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Mitoxantrone triggers immunogenic prostate cancer cell death via p53-dependent PERK expression

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Abstract

Background

Mitoxantrone (MTX) is a synthetic compound used as a second line chemotherapeutic drug for prostate cancer. It has been reported to trigger immunogenic cell death (ICD) in animal model studies, but the underlying mechanism is not fully understood yet, especially not in prostate cancer cells.

Methods

ICD was determined by assessing the release of damage-associated molecular patterns (DAMPs) in the prostate cancer-derived cell lines LNCaP, 22RV1 and PC-3. Short hairpin RNAs (shRNAs) were used to knock down target gene expression. Phagocytosis was assessed using a dual labeling technology in dendric cells co-cultured with cancer cells. The PERK gene promoter was cloned for dual luciferase assays. Chromatin immunoprecipitation (ChIP) was used to determine p53 protein-DNA binding activity. Immunocompetent mice and murine RM-1 prostate cancer cells were used for vaccination experiments.

Results

MTX treatment induced typical characteristics of DAMP release, including increased cell surface exposure of calreticulin (CALR), and extracellular release of ATP and high mobility group box-1 (HMGB1) protein. MTX also enhanced phagocytosis by dendritic cells. Moreover, MTX treatment increased eukaryotic initiation factor 2α (eIF2α) S51 phosphorylation, which was reduced when PERK and GCN2 were silenced using shRNAs. In addition, PERK or GCN2 silencing significantly reduced MTX-induced release of DAMPs in vitro and anti-tumor immunity in vivo. MTX treatment also resulted in dendritic cell activation in mice, which was attenuated when PERK or GCN2 were silenced in cancer cells used for vaccination. Further analysis revealed that PERK and GCN2 expression was enhanced by MTX treatment, of which PERK, but not GCN2, was enhanced via a p53-dependent mechanism.

Conclusion

MTX triggers ICD by activating eIF2α via PERK/GCN2 upregulation in prostate cancer cells. MTX-induced PERK expression upregulation depends on the p53 pathway, while that of GCN2 requires further investigation.

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References

  1. L. Galluzzi, I. Vitale, S.A. Aaronson, J.M. Abrams, D. Adam, P. Agostinis, E.S. Alnemri, L. Altucci, I. Amelio, D.W. Andrews, M. Annicchiarico-Petruzzelli, A.V. Antonov, E. Arama, E.H. Baehrecke, N.A. Barlev, N.G. Bazan, F. Bernassola, M.J.M. Bertrand, K. Bianchi, M.V. Blagosklonny, K. Blomgren, C. Borner, P. Boya, C. Brenner, M. Campanella, E. Candi, D. Carmona-Gutierrez, F. Cecconi, F.K. Chan, N.S. Chandel, E.H. Cheng, J.E. Chipuk, J.A. Cidlowski, A. Ciechanover, G.M. Cohen, M. Conrad, J.R. Cubillos-Ruiz, P.E. Czabotar, V. D'Angiolella, T.M. Dawson, V.L. Dawson, V. De Laurenzi, R. De Maria, K.M. Debatin, R.J. DeBerardinis, M. Deshmukh, N. Di Daniele, F. Di Virgilio, V.M. Dixit, S.J. Dixon, C.S. Duckett, B.D. Dynlacht, W.S. El-Deiry, J.W. Elrod, G.M. Fimia, S. Fulda, A.J. Garcia-Saez, A.D. Garg, C. Garrido, E. Gavathiotis, P. Golstein, E. Gottlieb, D.R. Green, L.A. Greene, H. Gronemeyer, A. Gross, G. Hajnoczky, J.M. Hardwick, I.S. Harris, M.O. Hengartner, C. Hetz, H. Ichijo, M. Jaattela, B. Joseph, P.J. Jost, P.P. Juin, W.J. Kaiser, M. Karin, T. Kaufmann, O. Kepp, A. Kimchi, R.N. Kitsis, D.J. Klionsky, R.A. Knight, S. Kumar, S.W. Lee, J.J. Lemasters, B. Levine, A. Linkermann, S.A. Lipton, R.A. Lockshin, C. Lopez-Otin, S.W. Lowe, T. Luedde, E. Lugli, M. MacFarlane, F. Madeo, M. Malewicz, W. Malorni, G. Manic, J.C. Marine, S.J. Martin, J.C. Martinou, J.P. Medema, P. Mehlen, P. Meier, S. Melino, E.A. Miao, J.D. Molkentin, U.M. Moll, C. Munoz-Pinedo, S. Nagata, G. Nunez, A. Oberst, M. Oren, M. Overholtzer, M. Pagano, T. Panaretakis, M. Pasparakis, J.M. Penninger, D.M. Pereira, S. Pervaiz, M.E. Peter, M. Piacentini, P. Pinton, J.H.M. Prehn, H. Puthalakath, G.A. Rabinovich, M. Rehm, R. Rizzuto, C.M.P. Rodrigues, D.C. Rubinsztein, T. Rudel, K.M. Ryan, E. Sayan, L. Scorrano, F. Shao, Y. Shi, J. Silke, H.U. Simon, A. Sistigu, B.R. Stockwell, A. Strasser, G. Szabadkai, S.W.G. Tait, D. Tang, N. Tavernarakis, A. Thorburn, Y. Tsujimoto, B. Turk, T. Vanden Berghe, P. Vandenabeele, M.G.V. Heiden, A. Villunger, H.W. Virgin, K.H. Vousden, D. Vucic, E.F. Wagner, H. Walczak, D. Wallach, Y. Wang, J.A. Wells, W. Wood, J. Yuan, Z. Zakeri, B. Zhivotovsky, L. Zitvogel, G. Melino, G. Kroemer, Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ 25, 486–541 (2018). https://doi.org/10.1038/s41418-017-0012-4

