Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma
Mucin 1 is a transmembrane glycoprotein which overexpresses cancer-specific epitopes (MUC1-CE) on pancreatic ductal adenocarcinoma (PDAC) cells. As PDAC is a low survival and highly aggressive malignancy, developing radioimmunoconjugates capable of targeting MUC1-CE could lead to improvements in PDA...
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MDPI AG
2022-09-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/11/19/2983 |
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| author | Ashleigh Hull Yanrui Li Dylan Bartholomeusz William Hsieh William Tieu Tara L. Pukala Alexander H. Staudacher Eva Bezak |
| author_facet | Ashleigh Hull Yanrui Li Dylan Bartholomeusz William Hsieh William Tieu Tara L. Pukala Alexander H. Staudacher Eva Bezak |
| author_sort | Ashleigh Hull |
| collection | DOAJ |
| description | Mucin 1 is a transmembrane glycoprotein which overexpresses cancer-specific epitopes (MUC1-CE) on pancreatic ductal adenocarcinoma (PDAC) cells. As PDAC is a low survival and highly aggressive malignancy, developing radioimmunoconjugates capable of targeting MUC1-CE could lead to improvements in PDAC outcomes. The aim of this study was to develop and perform preliminary testing of diagnostic and therapeutic radioimmunoconjugates for PDAC using an anti-MUC1 antibody, C595. Firstly, p-SCN-Bn-DOTA was conjugated to the C595 antibody to form a DOTA-C595 immunoconjugate. The stability and binding affinity of the DOTA-C595 conjugate was evaluated using mass spectrometry and ELISA. DOTA-C595 was radiolabelled to Copper-64, Lutetium-177, Gallium-68 and Technetium-99m to form novel radioimmunoconjugates. Cell binding assays were performed in PANC-1 (strong MUC1-CE expression) and AsPC-1 (weak MUC1-CE expression) cell lines using <sup>64</sup>Cu-DOTA-C595 and <sup>177</sup>Lu-DOTA-C595. An optimal molar ratio of 4:1 DOTA groups per C595 molecule was obtained from the conjugation process. DOTA-C595 labelled to Copper-64, Lutetium-177, and Technetium-99m with high efficiency, although the Gallium-68 labelling was low. <sup>177</sup>Lu-DOTA-C595 demonstrated high cellular binding to the PANC-1 cell lines which was significantly greater than AsPC-1 binding at concentrations exceeding 100 nM (<i>p</i> < 0.05). <sup>64</sup>Cu-DOTA-C595 showed similar binding to the PANC-1 and AsPC-1 cells with no significant differences observed between cell lines (<i>p</i> > 0.05). The high cellular binding of <sup>177</sup>Lu-DOTA-C595 to MUC1-CE positive cell lines suggests promise as a therapeutic radioimmunoconjugate against PDAC while further work is required to harness the potential of <sup>64</sup>Cu-DOTA-C595 as a diagnostic radioimmunoconjugate. |
| first_indexed | 2024-03-09T21:55:06Z |
| format | Article |
| id | doaj.art-c5b299e7a4b949ae817f2f2c43b6eb11 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T21:55:06Z |
| publishDate | 2022-09-01 |
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| spelling | doaj.art-c5b299e7a4b949ae817f2f2c43b6eb112023-11-23T20:00:35ZengMDPI AGCells2073-44092022-09-011119298310.3390/cells11192983Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal AdenocarcinomaAshleigh Hull0Yanrui Li1Dylan Bartholomeusz2William Hsieh3William Tieu4Tara L. Pukala5Alexander H. Staudacher6Eva Bezak7Allied Health and Human Performance Academic Unit, Cancer Research Institute, University of South Australia, Adelaide, SA 5001, AustraliaAllied Health and Human Performance Academic Unit, Cancer Research Institute, University of South Australia, Adelaide, SA 5001, AustraliaDepartment of PET, Nuclear Medicine & Bone Densitometry, Royal Adelaide Hospital, SA Medical Imaging, Adelaide, SA 5000, AustraliaAllied Health and Human Performance Academic Unit, Cancer Research Institute, University of South Australia, Adelaide, SA 5001, AustraliaMolecular Imaging and Therapy Research Unit, South Australian Health and Medical Research Institute, Adelaide, SA 5000, AustraliaSchool of Physical Sciences, The University of Adelaide, Adelaide, SA 5000, AustraliaAdelaide Medical School, The University of Adelaide, Adelaide, SA 5000, AustraliaAllied Health and Human Performance Academic Unit, Cancer Research Institute, University of South Australia, Adelaide, SA 5001, AustraliaMucin 1 is a transmembrane glycoprotein which overexpresses cancer-specific epitopes (MUC1-CE) on pancreatic ductal adenocarcinoma (PDAC) cells. As PDAC is a low survival and highly aggressive malignancy, developing radioimmunoconjugates capable of targeting MUC1-CE could lead to improvements in PDAC outcomes. The aim of this study was to develop and perform preliminary testing of diagnostic and therapeutic radioimmunoconjugates for PDAC using an anti-MUC1 antibody, C595. Firstly, p-SCN-Bn-DOTA was conjugated to the C595 antibody to form a DOTA-C595 immunoconjugate. The stability and binding affinity of the DOTA-C595 conjugate was evaluated using mass spectrometry and ELISA. DOTA-C595 was radiolabelled to Copper-64, Lutetium-177, Gallium-68 and Technetium-99m to form novel radioimmunoconjugates. Cell binding assays were performed in PANC-1 (strong MUC1-CE expression) and AsPC-1 (weak MUC1-CE expression) cell lines using <sup>64</sup>Cu-DOTA-C595 and <sup>177</sup>Lu-DOTA-C595. An optimal molar ratio of 4:1 DOTA groups per C595 molecule was obtained from the conjugation process. DOTA-C595 labelled to Copper-64, Lutetium-177, and Technetium-99m with high efficiency, although the Gallium-68 labelling was low. <sup>177</sup>Lu-DOTA-C595 demonstrated high cellular binding to the PANC-1 cell lines which was significantly greater than AsPC-1 binding at concentrations exceeding 100 nM (<i>p</i> < 0.05). <sup>64</sup>Cu-DOTA-C595 showed similar binding to the PANC-1 and AsPC-1 cells with no significant differences observed between cell lines (<i>p</i> > 0.05). The high cellular binding of <sup>177</sup>Lu-DOTA-C595 to MUC1-CE positive cell lines suggests promise as a therapeutic radioimmunoconjugate against PDAC while further work is required to harness the potential of <sup>64</sup>Cu-DOTA-C595 as a diagnostic radioimmunoconjugate.https://www.mdpi.com/2073-4409/11/19/2983pancreatic cancerradioimmunoconjugatesmucin 1C595 |
| spellingShingle | Ashleigh Hull Yanrui Li Dylan Bartholomeusz William Hsieh William Tieu Tara L. Pukala Alexander H. Staudacher Eva Bezak Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma Cells pancreatic cancer radioimmunoconjugates mucin 1 C595 |
| title | Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma |
| title_full | Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma |
| title_fullStr | Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma |
| title_full_unstemmed | Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma |
| title_short | Preliminary Development and Testing of C595 Radioimmunoconjugates for Targeting MUC1 Cancer Epitopes in Pancreatic Ductal Adenocarcinoma |
| title_sort | preliminary development and testing of c595 radioimmunoconjugates for targeting muc1 cancer epitopes in pancreatic ductal adenocarcinoma |
| topic | pancreatic cancer radioimmunoconjugates mucin 1 C595 |
| url | https://www.mdpi.com/2073-4409/11/19/2983 |
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