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Identification of pimavanserin tartrate as a potent Ca2+-calcineurin-NFAT pathway inhibitor for glioblastoma therapy

Abstract

Glioblastoma multiforme (GBM) is the most common and malignant type of primary brain tumor, and 95% of patients die within 2 years after diagnosis. In this study, aiming to overcome chemoresistance to the first-line drug temozolomide (TMZ), we carried out research to discover a novel alternative drug targeting the oncogenic NFAT signaling pathway for GBM therapy. To accelerate the drug’s clinical application, we took advantage of a drug repurposing strategy to identify novel NFAT signaling pathway inhibitors. After screening a set of 93 FDA-approved drugs with simple structures, we identified pimavanserin tartrate (PIM), an effective 5-HT2A receptor inverse agonist used for the treatment of Parkinson’s disease-associated psychiatric symptoms, as having the most potent inhibitory activity against the NFAT signaling pathway. Further study revealed that PIM suppressed STIM1 puncta formation to inhibit store-operated calcium entry (SOCE) and subsequent NFAT activity. In cellula, PIM significantly suppressed the proliferation, migration, division, and motility of U87 glioblastoma cells, induced G1/S phase arrest and promoted apoptosis. In vivo, the growth of subcutaneous and orthotopic glioblastoma xenografts was markedly suppressed by PIM. Unbiased omics studies revealed the novel molecular mechanism of PIM’s antitumor activity, which included suppression of the ATR/CDK2/E2F axis, MYC, and AuroraA/B signaling. Interestingly, the genes upregulated by PIM were largely associated with cholesterol homeostasis, which may contribute to PIM’s side effects and should be given more attention. Our study identified store-operated calcium channels as novel targets of PIM and was the first to systematically highlight the therapeutic potential of pimavanserin tartrate for glioblastoma.

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Fig. 1: Identification of pimavanserin tartrate as a potent NFAT pathway inhibitor.
Fig. 2: Pimavanserin tartrate inhibits TG-induced NFAT nuclear translocation independent of the 5-HT2A receptor.
Fig. 3: Pimavanserin tartrate inhibits NFAT nuclear translocation by targeting store-operated calcium entry.
Fig. 4: Pimavanserin tartrate prevents STIM1 puncta formation to inhibit store-operated calcium entry.
Fig. 5: Pimavanserin tartrate impedes the growth, migration and mobility of glioblastoma cells in vitro.
Fig. 6: Molecular signature of pimavanserin tartrate treatment in glioblastoma U87 cells, as assessed by transcriptomic and proteomic profiling.
Fig. 7: Pimavanserin tartrate inhibits the growth of subcutaneous and orthotopic glioblastoma xenografts in vivo.

Data availability

Additional experimental data are provided as Supplementary Information and are available from the corresponding author upon request. Raw RNA-Seq data are available in NCBI’s Sequence Read Archive (SRA) database under accession number PRJNA689001. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the iProX partner repository with the dataset identifier PXD023207 [51].

References

  1. 1.

    Stylli SS. Novel treatment strategies for glioblastoma. Cancers. 2020;12:2883.

    PubMed Central  CAS  Google Scholar 

  2. 2.

    Neftel C, Laffy J, Filbin MG, Hara T, Shore ME, Rahme GJ, et al. An integrative model of cellular states, plasticity, and genetics for glioblastoma. Cell. 2019;178:835–49. e21.

    PubMed  PubMed Central  CAS  Google Scholar 

  3. 3.

    Recurrent MET fusion genes represent a drug target in pediatric glioblastoma. Nat Med. 2016;22:1314–20.

  4. 4.

    Hu B, Wang Q, Wang YA, Hua S, Sauvé C-EG, Ong D, et al. Epigenetic activation of WNT5A drives glioblastoma stem cell differentiation and invasive growth. Cell. 2016;167:1281–95. e18.

    PubMed  PubMed Central  CAS  Google Scholar 

  5. 5.

    Qin JJ, Nag S, Wang W, Zhou J, Zhang WD, Wang H, et al. NFAT as cancer target: mission possible? Biochim Biophys Acta. 2014;1846:297–311.

    PubMed  PubMed Central  CAS  Google Scholar 

  6. 6.

    Muppidi JR. A role for NFAT signaling in ABC-DLBCL. Blood. 2020;135:81.

    PubMed  PubMed Central  CAS  Google Scholar 

  7. 7.

    Kaunisto A, Henry WS, Montaser-Kouhsari L, Jaminet SC, Oh EY, Zhao L, et al. NFAT1 promotes intratumoral neutrophil infiltration by regulating IL8 expression in breast cancer. Mol Oncol. 2015;9:1140–54.