  2. D.R. Green, The coming decade of cell death research: Five riddles. Cell 177, 1094–1107 (2019). https://doi.org/10.1016/j.cell.2019.04.024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. T.L. Aaes, A. Kaczmarek, T. Delvaeye, B. De Craene, S. De Koker, L. Heyndrickx, I. Delrue, J. Taminau, B. Wiernicki, P. De Groote, A.D. Garg, L. Leybaert, J. Grooten, M.J. Bertrand, P. Agostinis, G. Berx, W. Declercq, P. Vandenabeele, D.V. Krysko, Vaccination with Necroptotic Cancer cells induces efficient anti-tumor immunity. Cell Rep 15, 274–287 (2016). https://doi.org/10.1016/j.celrep.2016.03.037

    Article  CAS  PubMed  Google Scholar 

  4. N. Casares, M.O. Pequignot, A. Tesniere, F. Ghiringhelli, S. Roux, N. Chaput, E. Schmitt, A. Hamai, S. Hervas-Stubbs, M. Obeid, F. Coutant, D. Metivier, E. Pichard, P. Aucouturier, G. Pierron, C. Garrido, L. Zitvogel, G. Kroemer, Caspase-dependent immunogenicity of doxorubicin-induced tumor cell death. J Exp Med 202, 1691–1701 (2005). https://doi.org/10.1084/jem.20050915

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. H. Yang, Y. Ma, G. Chen, H. Zhou, T. Yamazaki, C. Klein, F. Pietrocola, E. Vacchelli, S. Souquere, A. Sauvat, L. Zitvogel, O. Kepp, G. Kroemer, Contribution of RIP3 and MLKL to immunogenic cell death signaling in cancer chemotherapy. Oncoimmunology 5, e1149673 (2016). https://doi.org/10.1080/2162402X.2016.1149673

  6. L. Galluzzi, A. Buque, O. Kepp, L. Zitvogel, G. Kroemer, Immunogenic cell death in cancer and infectious disease. Nat Rev Immunol 17, 97–111 (2017). https://doi.org/10.1038/nri.2016.107

    Article  CAS  PubMed  Google Scholar 

  7. O. Kepp, L. Senovilla, I. Vitale, E. Vacchelli, S. Adjemian, P. Agostinis, L. Apetoh, F. Aranda, V. Barnaba, N. Bloy, L. Bracci, K. Breckpot, D. Brough, A. Buque, M.G. Castro, M. Cirone, M.I. Colombo, I. Cremer, S. Demaria, L. Dini, A.G. Eliopoulos, A. Faggioni, S.C. Formenti, J. Fucikova, L. Gabriele, U.S. Gaipl, J. Galon, A. Garg, F. Ghiringhelli, N.A. Giese, Z.S. Guo, A. Hemminki, M. Herrmann, J.W. Hodge, S. Holdenrieder, J. Honeychurch, H.M. Hu, X. Huang, T.M. Illidge, K. Kono, M. Korbelik, D.V. Krysko, S. Loi, P.R. Lowenstein, E. Lugli, Y. Ma, F. Madeo, A.A. Manfredi, I. Martins, D. Mavilio, L. Menger, N. Merendino, M. Michaud, G. Mignot, K.L. Mossman, G. Multhoff, R. Oehler, F. Palombo, T. Panaretakis, J. Pol, E. Proietti, J.E. Ricci, C. Riganti, P. Rovere-Querini, A. Rubartelli, A. Sistigu, M.J. Smyth, J. Sonnemann, R. Spisek, J. Stagg, A.Q. Sukkurwala, E. Tartour, A. Thorburn, S.H. Thorne, P. Vandenabeele, F. Velotti, S.T. Workenhe, H. Yang, W.X. Zong, L. Zitvogel, G. Kroemer, L. Galluzzi, Consensus guidelines for the detection of immunogenic cell death. Oncoimmunology 3, e955691 (2014). https://doi.org/10.4161/21624011.2014.955691

    Article  PubMed  PubMed Central  Google Scholar 

  8. M. Diederich, Natural compound inducers of immunogenic cell death. Arch Pharm Res 42, 629–645 (2019). https://doi.org/10.1007/s12272-019-01150-z

    Article  CAS  PubMed  Google Scholar 

  9. J. Zhou, G. Wang, Y. Chen, H. Wang, Y. Hua, Z. Cai, Immunogenic cell death in cancer therapy: Present and emerging inducers. J Cell Mol Med 23, 4854–4865 (2019). https://doi.org/10.1111/jcmm.14356

    Article  PubMed  PubMed Central  Google Scholar 

  10. A.D. Garg, P. Agostinis, Cell death and immunity in cancer: From danger signals to mimicry of pathogen defense responses. Immunol Rev 280, 126–148 (2017). https://doi.org/10.1111/imr.12574

    Article  CAS  PubMed  Google Scholar 

  11. M. Obeid, A. Tesniere, F. Ghiringhelli, G.M. Fimia, L. Apetoh, J.L. Perfettini, M. Castedo, G. Mignot, T. Panaretakis, N. Casares, D. Metivier, N. Larochette, P. van Endert, F. Ciccosanti, M. Piacentini, L. Zitvogel, G. Kroemer, Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med 13, 54–61 (2007). https://doi.org/10.1038/nm1523

    Article  CAS  PubMed  Google Scholar 

  12. I. Martins, Y. Wang, M. Michaud, Y. Ma, A.Q. Sukkurwala, S. Shen, O. Kepp, D. Metivier, L. Galluzzi, J.L. Perfettini, L. Zitvogel, G. Kroemer, Molecular mechanisms of ATP secretion during immunogenic cell death. Cell Death Differ 21, 79–91 (2014). https://doi.org/10.1038/cdd.2013.75

    Article  CAS  PubMed  Google Scholar 

  13. A.D. Garg, D.V. Krysko, P. Vandenabeele, P. Agostinis, Extracellular ATP and P(2)X(7) receptor exert context-specific immunogenic effects after immunogenic cancer cell death. Cell Death Dis 7, e2097 (2016). https://doi.org/10.1038/cddis.2015.411

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. P. Scaffidi, T. Misteli, M.E. Bianchi, Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 418, 191–195 (2002). https://doi.org/10.1038/nature00858

