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. 2025 Feb 27;26(5):2145.
doi: 10.3390/ijms26052145.

Expression Profiles of Five Common Cancer Membrane Protein Antigens Collected for the Development of Cocktail CAR-T Cell Therapies Applicable to Most Solid Cancer Patients

Tetsuya Nakatsura  1 , Kazumasa Takenouchi  1 , Jun Kataoka  1 , Yusuke Ito  1 , Sae Kikuchi  1 , Hiroki Kinoshita  1 , Kazunobu Ohnuki  1 , Toshihiro Suzuki  1 , Nobuo Tsukamoto  1
Affiliations

Expression Profiles of Five Common Cancer Membrane Protein Antigens Collected for the Development of Cocktail CAR-T Cell Therapies Applicable to Most Solid Cancer Patients

Tetsuya Nakatsura et al. Int J Mol Sci. .

Abstract

Although CD19 CAR-T has been highly effective against B-cell blood cancers, there are few reports of successful treatments for solid cancers, probably because there are few protein antigens specifically expressed on the surface of the cancer cell membrane. The key to developing a groundbreaking CAR-T cell therapy effective against solid cancers is to "overcome the heterogeneity of cancer antigens". For this purpose, it is necessary to target multiple cancer antigens simultaneously. In this study, we performed immunohistochemical analysis of various solid cancer specimens using antibodies against ROBO1, EphB4, CLDN1, and LAT1 in addition to GPC3, which we have previously studied. These antigens were frequently expressed in various solid cancers but shown to be rarely expressed, with some exceptions, in non-cancerous normal organs adjacent to the cancer. Although ROBO1 and GPC3 are often expressed in cytoplasm, there are also cases in which they are expressed on the cell membrane depending on the type of cancer. On the other hand, it has been revealed that three antigens-EphB4, CLDN1, and LAT1-are frequently expressed only on the cell membrane of cancer cells in various solid cancers, suggesting that they may be ideal targets for CAR-T cell therapy.

Keywords: CAR-T cell therapy; CLDN1; EphB4; LAT1; ROBO1; cocktail CAR-T; common cancer antigens; glypican-3; membrane protein; solid cancer.

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Conflict of interest statement

Author Tetsuya Nakatsura (T.N.) has received research grants from BrightPath Biotherapeutics Co., Ltd., Thyas Co., Ltd., ONO PHARMACEUTICAL Co., Ltd., Resonac Corporation, MEDINET Co., Ltd., NapaJen Pharma Inc., Heartseed Inc., Takara BIO Inc., DAICEL CORPORATION, NA Vaccine Institute Co., Ltd., Logomix Inc., Optieum Biotechnologies Inc., and MaxCyte, Inc. T.N. hold stock ownership, stock options, or profits from Noile-Immune Biotech Inc., Logomix Inc., and Optieum Biotechnologies Inc. T.N. have royalties from OncoTherapyScience, Inc. The remaining authors declare that the research was conducted without commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Representative examples of plasma membrane expression of five common cancer antigens in cancer cells. Representative examples of five common cancer antigens, GPC3, ROBO1, CLDN1, EphB4, and LAT1, expressed on the cell membrane of cancer cells are shown.
Figure 2
Figure 2
Expression of 5 common cancer-specific antigens in various solid cancers and their expression as cell membrane proteins. Representative examples of the expression patterns of 5 common cancer-specific antigens in each cancer type by their intracellular localization (cell membrane, cytoplasm) in cancer tissues are shown.
Figure 3
Figure 3
Heatmap of expression frequency of five common cancer antigens on the cell membrane in various solid cancers. The expression frequency values in Table 1 are shown in a heatmap, with 1% in blue and 100% in red. The closer to red, the higher the expression frequency, and the closer to blue, the lower the expression frequency. Black is 0%, meaning no expression with no positive cases. White is not tested: NT.
Figure 4
Figure 4
Expression of 5 common cancer-specific antigens in non-cancerous normal organs adjacent to various cancers. Representative examples of the expression patterns of 5 common cancer-specific antigens in non-cancerous normal organs (pharynx, mammary gland, esophagus, lung, stomach, liver, gallbladder, pancreas, colon, kidney, ovary, uterus, and skin) adjacent to the cancer lesions of a total of 385 cases of surgically resected advanced cancer are shown.
Figure 5
Figure 5
Development of a companion diagnostic method for simultaneously determining the expression of common cancer-specific antigens and HLA class I using a multiplexed fluorescent immunostaining system. Using a multiplexed fluorescent immunostaining system, we attempted to establish a companion diagnostic method for simultaneously determining the expression of 5 common cancer-specific antigens and HLA class I. We established a system for simultaneously staining five membrane protein common cancer antigens EphB4, ROBO1, LAT1, CLDN1, GPC3, and HLA class I in six colors (A). We evaluated the expression of these six antigens on the cell membrane using formalin-fixed paraffin sections of surgically resected specimens from actual cases of hepatocellular carcinoma, cholangiocarcinoma, and oropharyngeal carcinoma (B).
Figure 6
Figure 6
A scheme for cocktail CAR-T cell therapy to overcome the diversity of solid tumors. If cocktail CAR-T cell therapy is developed that targets multiple of the five common membrane-expressed cancer antigens, it will likely be usable against most solid tumors.
See this image and copyright information in PMC

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