Intratumoural expression of TNF‐R1 and EMAP‐II in relation to response of patients treated with TNF‐based isolated limb perfusion

Tumour necrosis factor‐alpha (TNF) has been used in the clinic for more than 10 years in an isolated limb perfusion (ILP). However, intra‐tumoural expression of TNF receptor‐1 (TNF‐R1) and TNF‐R1 upregulating factors are unknown. We determined the expression of TNF‐R1, proEMAP and endothelial monocyte‐activating polypeptide‐II (EMAP‐II) before and after ILP and evaluated this against clinical response. Tumour biopsies were taken before and after ILP of patients (n = 27) with advanced sarcoma or metastatic melanoma. Biopsies were randomly analysed by western blotting for proEMAP/EMAP‐II and TNF‐R1 expression. Appropriate melanoma biopsies were stained for EMAP‐II, TNF‐R1, CD31 and CD68. For melanomas we found that an up‐regulation of EMAP‐II, in contrast to proEMAP or TNF‐R1, directly after ILP significantly correlated with a complete tumour response. No correlation was found for sarcoma patients. In a comparative analysis we found that the overall proEMAP and EMAP‐II expression was higher in melanoma as compared to sarcoma cases and measurements in cell lines revealed high proEMAP expression by melanoma cells. We report high EMAP‐II expression by endothelial cells and association with macrophages. In addition, macrophages are recruited to vessel‐remnants after ILP. An upregulation of EMAP‐II directly after ILP of melanoma patients correlates with and might predict a complete response to TNF‐based ILP. The association of macrophages with EMAP‐II expression and vascular damage suggests a role for EMAP‐II in regulating the TNF‐based anti‐tumour effects observed with an ILP. Analysis of EMAP‐II expression in melanoma biopsies should be implemented in the ILP procedure. © 2006 Wiley‐Liss, Inc.

[1]  S. Libutti,et al.  Endothelial monocyte activating polypeptide-II induced gene expression changes in endothelial cells. , 2005, Cytokine.

[2]  Sunghoon Kim,et al.  The novel cytokine p43 stimulates dermal fibroblast proliferation and wound repair. , 2005, The American journal of pathology.

[3]  A. Eggermont,et al.  One Hundred Consecutive Isolated Limb Perfusions With TNF-α and Melphalan in Melanoma Patients With Multiple In-Transit Metastases , 2004, Annals of surgery.

[4]  M. Sarker,et al.  SPOTS: signaling protein oligomeric transduction structures are early mediators of death receptor-induced apoptosis at the plasma membrane. , 2004, The Journal of cell biology.

[5]  P. Symonds,et al.  Endothelial monocyte‐activating polypeptide‐II (EMAP‐II): a novel inducer of lymphocyte apoptosis , 2004, Journal of leukocyte biology.

[6]  P. Symonds,et al.  Colorectal Cancer Cells Induce Lymphocyte Apoptosis by an Endothelial Monocyte-Activating Polypeptide-II-Dependent Mechanism 1 , 2004, The Journal of Immunology.

[7]  A. Eggermont,et al.  Current uses of isolated limb perfusion in the clinic and a model system for new strategies. , 2003, The Lancet. Oncology.

[8]  D. Ruiter,et al.  EMAP-II expression is associated with macrophage accumulation in primary uveal melanoma. , 2003, Investigative ophthalmology & visual science.

[9]  Noam Brown,et al.  The role of tumour‐associated macrophages in tumour progression: implications for new anticancer therapies , 2002, The Journal of pathology.

[10]  P. Vandenabeele,et al.  The EMAPII Cytokine Is Released from the Mammalian Multisynthetase Complex after Cleavage of Its p43/proEMAPII Component* , 2001, The Journal of Biological Chemistry.

[11]  D. Powe,et al.  Immunohistochemical analysis of endothelial-monocyte-activating polypeptide-II expression in vivo. , 2000, The American journal of pathology.

[12]  S. Libutti,et al.  Endothelial monocyte-activating polypeptide II, a tumor-derived cytokine that plays an important role in inflammation, apoptosis, and angiogenesis. , 2000, Journal of immunotherapy.

[13]  S. Libutti,et al.  Tumour necrosis factor receptor I (p55) is upregulated on endothelial cells by exposure to the tumour-derived cytokine endothelial monocyte- activating polypeptide II (EMAP-II). , 2000, Cytokine.