  15. A.D. Garg, L. Galluzzi, L. Apetoh, T. Baert, R.B. Birge, J.M.B.-S. Pedro, K. Breckpot, D. Brough, R. Chaurio, M. Cirone, A. Coosemans, P.G. Coulie, D. De Ruysscher, L. Dini, P. de Witte, A.M. Dudek-Peric, A. Faggioni, J. Fucikova, U.S. Gaipl, J. Golab, M.L. Gougeon, M.R. Hamblin, A. Hemminki, M. Herrmann, J.W. Hodge, O. Kepp, G. Kroemer, D.V. Krysko, W.G. Land, F. Madeo, A.A. Manfredi, S.R. Mattarollo, C. Maueroder, N. Merendino, G. Multhoff, T. Pabst, J.E. Ricci, C. Riganti, E. Romano, N. Rufo, M.J. Smyth, J. Sonnemann, R. Spisek, J. Stagg, E. Vacchelli, P. Vandenabeele, L. Vandenberk, B.J. Van den Eynde, S. Van Gool, F. Velotti, L. Zitvogel, P. Agostinis, Molecular and Translational Classifications of DAMPs in Immunogenic Cell Death. Front Immunol 6, 588 (2015). https://doi.org/10.3389/fimmu.2015.00588

    Article  PubMed  PubMed Central  Google Scholar 

  16. A.D. Garg, A.M. Dudek-Peric, E. Romano, P. Agostinis, Immunogenic cell death. Int J Dev Biol 59, 131–140 (2015). https://doi.org/10.1387/ijdb.150061pa

    Article  CAS  PubMed  Google Scholar 

  17. A. Almanza, A. Carlesso, C. Chintha, S. Creedican, D. Doultsinos, B. Leuzzi, A. Luis, N. McCarthy, L. Montibeller, S. More, A. Papaioannou, F. Puschel, M.L. Sassano, J. Skoko, P. Agostinis, J. de Belleroche, L.A. Eriksson, S. Fulda, A.M. Gorman, S. Healy, A. Kozlov, C. Munoz-Pinedo, M. Rehm, E. Chevet, A. Samali, Endoplasmic reticulum stress signalling - from basic mechanisms to clinical applications. FEBS J 286, 241–278 (2019). https://doi.org/10.1111/febs.14608

    Article  CAS  PubMed  Google Scholar 

  18. C. Hetz, F.R. Papa, The unfolded protein response and cell fate control. Mol Cell 69, 169–181 (2018). https://doi.org/10.1016/j.molcel.2017.06.017

    Article  CAS  PubMed  Google Scholar 

  19. L. Bezu, A. Sauvat, J. Humeau, L.C. Gomes-da-Silva, K. Iribarren, S. Forveille, P. Garcia, L. Zhao, P. Liu, L. Zitvogel, L. Senovilla, O. Kepp, G. Kroemer, eIF2alpha phosphorylation is pathognomonic for immunogenic cell death. Cell Death Differ 25, 1375–1393 (2018). https://doi.org/10.1038/s41418-017-0044-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. T. Panaretakis, O. Kepp, U. Brockmeier, A. Tesniere, A.C. Bjorklund, D.C. Chapman, M. Durchschlag, N. Joza, G. Pierron, P. van Endert, J. Yuan, L. Zitvogel, F. Madeo, D.B. Williams, G. Kroemer, Mechanisms of pre-apoptotic calreticulin exposure in immunogenic cell death. EMBO J 28, 578–590 (2009). https://doi.org/10.1038/emboj.2009.1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. P. Giglio, M. Gagliardi, N. Tumino, F. Antunes, S. Smaili, D. Cotella, C. Santoro, R. Bernardini, M. Mattei, M. Piacentini, M. Corazzari, PKR and GCN2 stress kinases promote an ER stress-independent eIF2alpha phosphorylation responsible for calreticulin exposure in melanoma cells. Oncoimmunology 7, e1466765 (2018). https://doi.org/10.1080/2162402X.2018.1466765