    PubMed  PubMed Central  CAS  Google Scholar 

  8. 8.

    Tie X, Han S, Meng L, Wang Y, Wu A. NFAT1 is highly expressed in, and regulates the invasion of, glioblastoma multiforme cells. PLoS One. 2013;8:e66008.

    PubMed  PubMed Central  CAS  Google Scholar 

  9. 9.

    Lee JV, Berry CT, Kim K, Sen P, Kim T, Carrer A, et al. Acetyl-CoA promotes glioblastoma cell adhesion and migration through Ca2+-NFAT signaling. Genes Dev. 2018;32:497–511.

    PubMed  PubMed Central  CAS  Google Scholar 

  10. 10.

    Chigurupati S, Venkataraman R, Barrera D, Naganathan A, Madan M, Paul L, et al. Receptor channel TRPC6 is a key mediator of Notch-driven glioblastoma growth and invasiveness. Cancer Res. 2010;70:418–27.

    PubMed  CAS  Google Scholar 

  11. 11.

    Robbs BK, Cruz AL, Werneck MB, Mognol GP, Viola JP. Dual roles for NFAT transcription factor genes as oncogenes and tumor suppressors. Mol Cell Biol. 2008;28:7168–81.

    PubMed  PubMed Central  CAS  Google Scholar 

  12. 12.

    Pushpakom S, Iorio F, Eyers PA, Escott KJ, Hopper S, Wells A, et al. Drug repurposing: progress, challenges and recommendations. Nat Rev Drug Discov. 2019;18:41–58.

    PubMed  CAS  Google Scholar 

  13. 13.

    Liu Z, Li H, He L, Xiang Y, Tian C, Li C, et al. Discovery of small-molecule inhibitors of the HSP90-calcineurin-NFAT pathway against glioblastoma. Cell Chem Biol. 2019;26:352–65. e7.

    PubMed  PubMed Central  CAS  Google Scholar 

  14. 14.

    Abbas A, Roth BL. Pimavanserin tartrate: a 5-HT2A inverse agonist with potential for treating various neuropsychiatric disorders. Expert Opin Pharmacother. 2008;9:3251–9.

    PubMed  CAS  Google Scholar 

  15. 15.

    Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, et al. Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP. Nat Commun. 2018;9:1504.

    PubMed  PubMed Central  Google Scholar 

  16. 16.

    Nguyen NT, Han W, Cao WM, Wang Y, Wen S, Huang Y, et al. Store-operated calcium entry mediated by ORAI and STIM. Compr Physiol. 2018;8:981–1002.

    PubMed  Google Scholar 

  17. 17.

    Liou J, Fivaz M, Inoue T, Meyer T. Live-cell imaging reveals sequential oligomerization and local plasma membrane targeting of stromal interaction molecule 1 after Ca2+ store depletion. Proc Natl Acad Sci USA. 2007;104:9301–6.

    PubMed  PubMed Central  CAS  Google Scholar 

  18. 18.

    Zhang SL, Yu Y, Roos J, Kozak JA, Deerinck TJ, Ellisman MH, et al. STIM1 is a Ca2+ sensor that activates CRAC channels and migrates from the Ca2+ store to the plasma membrane. Nature. 2005;437:902–5.

    PubMed  PubMed Central  CAS  Google Scholar 

  19. 19.

    He L, Zhang Y, Ma G, Tan P, Li Z, Zang S, et al. Near-infrared photoactivatable control of Ca2+ signaling and optogenetic immunomodulation. Elife. 2015;4:e10024.

  20. 20.

    Morgan AJ, Jacob R. Ionomycin enhances Ca2+ influx by stimulating store-regulated cation entry and not by a direct action at the plasma membrane. Biochem J. 1994;300:665–72.

    PubMed  PubMed Central  CAS  Google Scholar 

  21. 21.

    Kang F, Zhou M, Huang X, Fan J, Wei L, Boulanger J, et al. E-syt1 re-arranges STIM1 clusters to stabilize ring-shaped ER-PM contact sites and accelerate Ca2+ store replenishment. Sci Rep. 2019;9:3975.

    PubMed  PubMed Central  Google Scholar 

  22. 22.

    Woo JS, Sun Z, Srikanth S, Gwack Y. The short isoform of extended synaptotagmin-2 controls Ca2+ dynamics in T cells via interaction with STIM1. Sci Rep. 2020;10:14433.

    PubMed  PubMed Central  CAS  Google Scholar 

  23. 23.

