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hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor for immunotherapy of uterine serous papillary carcinoma.

Cocco E, Hu Z, Richter CE, Bellone S, Casagrande F, Bellone M, Todeschini P, Krikun G, Silasi DA, Azodi M, Schwartz PE, Rutherford TJ, Buza N, Pecorelli S, Lockwood CJ, Santin AD - Br. J. Cancer (2010)

Bottom Line: Cytoplasmic and/or membrane TF expression was observed in all 16 (100%) USPC samples tested by IHC, but not in normal endometrium.The addition of low doses of IL-2 further increased the cytotoxic effect induced by hI-con1 against chemotherapy-resistant USPC. hI-con1 induces strong cytotoxicity against primary chemotherapy-resistant USPC cell lines overexpressing TF.The hI-con1 may represent a novel therapeutic agent for the treatment of patients harbouring advanced, recurrent and/or metastatic USPC refractory to standard treatment modalities.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, Room 305 LSOG, 333 Cedar Street, PO Box 208063, New Haven, CT 06520-8063, USA.

ABSTRACT

Background: Uterine serous papillary adenocarcinoma (USPC) is a highly aggressive variant of endometrial cancer. Human immuno-conjugate molecule (hI-con1) is an antibody-like molecule targeted against tissue factor (TF), composed of two human Factor VII (fVII) as the targeting domain, fused to human immunoglobulin (Ig) G1 Fc as an effector domain. We evaluated hI-con1 potential activity against primary chemotherapy-resistant USPC cell lines expressing different levels of TF.

Methods: A total of 16 formalin-fixed, paraffin-embedded USPC samples were evaluated by immunohistochemistry (IHC) for TF expression. Six primary USPC cell lines, half of which overexpress the epidermal growth factor type II (HER2/neu) receptor at 3+ levels, were assessed by flow cytometry and real-time PCR for TF expression. Sensitivity to hI-con1-dependent cell-mediated cytotoxicity (IDCC) was evaluated in 5-hour-chromium release assays. Finally, to investigate the effect of interleukin-2 (IL-2) on IDCC, 5-h (51)Cr assays were also conducted in the presence of low doses of IL-2 (i.e., 50-100 IU ml(-1)).

Results: Cytoplasmic and/or membrane TF expression was observed in all 16 (100%) USPC samples tested by IHC, but not in normal endometrium. High expression of TF was found in 50% (three out of six) of the USPC cell lines tested by real-time PCR and flow cytometry when compared with normal endometrial cells (NECs; P<0.001). Uterine serous papillary adenocarcinoma cell lines overexpressing TF, regardless of their high or low HER2/neu expression, were highly sensitive to IDCC (mean killing+/-s.d., 65.6+/-3.7%, range 57.5-77.0%, P<0.001), although negligible cytotoxicity against USPC was seen in the absence of hI-con1 or in the presence of Rituximab control antibody. The addition of low doses of IL-2 further increased the cytotoxic effect induced by hI-con1 against chemotherapy-resistant USPC.

Conclusion: hI-con1 induces strong cytotoxicity against primary chemotherapy-resistant USPC cell lines overexpressing TF. The hI-con1 may represent a novel therapeutic agent for the treatment of patients harbouring advanced, recurrent and/or metastatic USPC refractory to standard treatment modalities.

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Related in: MedlinePlus

Representative immunohistochemistry localisation analyses of tissue factor (TF) in uterine serous papillary adenocarcinoma (USPC) specimens. Upper left panel: normal endometrium, which is negative for TF. Lower left panel: USPC specimen showing cytoplasmic expression of TF. Upper right panel: USPC specimen showing membrane expression of TF. Lower right panel: USPC specimen showing both cytoplasmic and membrane expressions of TF. Original magnification: × 200.
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fig1: Representative immunohistochemistry localisation analyses of tissue factor (TF) in uterine serous papillary adenocarcinoma (USPC) specimens. Upper left panel: normal endometrium, which is negative for TF. Lower left panel: USPC specimen showing cytoplasmic expression of TF. Upper right panel: USPC specimen showing membrane expression of TF. Lower right panel: USPC specimen showing both cytoplasmic and membrane expressions of TF. Original magnification: × 200.

Mentions: We performed immunohistochemical analysis of TF protein expression on formalin-fixed tumour tissues from 16 paraffin-embedded uterine serous carcinoma specimens. As representatively shown in Figure 1, with no exception, all USPC samples that were tested showed either membrane and/or cytoplasmic immunoreactivity for TF (i.e., all 16 samples=100%), whereas the non-neoplastic endometrial controls were negative. The staining intensity was focal (1+) in nine cases, moderately positive (2+) in four cases and strongly positive (3+) in two cases.


hI-con1, a factor VII-IgGFc chimeric protein targeting tissue factor for immunotherapy of uterine serous papillary carcinoma.

