Dai Shi, Yiqiu Zhang, Zhan Xu, Zhan Si, Yuan Cheng, Dengfeng Cheng, Guobing Liu
{"title":"使用基于99mTc-MAG3-Cet-F(ab')2的SPECT/CT成像无创评估消化道肿瘤EGFR表达","authors":"Dai Shi, Yiqiu Zhang, Zhan Xu, Zhan Si, Yuan Cheng, Dengfeng Cheng, Guobing Liu","doi":"10.1155/2022/3748315","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>This study is aimed at investigating the feasibility of cetuximab (Cet) F(ab')<sub>2</sub> fragment- (Cet-F(ab')<sub>2</sub>-) based single photon emission tomography/computed tomography (SPECT/CT) for assessing the epidermal growth factor receptor (EGFR) expression in digestive tumor mouse models.</p><p><strong>Methods: </strong>Cet-F(ab')<sub>2</sub> was synthesized using immunoglobulin G-degrading enzyme of <i>Streptococcus pyogenes</i> (IdeS) protease and purified with protein A beads. The product and its in vitro stability in normal saline and 1% bovine serum albumin were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The EGFR expression in the human colon tumor cell line HT29 and the human stomach tumor cell line MGC803 were verified using western blotting and immunocytochemistry. Cet-F(ab')<sub>2</sub> was conjugated with 5(6)-carboxytetramethylrhodamine succinimidyl ester to demonstrate its binding ability to the MGC803 and HT29 cells. Cet-F(ab')<sub>2</sub> was conjugated with NHS-MAG<sub>3</sub> for <sup>99m</sup>Tc radiolabeling. The best imaging time was determined using a biodistribution assay at 1, 4, 16, and 24 h after injection of the <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> tracer. Furthermore, <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> SPECT/CT was performed on MGC803 and HT29 tumor-bearing nude mice.</p><p><strong>Results: </strong>HT29 cells had low EGFR expression while MGC803 cell exhibited the high EGFR expression. Cet-F(ab')<sub>2</sub> and intact cetuximab showed similar high binding ability to MGC803 cells but not to HT29 cells. Cet-F(ab')<sub>2</sub> and <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> showed excellent in vitro stability. The biodistribution assay showed that the target to nontarget ratio was the highest at 16 h (17.29 ± 5.72, <i>n</i> = 4) after tracer injection. The <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub>-based SPECT/CT imaging revealed rapid and sustained tracer uptake in MGC803 tumors rather than in HT29 tumors with high image contrast, which was consistent with the results in vitro.</p><p><strong>Conclusion: </strong>SPECT/CT imaging using <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> enables the evaluation of the EGFR expression in murine EGFR-positive tumors, indicating the potential utility for noninvasive evaluation of the EGFR expression in tumors.</p>","PeriodicalId":18855,"journal":{"name":"Molecular Imaging","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2022-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9281432/pdf/","citationCount":"1","resultStr":"{\"title\":\"Noninvasive Evaluation of EGFR Expression of Digestive Tumors Using <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub>-Based SPECT/CT Imaging.\",\"authors\":\"Dai Shi, Yiqiu Zhang, Zhan Xu, Zhan Si, Yuan Cheng, Dengfeng Cheng, Guobing Liu\",\"doi\":\"10.1155/2022/3748315\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>This study is aimed at investigating the feasibility of cetuximab (Cet) F(ab')<sub>2</sub> fragment- (Cet-F(ab')<sub>2</sub>-) based single photon emission tomography/computed tomography (SPECT/CT) for assessing the epidermal growth factor receptor (EGFR) expression in digestive tumor mouse models.</p><p><strong>Methods: </strong>Cet-F(ab')<sub>2</sub> was synthesized using immunoglobulin G-degrading enzyme of <i>Streptococcus pyogenes</i> (IdeS) protease and purified with protein A beads. The product and its in vitro stability in normal saline and 1% bovine serum albumin were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The EGFR expression in the human colon tumor cell line HT29 and the human stomach tumor cell line MGC803 were verified using western blotting and immunocytochemistry. Cet-F(ab')<sub>2</sub> was conjugated with 5(6)-carboxytetramethylrhodamine succinimidyl ester to demonstrate its binding ability to the MGC803 and HT29 cells. Cet-F(ab')<sub>2</sub> was conjugated with NHS-MAG<sub>3</sub> for <sup>99m</sup>Tc radiolabeling. The best imaging time was determined using a biodistribution assay at 1, 4, 16, and 24 h after injection of the <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> tracer. Furthermore, <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> SPECT/CT was performed on MGC803 and HT29 tumor-bearing nude mice.