  22. Global Burden of Disease Cancer Collaboration, C. Fitzmaurice, D. Abate, N. Abbasi, H. Abbastabar, F. Abd-Allah, O. Abdel-Rahman, A. Abdelalim, A. Abdoli, I. Abdollahpour, A.S.M. Abdulle, N.D. Abebe, H.N. Abraha, L.J. Abu-Raddad, A. Abualhasan, I.A. Adedeji, S.M. Advani, M. Afarideh, M. Afshari, M. Aghaali, D. Agius, S. Agrawal, A. Ahmadi, E. Ahmadian, E. Ahmadpour, M.B. Ahmed, M.E. Akbari, T. Akinyemiju, Z. Al-Aly, A.M. AlAbdulKader, F. Alahdab, T. Alam, G.M. Alamene, B.T.T. Alemnew, K.A. Alene, C. Alinia, V. Alipour, S.M. Aljunid, F.A. Bakeshei, M.A.H. Almadi, A. Almasi-Hashiani, U. Alsharif, S. Alsowaidi, N. Alvis-Guzman, E. Amini, S. Amini, Y.A. Amoako, Z. Anbari, N.H. Anber, C.L. Andrei, M. Anjomshoa, F. Ansari, A. Ansariadi, S.C.Y. Appiah, M. Arab-Zozani, J. Arabloo, Z. Arefi, O. Aremu, H.A. Areri, A. Artaman, H. Asayesh, E.T. Asfaw, A.F. Ashagre, R. Assadi, B. Ataeinia, H.T. Atalay, Z. Ataro, S. Atique, M. Ausloos, L. Avila-Burgos, E. Avokpaho, A. Awasthi, N. Awoke, B.P. Ayala Quintanilla, M.A. Ayanore, H.T. Ayele, E. Babaee, U. Bacha, A. Badawi, M. Bagherzadeh, E. Bagli, S. Balakrishnan, A. Balouchi, T.W. Barnighausen, R.J. Battista, M. Behzadifar, M. Behzadifar, B.B. Bekele, Y.B. Belay, Y.M. Belayneh, K.K.S. Berfield, A. Berhane, E. Bernabe, M. Beuran, N. Bhakta, K. Bhattacharyya, B. Biadgo, A. Bijani, M.S. Bin Sayeed, C. Birungi, C. Bisignano, H. Bitew, T. Bjorge, A. Bleyer, K.A. Bogale, H.A. Bojia, A.M. Borzi, C. Bosetti, I.R. Bou-Orm, H. Brenner, J.D. Brewer, A.N. Briko, N.I. Briko, M.T. Bustamante-Teixeira, Z.A. Butt, G. Carreras, J.J. Carrero, F. Carvalho, C. Castro, F. Castro, F. Catala-Lopez, E. Cerin, Y. Chaiah, W.F. Chanie, V.K. Chattu, P. Chaturvedi, N.S. Chauhan, M. Chehrazi, P.P. Chiang, T.Y. Chichiabellu, O.G. Chido-Amajuoyi, O. Chimed-Ochir, J.J. Choi, D.J. Christopher, D.T. Chu, M.M. Constantin, V.M. Costa, E. Crocetti, C.S. Crowe, M.P. Curado, S.M.A. Dahlawi, G. Damiani, A.H. Darwish, A. Daryani, J. das Neves, F.M. Demeke, A.B. Demis, B.W. Demissie, G.T. Demoz, E. Denova-Gutierrez, A. Derakhshani, K.S. Deribe, R. Desai, B.B. Desalegn, M. Desta, S. Dey, S.D. Dharmaratne, M. Dhimal, D. Diaz, M.T.T. Dinberu, S. Djalalinia, D.T. Doku, T.M. Drake, M. Dubey, E. Dubljanin, E.E. Duken, H. Ebrahimi, A. Effiong, A. Eftekhari, I. El Sayed, M.E.S. Zaki, S.I. El-Jaafary, Z. El-Khatib, D.A. Elemineh, H. Elkout, R.G. Ellenbogen, A. Elsharkawy, M.H. Emamian, D.A. Endalew, A.Y. Endries, B. Eshrati, I. Fadhil, V. Fallah, M. Faramarzi, M.A. Farhangi, A. Farioli, F. Farzadfar, N. Fentahun, E. Fernandes, G.T. Feyissa, I. Filip, F. Fischer, J.L. Fisher, L.M. Force, M. Foroutan, M. Freitas, T. Fukumoto, N.D. Futran, S. Gallus, F.G. Gankpe, R.T. Gayesa, T.T. Gebrehiwot, G.G. Gebremeskel, G.A. Gedefaw, B.K. Gelaw, B. Geta, S. Getachew, K.E. Gezae, M. Ghafourifard, A. Ghajar, A. Ghashghaee, A. Gholamian, P.S. Gill, T.T.G. Ginindza, A. Girmay, M. Gizaw, R.S. Gomez, S.V. Gopalani, G. Gorini, B.N.G. Goulart, A. Grada, M. Ribeiro Guerra, A.L.S. Guimaraes, P.C. Gupta, R. Gupta, K. Hadkhale, A. Haj-Mirzaian, A. Haj-Mirzaian, R.R. Hamadeh, S. Hamidi, L.K. Hanfore, J.M. Haro, M. Hasankhani, A. Hasanzadeh, H.Y. Hassen, R.J. Hay, S.I. Hay, A. Henok, N.J. Henry, C. Herteliu, H.D. Hidru, C.L. Hoang, M.K. Hole, P. Hoogar, N. Horita, H.D. Hosgood, M. Hosseini, M. Hosseinzadeh, M. Hostiuc, S. Hostiuc, M. Househ, M.M. Hussen, B. Ileanu, M.D. Ilic, K. Innos, S.S.N. Irvani, K.R. Iseh, S.M.S. Islam, F. Islami, N. Jafari Balalami, M. Jafarinia, L. Jahangiry, M.A. Jahani, N. Jahanmehr, M. Jakovljevic, S.L. James, M. Javanbakht, S. Jayaraman, S.H. Jee, E. Jenabi, R.P. Jha, J.B. Jonas, J. Jonnagaddala, T. Joo, S.B. Jungari, M. Jurisson, A. Kabir, F. Kamangar, A. Karch, N. Karimi, A. Karimian, A. Kasaeian, G.G. Kasahun, B. Kassa, T.D. Kassa, M.W. Kassaw, A. Kaul, P.N. Keiyoro, A.G. Kelbore, A.A. Kerbo, Y.S. Khader, M. Khalilarjmandi, E.A. Khan, G. Khan, Y.H. Khang, K. Khatab, A. Khater, M. Khayamzadeh, M. Khazaee-Pool, S. Khazaei, A.T. Khoja, M.H. Khosravi, J. Khubchandani, N. Kianipour, D. Kim, Y.J. Kim, A. Kisa, S. Kisa, K. Kissimova-Skarbek, H. Komaki, A. Koyanagi, K.J. Krohn, B.K. Bicer, N. Kugbey, V. Kumar, D. Kuupiel, C. La