Allosteric Antagonist Modulation of TRPV2 by Piperlongumine Impairs Glioblastoma Progression.

João Conde,Sandra H Vaz,Sohyon Lee,Marc Remke,Daniel Picard,Amrita Samanta,Michael Weller,Florian Sieglitz,Ruth A Pumroy,Berend Snijder,Ana Guerreiro,Tobias Weiss,Francisco Corzana,Tiago Rodrigues,Vanessa A Morais,Bernardo P De Almeida,Vera Y Moiseenkova-Bell,Yuhang Liu,Rafael Roque,Maike Langini,Marta C Marques,Gonçalo J L Bernardes,Charlotte Baker,Seungil Han,Tânia Carvalho,Cláudia C Faria,Nuno L Barbosa‐Morais ,Bárbara Sousa ,Elvira Pequeno Leites ,P Filippakopoulos

doi:10.1021/acscentsci.1c00070

Abstract

The use of computational tools to identify biological targets of natural products with anticancer properties and unknown modes of action is gaining momentum. We employed self-organizing maps to deconvolute the phenotypic effects of piperlongumine (PL) and establish a link to modulation of the human transient receptor potential vanilloid 2 (hTRPV2) channel. The structure of the PL-bound full-length rat TRPV2 channel was determined by cryo-EM. PL binds to a transient allosteric pocket responsible for a new mode of anticancer activity against glioblastoma (GBM) in which hTRPV2 is overexpressed. Calcium imaging experiments revealed the importance of Arg539 and Thr522 residues on the antagonistic effect of PL and calcium influx modulation of the TRPV2 channel. Downregulation of hTRPV2 reduces sensitivity to PL and decreases ROS production. Analysis of GBM patient samples associates hTRPV2 overexpression with tumor grade, disease progression, and poor prognosis. Extensive tumor abrogation and long term survival was achieved in two murine models of orthotopic GBM by formulating PL in an implantable scaffold/hydrogel for sustained local therapy. Furthermore, in primary tumor samples derived from GBM patients, we observed a selective reduction of malignant cells in response to PL ex vivo. Our results establish a broadly applicable strategy, leveraging data-motivated research hypotheses for the discovery of novel means tackling cancer.

Highlights

Natural products provide ample opportunities to develop innovative medicines.[1−4] understanding their mechanisms of action remains a bottleneck to unlock their promise in drug discovery.[5−9] Chemoproteomics is a privileged approach to unveil new biology for molecules of therapeutic interest
We report on an unprecedented link between PL and the human transient receptor potential vanilloid 2 channel
Our studies reveal a new, transient binding site in TRPV2, whose recognition results in selective, potent allosteric antagonism by PL

Summary

■ INTRODUCTION

Natural products provide ample opportunities to develop innovative medicines.[1−4] understanding their mechanisms of action remains a bottleneck to unlock their promise in drug discovery.[5−9] Chemoproteomics is a privileged approach to unveil new biology for molecules of therapeutic interest. We minimized leakage of PL to healthy tissue by decorating the hydrogel with aldehyde groups from oxidized dextran to interact with cancer tissue amines and form adhesive bonds, as previously reported.[41−44] Profiling of our drug delivery system showed a significant discharge of PL in the first 4 h followed by steady release (3% of total) for at least 192 h under physiological conditions (Figure S28) These data provide a rationale to the in vitro toxicity experiments in which PL-doped hydrogels were able to induce near complete cell death after incubation with U251 cells for 24 h. As our tested cohort is relatively small, we hope future studies will further investigate the clinical potential that PL might hold as a putative GBM therapy

■ CONCLUSIONS

■ ACKNOWLEDGMENTS

■ REFERENCES