Limits...
A monoclonal antibody targeting ErbB2 domain III inhibits ErbB2 signaling and suppresses the growth of ErbB2-overexpressing breast tumors.

Meng Y, Zheng L, Yang Y, Wang H, Dong J, Wang C, Zhang Y, Yu X, Wang L, Xia T, Zhang D, Guo Y, Li B - Oncogenesis (2016)

Bottom Line: The anti-ErbB2 antibodies trastuzumab and pertuzumab in combination have recently been approved for the treatment of patients with ErbB2-positive metastatic breast cancer.Compared with trastuzumab plus pertuzumab, the combination of trastuzumab, pertuzumab and 3E10 provides a more potent blockade of ErbB2 signaling.Consistent with this, trastuzumab plus pertuzumab plus 3E10 results in greater in vitro and in vivo antitumor activity in ErbB2-overexpressing breast tumor models, suggesting its potential use for treating ErbB2-overexpressing breast cancer.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, Nankai University, Tianjin, People's Republic of China.

ABSTRACT
The anti-ErbB2 antibodies trastuzumab and pertuzumab in combination have recently been approved for the treatment of patients with ErbB2-positive metastatic breast cancer. Pertuzumab, which binds to ErbB2 near the center of domain II, and trastuzumab, which binds to the juxtamembrane region of ErbB2 domain IV, directly interfere with domain II- and domain IV-mediated heterodimerization contacts, respectively. In this study, we report a novel anti-ErbB2 antibody, 3E10, which binds to an epitope in domain III that appears to be located opposite to the dimerization interfaces in domain II and domain IV of ErbB2. Our data show that the 3E10 antibody inhibits ErbB2 heterodimerization via a mechanism that strikingly differs from trastuzumab and pertuzumab. It could be speculated that the 3E10 antibody may affect ErbB2 heterodimerization by causing major conformational changes of ErbB2. Furthermore, 3E10 provides synergistic inhibition of ErbB2 heterodimerization and signaling in combination with either trastuzumab or pertuzumab. The combination of these three anti-ErbB2 antibodies that have complementary mechanisms of action appears to be an extremely potent ErbB2 heterodimerization blocker. Compared with trastuzumab plus pertuzumab, the combination of trastuzumab, pertuzumab and 3E10 provides a more potent blockade of ErbB2 signaling. Consistent with this, trastuzumab plus pertuzumab plus 3E10 results in greater in vitro and in vivo antitumor activity in ErbB2-overexpressing breast tumor models, suggesting its potential use for treating ErbB2-overexpressing breast cancer.

No MeSH data available.


Related in: MedlinePlus

The combination of trastuzumab, pertuzumab and C3E10 potently blocks ErbB2 heterodimerization and inhibits breast cancer cell growth. (a) Coimmunoprecipitation assay examining the ability of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 to disrupt the ligand-independent association of ErbB2 with EGFR or ErbB3 in BT-474 cells. (b) Coimmunoprecipitation assay assessing the effects of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 pretreatment on EGF-induced ErbB2/EGFR and HRG-induced ErbB2/ErbB3 heterodimerization in BT-474 cells. (c) MTS assay evaluating the effects of recombinant anti-ErbB2 mAbs on breast cancer cell proliferation in the absence or presence of ErbB ligand (EGF or HRG). Cells were incubated with 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab or trastuzumab plus pertuzumab plus C3E10 for 2 h, followed by the addition of ErbB ligands or not. Recombinant human EGF and HRG were added at a final concentration of 5 and 1 nM, respectively. After an additional 4-day incubation, cell proliferation was determined by MTS assay. Results are shown as percentage of control cell proliferation. Error bars, s.d. *P<0.05; **P<0.001. (d) Tumor volume of BT-474 breast tumor xenografts after treatment with control IgG (5 mg/kg), trastuzumab (5 mg/kg), pertuzumab (5 mg/kg), C3E10 (5 mg/kg), trastuzumab plus pertuzumab (5 mg/kg each) or trastuzumab plus pertuzumab plus C3E10 (5 mg/kg each). Treatments consisted of twice weekly intravenous injection of different anti-ErbB2 mAbs for four consecutive weeks. Data are shown as means±s.e.m. *P<0.05; **P<0.001; Mann–Whitney test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4815051&req=5

