Soluble aminopeptidase N/CD13 in malignant and nonmalignant effusions and intratumoral fluid.

PURPOSE On the basis of the finding of marked overexpression in angiogenic microvessels, aminopeptidase N/CD13 has recently been suggested to play a prominent role in tumor angiogenesis. A soluble form of CD13 (sCD13) is present in human plasma, but its role in cancer has not been addressed. We hypothesized that sCD13 would be shed by tumor cells and/or endothelial cells lining tumor vessels, giving high levels of sCD13 in intratumoral fluid (TF) deposits and in malignant effusions. If so, sCD13 could be a convenient potential marker for tumor load and/or activated tumor endothelium. EXPERIMENTAL DESIGN We have measured the specific sCD13 activity in effusions from 90 cancer patients and 12 patients with a nonmalignant condition, and studied its relationship with other major (anti-)angiogenic factors. In a separate group of patients (n = 41), the relationship of sCD13 activity in plasma with tumor load was studied. RESULTS The sCD13 activity was highest in plasma from cancer patients 71.9 (fmol/ml/s hydrolyzed substrate) versus 42.4 for healthy subjects. In TF, malignant effusions, and nonmalignant effusions, the activities were 52.8, 33.5, and 18.6, respectively. We further studied the relationship of sCD13 with tumor load as well as with vascular endothelial growth factor (VEGF), endostatin, matrix metalloproteinase (MMP)-2, MMP-9, urokinase-type plasminogen activator, and plasmin. A significant correlation of sCD13 activity in plasma was found with tumor load (r = 0.68; P = 0.01), suggesting that plasma sCD13 is, at least, partly originating from tumor(-endothelium). The concentrations of VEGF and endostatin and the activities of urokinase-type plasminogen activator and MMP-9, but not MMP-2, were significantly higher in TF compared with all other effusions. In TF, a correlation between sCD13 and VEGF was found (r = 0.67; P = 0.03). No correlation of sCD13 with the other protease activities was found. CONCLUSION The sCD13 activity is elevated in plasma and effusions of cancer patients. A strong correlation of plasma sCD13 with tumor load was found. On the basis of these results, the potential of sCD13 activity as a tumor and/or angiogenesis marker warrants further investigation.

[1]  E. Favaloro,et al.  The hepatobiliary disease marker serum alanine aminopeptidase predominantly comprises an isoform of the haematological myeloid differentiation antigen and leukaemia marker CD-13/gp150. , 1993, Clinica chimica acta; international journal of clinical chemistry.

[2]  P. Koolwijk,et al.  A novel and simple immunocapture assay for determination of gelatinase-B (MMP-9) activities in biological fluids: saliva from patients with Sjögren's syndrome contain increased latent and active gelatinase-B levels. , 1998, Matrix biology : journal of the International Society for Matrix Biology.

[3]  N. Ferrara,et al.  The biology of vascular endothelial growth factor. , 1997, Endocrine reviews.

[4]  H. Dvorak,et al.  Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. , 1995, The American journal of pathology.

[5]  J. Verheijen,et al.  Determination of Gelatinase‐A (MMP‐2) Activity Using a Novel Immunocapture Assay , 1999, Annals of the New York Academy of Sciences.

[6]  R Pasqualini,et al.  Aminopeptidase N is a receptor for tumor-homing peptides and a target for inhibiting angiogenesis. , 2000, Cancer research.

[7]  M. Dewhirst,et al.  Available volume fraction of macromolecules in the extravascular space of a fibrosarcoma: implications for drug delivery. , 1999, Cancer research.

[8]  J. Richardson,et al.  Diagnosis and management of malignant pleural effusions. , 1995, American journal of surgery.

[9]  R K Jain,et al.  Vascular permeability in a human tumor xenograft: molecular size dependence and cutoff size. , 1995, Cancer research.

[10]  S. Zucker,et al.  Human pleural effusions are rich in matrix metalloproteinases. , 1992, Chest.

[11]  R. Light,et al.  Vascular endothelial growth factor in pleural fluid. , 1999, Chest.

[12]  H. Verheul,et al.  Platelet and coagulation activation with vascular endothelial growth factor generation in soft tissue sarcomas. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[13]  A. Feldman,et al.  Serum endostatin levels are elevated and correlate with serum vascular endothelial growth factor levels in patients with stage IV clear cell renal cancer. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[14]  T. Tsuruo,et al.  Role of aminopeptidase N (CD13) in tumor‐cell invasion and extracellular matrix degradation , 1993, International journal of cancer.

