Deconstructing the Checkpoints of Necroptosis

解构坏死性凋亡的检查点

• 批准号: BB/X007383/1
• 负责人: Pascal Meier
• 金额: $ 76.48万
• 依托单位: Institute of Cancer Research
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX007383%2F1
• 关键词: Deconstructing; Checkpoints; Necroptosis

For a long time, apoptosis was considered the sole form of programmed cell death during development, homeostasis, and disease, whereas necrosis was regarded as an unregulated and uncontrollable process. Evidence now reveals that necrosis can also occur in a regulated manner. Programmed necrosis, 'necroptosis' plays vital roles during host-pathogen interactions where it is triggered as host-defence mechanism for the elimination of pathogen-infected cells. However, necroptosis also participates in the pathogenesis of diseases, including ischaemic injury, neuro-degeneration, and viral infection. Moreover, necroptosis has also sparked considerable interest among cancer researchers for its potential to overcome tumour resistance to apoptosis, and because it is more immunogenic than apoptosis, flagging up tumours for immunological attack. For these reasons, there has been much interest in obtaining a better understanding of how necroptosis is activated and how this potentially catastrophic event is regulated. Necroptosis is mediated by MLKL, a membrane permeabilizing pseudo-kinase that translocates to the plasma membrane upon its activation. While necroptosis signalling has attracted much attention for its therapeutic potential, little is known how necroptosis is regulated, and how MLKL translocates to hotspots at the plasma membrane to trigger necroptosis.We now have identified that the Ubiquitin (Ub)-signalling system critically regulates necroptosis, and that ubiquitylation of MLKL is required for MLKL to traffic to the plasma membrane. Moreover, we have identified several putative MLKL-regulatory Ub-E3 ligases and deubiquitylating enzymes that might operate as decisive necroptotic checkpoint. Aim:The aim of this proposal is to identify the mechanism through which active MLKL translocates to the plasma membrane where it accumulates at hotspots to cause lytic cell death. We will also identify the intercellular structures at which MLKL accumulates and characterise their contribution to necroptosis signalling and membrane rupture.To achieve this, we will characterise the molecular players of the Ub signalling system (E3 ligases, deubiquitylating enzymes and Ub-receptors) that underpin MLKL ubiquitylation, trafficking and accumulation at intercellular contact sites. Moreover, we will study whether the identified E3 ligases, deubiquitylating enzymes and Ub-receptors contribute to antiviral host defence.Methods:Using biochemical, single-cell imaging and in vivo approaches, we will elucidate how MLKL is ubiquitylated by E3 ligases, how the ubiquitylation status of MLKL is edited by deubiquitylating enzymes, and how such signalling chains are detected by Ub-binding proteins (Ub-receptors) to shuttle active MLKL to intercellular hotspots at the plasma membrane. Moreover, we will unravel the role of the identified Ub-E3 ligases, deubiquitylating enzymes and Ub-receptors in modulating anti-viral host defences. Further, we will evaluate the contribution of desmosomes and Flotillins in necroptosis signalling.How the results will be usedA better understanding of necroptosis signalling will be of enormous interest to basic scientists as well as clinical researchers because it will lay the foundation for the design of future therapeutic strategies aimed at boosting antiviral defence, fighting cancer and suppressing inflammatory diseases.