Vecchia, D.P. Lad, E.A. Lake, A.M. Lakew, D.K. Lal, F.H. Lami, Q. Lan, S. Lasrado, P. Lauriola, J.V. Lazarus, J. Leigh, C.T. Leshargie, Y. Liao, M.A. Limenih, S. Listl, A.D. Lopez, P.D. Lopukhov, R. Lunevicius, M. Madadin, S. Magdeldin, H.M.A. El Razek, A. Majeed, A. Maleki, R. Malekzadeh, A. Manafi, N. Manafi, W.A. Manamo, M. Mansourian, M.A. Mansournia, L.G. Mantovani, S. Maroufizadeh, S.M.S. Martini, T.P. Mashamba-Thompson, B.B. Massenburg, M.T. Maswabi, M.R. Mathur, C. McAlinden, M. McKee, H.A.A. Meheretu, R. Mehrotra, V. Mehta, T. Meier, Y.A. Melaku, G.G. Meles, H.G. Meles, A. Melese, M. Melku, P.T.N. Memiah, W. Mendoza, R.G. Menezes, S. Merat, T.J. Meretoja, T. Mestrovic, B. Miazgowski, T. Miazgowski, K.M.M. Mihretie, T.R. Miller, E.J. Mills, S.M. Mir, H. Mirzaei, H.R. Mirzaei, R. Mishra, B. Moazen, D.K. Mohammad, K.A. Mohammad, Y. Mohammad, A.M. Darwesh, A. Mohammadbeigi, H. Mohammadi, M. Mohammadi, M. Mohammadian, A. Mohammadian-Hafshejani, M. Mohammadoo-Khorasani, R. Mohammadpourhodki, A.S. Mohammed, J.A. Mohammed, S. Mohammed, F. Mohebi, A.H. Mokdad, L. Monasta, Y. Moodley, M. Moosazadeh, M. Moossavi, G. Moradi, M. Moradi-Joo, M. Moradi-Lakeh, F. Moradpour, L. Morawska, J. Morgado-da-Costa, N. Morisaki, S.D. Morrison, A. Mosapour, S.M. Mousavi, A.A. Muche, O.S.S. Muhammed, J. Musa, A.R. Nabhan, M. Naderi, A.J. Nagarajan, G. Nagel, A. Nahvijou, G. Naik, F. Najafi, L. Naldi, H.S. Nam, N. Nasiri, J. Nazari, I. Negoi, S. Neupane, P.A. Newcomb, H.A. Nggada, J.W. Ngunjiri, C.T. Nguyen, L. Nikniaz, D.N.A. Ningrum, Y.L. Nirayo, M.R. Nixon, C.A. Nnaji, M. Nojomi, S. Nosratnejad, M.N. Shiadeh, M.S. Obsa, R. Ofori-Asenso, F.A. Ogbo, I.H. Oh, A.T. Olagunju, T.O. Olagunju, M.M. Oluwasanu, A.E. Omonisi, O.E. Onwujekwe, A.M. Oommen, E. Oren, D.D.V. Ortega-Altamirano, E. Ota, S.S. Otstavnov, M.O. Owolabi, A. M. P, J.R. Padubidri, S. Pakhale, A.H. Pakpour, A. Pana, E.K. Park, H. Parsian, T. Pashaei, S. Patel, S.T. Patil, A. Pennini, D.M. Pereira, C. Piccinelli, J.D. Pillay, M. Pirestani, F. Pishgar, M.J. Postma, H. Pourjafar, F. Pourmalek, A. Pourshams, S. Prakash, N. Prasad, M. Qorbani, M. Rabiee, N. Rabiee, A. Radfar, A. Rafiei, F. Rahim, M. Rahimi, M.A. Rahman, F. Rajati, S.M. Rana, S. Raoofi, G.K. Rath, D.L. Rawaf, S. Rawaf, R.C. Reiner, A.M.N. Renzaho, N. Rezaei, A. Rezapour, A.I. Ribeiro, D. Ribeiro, L. Ronfani, E.M. Roro, G. Roshandel, A. Rostami, R.S. Saad, P. Sabbagh, S. Sabour, B. Saddik, S. Safiri, A. Sahebkar, M.R. Salahshoor, F. Salehi, H. Salem, M.R. Salem, H. Salimzadeh, J.A. Salomon, A.M. Samy, J. Sanabria, M.M. Santric Milicevic, B. Sartorius, A. Sarveazad, B. Sathian, M. Satpathy, M. Savic, M. Sawhney, M. Sayyah, I.J.C. Schneider, B. Schottker, M. Sekerija, S.G. Sepanlou, M. Sepehrimanesh, S. Seyedmousavi, F. Shaahmadi, H. Shabaninejad, M. Shahbaz, M.A. Shaikh, A. Shamshirian, M. Shamsizadeh, H. Sharafi, Z. Sharafi, M. Sharif, A. Sharifi, H. Sharifi, R. Sharma, A. Sheikh, R. Shirkoohi, S.R. Shukla, S. Si, S. Siabani, D.A.S. Silva, D.G.A. Silveira, A. Singh, J.A. Singh, S. Sisay, F. Sitas, E. Sobngwi, M. Soofi, J.B. Soriano, V. Stathopoulou, M.B. Sufiyan, R. Tabares-Seisdedos, T. Tabuchi, K. Takahashi, O.R. Tamtaji, M.R. Tarawneh, S.G. Tassew, P. Taymoori, A. Tehrani-Banihashemi, M.H. Temsah, O. Temsah, B.E. Tesfay, F.H. Tesfay, M.Y. Teshale, G.A. Tessema, S. Thapa, K.G. Tlaye, R. Topor-Madry, M.R. Tovani-Palone, E. Traini, B.X. Tran, K.B. Tran, A.G. Tsadik, I. Ullah, O.A. Uthman, M. Vacante, M. Vaezi, P. Varona Perez, Y. Veisani, S. Vidale, F.S. Violante, V. Vlassov, S.E. Vollset, T. Vos, K. Vosoughi, G.T. Vu, I.S. Vujcic, H. Wabinga, T.M. Wachamo, F.S. Wagnew, Y. Waheed, F. Weldegebreal, G.T. Weldesamuel, T. Wijeratne, D.Z. Wondafrash, T.E. Wonde, A.B. Wondmieneh, H.M. Workie, R. Yadav, A. Yadegar, A. Yadollahpour, M. Yaseri, V. Yazdi-Feyzabadi, A. Yeshaneh, M.A. Yimam, E.M. Yimer, E. Yisma, N. Yonemoto, M.Z. Younis, B. Yousefi, M. Yousefifard, C. Yu, E. Zabeh, V. Zadnik, T.Z. Moghadam, Z. Zaidi, M. Zamani, H. Zandian, A. Zangeneh, L. Zaki, K. Zendehdel, Z.M. Zenebe, T.A. Zewale, A. Ziapour, S. Zodpey and C.J.L. Murray, Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-Years for 29 Cancer Groups, 1990 to 2017: A Systematic Analysis for the Global Burden of Disease Study. JAMA Oncol 5, 1749–1768 (2019). doi: https://doi.org/10.1001/jamaoncol.2019.2996

