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Area of Science:

  • Biochemistry
  • Gastroenterology
  • Oncology

Background:

  • Advanced adenomas (AA) and colorectal cancer (CRC) screening is crucial for reducing incidence and mortality.
  • Current fecal immunochemical testing has limitations in sensitivity for AA and high false-positivity rates.
  • Proteases are implicated in CRC development via angiogenesis and immunoregulation.

Purpose of the Study:

  • To identify proteases and their substrates associated with colorectal neoplasia for a novel noninvasive screening test.
  • To apply fecal protease profiling, an innovative approach previously limited to tissue samples.

Main Methods:

  • Eighteen fluorogenic substrates were designed and tested in fecal samples from patients with CRC, AA, nonadvanced adenomas, and controls.
  • Substrate degradation was correlated with proteome data to identify proteases.
  • Protease inhibitors and ZnCl2 were used for further characterization.

Main Results:

  • Seven of 18 substrates showed significantly decreased proteolytic degradation in patients with colorectal neoplasia.
  • The l-aspartic acid-l-glutamic acid (ED) substrate exhibited significantly decreased degradation in AA and CRC patients.
  • ED degradation was further reduced by ZnCl2 and the cysteine protease inhibitor NEM.

Conclusions:

  • Colorectal neoplasia-specific fluorogenic substrates were successfully developed.
  • The ED substrate shows potential for detecting advanced adenomas and colorectal cancer.
  • Results suggest a link to calcium-dependent cysteine proteases, though further identification is needed.