    Giordano F, Saheki Y, Idevall-Hagren O, Colombo SF, Pirruccello M, Milosevic I, et al. PI4,5P2-dependent and Ca2+-regulated ER-PM interactions mediated by the extended synaptotagmins. Cell. 2013;153:1494–509.

    PubMed  PubMed Central  CAS  Google Scholar 

  24. 24.

    Schmidt JH, Pietkiewicz S, Naumann M, Lavrik IN. Quantification of CD95-induced apoptosis and NF-κB activation at the single cell level. J Immunol Methods. 2015;423:12–7.

    PubMed  CAS  Google Scholar 

  25. 25.

    Attwooll C, Lazzerini Denchi E, Helin K. The E2F family: specific functions and overlapping interests. EMBO J. 2004;23:4709–16.

    PubMed  PubMed Central  CAS  Google Scholar 

  26. 26.

    Chen H, Liu H, Qing G. Targeting oncogenic Myc as a strategy for cancer treatment. Signal Transduct Target Ther. 2018;3:5.

    PubMed  PubMed Central  Google Scholar 

  27. 27.

    Chen HZ, Tsai SY, Leone G. Emerging roles of E2Fs in cancer: an exit from cell cycle control. Nat Rev Cancer. 2009;9:785–97.

    PubMed  PubMed Central  CAS  Google Scholar 

  28. 28.

    Mei L, Zhang J, He K, Zhang J. Ataxia telangiectasia and Rad3-related inhibitors and cancer therapy: where we stand. J Hematol Oncol. 2019;12:43.

    PubMed  PubMed Central  Google Scholar 

  29. 29.

    Harbour JW, Luo RX, Santi AD, Postigo AA, Dean DC. Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through G1. Cell. 1999;98:859–69.

    PubMed  CAS  Google Scholar 

  30. 30.

    Laoukili J, Alvarez M, Meijer LA, Stahl M, Mohammed S, Kleij L, et al. Activation of FoxM1 during G2 requires cyclin A/Cdk-dependent relief of autorepression by the FoxM1 N-terminal domain. Mol Cell Biol. 2008;28:3076–87.

    PubMed  PubMed Central  CAS  Google Scholar 

  31. 31.

    Carmena M, Wheelock M, Funabiki H, Earnshaw WC. The chromosomal passenger complex (CPC): from easy rider to the godfather of mitosis. Nat Rev Mol Cell Biol. 2012;13:789–803.

    PubMed  PubMed Central  CAS  Google Scholar 

  32. 32.

    Bayliss R, Sardon T, Vernos I, Conti E. Structural basis of aurora-A activation by TPX2 at the mitotic spindle. Mol Cell. 2003;12:851–62.

    PubMed  CAS  Google Scholar 

  33. 33.

    Wood MA, McMahon SB, Cole MD. An ATPase/helicase complex is an essential cofactor for oncogenic transformation by c-Myc. Mol Cell. 2000;5:321–30.

    PubMed  CAS  Google Scholar 

  34. 34.

    Park J, Wood MA, Cole MD. BAF53 forms distinct nuclear complexes and functions as a critical c-Myc-interacting nuclear cofactor for oncogenic transformation. Mol Cell Biol. 2002;22:1307–16.

    PubMed  PubMed Central  CAS  Google Scholar 

  35. 35.

    Luo J, Yang H, Song BL. Mechanisms and regulation of cholesterol homeostasis. Nat Rev Mol Cell Biol. 2020;21:225–45.

    PubMed  CAS  Google Scholar 

  36. 36.

    Rosenbaum AI, Maxfield FR. Niemann-Pick type C disease: molecular mechanisms and potential therapeutic approaches. J Neurochem. 2011;116:789–95.

    PubMed  PubMed Central  CAS  Google Scholar 

  37. 37.

    Horton JD, Goldstein JL, Brown MS. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. J Clin Invest. 2002;109:1125–31.

    PubMed  PubMed Central  CAS  Google Scholar 

  38. 38.

    Laplante M, Sabatini DM. An emerging role of mTOR in lipid biosynthesis. Curr Biol. 2009;19:R1046–52.

    PubMed  PubMed Central  CAS  Google Scholar 

  39. 39.

    Eid W, Dauner K, Courtney KC, Gagnon A, Parks RJ, Sorisky A, et al. mTORC1 activates SREBP-2 by suppressing cholesterol trafficking to lysosomes in mammalian cells. Proc Natl Acad Sci USA. 2017;114:7999–8004.

    PubMed  PubMed Central  CAS  Google Scholar 

  40. 40.