[15]  A. Harris,et al.  A phase II study of razoxane, an antiangiogenic topoisomerase II inhibitor, in renal cell cancer with assessment of potential surrogate markers of angiogenesis. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[16]  V. V. van Hinsbergh,et al.  Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity , 1996, The Journal of cell biology.

[17]  W. Arap,et al.  CD13/APN is activated by angiogenic signals and is essential for capillary tube formation. , 2001, Blood.

[18]  T. Tsuruo,et al.  Human melanoma invasion and metastasis enhancement by high expression of aminopeptidase N/CD13 , 1995, Clinical & Experimental Metastasis.

[19]  E. Favaloro,et al.  CD13 (GP150; aminopeptidase-N): predominant functional activity in blood is localized to plasma and is not cell-surface associated. , 1993, Experimental hematology.

[20]  Gordon B. Mills,et al.  Markedly Elevated Levels of Vascular Endothelial Growth Factor in Malignant Ascites , 1999, Annals of Surgical Oncology.

[21]  A. Harris,et al.  Expression of aminopeptidase-n (CD 13) in normal tissues and malignant neoplasms of epithelial and lymphoid origin. , 1994, Journal of clinical pathology.

[22]  A. Harris,et al.  Phase I study of intrapleural batimastat (BB-94), a matrix metalloproteinase inhibitor, in the treatment of malignant pleural effusions. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[23]  D. Marmé,et al.  Vascular endothelial growth factor in the sera and effusions of patients with malignant and nonmalignant disease , 1999, Cancer.

[24]  D. Thickett,et al.  Vascular endothelial growth factor (VEGF) in inflammatory and malignant pleural effusions , 1999, Thorax.

[25]  R K Jain,et al.  Augmentation of transvascular transport of macromolecules and nanoparticles in tumors using vascular endothelial growth factor. , 1999, Cancer research.

[26]  T. Salo,et al.  The levels of trypsinogen isoenzymes in ovarian tumour cyst fluids are associated with promatrix metalloproteinase-9 but not promatrix metalloproteinase-2 activation , 2001, British Journal of Cancer.

[27]  F. Reichenberger,et al.  MMP and TIMP expression pattern in pleural effusions of different origins. , 1997, American journal of respiratory and critical care medicine.

[28]  D. Sane,et al.  Vascular Origin of a Soluble Truncated Form of the Hepatocyte Growth Factor Receptor (c-met) , 2002, Circulation research.

[29]  William Arbuthnot Sir Lane,et al.  Endostatin: An Endogenous Inhibitor of Angiogenesis and Tumor Growth , 1997, Cell.

[30]  J. Riordan,et al.  Membrane proteins with soluble counterparts: role of proteolysis in the release of transmembrane proteins. , 1991, Biochemistry.

[31]  V. Gerkis,et al.  Variable O-glycosylation of CD13 (aminopeptidase N). , 1991, The Journal of biological chemistry.

[32]  J. Newcombe,et al.  Plasminogen activators in multiple sclerosis lesions: implications for the inflammatory response and axonal damage. , 2001, Brain : a journal of neurology.

[33]  S. Sahn.  Pleural diseases related to metastatic malignancies. , 1997, The European respiratory journal.

[34]  D. Hicklin,et al.  Vascular endothelial growth factor levels and induction of permeability in malignant pleural effusions. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[35]  N. Sakuragi,et al.  Expression of matrix metalloproteinases and related tissue inhibitors in the cyst fluids of ovarian mucinous neoplasms. , 2000, Gynecologic oncology.

[36]  V. Bertone,et al.  Tumor interstitial fluid: misconsidered component of the internal milieu of a solid tumor. , 1997, Anticancer research.

[37]  E. Favaloro.  CD‐13 (‘gp150’; aminopeptidase‐N): Co‐expression on endothelial and haemopoietic cells with conservation of functional activity , 1991, Immunology and cell biology.

[38]  H. Höfler,et al.  Ovarian Prognostic Significance of Urokinase (upa) and Its Inhibitor Pai-1 for Survival in Advanced Ovarian Carcinoma Stage Figo Iiic , 2022 .

[39]  R. Ashmun,et al.  Human myeloid plasma membrane glycoprotein CD13 (gp150) is identical to aminopeptidase N. , 1989, The Journal of clinical investigation.

[40]  S. Nicosia,et al.  Vascular endothelial growth factor levels in ovarian cyst fluid correlate with malignancy. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.