Nano-targeted induction of dual ferroptotic mechanisms eradicates high-risk neuroblastoma
Behrouz Hassannia, … , Peter Vandenabeele, Tom Vanden Berghe
Behrouz Hassannia, … , Peter Vandenabeele, Tom Vanden Berghe
Published June 25, 2018
Citation Information: J Clin Invest. 2018;128(8):3341-3355. https://doi.org/10.1172/JCI99032.
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Research Article Neuroscience Oncology

Nano-targeted induction of dual ferroptotic mechanisms eradicates high-risk neuroblastoma

Abstract

High-risk neuroblastoma is a devastating malignancy with very limited therapeutic options. Here, we identify withaferin A (WA) as a natural ferroptosis-inducing agent in neuroblastoma, which acts through a novel double-edged mechanism. WA dose-dependently either activates the nuclear factor–like 2 pathway through targeting of Kelch-like ECH-associated protein 1 (noncanonical ferroptosis induction) or inactivates glutathione peroxidase 4 (canonical ferroptosis induction). Noncanonical ferroptosis induction is characterized by an increase in intracellular labile Fe(II) upon excessive activation of heme oxygenase-1, which is sufficient to induce ferroptosis. This double-edged mechanism might explain the superior efficacy of WA as compared with etoposide or cisplatin in killing a heterogeneous panel of high-risk neuroblastoma cells, and in suppressing the growth and relapse rate of neuroblastoma xenografts. Nano-targeting of WA allows systemic application and suppressed tumor growth due to an enhanced accumulation at the tumor site. Collectively, our data propose a novel therapeutic strategy to efficiently kill cancer cells by ferroptosis.

Authors

Behrouz Hassannia, Bartosz Wiernicki, Irina Ingold, Feng Qu, Simon Van Herck, Yulia Y. Tyurina, Hülya Bayır, Behnaz A. Abhari, Jose Pedro Friedmann Angeli, Sze Men Choi, Eline Meul, Karen Heyninck, Ken Declerck, Chandra Sekhar Chirumamilla, Maija Lahtela-Kakkonen, Guy Van Camp, Dmitri V. Krysko, Paul G. Ekert, Simone Fulda, Bruno G. De Geest, Marcus Conrad, Valerian E. Kagan, Wim Vanden Berghe, Peter Vandenabeele, Tom Vanden Berghe

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Figure 3

WA-eradicated high-risk neuroblastoma tumors show decreased GPX4 expression, increased HMOX1 expression, and decreased relapse rates.

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WA-eradicated high-risk neuroblastoma tumors show decreased GPX4 express...
(A and B) Light microscopic pictures of H&E staining of tumor sections (A) and quantification of tumor size area (B) representing effect of WA and vehicle (Ctrl) on SH-EP–derived tumors in a chicken chorioallantoic membrane (CAM) model. The combined results of 2 independent experiments are shown. Error bars represent SEM; n = 10 (Ctrl), n = 10 (WA). (C) Quantification of tumor growth rates after therapeutic treatment regime (100–200 mm3 at the start of treatment) with vehicle (Ctrl), WA, or etoposide (Eto). The results of 1 experiment for etoposide and combined results of 2 independent experiments for WA are shown. Each point indicates an individual mouse. Error bars represent SEM; n = 11 (Ctrl), n = 13 (WA), n = 6 (Eto). (D) Quantification of tumor growth rates in mice after termination of treatment with WA and Eto (tumor size cutoff below 180 mm3 at time of termination of the treatment). (E–I) Immunohistochemical staining revealing proliferation using Ki67 antibody (E), DNA fragmentation using TUNEL staining (F), caspase-3 activation using anti–active caspase-3 antibodies (G), GPX4 (H), and HMOX1 expression level (I) in tumor sections after therapeutic treatment regime with vehicle (Ctrl) or WA. (J) Western blot revealing GPX4 and HMOX1 in lysates of tumor sections after therapeutic treatment regime with vehicle (Ctrl) or WA. ****P < 0.0001, 2-tailed t test (B), 1-way ANOVA (C).