Identification of a small peptide that inhibits the phosphorylation of ErbB2 and proliferation of ErbB2 overexpressing breast cancer cells

Pero, Stephanie C.; Shukla, Girja S.; Armstrong, Amy; Peterson, Daniel L.; Fuller, Susan P.; Godin, Katherine S.; Kingsley-Richards, S.L.; Weaver, Donald L.; Bond, Jeffrey P.; Krag, David N.

doi:10.1002/ijc.20306

Cited by 38 publications

(48 citation statements)

References 36 publications

(45 reference statements)

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“…Though other groups have reported peptides derived from phage display that bind EGFR-2, 13,34,35 our studies are the first to demonstrate in vivo in a animal model that an EGFR-2 targeting peptide selected by phage display can be exploited as an SPECT or PET radioimaging agent.…”

Section: Discussionmentioning

confidence: 77%

In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

Kumar

Gallazzi

Ferdani

et al. 2010

Cancer Biotherapy and Radiopharmaceuticals

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Epidermal growth factor receptor-2 (EGFR-2) has been implicated in the pathogenesis of breast and other carcinomas. In this report, we tested the ability of a bacteriophage selected peptide KCCYSL, radiolabeled with 64 Cu, to image EGFR-2 expressing breast tumors in vivo by positron emission tomography (PET). We evaluated and compared the in vivo tissue distribution and imaging properties of 64 Cu-X-(Gly-Ser-Gly)-KCCYSL peptide (X ¼ 1,4,7,10, tetraazacyclododecane-N,N',N'',N'''-tetracetic acid, [DOTA] 1,4,8, hexadecane-4,11-diacetic acid [CB-TE2A], and 1,4,7-triazacyclononane-1,4,7-triacetic acid [NOTA] chelators) in a human breast cancer xenograft mouse model using dual modality PET and computed tomography (CT). The synthesized peptides DO3A-GSG-KCCYSL, CB-TE2A-GSG-KCCYSL, and NO2A-GSG-KCCYSL were purified, radiolabeled with 64 Cu, and evaluated for human breast cancer cell (MDA-MB-435) binding, 50% inhibitory concentration, and serum stability. In vivo pharmacokinetic and small animal PET/CT imaging studies were performed using SCID mice bearing MDA-MB-435 xenografts. The radiolabeled peptides bound with an 50% inhibitory concentration of 42.0 AE 10.2 nM (DO3A), 31 AE 7.9 nM (CB-TE2A), and 44.2 AE 6.6 nM (NO2A) to cultured MDA-MB-435 cells. All of the radiolabeled peptides were stable in vitro. The tumor uptake of DO3A, CB-TE2A, and NO2A peptides were 0.73 AE 0.15 percent injected dose per gram (%ID/g), 0.64 AE 0.08%ID/g, and 0.52 AE 0.04%ID/g, respectively at 1 hour. Liver uptake for the 64 Cu-DO3A-peptide (1.68 AE 0.42%ID/g) was more than that of the 64 Cu-CB-TE2A-peptide (0.52 AE 0.02% ID/g) and 64 Cu-NO2A-peptide (0.48 AE 0.05%ID/g) at 2 hours. PET/CT studies revealed successful tumor uptake of 64 Cu-peptides at 2 hours postinjection. In vivo kidney retention was observed with all of the radiolabeled peptides. The optimization of bifunctional chelators improves the pharmacokinetic properties of the 64 Cu-labeled GSG-KCCYSL peptide, which enables the selection of a suitable peptide homolog as a PET imaging agent for EGFR-2 expressing breast carcinomas.

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Section: Discussionmentioning

confidence: 77%

In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

Kumar

Gallazzi

Ferdani

et al. 2010

Cancer Biotherapy and Radiopharmaceuticals

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“…The complexities of the random 12-and 20-mer libraries were 1.26 Â 10 7 and 1.8 Â 10 7 different peptides, respectively. The details of library preparation, titration, elution of cell-bound phage, phage plating, and amplification have been described in detail in our earlier publications (19,32,34,35).…”

Section: Phage Librariesmentioning

confidence: 99%

“…The rinsed tissue and cells were homogenized to release cell-internalized phage. The bacterial host infection, plating, titration, and amplification were done by the methods as described earlier for peptide (19,32,34,35) and scFv (36,37) libraries. Phage administration, tumor harvest, and amplification were repeated twice for a maximum of three infusions.…”