长期以来，凋亡被认为是在发育，体内平衡和疾病期间编程细胞死亡的唯一形式，而坏死被认为是不受监管且无法控制的过程。现在的证据表明，坏死也可以以受调节的方式发生。编程坏死，“坏死”在宿主病原体相互作用中起着至关重要的作用，在宿主 - 病原体相互作用中，它被触发为消除病原体感染细胞的宿主防御机制。然而，坏死性还参与疾病的发病机理，包括缺血性损伤，神经脱生和病毒感染。此外，坏死性还引起了癌症研究人员克服肿瘤对凋亡的潜力的极大兴趣，并且因为它比凋亡更具有免疫原性，因此为免疫学攻击增加了肿瘤。由于这些原因，人们有很大的兴趣更好地了解如何激活坏死性，以及如何调节这种潜在的灾难性事件。坏死作用是由MLKL介导的，MLKL是一种膜通透性伪激酶，在其激活后将其转移到质膜。虽然坏死性信号传导因其治疗潜力引起了很多关注，但几乎几乎不知道坏死性如何受到调节，以及MLKL如何转移到质膜处的热点以触发坏死。 MLKL到达质膜。此外，我们已经确定了一些推定的MLKL调节性UB-E3连接酶和去偶联酶，这些酶可能作为决定性的坏死性检查点。目的：该提案的目的是确定活性MLKL转移到质膜上的机制，在该机制在热点积聚以引起裂解细胞死亡。 We will also identify the intercellular structures at which MLKL accumulates and characterise their contribution to necroptosis signalling and membrane rupture.To achieve this, we will characterise the molecular players of the Ub signalling system (E3 ligases, deubiquitylating enzymes and Ub-receptors) that underpin MLKL ubiquitylation, trafficking and accumulation at intercellular contact sites.此外，我们将研究确定的E3连接酶，去泛素化酶和UB受体受益者是否有助于抗病毒宿主的防御。酶，以及如何通过UB结合蛋白（UB受体）检测到这种信号链，以将活性MLKL驶向质膜的细胞间热点。此外，我们将揭示已鉴定的UB-E3连接酶，去偶联酶和UB受体在调节抗病毒宿主防御方面的作用。此外，我们将评估脱糖体和鞭毛素在坏死性信号中的贡献。结果将如何更好地理解坏死性信号，这将引起基础科学家以及临床研究人员的极大兴趣，因为它将为未来的抗抗病性抗议和抗抑郁抗拒的抗癌症和抗击癌症和抗击癌症的设计奠定基础。

项目成果

Apoptotic cell death in disease-Current understanding of the NCCD 2023.