  23. J. Zhou, T. Yang, L. Liu, B. Lu, Chemotherapy oxaliplatin sensitizes prostate cancer to immune checkpoint blockade therapies via stimulating tumor immunogenicity. Mol Med Rep 16, 2868–2874 (2017). https://doi.org/10.3892/mmr.2017.6908

    Article  CAS  PubMed  Google Scholar 

  24. J. Kapuscinski, Z. Darzynkiewicz, Interactions of antitumor agents Ametantrone and Mitoxantrone (Novatrone) with double-stranded DNA. Biochem Pharmacol 34, 4203–4213 (1985). https://doi.org/10.1016/0006-2952(85)90275-8

    Article  CAS  PubMed  Google Scholar 

  25. J.S. de Bono, S. Oudard, M. Ozguroglu, S. Hansen, J.P. Machiels, I. Kocak, G. Gravis, I. Bodrogi, M.J. Mackenzie, L. Shen, M. Roessner, S. Gupta, A.O. Sartor, T. Investigators, Prednisone plus cabazitaxel or mitoxantrone for metastatic castration-resistant prostate cancer progressing after docetaxel treatment: A randomised open-label trial. Lancet 376, 1147–1154 (2010). https://doi.org/10.1016/S0140-6736(10)61389-X

  26. G. Gravis, Systemic treatment for metastatic prostate cancer. Asian J Urol 6, 162–168 (2019). https://doi.org/10.1016/j.ajur.2019.02.002

    Article  PubMed  PubMed Central  Google Scholar 

  27. J. Qin, N.M. Kunda, G. Qiao, K. Tulla, B.S. Prabhakar, A.V. Maker, Vaccination with Mitoxantrone-treated primary Colon Cancer cells enhances tumor-infiltrating lymphocytes and clinical responses in colorectal liver metastases. J Surg Res 233, 57–64 (2019). https://doi.org/10.1016/j.jss.2018.07.068

    Article  CAS  PubMed  Google Scholar 

  28. T. Colangelo, G. Polcaro, P. Ziccardi, L. Muccillo, M. Galgani, B. Pucci, M.R. Milone, A. Budillon, M. Santopaolo, G. Mazzoccoli, G. Matarese, L. Sabatino, V. Colantuoni, The miR-27a-calreticulin axis affects drug-induced immunogenic cell death in human colorectal cancer cells. Cell Death Dis 7, e2108 (2016). https://doi.org/10.1038/cddis.2016.29

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. C. Li, C. He, Y. Xu, H. Xu, Y. Tang, H. Chavan, S. Duan, A. Artigues, M.L. Forrest, P. Krishnamurthy, S. Han, J.M. Holzbeierlein, B. Li, Alternol eliminates excessive ATP production by disturbing Krebs cycle in prostate cancer. Prostate 79, 628–639 (2019). https://doi.org/10.1002/pros.23767

  30. C. Li, W. Jiang, Q. Hu, L.C. Li, L. Dong, R. Chen, Y. Zhang, Y. Tang, J.B. Thrasher, C.B. Liu, B. Li, Enhancing DPYSL3 gene expression via a promoter-targeted small activating RNA approach suppresses cancer cell motility and metastasis, Oncotarget 7, 22893–22910 (2016). https://doi.org/10.18632/oncotarget.8290

  31. K. Berns, E.M. Hijmans, J. Mullenders, T.R. Brummelkamp, A. Velds, M. Heimerikx, R.M. Kerkhoven, M. Madiredjo, W. Nijkamp, B. Weigelt, R. Agami, W. Ge, G. Cavet, P.S. Linsley, R.L. Beijersbergen, R. Bernards, A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428, 431–437 (2004). https://doi.org/10.1038/nature02371

  32. M. Soto, J.A. Raaijmakers, B. Bakker, D.C.J. Spierings, P.M. Lansdorp, F. Foijer, R.H. Medema, p53 prohibits propagation of chromosome segregation errors that produce structural aneuploidies. Cell Rep 19, 2423–2431 (2017). https://doi.org/10.1016/j.celrep.2017.05.055

    Article  CAS  PubMed  Google Scholar 

  33. C. Li, H. Xu, L. Xiao, H. Zhu, G. Zhang, W. Wei, K. Li, X. Cao, D. Shen, J. Holzbeierlein, B. Li, CRMP4a suppresses cell motility by sequestering RhoA activity in prostate cancer cells. Cancer Biol Ther 19, 1193–1203 (2018). https://doi.org/10.1080/15384047.2018.1491507

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. C. Pozzi, A. Cuomo, I. Spadoni, E. Magni, A. Silvola, A. Conte, S. Sigismund, P.S. Ravenda, T. Bonaldi, M.G. Zampino, C. Cancelliere, P.P. Di Fiore, A. Bardelli, G. Penna, M. Rescigno, The EGFR-specific antibody cetuximab combined with chemotherapy triggers immunogenic cell death. Nat Med 22, 624–631 (2016). https://doi.org/10.1038/nm.4078

    Article  CAS  PubMed  Google Scholar 

  35. G.H. Nam, E.J. Lee, Y.K. Kim, Y. Hong, Y. Choi, M.J. Ryu, J. Woo, Y. Cho, D.J. Ahn, Y. Yang, I.C. Kwon, S.Y. Park, I.S. Kim, Combined rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer. Nat Commun 9, 2165 (2018). https://doi.org/10.1038/s41467-018-04607-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. A. Khan, O. Fornes, A. Stigliani, M. Gheorghe, J.A. Castro-Mondragon, R. van der Lee, A. Bessy, J. Cheneby, S.R. Kulkarni, G. Tan, D. Baranasic, D.J. Arenillas, A. Sandelin, K. Vandepoele, B. Lenhard, B. Ballester, W.W. Wasserman, F. Parcy, A. Mathelier, JASPAR 2018: Update of the open-access database of transcription factor binding profiles and its web framework. Nucleic Acids Res 46, D1284 (2018). https://doi.org/10.1093/nar/gkx1188

    Article  PubMed  Google Scholar 

  37. D. Schmidt, M.D. Wilson, C. Spyrou, G.D. Brown, J. Hadfield, D.T. Odom, ChIP-seq: Using high-throughput sequencing to discover protein-DNA interactions. Methods 48, 240–248 (2009). https://doi.org/10.1016/j.ymeth.2009.03.001

