Circulating proteolytic products of carboxypeptidase N for early detection of breast cancer

Yaojun Li, Yueguo Li, Tao Chen, Anna S. Kuklina, Paul Bernard, Francisco J. Esteva, Haifa Shen, Mauro Ferrari, Ye Hu

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

BACKGROUND: CarboxypeptidaseN(CPN) is important in regulating vasoactive peptide hormones, growth factors, and cytokines by specifically cleaving their C-terminal basic residues. We investigated whether circulating peptides specifically cleaved by CPN in the tumor microenvironment can be stage-specific indicators of breast cancer. METHODS: CPN activity was measured using an ex vivo peptide cleavage assay by incubating synthesized C3f peptide (His 6-C3f-S1304-R1320-His6) in interstitial fluids of breast tumors and adjacent normal breast tissues in mice with orthotopic implantation of the human cell line MDA-MB-231. The nature and extent of peptide cleavage by CPN was investigated by fragment profiling using nanopore fractionation and mass spectrometry. The fragment profiles in interstitial fluid correlated with concentrations of CPN-catalyzed peptides in blood samples taken from the tumor-bearing mice, healthy women, and breast cancer patients. CPN expression in the same set of samples was further examined by immunohistochemistry and immunoblotting. RESULTS: We showed that generation of C3f-R1310- L1319 specifically correlated with the CPN expression level. In both the mouse and clinical patient samples, CPN was clearly increased in tumor tissues compared with normal breast tissue, whereas corresponding CPN abundance in blood remained constant. Concentrations of 6 CPN-catalyzed peptides predominantly increased in sera taken from the mice (n = 8) at 2 weeks after orthotopic implantation. Six homologous peptides displayed significantly higher expression in the patients' plasma as early as the first pathologic stage of breast cancer. CONCLUSIONS: Circulating CPN-catalyzed peptide concentrations reflect the CPN activity in tumors. These biomarkers show strong potential for the noninvasive and early diagnosis of breast cancer.

Original languageEnglish (US)
Pages (from-to)233-242
Number of pages10
JournalClinical Chemistry
Volume60
Issue number1
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

Fingerprint

Lysine Carboxypeptidase
Early Detection of Cancer
Breast Neoplasms
Peptides
Tumors
His-His-His-His-His-His
Extracellular Fluid
Tissue
Breast
Blood
Bearings (structural)
Nanopores
Neoplasms
Tumor Microenvironment
Fluids
Peptide Hormones
Biomarkers
Fractionation
Immunoblotting
Mass spectrometry

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Circulating proteolytic products of carboxypeptidase N for early detection of breast cancer. / Li, Yaojun; Li, Yueguo; Chen, Tao; Kuklina, Anna S.; Bernard, Paul; Esteva, Francisco J.; Shen, Haifa; Ferrari, Mauro; Hu, Ye.

In: Clinical Chemistry, Vol. 60, No. 1, 01.01.2014, p. 233-242.

Research output: Contribution to journalArticle

Li, Y, Li, Y, Chen, T, Kuklina, AS, Bernard, P, Esteva, FJ, Shen, H, Ferrari, M & Hu, Y 2014, 'Circulating proteolytic products of carboxypeptidase N for early detection of breast cancer' Clinical Chemistry, vol. 60, no. 1, pp. 233-242. https://doi.org/10.1373/clinchem.2013.211953
Li, Yaojun ; Li, Yueguo ; Chen, Tao ; Kuklina, Anna S. ; Bernard, Paul ; Esteva, Francisco J. ; Shen, Haifa ; Ferrari, Mauro ; Hu, Ye. / Circulating proteolytic products of carboxypeptidase N for early detection of breast cancer. In: Clinical Chemistry. 2014 ; Vol. 60, No. 1. pp. 233-242.
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AU - Bernard, Paul

AU - Esteva, Francisco J.

AU - Shen, Haifa

AU - Ferrari, Mauro

AU - Hu, Ye

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N2 - BACKGROUND: CarboxypeptidaseN(CPN) is important in regulating vasoactive peptide hormones, growth factors, and cytokines by specifically cleaving their C-terminal basic residues. We investigated whether circulating peptides specifically cleaved by CPN in the tumor microenvironment can be stage-specific indicators of breast cancer. METHODS: CPN activity was measured using an ex vivo peptide cleavage assay by incubating synthesized C3f peptide (His 6-C3f-S1304-R1320-His6) in interstitial fluids of breast tumors and adjacent normal breast tissues in mice with orthotopic implantation of the human cell line MDA-MB-231. The nature and extent of peptide cleavage by CPN was investigated by fragment profiling using nanopore fractionation and mass spectrometry. The fragment profiles in interstitial fluid correlated with concentrations of CPN-catalyzed peptides in blood samples taken from the tumor-bearing mice, healthy women, and breast cancer patients. CPN expression in the same set of samples was further examined by immunohistochemistry and immunoblotting. RESULTS: We showed that generation of C3f-R1310- L1319 specifically correlated with the CPN expression level. In both the mouse and clinical patient samples, CPN was clearly increased in tumor tissues compared with normal breast tissue, whereas corresponding CPN abundance in blood remained constant. Concentrations of 6 CPN-catalyzed peptides predominantly increased in sera taken from the mice (n = 8) at 2 weeks after orthotopic implantation. Six homologous peptides displayed significantly higher expression in the patients' plasma as early as the first pathologic stage of breast cancer. CONCLUSIONS: Circulating CPN-catalyzed peptide concentrations reflect the CPN activity in tumors. These biomarkers show strong potential for the noninvasive and early diagnosis of breast cancer.

AB - BACKGROUND: CarboxypeptidaseN(CPN) is important in regulating vasoactive peptide hormones, growth factors, and cytokines by specifically cleaving their C-terminal basic residues. We investigated whether circulating peptides specifically cleaved by CPN in the tumor microenvironment can be stage-specific indicators of breast cancer. METHODS: CPN activity was measured using an ex vivo peptide cleavage assay by incubating synthesized C3f peptide (His 6-C3f-S1304-R1320-His6) in interstitial fluids of breast tumors and adjacent normal breast tissues in mice with orthotopic implantation of the human cell line MDA-MB-231. The nature and extent of peptide cleavage by CPN was investigated by fragment profiling using nanopore fractionation and mass spectrometry. The fragment profiles in interstitial fluid correlated with concentrations of CPN-catalyzed peptides in blood samples taken from the tumor-bearing mice, healthy women, and breast cancer patients. CPN expression in the same set of samples was further examined by immunohistochemistry and immunoblotting. RESULTS: We showed that generation of C3f-R1310- L1319 specifically correlated with the CPN expression level. In both the mouse and clinical patient samples, CPN was clearly increased in tumor tissues compared with normal breast tissue, whereas corresponding CPN abundance in blood remained constant. Concentrations of 6 CPN-catalyzed peptides predominantly increased in sera taken from the mice (n = 8) at 2 weeks after orthotopic implantation. Six homologous peptides displayed significantly higher expression in the patients' plasma as early as the first pathologic stage of breast cancer. CONCLUSIONS: Circulating CPN-catalyzed peptide concentrations reflect the CPN activity in tumors. These biomarkers show strong potential for the noninvasive and early diagnosis of breast cancer.

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