Doxorubicin-Induced TrkAIII Activation: A Selection Mechanism for Resistant Dormant Neuroblastoma Cells

• Published in: International journal of molecular sciences Vol. 23; no. 18; p. 10895
• Main Authors: Cappabianca, Lucia, Sebastiano, Michela, Ruggieri, Marianna, Sbaffone, Maddalena, Zelli, Veronica, Farina, Antonietta Rosella, Mackay, Andrew Reay
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 17-09-2022; MDPI
• Subjects: AKT protein; Alternative splicing; Anthracycline; Calcium ions; Calcium-binding protein; Calmodulin; Cardiotoxicity; Cell cycle; Chemotherapy; Doxorubicin; doxorubicin-resistance; Drug dosages; Gene expression; Genotype & phenotype; Hsp90 protein; Hypoxia; Immunoprecipitation; Kinases; Ligands; Medical prognosis; Metastasis; Neuroblastoma; Neuroblasts; Oxidation; Phosphorylation; Protein-tyrosine kinase; Ryanodine receptors; Senescence; TrkA; TrkA protein; TrkA receptors; TrkAIII; Tyrosine; tyrosine kinase inhibitors; Western blotting
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms231810895

Patients with advanced neuroblastoma (NB) receive multimodal clinical therapy, including the potent anthracycline chemotherapy drug doxorubicin (Dox). The acquisition of Dox resistance, however, is a major barrier to a sustained response and leads to a poor prognosis in advanced disease states, reinforcing the need to identify and inhibit Dox resistance mechanisms. In this context, we report on the identification and inhibition of a novel Dox resistance mechanism. This mechanism is characterized by the Dox-induced activation of the oncogenic TrkAIII alternative splice variant, resulting in increased Dox resistance, and is blocked by lestaurtinib, entrectinib, and crizotinib tyrosine kinase and LY294002 IP3-K inhibitors. Using time lapse live cell imaging, conventional and co-immunoprecipitation Western blots, RT-PCR, and inhibitor studies, we report that the Dox-induced TrkAIII activation correlates with proliferation inhibition and is CDK1- and Ca2+-uniporter-independent. It is mediated by ryanodine receptors; involves Ca2+-dependent interactions between TrkAIII, calmodulin and Hsp90; requires oxygen and oxidation; occurs within assembled ERGICs; and does not occur with fully spliced TrkA. The inhibitory effects of lestaurtinib, entrectinib, crizotinib, and LY294002 on the Dox-induced TrkAIII and Akt phosphorylation and resistance confirm roles for TrkAIII and IP3-K consistent with Dox-induced, TrkAIII-mediated pro-survival IP3K/Akt signaling. This mechanism has the potential to select resistant dormant TrkAIII-expressing NB cells, supporting the use of Trk inhibitors during Dox therapy in TrkAIII-expressing NBs.