title: Expression of tissue factor in adenocarcinoma and squamous cell carcinoma of the uterine cervix: implications for immunotherapy with hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor.
abstract: BACKGROUND
  Cervical cancer continues to be an important worldwide health problem for women. Up to 35% of patients who are diagnosed with and appropriately treated for cervical cancer will recur and treatment results are poor for recurrent disease. Given these sobering statistics, development of novel therapies for cervical cancer remains a high priority. We evaluated the expression of Tissue Factor (TF) in cervical cancer and the potential of hI-con1, an antibody-like-molecule targeted against TF, as a novel form of immunotherapy against multiple primary cervical carcinoma cell lines with squamous- and adenocarcinoma histology.
  
  
  METHODS
  Because TF is a transmembrane receptor for coagulation factor VII/VIIa (fVII), in this study we evaluated the in vitro expression of TF in cervical carcinoma cell lines by immunohistochemistry (IHC), real time-PCR (qRT-PCR) and flow cytometry. Sensitivity to hI-con1-dependent cell-mediated-cytotoxicity (IDCC) was evaluated in 5-hrs-51chromium-release-assays against cervical cancer cell lines in vitro.
  
  
  RESULTS
  Cytoplasmic and/or membrane TF expression was observed in 8 out of 8 (100%) of the tumor tissues tested by IHC and in 100% (11 out of 11) of the cervical carcinoma cell lines tested by real-time-PCR and flow cytometry but not in normal cervical keratinocytes (p=0.0023 qRT-PCR; p=0.0042 flow cytometry). All primary cervical cancer cell lines tested overexpressing TF, regardless of their histology, were highly sensitive to IDCC (mean killing±SD, 56.2%±15.9%, range, 32.4%-76.9%, p<0.001), while negligible cytotoxicity was seen in the absence of hI-con1 or in the presence of rituximab-control-antibody. Low doses of interleukin-2 further increased the cytotoxic effect induced by hI-con1 (p=0.025) while human serum did not significantly decrease IDCC against cervical cancer cell lines (p=0.597).
  
  
  CONCLUSIONS
  TF is highly expressed in squamous and adenocarcinoma of the uterine cervix. hI-con1 induces strong cytotoxicity against primary cervical cancer cell lines overexpressing TF and may represent a novel therapeutic agent for the treatment of cervical cancer refractory to standard treatment modalities.
authors.affiliation: Department of Obstetrics, Gynecology & Reproductive Sciences, Yale University School of Medicine, 333 Cedar street, New Haven, CT 06519, USA.
authors.forename: Emiliano
authors.initials: E
authors.lastname: Cocco
authors.forename: Joyce
authors.initials: J
authors.lastname: Varughese
authors.forename: Natalia
authors.initials: N
authors.lastname: Buza
authors.forename: Stefania
authors.initials: S
authors.lastname: Bellone
authors.forename: Michelle
authors.initials: M
authors.lastname: Glasgow
authors.forename: Marta
authors.initials: M
authors.lastname: Bellone
authors.forename: Paola
authors.initials: P
authors.lastname: Todeschini
authors.forename: Luisa
authors.initials: L
authors.lastname: Carrara
authors.forename: Dan-Arin
authors.initials: DA
authors.lastname: Silasi
authors.forename: Masoud
authors.initials: M
authors.lastname: Azodi
authors.forename: Peter E
authors.initials: PE
authors.lastname: Schwartz
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Rutherford
authors.forename: Sergio
authors.initials: S
authors.lastname: Pecorelli
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Lockwood
authors.forename: Alessandro D
authors.initials: AD
authors.lastname: Santin
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy* / Q000821:virology; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy* / Q000821:virology; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism; D003165:Complement System Proteins / Q000494:pharmacology; D003601:Cytotoxicity Tests, Immunologic; D004354:Drug Screening Assays, Antitumor; D005260:Female; D052162:Human papillomavirus 16 / Q000302:isolation & purification; D052161:Human papillomavirus 18 / Q000302:isolation & purification; D006801:Humans; D018796:Immunoconjugates / Q000494:pharmacology / Q000627:therapeutic use*; D007074:Immunoglobulin G / Q000494:pharmacology; D007167:Immunotherapy; D007376:Interleukin-2 / Q000494:pharmacology; D015603:Keratinocytes / Q000378:metabolism; D058990:Molecular Targeted Therapy; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000188:drug therapy; D030361:Papillomavirus Infections / Q000821:virology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D013925:Thromboplastin / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy* / Q000821:virology
chemical_list: D018796:Immunoconjugates; D007074:Immunoglobulin G; D007376:Interleukin-2; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C562041:hI-con1; D003165:Complement System Proteins; D013925:Thromboplastin