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. 2025 Mar;116(3):744-752.
doi: 10.1111/cas.16433. Epub 2024 Dec 20.

Evaluation of MAGE-A4 expression in breast cancer and its impact on prognosis

Kaho Nakamura  1 , Kanako Saito  2 , Chihiro Higashi  1 , Yuji Kozuka  3 , Hiroto Yuasa  3 , Yurina Nishimura  2 , Makoto Ishitobi  1   4 , Mikiya Ishihara  5 , Toshiro Mizuno  2 , Isao Tawara  6 , Taizo Shiraishi  7 , Tomoko Ogawa  1 , Shinichi Kageyama  8 , Yoshihiro Miyahara  9
Affiliations

Evaluation of MAGE-A4 expression in breast cancer and its impact on prognosis

Kaho Nakamura et al. Cancer Sci. 2025 Mar.

Abstract

Melanoma-associated antigen (MAGE)-A4, a cancer testis antigen, presents a promising target for chimeric antigen receptor T cell therapy in refractory solid tumors, including breast cancer (BC). However, the lack of highly specific Abs against MAGE-A4 is a major challenge for the development of MAGE-A4-targeted immunotherapies. This study aimed to validate the specificity of a novel MAGE-A4 Ab (E701U) and examine MAGE-A4 expression in clinical BC samples. MAGE-A1, -A2B, -A3, -A4, -A6, -A9, -A10, and -A12 genes were transfected into HEK293 cells. MAGE-A4 expression in each inserted cell block was evaluated using an E701U Ab. Subsequently, we evaluated MAGE-A4 expression in 403 primary BC tissue samples by immunohistochemistry using E701U and analyzed the clinical impact of MAGE-A4 in patients with early BC. The results showed that MAGE-A4 expression was limited to cells transduced with the MAGE-A4 gene. MAGE-A4 expression was observed in 5.7% of the BC samples. Positivity in triple-negative BC was significantly higher than in the other subtypes. The 5-year overall survival rate of patients with MAGE-A4(+) was significantly worse than those with MAGE-A4(-) BC. Moreover, the 5-year recurrence-free survival (RFS) rate of patients with MAGE-A4(+) BC was significantly lower than that of patients with MAGE-A4(-) BC. MAGE-A4 expression was an independent prognostic factor for RFS. In conclusion, the E701U Ab showed reliable specificity for MAGE-A4 expression among MAGE family genes. Patients with MAGE-A4(+) BC have an unfavorable prognosis and represent potential candidates for MAGE-A4-specific immunotherapy.

Keywords: MAGE‐A4; breast cancer; immunotherapy; prognosis; triple‐negative breast cancer.

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Conflict of interest statement

Y.M. belongs to the Department of Personalized Cancer Immunotherapy at the Mie University Graduate School of Medicine. This is an endowment department supported by a grant from T Cell Nouveau. Y.M. received research funds from ASAHI KASEI Corporation (Japan), Kohjin Bio Co. Ltd. (Japan), and T cell Nouveau Inc. (Japan). The other authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Validation of the specificity of E701U Ab for MAGE‐A4 expression (HEK293 cells). Each cell block of HEK293 cells, in which MAGE family genes were inserted, was evaluated using E701U (left panels, H&E staining; right panels, E701U Ab).
FIGURE 2
FIGURE 2
Representative images of MAGE‐A4 expression in invasive breast cancer. Positive expression of MAGE‐A4 was observed in (A) invasive ductal carcinoma (IDC) and (B) invasive lobular carcinoma. (C) Negative expression in IDC is shown.
FIGURE 3
FIGURE 3
Overall survival (OS) of patients in 403 patients with invasive breast cancer (Cohort 1) according to MAGE‐A4 expression. The Kaplan–Meier curves show the OS of MAGE‐A4‐negative (blue) and ‐positive (red) patients in Cohort 1.
FIGURE 4
FIGURE 4
Recurrence‐free survival (RFS) of 382 patients with invasive breast cancer excluding patients who received neoadjuvant chemotherapy (Cohort 2) according to melanoma‐associated antigen (MAGE)‐A4 expression. Kaplan–Meier curves show the RFS of MAGE‐A4‐negative (blue) and ‐positive (red) patients in Cohort 2.
FIGURE 5
FIGURE 5
Overall survival (OS) of patients with triple‐negative breast cancer (TNBC) among 403 patients with invasive breast cancer (Cohort 1), according to MAGE‐A4 expression. Kaplan–Meier curves showing the OS of patients with MAGE‐A4‐negative (blue) and MAGE‐A4‐positive (red) TNBC in Cohort 1.
FIGURE 6
FIGURE 6
Recurrence‐free survival (RFS) of patients with triple‐negative breast cancer (TNBC) among 382 patients with invasive breast cancer excluding patients who received neoadjuvant chemotherapy (Cohort 2), according to MAGE‐A4 expression. Kaplan–Meier curves showing the RFS of patients with MAGE‐A4‐negative (blue) and MAGE‐A4‐positive (red) TNBC in Cohort 2.
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