Cocco E, Hu Z, Richter CE, Bellone S, Casagrande F, Bellone M, Todeschini P, Krikun G, Silasi DA, Azodi M, Schwartz PE, Rutherford TJ, Buza N, Pecorelli S, Lockwood CJ, Santin AD - Br. J. Cancer (2010)

Representative immunohistochemistry localisation analyses of tissue factor (TF) in uterine serous papillary adenocarcinoma (USPC) specimens. Upper left panel: normal endometrium, which is negative for TF. Lower left panel: USPC specimen showing cytoplasmic expression of TF. Upper right panel: USPC specimen showing membrane expression of TF. Lower right panel: USPC specimen showing both cytoplasmic and membrane expressions of TF. Original magnification: × 200.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2966612&req=5

fig1: Representative immunohistochemistry localisation analyses of tissue factor (TF) in uterine serous papillary adenocarcinoma (USPC) specimens. Upper left panel: normal endometrium, which is negative for TF. Lower left panel: USPC specimen showing cytoplasmic expression of TF. Upper right panel: USPC specimen showing membrane expression of TF. Lower right panel: USPC specimen showing both cytoplasmic and membrane expressions of TF. Original magnification: × 200.
Mentions: We performed immunohistochemical analysis of TF protein expression on formalin-fixed tumour tissues from 16 paraffin-embedded uterine serous carcinoma specimens. As representatively shown in Figure 1, with no exception, all USPC samples that were tested showed either membrane and/or cytoplasmic immunoreactivity for TF (i.e., all 16 samples=100%), whereas the non-neoplastic endometrial controls were negative. The staining intensity was focal (1+) in nine cases, moderately positive (2+) in four cases and strongly positive (3+) in two cases.

Bottom Line: Cytoplasmic and/or membrane TF expression was observed in all 16 (100%) USPC samples tested by IHC, but not in normal endometrium.The addition of low doses of IL-2 further increased the cytotoxic effect induced by hI-con1 against chemotherapy-resistant USPC. hI-con1 induces strong cytotoxicity against primary chemotherapy-resistant USPC cell lines overexpressing TF.The hI-con1 may represent a novel therapeutic agent for the treatment of patients harbouring advanced, recurrent and/or metastatic USPC refractory to standard treatment modalities.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, Room 305 LSOG, 333 Cedar Street, PO Box 208063, New Haven, CT 06520-8063, USA.

ABSTRACT

Background: Uterine serous papillary adenocarcinoma (USPC) is a highly aggressive variant of endometrial cancer. Human immuno-conjugate molecule (hI-con1) is an antibody-like molecule targeted against tissue factor (TF), composed of two human Factor VII (fVII) as the targeting domain, fused to human immunoglobulin (Ig) G1 Fc as an effector domain. We evaluated hI-con1 potential activity against primary chemotherapy-resistant USPC cell lines expressing different levels of TF.

Methods: A total of 16 formalin-fixed, paraffin-embedded USPC samples were evaluated by immunohistochemistry (IHC) for TF expression. Six primary USPC cell lines, half of which overexpress the epidermal growth factor type II (HER2/neu) receptor at 3+ levels, were assessed by flow cytometry and real-time PCR for TF expression. Sensitivity to hI-con1-dependent cell-mediated cytotoxicity (IDCC) was evaluated in 5-hour-chromium release assays. Finally, to investigate the effect of interleukin-2 (IL-2) on IDCC, 5-h (51)Cr assays were also conducted in the presence of low doses of IL-2 (i.e., 50-100 IU ml(-1)).

Results: Cytoplasmic and/or membrane TF expression was observed in all 16 (100%) USPC samples tested by IHC, but not in normal endometrium. High expression of TF was found in 50% (three out of six) of the USPC cell lines tested by real-time PCR and flow cytometry when compared with normal endometrial cells (NECs; P<0.001). Uterine serous papillary adenocarcinoma cell lines overexpressing TF, regardless of their high or low HER2/neu expression, were highly sensitive to IDCC (mean killing+/-s.d., 65.6+/-3.7%, range 57.5-77.0%, P<0.001), although negligible cytotoxicity against USPC was seen in the absence of hI-con1 or in the presence of Rituximab control antibody. The addition of low doses of IL-2 further increased the cytotoxic effect induced by hI-con1 against chemotherapy-resistant USPC.

Conclusion: hI-con1 induces strong cytotoxicity against primary chemotherapy-resistant USPC cell lines overexpressing TF. The hI-con1 may represent a novel therapeutic agent for the treatment of patients harbouring advanced, recurrent and/or metastatic USPC refractory to standard treatment modalities.

Show MeSH
Related in: MedlinePlus