</p><p><strong>Results: </strong>HT29 cells had low EGFR expression while MGC803 cell exhibited the high EGFR expression. Cet-F(ab')<sub>2</sub> and intact cetuximab showed similar high binding ability to MGC803 cells but not to HT29 cells. Cet-F(ab')<sub>2</sub> and <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> showed excellent in vitro stability. The biodistribution assay showed that the target to nontarget ratio was the highest at 16 h (17.29 ± 5.72, <i>n</i> = 4) after tracer injection. The <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub>-based SPECT/CT imaging revealed rapid and sustained tracer uptake in MGC803 tumors rather than in HT29 tumors with high image contrast, which was consistent with the results in vitro.</p><p><strong>Conclusion: </strong>SPECT/CT imaging using <sup>99m</sup>Tc-MAG<sub>3</sub>-Cet-F(ab')<sub>2</sub> enables the evaluation of the EGFR expression in murine EGFR-positive tumors, indicating the potential utility for noninvasive evaluation of the EGFR expression in tumors.</p>\",\"PeriodicalId\":18855,\"journal\":{\"name\":\"Molecular Imaging\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2022-06-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9281432/pdf/\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular Imaging\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1155/2022/3748315\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Imaging","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1155/2022/3748315","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/1/1 0:00:00","PubModel":"eCollection","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Noninvasive Evaluation of EGFR Expression of Digestive Tumors Using 99mTc-MAG3-Cet-F(ab')2-Based SPECT/CT Imaging.
Purpose: This study is aimed at investigating the feasibility of cetuximab (Cet) F(ab')2 fragment- (Cet-F(ab')2-) based single photon emission tomography/computed tomography (SPECT/CT) for assessing the epidermal growth factor receptor (EGFR) expression in digestive tumor mouse models.
Methods: Cet-F(ab')2 was synthesized using immunoglobulin G-degrading enzyme of Streptococcus pyogenes (IdeS) protease and purified with protein A beads. The product and its in vitro stability in normal saline and 1% bovine serum albumin were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The EGFR expression in the human colon tumor cell line HT29 and the human stomach tumor cell line MGC803 were verified using western blotting and immunocytochemistry. Cet-F(ab')2 was conjugated with 5(6)-carboxytetramethylrhodamine succinimidyl ester to demonstrate its binding ability to the MGC803 and HT29 cells. Cet-F(ab')2 was conjugated with NHS-MAG3 for 99mTc radiolabeling. The best imaging time was determined using a biodistribution assay at 1, 4, 16, and 24 h after injection of the 99mTc-MAG3-Cet-F(ab')2 tracer. Furthermore, 99mTc-MAG3-Cet-F(ab')2 SPECT/CT was performed on MGC803 and HT29 tumor-bearing nude mice.
Results: HT29 cells had low EGFR expression while MGC803 cell exhibited the high EGFR expression. Cet-F(ab')2 and intact cetuximab showed similar high binding ability to MGC803 cells but not to HT29 cells. Cet-F(ab')2 and 99mTc-MAG3-Cet-F(ab')2 showed excellent in vitro stability. The biodistribution assay showed that the target to nontarget ratio was the highest at 16 h (17.29 ± 5.72, n = 4) after tracer injection. The 99mTc-MAG3-Cet-F(ab')2-based SPECT/CT imaging revealed rapid and sustained tracer uptake in MGC803 tumors rather than in HT29 tumors with high image contrast, which was consistent with the results in vitro.
Conclusion: SPECT/CT imaging using 99mTc-MAG3-Cet-F(ab')2 enables the evaluation of the EGFR expression in murine EGFR-positive tumors, indicating the potential utility for noninvasive evaluation of the EGFR expression in tumors.
Molecular ImagingBiochemistry, Genetics and Molecular Biology-Biotechnology
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3.60%
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21
期刊介绍：
Molecular Imaging is a peer-reviewed, open access journal highlighting the breadth of molecular imaging research from basic science to preclinical studies to human applications. This serves both the scientific and clinical communities by disseminating novel results and concepts relevant to the biological study of normal and disease processes in both basic and translational studies ranging from mice to humans.
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