    Lim M, Xia Y, Bettegowda C, Weller M. Current state of immunotherapy for glioblastoma. Nat Rev Clin Oncol. 2018;15:422–42.

    PubMed  CAS  Google Scholar 

  41. 41.

    Varalda M, Antona A, Bettio V, Roy K, Vachamaram A, Yellenki V, et al. Psychotropic drugs show anticancer activity by disrupting mitochondrial and lysosomal function. Front Oncol. 2020;10:562196.

    PubMed  PubMed Central  Google Scholar 

  42. 42.

    Pappas SG, Jordan VC. Raloxifene for the treatment and prevention of breast cancer? Expert Rev Anticancer Ther. 2001;1:334–40.

    PubMed  CAS  Google Scholar 

  43. 43.

    Chang CL, Chen YJ, Quintanilla CG, Hsieh TS, Liou J. EB1 binding restricts STIM1 translocation to ER-PM junctions and regulates store-operated Ca2+ entry. J Cell Biol. 2018;217:2047–58.

    PubMed  PubMed Central  CAS  Google Scholar 

  44. 44.

    Jing J, He L, Sun A, Quintana A, Ding Y, Ma G, et al. Proteomic mapping of ER–PM junctions identifies STIMATE as a regulator of Ca2+ influx. Nat Cell Biol. 2015;17:1339–47.

    PubMed  PubMed Central  CAS  Google Scholar 

  45. 45.

    Wiklund ED, Catts VS, Catts SV, Ng TF, Whitaker NJ, Brown AJ, et al. Cytotoxic effects of antipsychotic drugs implicate cholesterol homeostasis as a novel chemotherapeutic target. Int J Cancer. 2010;126:28–40.

    PubMed  CAS  Google Scholar 

  46. 46.

    Barak Y, Achiron A, Mandel M, Mirecki I, Aizenberg D. Reduced cancer incidence among patients with schizophrenia. Cancer. 2005;104:2817–21.

    PubMed  Google Scholar 

  47. 47.

    Ramachandran S, Srivastava SK. Repurposing pimavanserin, an anti-Parkinson drug for pancreatic cancer therapy. Mol Ther Oncolytics. 2020;19:19–32.

    PubMed  PubMed Central  CAS  Google Scholar 

  48. 48.

    Breen DP, Vuono R, Nawarathna U, Fisher K, Shneerson JM, Reddy AB, et al. Sleep and circadian rhythm regulation in early Parkinson disease. JAMA Neurol. 2014;71:589–95.

    PubMed  PubMed Central  Google Scholar 

  49. 49.

    Canfrán-Duque A, Casado ME, Pastor O, Sánchez-Wandelmer J, de la Peña G, Lerma M, et al. Atypical antipsychotics alter cholesterol and fatty acid metabolism in vitro. J Lipid Res. 2013;54:310–24.

    PubMed  PubMed Central  Google Scholar 

  50. 50.

    Fernø J, Skrede S, Vik-Mo AO, Håvik B, Steen VM. Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines: marked differences between various antipsychotic drugs. BMC Neurosci. 2006;7:69.

    PubMed  PubMed Central  Google Scholar 

  51. 51.

    Ma J, Chen T, Wu S, Yang C, Bai M, Shu K, et al. iProX: an integrated proteome resource. Nucleic Acids Res. 2019;47:D1211–D7.

    PubMed  Google Scholar 

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21927811, 91753111, 21907061, 62006144), the Key Research and Development Program of Shandong Province (2018YFJH0502), the Postdoctoral Research Foundation of China (2017M622225), Postdoctoral Innovation Foundation of Shandong Province (201703009) and Jinan Innovative Team Project (2019GXRC039).

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ZZL, XNL, PL and BT designed the research. ZZL, XNL, HLL, YPJ, RCF, LZJ, and YQZ conducted experiments and acquired the data. ZZL, XNL, HLL, YPJ, RCF, QKZ, LZJ, XQG, YQW, and MQY performed data analysis. ZZL, XNL, HLL, PL, and BT wrote and revised the manuscript.

Corresponding authors

Correspondence to Hong-li Li, Ping Li or Bo Tang.

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The authors declare no competing interests.

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Liu, Zz., Liu, Xn., Fan, Rc. et al. Identification of pimavanserin tartrate as a potent Ca2+-calcineurin-NFAT pathway inhibitor for glioblastoma therapy. Acta Pharmacol Sin 42, 1860–1874 (2021). https://doi.org/10.1038/s41401-021-00724-2

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Keywords

  • pimavanserin tartrate
  • drug repurposing
  • SOCE
  • NFAT signaling pathway
  • glioblastoma

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