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A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus

Biodistribution of 64Cu-DOTA-mAbs in mice bearing BxPC-3, CFPAC-1, and PANC-1 tumors at 24 hours (a) and 48 hours (b) after intravenous injection. The mice were sacrificed at 24 hours and 48 hours after intravenous injection of 11 MBq of 64Cu-DOTA-11-25 mAb (black bars) or 64Cu-DOTA-anti-KLH mAb (white bars). The organs were collected and weighed and radioactivity was measured by γ-counter. ∗P < 0.05 versus the accumulation of 64Cu-DOTA-anti-KLH mAb. Tumor to blood ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). Tumor to muscle ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). The data were calculated as percentage of injected dose per gram of tissue (%ID/g). Mean and standard deviation have been corrected for physical decay of 64Cu. ∗P < 0.05, ∗∗P < 0.01 versus the ratio of PANC-1 tumor.
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fig8: Biodistribution of 64Cu-DOTA-mAbs in mice bearing BxPC-3, CFPAC-1, and PANC-1 tumors at 24 hours (a) and 48 hours (b) after intravenous injection. The mice were sacrificed at 24 hours and 48 hours after intravenous injection of 11 MBq of 64Cu-DOTA-11-25 mAb (black bars) or 64Cu-DOTA-anti-KLH mAb (white bars). The organs were collected and weighed and radioactivity was measured by γ-counter. ∗P < 0.05 versus the accumulation of 64Cu-DOTA-anti-KLH mAb. Tumor to blood ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). Tumor to muscle ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). The data were calculated as percentage of injected dose per gram of tissue (%ID/g). Mean and standard deviation have been corrected for physical decay of 64Cu. ∗P < 0.05, ∗∗P < 0.01 versus the ratio of PANC-1 tumor.

Mentions: In the biodistribution study, relatively high accumulation of 64Cu-DOTA-11-25 mAb was observed in the blood, liver, and MSLN-positive tumors (Figure 8). In CFPAC-1 xenografts, the accumulation of 64Cu-DOTA-11-25 mAb was significantly higher than that of 64Cu-DOTA-anti-KLH mAb at both time points. In BxPC-3 xenografts, the accumulation of 64Cu-DOTA-11-25 mAb was significantly higher than that of 64Cu-DOTA-anti-KLH mAb at 48 hours after injection (Figures 8(a) and 8(b)). At both 24 and 48 hours after injection, the tumor to blood ratio and tumor to muscle ratio of 64Cu-DOTA-11-25 mAb in BxPC-3 tumor and in CFPAC-1 tumor were significantly higher than those in PANC-1 tumor (P < 0.05 and P < 0.01, resp., Figures 8(c)–8(f)). The average weight of individual PANC-1, BxPC-3, or CFPAC-1 tumor was 77 ± 49 mg, 50 ± 38/mg, or 207 ± 78 mg, respectively. In vivo stability of 64Cu-DOTA-11-25 mAb and 64Cu-DOTA-aKLH mAb was measured by TLC.


A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Biodistribution of 64Cu-DOTA-mAbs in mice bearing BxPC-3, CFPAC-1, and PANC-1 tumors at 24 hours (a) and 48 hours (b) after intravenous injection. The mice were sacrificed at 24 hours and 48 hours after intravenous injection of 11 MBq of 64Cu-DOTA-11-25 mAb (black bars) or 64Cu-DOTA-anti-KLH mAb (white bars). The organs were collected and weighed and radioactivity was measured by γ-counter. ∗P < 0.05 versus the accumulation of 64Cu-DOTA-anti-KLH mAb. Tumor to blood ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). Tumor to muscle ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). The data were calculated as percentage of injected dose per gram of tissue (%ID/g). Mean and standard deviation have been corrected for physical decay of 64Cu. ∗P < 0.05, ∗∗P < 0.01 versus the ratio of PANC-1 tumor.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4390102&req=5

fig8: Biodistribution of 64Cu-DOTA-mAbs in mice bearing BxPC-3, CFPAC-1, and PANC-1 tumors at 24 hours (a) and 48 hours (b) after intravenous injection. The mice were sacrificed at 24 hours and 48 hours after intravenous injection of 11 MBq of 64Cu-DOTA-11-25 mAb (black bars) or 64Cu-DOTA-anti-KLH mAb (white bars). The organs were collected and weighed and radioactivity was measured by γ-counter. ∗P < 0.05 versus the accumulation of 64Cu-DOTA-anti-KLH mAb. Tumor to blood ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). Tumor to muscle ratio of 64Cu-lableld 11-25 mAb and 64Cu-DOTA-anti-KLH mAb in the tumors at 24 hours (c) and 48 hours (d) after injection of PANC-1 (white bars), BxPC-3 (gray bars), and CFPAC-1 (black bars). The data were calculated as percentage of injected dose per gram of tissue (%ID/g). Mean and standard deviation have been corrected for physical decay of 64Cu. ∗P < 0.05, ∗∗P < 0.01 versus the ratio of PANC-1 tumor.
Mentions: In the biodistribution study, relatively high accumulation of 64Cu-DOTA-11-25 mAb was observed in the blood, liver, and MSLN-positive tumors (Figure 8). In CFPAC-1 xenografts, the accumulation of 64Cu-DOTA-11-25 mAb was significantly higher than that of 64Cu-DOTA-anti-KLH mAb at both time points. In BxPC-3 xenografts, the accumulation of 64Cu-DOTA-11-25 mAb was significantly higher than that of 64Cu-DOTA-anti-KLH mAb at 48 hours after injection (Figures 8(a) and 8(b)). At both 24 and 48 hours after injection, the tumor to blood ratio and tumor to muscle ratio of 64Cu-DOTA-11-25 mAb in BxPC-3 tumor and in CFPAC-1 tumor were significantly higher than those in PANC-1 tumor (P < 0.05 and P < 0.01, resp., Figures 8(c)–8(f)). The average weight of individual PANC-1, BxPC-3, or CFPAC-1 tumor was 77 ± 49 mg, 50 ± 38/mg, or 207 ± 78 mg, respectively. In vivo stability of 64Cu-DOTA-11-25 mAb and 64Cu-DOTA-aKLH mAb was measured by TLC.

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus