This study introduces a novel method for detecting four key mismatch repair (MMR) proteins simultaneously in cancer tissues, crucial for identifying microsatellite instability (MSI) phenotypes and guiding immunotherapy.

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Area of Science:

  • Biomedical Engineering
  • Molecular Oncology
  • Spectroscopy

Background:

  • Microsatellite instability (MSI) is a critical biomarker in cancer, particularly colorectal cancer, influencing immunotherapy response.
  • Accurate MSI phenotyping relies on the interdependent expression of four mismatch repair (MMR) proteins: MLH1, MSH2, MSH6, and PMS2.
  • Current detection methods often lack the ability for simultaneous, high-resolution analysis of these interdependent proteins.

Purpose of the Study:

  • To develop a sensitive and specific method for simultaneous detection of four MMR proteins in cancer tissues.
  • To classify cancer subtypes based on microsatellite stable (MSS) or high-frequency MSI (MSI-H) status using quantitative Raman imaging.
  • To establish correlations between MMR protein expression and clinicopathologic parameters, and explore MSI's role in Lynch syndrome screening and immunotherapy prediction.

Main Methods:

  • Fabrication of four orthogonal gap-enhanced Raman tag (O-GERT) flavors with distinct, interference-free spectral readouts.
  • Antibody functionalization of O-GERTs for specific, one-shot detection of MLH1, MSH2, MSH6, and PMS2 proteins.
  • Quantitative Raman imaging for classifying cancer tissues into MSS or MSI-H subtypes and analyzing MMR protein expression.

Main Results:

  • Achieved high sensitivity, photostability, and specificity in detecting the four MMR proteins.
  • Successfully classified colorectal, breast, and gastric cancer tissues into MSS or MSI-H subtypes with detected MSI-H ratios of 13.3%, 6.7%, and 3.3%, respectively.
  • Established novel correlations between MMR protein expression and clinicopathologic parameters in colorectal cancer subtypes.

Conclusions:

  • Quantitative multiplexed Raman imaging offers a fast and accurate approach for MSI detection in various cancers.
  • This technique facilitates MSI phenotyping, aiding in Lynch syndrome screening and identifying candidates for PD1/PD-L1 blockade immunotherapy.
  • The simultaneous detection of interdependent MMR proteins provides a more comprehensive understanding of MSI status in cancer.