Neuropilin-1 upregulation elicits adaptive resistance to oncogene-targeted therapies
Sabrina Rizzolio, Gabriella Cagnoni, Chiara Battistini, Stefano Bonelli, Claudio Isella, Jo A. Van Ginderachter, René Bernards, Federica Di Nicolantonio, Silvia Giordano, Luca Tamagnone
Sabrina Rizzolio, Gabriella Cagnoni, Chiara Battistini, Stefano Bonelli, Claudio Isella, Jo A. Van Ginderachter, René Bernards, Federica Di Nicolantonio, Silvia Giordano, Luca Tamagnone
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Research Article Oncology

Neuropilin-1 upregulation elicits adaptive resistance to oncogene-targeted therapies

Abstract

Cancer cell dependence on activated oncogenes is therapeutically targeted, but acquired resistance is virtually unavoidable. Here we show that the treatment of addicted melanoma cells with BRAF inhibitors, and of breast cancer cells with HER2-targeted drugs, led to an adaptive rise in neuropilin-1 (NRP1) expression, which is crucial for the onset of acquired resistance to therapy. Moreover, NRP1 levels dictated the efficacy of MET oncogene inhibitors in addicted stomach and lung carcinoma cells. Mechanistically, NRP1 induced a JNK-dependent signaling cascade leading to the upregulation of alternative effector kinases EGFR or IGF1R, which in turn sustained cancer cell growth and mediated acquired resistance to BRAF, HER2, or MET inhibitors. Notably, the combination with NRP1-interfering molecules improved the efficacy of oncogene-targeted drugs and prevented or even reversed the onset of resistance in cancer cells and tumor models. Our study provides the rationale for targeting the NRP1-dependent upregulation of tyrosine kinases, which are responsible for loss of responsiveness to oncogene-targeted therapies.

Authors

Sabrina Rizzolio, Gabriella Cagnoni, Chiara Battistini, Stefano Bonelli, Claudio Isella, Jo A. Van Ginderachter, René Bernards, Federica Di Nicolantonio, Silvia Giordano, Luca Tamagnone

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Figure 8

SOX2 and JUN transcription factors mediate NRP1/JNK-driven upregulation of EGFR and IGF1R, respectively.

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SOX2 and JUN transcription factors mediate NRP1/JNK-driven upregulation ...
(A) qPCR analysis of SOX2 expression in parental and drug-resistant A375 and EBC1 cells (shown in B), either in basal conditions or in the presence of 25 μM SP600125; values normalized to untreated controls (n = 5). (B) qPCR analysis of EGFR expression in parental and drug-resistant A375 cells subjected to SOX2 silencing by targeted siRNAs (validated in Supplemental Figure 7F) (n = 4). (C) EGFR expression in EBC1 cells (shown in B), subjected to SOX2 silencing (validated in Supplemental Figure 7F) (1 representative experiment of 4 repetitions). (D) qPCR analysis of JUN expression in parental and drug-resistant BT474 cells, either in basal conditions or in the presence of 25 μM SP600125; values normalized to untreated parental cells (n = 3). (E) IGF1R expression in BT474 cells (same as above), subjected to JUN silencing by RNAi (validated in Supplemental Figure 7G) (1 representative experiment of 4 repetitions). (F) Schematic working model of the proposed signaling cascade leading to the onset of resistance to oncogene-targeted therapy in different cancer cells. The upstream regulation of NRP1 levels by miR-338-3p (marked in red) was specifically observed in melanoma cells, whereas in carcinoma cells NRP1 expression might depend on RTK-Ras signaling cascade (marked in blue). (G) SK-MEL-28 cell viability was assessed upon treatment with 1 μM PLX-4720, alone or in association with the indicated concentrations of JNK-i SP600125. While naive cells were initially sensitive to PLX-4720, after 3–5 weeks in culture they progressively developed drug resistance, but this was prevented in the presence of JNK-i (n > 3). In all panels, statistical significance was assessed by Student’s t test with Bonferroni’s correction; *P < 0.01, **P < 0.001, ***P < 0.0001.