  38. C.P. Bergstrom, B. Ruffell, C.M. Ho, C.S. Higano, W.J. Ellis, M. Garzotto, T.M. Beer, J.N. Graff, Docetaxel and mitoxantrone before radical prostatectomy in men with high-risk prostate cancer: 10-year follow-up and immune correlates. Anti-Cancer Drugs 28, 120–126 (2017). https://doi.org/10.1097/CAD.0000000000000438

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. C. Sanchez, P. Mendoza, H.R. Contreras, J. Vergara, J.A. McCubrey, C. Huidobro, E.A. Castellon, Expression of multidrug resistance proteins in prostate cancer is related with cell sensitivity to chemotherapeutic drugs. Prostate 69, 1448–1459 (2009). https://doi.org/10.1002/pros.20991

  40. L. Bezu, L.C. Gomes-de-Silva, H. Dewitte, K. Breckpot, J. Fucikova, R. Spisek, L. Galluzzi, O. Kepp, G. Kroemer, Combinatorial strategies for the induction of immunogenic cell death. Front Immunol 6, 187 (2015). https://doi.org/10.3389/fimmu.2015.00187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. R.C. Wek, Role of eIF2alpha kinases in translational control and adaptation to cellular stress. Cold Spring Harb Perspect Biol 10 (2018). https://doi.org/10.1101/cshperspect.a032870

  42. A. Rashidi, J. Miska, C. Lee-Chang, D. Kanojia, W.K. Panek, A. Lopez-Rosas, P. Zhang, Y. Han, T. Xiao, K.C. Pituch, J.W. Kim, M. Talebian, J. Fares, M.S. Lesniak, GCN2 is essential for CD8(+) T cell survival and function in murine models of malignant glioma. Cancer Immunol Immunother 69, 81–94 (2020). https://doi.org/10.1007/s00262-019-02441-6

    Article  CAS  PubMed  Google Scholar 

  43. P.R. Romano, M.T. Garcia-Barrio, X. Zhang, Q. Wang, D.R. Taylor, F. Zhang, C. Herring, M.B. Mathews, J. Qin, A.G. Hinnebusch, Autophosphorylation in the activation loop is required for full kinase activity in vivo of human and yeast eukaryotic initiation factor 2alpha kinases PKR and GCN2. Mol Cell Biol 18, 2282–2297 (1998). https://doi.org/10.1128/mcb.18.4.2282

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. H.P. Harding, Y. Zhang, A. Bertolotti, H. Zeng, D. Ron, Perk is essential for translational regulation and cell survival during the unfolded protein response. Mol Cell 5, 897–904 (2000). https://doi.org/10.1016/s1097-2765(00)80330-5

    Article  CAS  PubMed  Google Scholar 

  45. G. Guo, M. Yu, W. Xiao, E. Celis, Y. Cui, Local activation of p53 in the tumor microenvironment overcomes immune suppression and enhances antitumor immunity. Cancer Res 77, 2292–2305 (2017). https://doi.org/10.1158/0008-5472.CAN-16-2832

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. S. Bottger, E. Jerszyk, B. Low, C. Walker, Genotoxic stress-induced expression of p53 and apoptosis in leukemic clam hemocytes with cytoplasmically sequestered p53. Cancer Res 68, 777–782 (2008). https://doi.org/10.1158/0008-5472.CAN-06-0968

    Article  CAS  PubMed  Google Scholar 

  47. A.G. Carroll, H.J. Voeller, L. Sugars, E.P. Gelmann, p53 oncogene mutations in three human prostate cancer cell lines. Prostate 23, 123–134 (1993). https://doi.org/10.1002/pros.2990230206

  48. M. Michaud, I. Martins, A.Q. Sukkurwala, S. Adjemian, Y. Ma, P. Pellegatti, S. Shen, O. Kepp, M. Scoazec, G. Mignot, S. Rello-Varona, M. Tailler, L. Menger, E. Vacchelli, L. Galluzzi, F. Ghiringhelli, F. di Virgilio, L. Zitvogel, G. Kroemer, Autophagy-dependent anticancer immune responses induced by chemotherapeutic agents in mice. Science 334, 1573–1577 (2011). https://doi.org/10.1126/science.1208347

  49. J.S. Park, F. Gamboni-Robertson, Q. He, D. Svetkauskaite, J.Y. Kim, D. Strassheim, J.W. Sohn, S. Yamada, I. Maruyama, A. Banerjee, A. Ishizaka, E. Abraham, High mobility group box 1 protein interacts with multiple toll-like receptors. Am J Physiol Cell Physiol 290, C917–C924 (2006). https://doi.org/10.1152/ajpcell.00401.2005

    Article  CAS  PubMed  Google Scholar 

  50. I. Martins, A. Tesniere, O. Kepp, M. Michaud, F. Schlemmer, L. Senovilla, C. Seror, D. Metivier, J.L. Perfettini, L. Zitvogel, G. Kroemer, Chemotherapy induces ATP release from tumor cells. Cell Cycle 8, 3723–3728 (2009). https://doi.org/10.4161/cc.8.22.10026

    Article  CAS  PubMed  Google Scholar 

  51. A.D. Garg, D.V. Krysko, T. Verfaillie, A. Kaczmarek, G.B. Ferreira, T. Marysael, N. Rubio, M. Firczuk, C. Mathieu, A.J. Roebroek, W. Annaert, J. Golab, P. de Witte, P. Vandenabeele, P. Agostinis, A novel pathway combining calreticulin exposure and ATP secretion in immunogenic cancer cell death. EMBO J 31, 1062–1079 (2012). https://doi.org/10.1038/emboj.2011.497

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. L. Bezu, A. Sauvat, J. Humeau, M. Leduc, O. Kepp, G. Kroemer, eIF2alpha phosphorylation: A hallmark of immunogenic cell death. Oncoimmunology 7, e1431089 (2018). https://doi.org/10.1080/2162402X.2018.1431089