Section: Tumor Processing and Phage Recoverymentioning

confidence: 99%

Selection of Tumor-binding Ligands in Cancer Patients with Phage Display Libraries

Krag

Shukla²,

Shen³

et al. 2006

Cancer Research

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185

145

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Phage display has been used extensively in vitro and in animal models to generate ligands and to identify cancer-relevant targets. We report here the use of phage-display libraries in cancer patients to identify tumor-targeting ligands. Eight patients with stage IV cancer, including breast, melanoma, and pancreas, had phage-displayed random peptide or scFv library (1.6 Â 10 8 -1 Â 10 11 transducing units/kg) administered i.v.; tumors were excised after 30 minutes; and tumor-homing phage were recovered. In three patients, repeat panning was possible using phage recovered and amplified from that same patient's tumor. No serious side effects, including allergic reactions, were observed with up to three infusions. Patients developed antiphage antibodies that reached a submaximal level within the 10-day protocol window for serial phage administration. Tumor phage were recoverable from all the patients. Using a filter-based ELISA, several clones from a subset of the patients were identified that bound to a tumor from the same patient in which clones were recovered. The clone-binding to tumor was confirmed by immunostaining, bioassay, and real-time PCR-based methods. Binding studies with noncancer and cancer cell lines of the same histology showed specificity of the tumor-binding clones. Analysis of insert sequences of tumor-homing peptide clones showed several motifs, indicating nonrandom accumulation of clones in human tumors. This is the first reported series of cancer patients to receive phage library for serial panning of tumor targeting ligands. The lack of toxicity and the ability to recover clones with favorable characteristics are a first step for further research with this technology in cancer patients. (Cancer Res 2006; 66(15): 7724-33)

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“…Surprisingly, however, Tyr845 phosphorylation seems not to affect ErbB1 kinase activity, as ErbB1 carrying a Y845F mutation is fully active (34). Phosphorylation, in tumor cells, on the analogous site in ErbB2, Tyr877, was reported recently (12,27), but the functional implication is not clear. In contrast to ErbB1, data reported for the oncogenic rat protein ErbB2/neu, which carries an activating point mutation in the transmembrane domain, suggest that A-loop phosphorylation is important for intrinsic kinase activity, as mutation of the analogous Tyr882 to phenylalanine compromised kinase activity (39).…”

mentioning

confidence: 95%

Loss of Hsp90 Association Up-Regulates Src-Dependent ErbB2 Activity

Yuan

Beebe

et al. 2007

Molecular and Cellular Biology

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The receptor tyrosine kinase ErbB2 plays a crucial role in tumorigenesis. We showed previously that the molecular chaperone Hsp90 protects ErbB2 from proteasome-mediated degradation by binding to a short loop structure in the N-lobe of the kinase domain. Here we show that loss of Hsp90 binding correlates with enhanced ErbB2 kinase activity and its transactivating potential, concomitant with constitutively increased phosphorylation of Tyr877, located in the activation loop of the kinase domain. We show further that Tyr877 phosphorylation is mediated by Src and that it is necessary for the enhanced kinase activity of ErbB2. Finally, computer modeling of the kinase domain suggests a phosphorylation-dependent reorientation of the activation loop, denoting the importance of Tyr877 phosphorylation for ErbB2 activity. These findings suggest that Hsp90 binding to ErbB2 participates in regulation of kinase activity as well as kinase stability.

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Identification of a small peptide that inhibits the phosphorylation of ErbB2 and proliferation of ErbB2 overexpressing breast cancer cells

Cited by 38 publications

References 36 publications

In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

Selection of Tumor-binding Ligands in Cancer Patients with Phage Display Libraries

Loss of Hsp90 Association Up-Regulates Src-Dependent ErbB2 Activity

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Identification of a small peptide that inhibits the phosphorylation of ErbB2 and proliferation of ErbB2 overexpressing breast cancer cells

Cited by 38 publications

References 36 publications

In VitroandIn VivoEvaluation of64Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

In VitroandIn VivoEvaluation of64Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

Selection of Tumor-binding Ligands in Cancer Patients with Phage Display Libraries

Loss of Hsp90 Association Up-Regulates Src-Dependent ErbB2 Activity

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In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas

In VitroandIn VivoEvaluation of⁶⁴Cu-Radiolabeled KCCYSL Peptides for Targeting Epidermal Growth Factor Receptor-2 in Breast Carcinomas