SH3BP5L triggers the RAB11A-regulated integrin recycling network implicated in breast cancer metastasis
Huayi Li, Maria Chiara De Santis, Francesco A. Tucci, Daniela Tosoni, Ping Zhang, Meredith L. Jenkins, Giulia Villari, Maria Grazia Filippone, Elisa Guerrera, Simone Tealdi, Luca Gozzelino, Federico Gulluni, Lorenzo Prever, Cristina Zanini, Marco Forni, Irene Franco, Miriam Martini, John E. Burke, Guido Serini, Carlo Cosimo Campa, Salvatore Pece, Jean Piero Margaria, Emilio Hirsch
Huayi Li, Maria Chiara De Santis, Francesco A. Tucci, Daniela Tosoni, Ping Zhang, Meredith L. Jenkins, Giulia Villari, Maria Grazia Filippone, Elisa Guerrera, Simone Tealdi, Luca Gozzelino, Federico Gulluni, Lorenzo Prever, Cristina Zanini, Marco Forni, Irene Franco, Miriam Martini, John E. Burke, Guido Serini, Carlo Cosimo Campa, Salvatore Pece, Jean Piero Margaria, Emilio Hirsch
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Research Article Cell biology Oncology

SH3BP5L triggers the RAB11A-regulated integrin recycling network implicated in breast cancer metastasis

Abstract

Metastatic progression in aggressive breast cancer (BC) depends on a tightly controlled vesicular recycling network regulated by RAB11, a small guanosine triphosphate enzyme (GTPase). In a cohort of more than 1,000 patients with BC, we identified SH3BP5L as the most highly expressed guanine nucleotide exchange factor (GEF) for RAB11A. High SH3BP5L expression marked an advanced tumor stage, distant metastasis, and poor prognosis, with significant associations in human epidermal growth factor receptor 2–positive (HER2+) and triple-negative breast cancer (TNBC). Using Förster resonance energy transfer (FRET) sensors and artificial intelligence– (AI-assisted) microscopy, we showed that cargo delivery to the plasma membrane required SH3BP5L-dependent activation of RAB11A and assembly of a complex with the anterograde motor KIF5B. This trafficking governed key metastatic features of TNBC, including β1 integrin recycling and α3β1 integrin surface exposure. Inhibition of SH3BP5L or its GEF activity reduced cell spreading in zebrafish and lung metastasis in mouse models, revealing a previously unidentified driver of BC dissemination and a potential therapeutic vulnerability.

Authors

Huayi Li, Maria Chiara De Santis, Francesco A. Tucci, Daniela Tosoni, Ping Zhang, Meredith L. Jenkins, Giulia Villari, Maria Grazia Filippone, Elisa Guerrera, Simone Tealdi, Luca Gozzelino, Federico Gulluni, Lorenzo Prever, Cristina Zanini, Marco Forni, Irene Franco, Miriam Martini, John E. Burke, Guido Serini, Carlo Cosimo Campa, Salvatore Pece, Jean Piero Margaria, Emilio Hirsch

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Figure 1

SH3BP5L expression in BC and association with patient outcomes.

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SH3BP5L expression in BC and association with patient outcomes. (A) Sche...
(A) Schematic representation of the RAB11 regulatory network. RAB11-GDP is activated by its GEFs, SH3BP5L and SH3BP5, and inactivated by its GAP, TBC1D9B. (B) Copy number alterations of SH3BP5L, SH3BP5, TBC1D9B, and RAB11A across cancer types in TCGA database. Adren., adrenal gland; Oesop., esophagus; Endom., endometrium; Stoma., stomach; Pancr., pancreas; UAT, upper aerodigestive tract; Urina., urinary tract; H&L, hematopoietic and lymphoid; Intes., large intestinal; Soft t., soft tissue. (C) Violin plot showing SH3BP5L mRNA expression across tumor grades in the METABRIC dataset (n = 1,893). **P = 0.002 and ***P < 0.001, by Wilcoxon–Mann-Whitney U test. (D and E) Association of SH3BP5L expression with RFS in a composite BC cohort. (D) Continuous analysis of the log relative hazard against SH3BP5L expression. Both a linear fit (red) and a penalized spline fit (gray) are shown. The HR and log-rank test P value refer to the linear term. Shaded areas represent 95% CIs. Dashed line indicates the upper tertile cutoff. (E) Kaplan-Meier analysis of patients stratified according to SH3BP5Lhi (upper tertile) and SH3BP5Llo expression. P denotes the log-rank P value; n = 4,005. (F) Representative immunoblots of SH3BP5L and SH3BP5 expression normalized to vinculin in luminal and TNBC cell lines. (G) Immunohistochemical assessment of SH3BP5L expression in patients with BC (n = 1590). Representative images of SH3BP5Llo tumors (n = 1,470, score 0/1) and SH3BP5Lhi tumors (n = 120, score 2/3). Scale bars: 100 μm. (H) DMFS analysis of patients with SH3BP5Lhi versus SH3BP5Llo BC. P denotes the log-rank test P value; n = 1,590.