  53. J. Han, S.H. Back, J. Hur, Y.H. Lin, R. Gildersleeve, J. Shan, C.L. Yuan, D. Krokowski, S. Wang, M. Hatzoglou, M.S. Kilberg, M.A. Sartor, R.J. Kaufman, ER-stress-induced transcriptional regulation increases protein synthesis leading to cell death. Nat Cell Biol 15, 481–490 (2013). https://doi.org/10.1038/ncb2738

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. S.M. Reed, D.E. Quelle, p53 acetylation: Regulation and consequences. Cancers (Basel) 7, 30–69 (2014). https://doi.org/10.3390/cancers7010030

    Article  CAS  Google Scholar 

  55. M.F. Lavin, N. Gueven, The complexity of p53 stabilization and activation. Cell Death Differ 13, 941–950 (2006). https://doi.org/10.1038/sj.cdd.4401925

    Article  CAS  PubMed  Google Scholar 

  56. Y. Xia, R.C. Padre, T.H. De Mendoza, V. Bottero, V.B. Tergaonkar, I.M. Verma, Phosphorylation of p53 by IkappaB kinase 2 promotes its degradation by beta-TrCP. Proc Natl Acad Sci U S A 106, 2629–2634 (2009). https://doi.org/10.1073/pnas.0812256106

    Article  PubMed  PubMed Central  Google Scholar 

  57. G.S. Ivanov, T. Ivanova, J. Kurash, A. Ivanov, S. Chuikov, F. Gizatullin, E.M. Herrera-Medina, F. Rauscher 3rd, D. Reinberg, N.A. Barlev, Methylation-acetylation interplay activates p53 in response to DNA damage. Mol Cell Biol 27, 6756–6769 (2007). https://doi.org/10.1128/MCB.00460-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. J. Huang, L. Perez-Burgos, B.J. Placek, R. Sengupta, M. Richter, J.A. Dorsey, S. Kubicek, S. Opravil, T. Jenuwein, S.L. Berger, Repression of p53 activity by Smyd2-mediated methylation. Nature 444, 629–632 (2006). https://doi.org/10.1038/nature05287

  59. S.F. Slovin, Immunotherapy for castration-resistant prostate cancer: Has its time arrived? Expert Opin Biol Ther 20, 481–487 (2020). https://doi.org/10.1080/14712598.2020.1735345

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. I. Vanmeerbeek, J. Sprooten, D. De Ruysscher, S. Tejpar, P. Vandenberghe, J. Fucikova, R. Spisek, L. Zitvogel, G. Kroemer, L. Galluzzi, A.D. Garg, Trial watch: Chemotherapy-induced immunogenic cell death in immuno-oncology. Oncoimmunology 9, 1703449 (2020). https://doi.org/10.1080/2162402X.2019.1703449

  61. J.W. Hodge, C.T. Garnett, B. Farsaci, C. Palena, K.Y. Tsang, S. Ferrone, S.R. Gameiro, Chemotherapy-induced immunogenic modulation of tumor cells enhances killing by cytotoxic T lymphocytes and is distinct from immunogenic cell death. Int J Cancer 133, 624–636 (2013). https://doi.org/10.1002/ijc.28070

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

This work was partially supported by the Shandong Provincial Natural Science Foundation of China (ZR2016HL25 and ZR2016CQ01), NSFC grant 81501770 and an internal grant from Jining Medical University (JYHL2018ZD02).

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Authors and Affiliations

  1. Institute of Precision Medicine, Jining Medical University, Jining, 272067, China

    Changlin Li, Qiuzi Liu, Ying Zhang, Fuming Lian, Chenchen Li, Kaicheng Ying & Hang Huo

  2. Department of Urology, The University of Kansas Medical Center, Kansas City, KS, 66160, USA

    Changlin Li & Benyi Li

  3. Shandong Institute of Parasitic Diseases, Shandong First Medical University & Shandong Academy of Medical Sciences, Jining, 272033, China

    Hui Sun

  4. Center for Experimental Medicine, School of Public Health, Jining Medical University, Jining, 272067, China

    Wei Wei

  5. Department of Urology, The Affiliated Hospital, Binzhou Medical University, Yantai, 264003, China

    Yinglei Wang

  6. Department of Urology, The Affiliated Hospital, Jining Medical University, Jining, 272067, China

    Fangchao Liu

  7. Department of Histology and Embryology, School of Medicine, Nankai University, Tianjin, 300071, China

    Zhi Qi

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  1. Changlin Li
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ESM 2

MTX induces apoptosis in prostate cancer cells. a. LNCaP and 22RV1 cells were treated with the vehicle or MTX (1 μM) for 20 h and then harvested for apoptosis analysis using Annexin-V/PI staining approach coupled with flow cytometry. b. Quantitative data of apoptosis were presented from three independent experiments. Early apoptotic was defined as Annexin positive population, later apoptotic as Annexin/PI positive and necrotic death as PI positive only. The asterisks indicate a significant difference compared to the vehicle control (Student t-test, *p < 0.05, **p < 0.01, ***p < 0.001). c. LNCaP and 22RV1 cells were treated with the vehicle or MTX (1 μM) for 20 h and then whole cell proteins were extracted for western blot assay with PARP antibodies. Actin blot served as protein loading control. Data were representative of two independent experiments. (PNG 70 kb)

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ESM 3

Effect of PERK or GCN2 Knockdown on MTX-induced apoptosis. LNCaP cells transfected with the shRNAs as indicated were treated with MTX (1 μM) or the vehicle for 20 h, followed by Annexin-V/PI staining, as described earlier. Representative plots of flow cytometry from three independent experiments were shown in panel a. Quantitative data of apoptotic population (%) were summarized in panel b. The asterisks indicate a significant difference compared to the shScr control (Student t-test, *p < 0.05, ***p < 0.001). (PNG 64 kb)

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Li, C., Sun, H., Wei, W. et al. Mitoxantrone triggers immunogenic prostate cancer cell death via p53-dependent PERK expression. Cell Oncol. 43, 1099–1116 (2020). https://doi.org/10.1007/s13402-020-00544-2

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  • DOI: https://doi.org/10.1007/s13402-020-00544-2

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