Noninvasive detection of breast cancer lymph node metastasis using carbonic anhydrases IX and XII targeted imaging probes

Clin Cancer Res. 2012 Jan 1;18(1):207-19. doi: 10.1158/1078-0432.CCR-11-0238. Epub 2011 Oct 20.

Abstract

Purpose: To develop targeted molecular imaging probes for the noninvasive detection of breast cancer lymph node metastasis.

Experimental design: Six cell surface or secreted markers were identified by expression profiling and from the literature as being highly expressed in breast cancer lymph node metastases. Two of these markers were cell surface carbonic anhydrase isozymes (CAIX and/or CAXII) and were validated for protein expression by immunohistochemistry of patient tissue samples on a breast cancer tissue microarray containing 47 normal breast tissue samples, 42 ductal carcinoma in situ, 43 invasive ductal carcinomas without metastasis, 46 invasive ductal carcinomas with metastasis, and 49 lymph node macrometastases of breast carcinoma. Targeted probes were developed by conjugation of CAIX- and CAXII-specific monoclonal antibodies to a near-infrared fluorescent dye.

Results: Together, these two markers were expressed in 100% of the lymph node metastases surveyed. Selectivity of the imaging probes were confirmed by intravenous injection into nude mice-bearing mammary fat pad tumors of marker-expressing cells and nonexpressing cells or by preinjection of unlabeled antibody. Imaging of lymph node metastases showed that peritumorally injected probes detected nodes harboring metastatic tumor cells. As few as 1,000 cells were detected, as determined by implanting, under ultrasound guidance, a range in number of CAIX- and CAXII-expressing cells into the axillary lymph nodes.

Conclusion: These imaging probes have potential for noninvasive staging of breast cancer in the clinic and elimination of unneeded surgery, which is costly and associated with morbidities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal* / immunology
  • Antibodies, Monoclonal* / pharmacokinetics
  • Antigens, Neoplasm / metabolism*
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism
  • Blotting, Western
  • Breast / immunology
  • Breast / metabolism
  • Breast / pathology
  • Breast Neoplasms / diagnosis*
  • Breast Neoplasms / immunology
  • Breast Neoplasms / metabolism
  • Carbonic Anhydrase IX
  • Carbonic Anhydrases / metabolism*
  • Carcinoma, Ductal, Breast / diagnosis*
  • Carcinoma, Ductal, Breast / immunology
  • Carcinoma, Ductal, Breast / metabolism
  • Carcinoma, Intraductal, Noninfiltrating / diagnosis*
  • Carcinoma, Intraductal, Noninfiltrating / immunology
  • Carcinoma, Intraductal, Noninfiltrating / metabolism
  • Diagnostic Imaging*
  • Female
  • Fluorescent Antibody Technique
  • Gene Expression Profiling
  • Humans
  • Luciferases / metabolism
  • Luminescent Measurements
  • Lymphatic Metastasis
  • Mice
  • Mice, Nude
  • Neoplasm Invasiveness
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Tissue Array Analysis
  • Tissue Distribution
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Antigens, Neoplasm
  • Biomarkers, Tumor
  • RNA, Messenger
  • Luciferases
  • CA9 protein, human
  • Carbonic Anhydrase IX
  • Carbonic Anhydrases
  • carbonic anhydrase XII