Double-positive T cells form heterotypic clusters with circulating tumor cells to foster cancer metastasis
David Scholten, Lamiaa El-Shennawy, Yuzhi Jia, Youbin Zhang, Elizabeth Hyun, Carolina Reduzzi, Andrew D. Hoffmann, Hannah F. Almubarak, Fangjia Tong, Nurmaa K. Dashzeveg, Yuanfei Sun, Joshua R. Squires, Janice Lu, Leonidas C. Platanias, Clive H. Wasserfall, William J. Gradishar, Massimo Cristofanilli, Deyu Fang, Huiping Liu
David Scholten, Lamiaa El-Shennawy, Yuzhi Jia, Youbin Zhang, Elizabeth Hyun, Carolina Reduzzi, Andrew D. Hoffmann, Hannah F. Almubarak, Fangjia Tong, Nurmaa K. Dashzeveg, Yuanfei Sun, Joshua R. Squires, Janice Lu, Leonidas C. Platanias, Clive H. Wasserfall, William J. Gradishar, Massimo Cristofanilli, Deyu Fang, Huiping Liu
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Research Article Clinical Research Immunology Oncology

Double-positive T cells form heterotypic clusters with circulating tumor cells to foster cancer metastasis

Abstract

The immune ecosystem is central to maintaining effective defensive responses. However, it remains largely understudied how immune cells in the peripheral blood interact with circulating tumor cells (CTCs) in metastasis. Here, blood analysis of patients with advanced breast cancer revealed that over 75% of CTC-positive blood specimens contained heterotypic CTC clusters with CD45+ white blood cells (WBCs), which correlates with breast cancer subtypes, racial groups, and decreased survival. CTC-WBC clusters included overrepresented T cells and underrepresented neutrophils. Specifically, a rare subset of CD4 and CD8 double-positive T (DPT) cells was 140-fold enriched in CTC clusters versus their frequency in WBCs. DPT cells shared properties with CD4+ and CD8+ T cells but exhibited unique features of T cell exhaustion and immune suppression. Mechanistically, the integrin heterodimer α4β1, also named very late antigen 4 (VLA-4), in DPT cells and its ligand, VCAM1, in tumor cells are essential mediators of DPT-CTC clusters. Neoadjuvant administration of anti-VLA-4 neutralizing antibodies markedly blocked CTC–DPT clusters, inhibited metastasis, and extended mouse survival. These findings highlight a pivotal role of rare DPT cells in fostering cancer dissemination through CTC clustering. It lays a foundation for developing innovative biomarker-guided therapeutic strategies to prevent and target cancer metastasis.

Authors

David Scholten, Lamiaa El-Shennawy, Yuzhi Jia, Youbin Zhang, Elizabeth Hyun, Carolina Reduzzi, Andrew D. Hoffmann, Hannah F. Almubarak, Fangjia Tong, Nurmaa K. Dashzeveg, Yuanfei Sun, Joshua R. Squires, Janice Lu, Leonidas C. Platanias, Clive H. Wasserfall, William J. Gradishar, Massimo Cristofanilli, Deyu Fang, Huiping Liu

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Figure 6

Targeting VLA-4 inhibits spontaneous lung metastasis and CTC-DPT clusters in vivo.

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Targeting VLA-4 inhibits spontaneous lung metastasis and CTC-DPT cluster...
(A) Schematic of anti–VLA-4 neutralizing antibody (αVLA-4 Ab) treatment for orthotopic 4T1 breast tumors on days 3, 5, 7, and 9. Blood/tissue harvests for primary tumors, CTCs, and lung metastasis analyses on day 10 (N = 6). The primary tumor was removed via survival surgeries with the rest of the mice (N = 7 per group) for extended survival analyses until 6 weeks. (B) Bar graphs of 4T1 tumor weight and photos of dissected tumors of IgG control and αVLA-4–treated groups on day 10. Unpaired 2-tailed t test (N = 6) between 2 groups. (C) Mouse survival after neoadjuvant treatment of IgG and αVLA-4 followed by a surgical removal of primary tumors on day 10 (N = 7). Log-rank P = 0.03846 for distinct survival of 2 groups of the mice by 6 weeks. Hazard ratio = 4.435 (0.9842–20.75, 95% CI of ratio) when the IgG group was compared with the αVLA-4–treated group. (D) Bar graphs of blood-detected CTC-WBC clusters (left) and CTC-DPT clusters (middle), and the burden of lung tumor cells (right) detected and quantified via flow cytometry on day 10 (N = 6). Unpaired, 2-tailed t test P values are shown.