Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8+ T cells in triple-negative breast cancer

Lim, B. J. W.; Liu, M.; Wang, L.; Kong, S. L. Y.; Yin, T.; Yan, C.; Xiang, K.; Cao, C.; Wu, H.; Mihai, A.; Tay, F.; Wang, E.; Jiang, Q.; Ma, Z.; Tan, L.; Chia, R. N.; Qin, D.; Pan, C. C.; Wang, X.-F.; Li, Q.-J.

doi:10.1101/2025.01.29.635356

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bioRxiv · 10.1101/2025.01.29.635356

Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8+ T cells in triple-negative breast cancer

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Abstract

Neoadjuvant immunotherapy seeks to harness the primary tumor as a source of relevant tumor antigens to enhance systemic anti-tumor immunity through improved immunological surveillance. Despite having revolutionized the treatment of patients with high-risk early-stage triple-negative breast cancer (TNBC), a significant portion of patients remain unresponsive and succumb to metastatic recurrence post-treatment. Here, we found that optimally scheduled neoadjuvant administration of anti-4-1BB monotherapy was able to counteract metastases and prolong survival following surgical resection. Phenotypic and transcriptional profiling revealed enhanced 4-1BB expression on tumor-infiltrating intermediate (Tint), relative to progenitor (Tprog) and terminally exhausted (Tterm) T cells. Furthermore, Tint was enriched in low-affinity T cells. Treatment with anti-4-1BB drove clonal expansion of Tint, with reduced expression of tissue-retention marker CD103 in Tprog. This was accompanied by increased TCR clonotype sharing between paired tumors and pre-metastatic lungs. Further interrogation of sorted intratumor T cells confirmed enhanced T cell egress into circulation following anti-4-1BB treatment. In addition, gene signature extracted from anti-4-1BB treated Tint was consistently associated with improved clinical outcomes in BRCA patients. Combinatorial neoadjuvant anti-4-1BB and ablation of tumor-derived CXCL16 resulted in enhanced therapeutic effect. These findings illustrate the intratumor changes underpinning the efficacy of neoadjuvant anti-4-1BB, highlighting the reciprocity between local tissue-retention and distant immunologic fortification, suggesting treatment can reverse the siphoning of intratumor T cells to primary tumor, enabling redistribution to distant tissues and subsequent protection against metastases.

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Lim, B. J. W., Liu, M., Wang, L., Kong, S. L. Y., Yin, T., Yan, C., Xiang, K., Cao, C., Wu, H., Mihai, A., Tay, F., Wang, E., Jiang, Q., Ma, Z., Tan, L., Chia, R. N., Qin, D., Pan, C. C., Wang, X.-F., Li, Q.-J.. 2025-02-02. Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8+ T cells in triple-negative breast cancer. https://doi.org/10.1101/2025.01.29.635356

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