• DOI: 10.1038/s41418-023-01153-w
• 发表时间: 2023-05
• 期刊: CELL DEATH AND DIFFERENTIATION
• 影响因子: 12.4
• 作者: Vitale, Ilio;Pietrocola, Federico;Guilbaud, Emma;Aaronson, Stuart A.;Abrams, John M.;Adam, Dieter;Agostini, Massimiliano;Agostinis, Patrizia;Alnemri, Emad S.;Altucci, Lucia;Amelio, Ivano;Andrews, David W.;Aqeilan, Rami, I;Arama, Eli;Baehrecke, Eric H.;Balachandran, Siddharth;Bano, Daniele;Barlev, Nickolai A.;Bartek, Jiri;Bazan, Nicolas G.;Becker, Christoph;Bernassola, Francesca;Bertrand, Mathieu J. M.;Bianchi, Marco E.;Blagosklonny, Mikhail V.;Blander, J. Magarian;Blandino, Giovanni;Blomgren, Klas;Borner, Christoph;Bortner, Carl D.;Bove, Pierluigi;Boya, Patricia;Brenner, Catherine;Broz, Petr;Brunner, Thomas;Damgaard, Rune Busk;Calin, George A.;Campanella, Michelangelo;Candi, Eleonora;Carbone, Michele;Carmona-Gutierrez, Didac;Cecconi, Francesco;Chan, Francis K-M;Chen, Guo-Qiang;Chen, Quan;Chen, Youhai H.;Cheng, Emily H.;Chipuk, Jerry E.;Cidlowski, John A.;Ciechanover, Aaron;Ciliberto, Gennaro;Conrad, Marcus;Cubillos-Ruiz, Juan R.;Czabotar, Peter E.;D'Angiolella, Vincenzo;Daugaard, Mads;Dawson, Ted M.;Dawson, Valina L.;De Maria, Ruggero;De Strooper, Bart;Debatin, Klaus-Michael;Deberardinis, Ralph J.;Degterev, Alexei;Del Sal, Giannino;Deshmukh, Mohanish;Di Virgilio, Francesco;Diederich, Marc;Dixon, Scott J.;Dynlacht, Brian D.;El-Deiry, Wafik S.;Elrod, John W.;Engeland, Kurt;Fimia, Gian Maria;Galassi, Claudia;Ganini, Carlo;Garcia-Saez, Ana J.;Garg, Abhishek D.;Garrido, Carmen;Gavathiotis, Evripidis;Gerlic, Motti;Ghosh, Sourav;Green, Douglas R.;Greene, Lloyd A.;Gronemeyer, Hinrich;Haecker, Georg;Hajnoczky, Gyorgy;Hardwick, J. Marie;Haupt, Ygal;He, Sudan;Heery, David M.;Hengartner, Michael O.;Hetz, Claudio;Hildeman, David A.;Ichijo, Hidenori;Inoue, Satoshi;Jaeaettelae, Marja;Janic, Ana;Joseph, Bertrand;Jost, Philipp J.;Kanneganti, Thirumala-Devi;Karin, Michael;Kashkar, Hamid;Kaufmann, Thomas;Kelly, Gemma L.;Kepp, Oliver;Kimchi, Adi;Kitsis, Richard N.;Klionsky, Daniel J.;Kluck, Ruth;Krysko, Dmitri, V;Kulms, Dagmar;Kumar, Sharad;Lavandero, Sergio;Lavrik, Inna N.;Lemasters, John J.;Liccardi, Gianmaria;Linkermann, Andreas;Lipton, Stuart A.;Lockshin, Richard A.;Lopez-Otin, Carlos;Luedde, Tom;MacFarlane, Marion;Madeo, Frank;Malorni, Walter;Manic, Gwenola;Mantovani, Roberto;Marchi, Saverio;Marine, Jean-Christophe;Martin, Seamus J.;Martinou, Jean-Claude;Mastroberardino, Pier G.;Medema, Jan Paul;Mehlen, Patrick;Meier, Pascal;Melino, Gerry;Melino, Sonia;Miao, Edward A.;Moll, Ute M.;Munoz-Pinedo, Cristina;Murphy, Daniel J.;Niklison-Chirou, Maria Victoria;Novelli, Flavia;Nunez, Gabriel;Oberst, Andrew;Ofengeim, Dimitry;Opferman, Joseph T.;Oren, Moshe;Pagano, Michele;Panaretakis, Theocharis;Pasparakis, Manolis;Penninger, Josef M.;Pentimalli, Francesca;Pereira, David M.;Pervaiz, Shazib;Peter, Marcus E.;Pinton, Paolo;Porta, Giovanni;Prehn, Jochen H. M.;Puthalakath, Hamsa;Rabinovich, Gabriel A.;Rajalingam, Krishnaraj;Ravichandran, Kodi S.;Rehm, Markus;Ricci, Jean-Ehrland;Rizzuto, Rosario;Robinson, Nirmal;Rodrigues, Cecilia M. P.;Rotblat, Barak;Rothlin, Carla, V;Rubinsztein, David C.;Rudel, Thomas;Rufini, Alessandro;Ryan, Kevin M.;Sarosiek, Kristopher A.;Sawa, Akira;Sayan, Emre;Schroder, Kate;Scorrano, Luca;Sesti, Federico;Shao, Feng;Shi, Yufang;Sica, Giuseppe S.;Silke, John;Simon, Hans-Uwe;Sistigu, Antonella;Stephanou, Anastasis;Stockwell, Brent R.;Strapazzon, Flavie;Strasser, Andreas;Sun, Liming;Sun, Erwei;Sun, Qiang;Szabadkai, Gyorgy;Tait, Stephen W. G.;Tang, Daolin;Tavernarakis, Nektarios;Troy, Carol M.;Turk, Boris;Urbano, Nicoletta;Vandenabeele, Peter;Vanden Berghe, Tom;Vander Heiden, Matthew G.;Vanderluit, Jacqueline L.;Verkhratsky, Alexei;Villunger, Andreas;von Karstedt, Silvia;Voss, Anne K.;Vousden, Karen H.;Vucic, Domagoj;Vuri, Daniela;Wagner, Erwin F.;Walczak, Henning;Wallach, David;Wang, Ruoning;Wang, Ying;Weber, Achim;Wood, Will;Yamazaki, Takahiro;Yang, Huang-Tian;Zakeri, Zahra;Zawacka-Pankau, Joanna E.;Zhang, Lin;Zhang, Haibing;Zhivotovsky, Boris;Zhou, Wenzhao;Piacentini, Mauro;Kroemer, Guido;Galluzzi, Lorenzo
• 通讯作者: Galluzzi, Lorenzo