fig4: The combination of trastuzumab, pertuzumab and C3E10 potently blocks ErbB2 heterodimerization and inhibits breast cancer cell growth. (a) Coimmunoprecipitation assay examining the ability of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 to disrupt the ligand-independent association of ErbB2 with EGFR or ErbB3 in BT-474 cells. (b) Coimmunoprecipitation assay assessing the effects of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 pretreatment on EGF-induced ErbB2/EGFR and HRG-induced ErbB2/ErbB3 heterodimerization in BT-474 cells. (c) MTS assay evaluating the effects of recombinant anti-ErbB2 mAbs on breast cancer cell proliferation in the absence or presence of ErbB ligand (EGF or HRG). Cells were incubated with 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab or trastuzumab plus pertuzumab plus C3E10 for 2 h, followed by the addition of ErbB ligands or not. Recombinant human EGF and HRG were added at a final concentration of 5 and 1 nM, respectively. After an additional 4-day incubation, cell proliferation was determined by MTS assay. Results are shown as percentage of control cell proliferation. Error bars, s.d. *P<0.05; **P<0.001. (d) Tumor volume of BT-474 breast tumor xenografts after treatment with control IgG (5 mg/kg), trastuzumab (5 mg/kg), pertuzumab (5 mg/kg), C3E10 (5 mg/kg), trastuzumab plus pertuzumab (5 mg/kg each) or trastuzumab plus pertuzumab plus C3E10 (5 mg/kg each). Treatments consisted of twice weekly intravenous injection of different anti-ErbB2 mAbs for four consecutive weeks. Data are shown as means±s.e.m. *P<0.05; **P<0.001; Mann–Whitney test.

Mentions: We investigated the effects of the combination of C3E10 with trastuzumab and pertuzumab on ErbB2 heterodimerization (Figures 4a and b). Our results showed that this combination appeared to be a much more potent ErbB2 heterodimerization blocker than trastuzumab plus pertuzumab. Consistent with this, the three mAbs in combination had a significantly greater ability to inhibit the in vitro breast cancer cell proliferation compared with trastuzumab plus pertuzumab (Figure 4c). Next, the therapeutic efficacy of anti-ErbB2 mAbs was compared in nude mice bearing established BT-474 xenograft tumors. We demonstrated that trastuzumab plus pertuzumab plus C3E10 was more efficient in inhibition of BT-474 tumors than combinatorial treatment with trastuzumab and pertuzumab (Figure 4d). Importantly, all tumors were completely eliminated in tumor-bearing mice treated with trastuzumab plus pertuzumab plus C3E10 (Figure 4d).


A monoclonal antibody targeting ErbB2 domain III inhibits ErbB2 signaling and suppresses the growth of ErbB2-overexpressing breast tumors.

Meng Y, Zheng L, Yang Y, Wang H, Dong J, Wang C, Zhang Y, Yu X, Wang L, Xia T, Zhang D, Guo Y, Li B - Oncogenesis (2016)

The combination of trastuzumab, pertuzumab and C3E10 potently blocks ErbB2 heterodimerization and inhibits breast cancer cell growth. (a) Coimmunoprecipitation assay examining the ability of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 to disrupt the ligand-independent association of ErbB2 with EGFR or ErbB3 in BT-474 cells. (b) Coimmunoprecipitation assay assessing the effects of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 pretreatment on EGF-induced ErbB2/EGFR and HRG-induced ErbB2/ErbB3 heterodimerization in BT-474 cells. (c) MTS assay evaluating the effects of recombinant anti-ErbB2 mAbs on breast cancer cell proliferation in the absence or presence of ErbB ligand (EGF or HRG). Cells were incubated with 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab or trastuzumab plus pertuzumab plus C3E10 for 2 h, followed by the addition of ErbB ligands or not. Recombinant human EGF and HRG were added at a final concentration of 5 and 1 nM, respectively. After an additional 4-day incubation, cell proliferation was determined by MTS assay. Results are shown as percentage of control cell proliferation. Error bars, s.d. *P<0.05; **P<0.001. (d) Tumor volume of BT-474 breast tumor xenografts after treatment with control IgG (5 mg/kg), trastuzumab (5 mg/kg), pertuzumab (5 mg/kg), C3E10 (5 mg/kg), trastuzumab plus pertuzumab (5 mg/kg each) or trastuzumab plus pertuzumab plus C3E10 (5 mg/kg each). Treatments consisted of twice weekly intravenous injection of different anti-ErbB2 mAbs for four consecutive weeks. Data are shown as means±s.e.m. *P<0.05; **P<0.001; Mann–Whitney test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4815051&req=5