[14]  J. Carmichael,et al.  Prostate adenocarcinoma cells release the novel proinflammatory polypeptide EMAP-II in response to stress. , 2000, Cancer research.

[15]  S. Libutti,et al.  Sensitization of tumor necrosis factor alpha-resistant human melanoma by tumor-specific in vivo transfer of the gene encoding endothelial monocyte-activating polypeptide II using recombinant vaccinia virus. , 1999, Cancer research.

[16]  Jun Li,et al.  Endothelial-Monocyte Activating Polypeptide Ii, a Novel Antitumor Cytokine That Suppresses Primary and Metastatic Tumor Growth and Induces Apoptosis in Growing Endothelial Cells , 1999, The Journal of experimental medicine.

[17]  S. Jones,et al.  TNF recruits TRADD to the plasma membrane but not the trans-Golgi network, the principal subcellular location of TNF-R1. , 1999, Journal of immunology.

[18]  A. Cope,et al.  Shedding kinetics of soluble tumor necrosis factor (TNF) receptors after systemic TNF leaking during isolated limb perfusion. Relevance to the pathophysiology of septic shock. , 1998, The Journal of clinical investigation.

[19]  A. Eggermont,et al.  Systemic release of soluble TNF receptors after high-dose TNF in isolated limb perfusion. , 1997, Cytokine.

[20]  P. Schlag,et al.  Isolated limb perfusion with tumor necrosis factor and melphalan for limb salvage in 186 patients with locally advanced soft tissue extremity sarcomas. The cumulative multicenter European experience. , 1996, Annals of surgery.

[21]  A. Eggermont,et al.  Isolated limb perfusion with high-dose tumor necrosis factor-alpha in combination with interferon-gamma and melphalan for nonresectable extremity soft tissue sarcomas: a multicenter trial. , 1996, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  D. Fraker,et al.  Treatment of patients with melanoma of the extremity using hyperthermic isolated limb perfusion with melphalan, tumor necrosis factor, and interferon gamma: results of a tumor necrosis factor dose-escalation study. , 1996, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[23]  S. Libutti,et al.  Characterization of a novel tumor-derived cytokine. Endothelial-monocyte activating polypeptide II. , 1994, The Journal of biological chemistry.

[24]  A. Eggermont,et al.  Rationale for using TNFα and chemotherapy in regional therapy of melanoma , 1994 .

[25]  W. Kisiel,et al.  A peptide derived from the amino terminus of endothelial-monocyte-activating polypeptide II modulates mononuclear and polymorphonuclear leukocyte functions, defines an apparently novel cellular interaction site, and induces an acute inflammatory response. , 1994, The Journal of biological chemistry.

[26]  P. Familletti,et al.  Endothelial monocyte-activating polypeptide II. A novel tumor-derived polypeptide that activates host-response mechanisms. , 1992, The Journal of biological chemistry.

[27]  J. Denekamp,et al.  Selective induction of endothelial cell tissue factor in the presence of a tumour‐derived mediator: A potential mechanism of flavone acetic acid action in tumour vasculature , 1991, International journal of cancer.

[28]  P. Familletti,et al.  A polypeptide factor produced by fibrosarcoma cells that induces endothelial tissue factor and enhances the procoagulant response to tumor necrosis factor/cachectin. , 1990, The Journal of biological chemistry.

[29]  M. Palmer,et al.  WHO Handbook for Reporting Results of Cancer Treatment , 1982, British Journal of Cancer.

[30]  S. Libutti,et al.  Improved antitumor response to isolated limb perfusion with tumor necrosis factor after upregulation of endothelial monocyte-activating polypeptide II in soft tissue sarcoma , 2007, Annals of Surgical Oncology.

[31]  S. Libutti,et al.  In vivo sensitivity of human melanoma to tumor necrosis factor (TNF)-alpha is determined by tumor production of the novel cytokine endothelial-monocyte activating polypeptide II (EMAPII). , 1999, Cancer research.

[32]  C. Wiltschke,et al.  Isolated limb perfusion with high-dose tumor necrosis factor-alpha in combination with interferon-gamma and melphalan for nonresectable extremity soft tissue sarcomas: a multicenter trial. , 1997, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[33]  J. Murray,et al.  Endothelial-monocyte-activating polypeptide II. , 1996, The international journal of biochemistry & cell biology.

[34]  R. D. Hunter,et al.  WHO Handbook for Reporting Results of Cancer Treatment , 1980 .