fig4: The combination of trastuzumab, pertuzumab and C3E10 potently blocks ErbB2 heterodimerization and inhibits breast cancer cell growth. (a) Coimmunoprecipitation assay examining the ability of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 to disrupt the ligand-independent association of ErbB2 with EGFR or ErbB3 in BT-474 cells. (b) Coimmunoprecipitation assay assessing the effects of 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab, trastuzumab plus C3E10, pertuzumab plus C3E10 or trastuzumab plus pertuzumab plus C3E10 pretreatment on EGF-induced ErbB2/EGFR and HRG-induced ErbB2/ErbB3 heterodimerization in BT-474 cells. (c) MTS assay evaluating the effects of recombinant anti-ErbB2 mAbs on breast cancer cell proliferation in the absence or presence of ErbB ligand (EGF or HRG). Cells were incubated with 100 nM of control IgG, trastuzumab, pertuzumab, C3E10, trastuzumab plus pertuzumab or trastuzumab plus pertuzumab plus C3E10 for 2 h, followed by the addition of ErbB ligands or not. Recombinant human EGF and HRG were added at a final concentration of 5 and 1 nM, respectively. After an additional 4-day incubation, cell proliferation was determined by MTS assay. Results are shown as percentage of control cell proliferation. Error bars, s.d. *P<0.05; **P<0.001. (d) Tumor volume of BT-474 breast tumor xenografts after treatment with control IgG (5 mg/kg), trastuzumab (5 mg/kg), pertuzumab (5 mg/kg), C3E10 (5 mg/kg), trastuzumab plus pertuzumab (5 mg/kg each) or trastuzumab plus pertuzumab plus C3E10 (5 mg/kg each). Treatments consisted of twice weekly intravenous injection of different anti-ErbB2 mAbs for four consecutive weeks. Data are shown as means±s.e.m. *P<0.05; **P<0.001; Mann–Whitney test.
Mentions: We investigated the effects of the combination of C3E10 with trastuzumab and pertuzumab on ErbB2 heterodimerization (Figures 4a and b). Our results showed that this combination appeared to be a much more potent ErbB2 heterodimerization blocker than trastuzumab plus pertuzumab. Consistent with this, the three mAbs in combination had a significantly greater ability to inhibit the in vitro breast cancer cell proliferation compared with trastuzumab plus pertuzumab (Figure 4c). Next, the therapeutic efficacy of anti-ErbB2 mAbs was compared in nude mice bearing established BT-474 xenograft tumors. We demonstrated that trastuzumab plus pertuzumab plus C3E10 was more efficient in inhibition of BT-474 tumors than combinatorial treatment with trastuzumab and pertuzumab (Figure 4d). Importantly, all tumors were completely eliminated in tumor-bearing mice treated with trastuzumab plus pertuzumab plus C3E10 (Figure 4d).

Bottom Line: The anti-ErbB2 antibodies trastuzumab and pertuzumab in combination have recently been approved for the treatment of patients with ErbB2-positive metastatic breast cancer.Compared with trastuzumab plus pertuzumab, the combination of trastuzumab, pertuzumab and 3E10 provides a more potent blockade of ErbB2 signaling.Consistent with this, trastuzumab plus pertuzumab plus 3E10 results in greater in vitro and in vivo antitumor activity in ErbB2-overexpressing breast tumor models, suggesting its potential use for treating ErbB2-overexpressing breast cancer.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, Nankai University, Tianjin, People's Republic of China.

ABSTRACT
The anti-ErbB2 antibodies trastuzumab and pertuzumab in combination have recently been approved for the treatment of patients with ErbB2-positive metastatic breast cancer. Pertuzumab, which binds to ErbB2 near the center of domain II, and trastuzumab, which binds to the juxtamembrane region of ErbB2 domain IV, directly interfere with domain II- and domain IV-mediated heterodimerization contacts, respectively. In this study, we report a novel anti-ErbB2 antibody, 3E10, which binds to an epitope in domain III that appears to be located opposite to the dimerization interfaces in domain II and domain IV of ErbB2. Our data show that the 3E10 antibody inhibits ErbB2 heterodimerization via a mechanism that strikingly differs from trastuzumab and pertuzumab. It could be speculated that the 3E10 antibody may affect ErbB2 heterodimerization by causing major conformational changes of ErbB2. Furthermore, 3E10 provides synergistic inhibition of ErbB2 heterodimerization and signaling in combination with either trastuzumab or pertuzumab. The combination of these three anti-ErbB2 antibodies that have complementary mechanisms of action appears to be an extremely potent ErbB2 heterodimerization blocker. Compared with trastuzumab plus pertuzumab, the combination of trastuzumab, pertuzumab and 3E10 provides a more potent blockade of ErbB2 signaling. Consistent with this, trastuzumab plus pertuzumab plus 3E10 results in greater in vitro and in vivo antitumor activity in ErbB2-overexpressing breast tumor models, suggesting its potential use for treating ErbB2-overexpressing breast cancer.

No MeSH data available.


Related in: MedlinePlus