Purpose: Alteration in cell death is a hallmark of cancer. A functional role regulating survival, apoptosis, and necroptosis has been attributed to RIP1/3 complexes.Experimental Design: We have investigated the role of RIP1 and the effects of MC2494 in cell death induction, using different methods as flow cytometry, transcriptome analysis, immunoprecipitation, enzymatic assays, transfections, mutagenesis, and in vivo studies with different mice models.Results: Here, we show that RIP1 is highly expressed in cancer, and we define a novel RIP1/3-SIRT1/2-HAT1/4 complex. Mass spectrometry identified five acetylations in the kinase and death domain of RIP1. The novel characterized pan-SIRT inhibitor, MC2494, increases RIP1 acetylation at two additional sites in the death domain. Mutagenesis of the acetylated lysine decreases RIP1-dependent cell death, suggesting a role for acetylation of the RIP1 complex in cell death modulation. Accordingly, MC2494 displays tumor-selective potential in vitro, in leukemic blasts ex vivo, and in vivo in both xenograft and allograft cancer models. Mechanistically, MC2494 induces bona fide tumor-restricted acetylated RIP1/caspase-8-mediated apoptosis. Excitingly, MC2494 displays tumor-preventive activity by blocking 7,12-dimethylbenz(α)anthracene-induced mammary gland hyperproliferation in vivoConclusions: These preventive features might prove useful in patients who may benefit from a recurrence-preventive approach with low toxicity during follow-up phases and in cases of established cancer predisposition. Thus, targeting the newly identified RIP1 complex may represent an attractive novel paradigm in cancer treatment and prevention.

RIP1-HAT1-SirT complex identification and targeting in treatment and prevention of cancer / Carafa, Vincenzo; Nebbioso, Angela; Cuomo, Francesca; Rotili, Dante; Cobellis, Gilda; Bontempo, Paola; Baldi, Alfonso; Spugnini, Enrico P.; Citro, Alfonso Gennaro; Chambery, Angela; Russo, Rosita; Ciana, Paolo; Maravigna, Luca; Shaik, Jani; Radaelli, Enrico; De Antonellis, Pasquale; Tarantino, Domenico; Pirolli, Adele; Ragno, Rino; Massimo Zollo, .; Stunnenberg, Hendrik G.; Mai, Antonello; Altucci, Lucia. - In: CLINICAL CANCER RESEARCH. - ISSN 1078-0432. - STAMPA. - 24:12(2018), pp. 2886-2900. [10.1158/1078-0432.CCR-17-3081]

RIP1-HAT1-SirT complex identification and targeting in treatment and prevention of cancer

Dante Rotili
Membro del Collaboration Group
;
Gennaro Citro
Membro del Collaboration Group
;
Domenico Tarantino
Membro del Collaboration Group
;
Adele Pirolli
Membro del Collaboration Group
;
Rino Ragno
Membro del Collaboration Group
;
Antonello Mai
Membro del Collaboration Group
;
2018

Abstract

Purpose: Alteration in cell death is a hallmark of cancer. A functional role regulating survival, apoptosis, and necroptosis has been attributed to RIP1/3 complexes.Experimental Design: We have investigated the role of RIP1 and the effects of MC2494 in cell death induction, using different methods as flow cytometry, transcriptome analysis, immunoprecipitation, enzymatic assays, transfections, mutagenesis, and in vivo studies with different mice models.Results: Here, we show that RIP1 is highly expressed in cancer, and we define a novel RIP1/3-SIRT1/2-HAT1/4 complex. Mass spectrometry identified five acetylations in the kinase and death domain of RIP1. The novel characterized pan-SIRT inhibitor, MC2494, increases RIP1 acetylation at two additional sites in the death domain. Mutagenesis of the acetylated lysine decreases RIP1-dependent cell death, suggesting a role for acetylation of the RIP1 complex in cell death modulation. Accordingly, MC2494 displays tumor-selective potential in vitro, in leukemic blasts ex vivo, and in vivo in both xenograft and allograft cancer models. Mechanistically, MC2494 induces bona fide tumor-restricted acetylated RIP1/caspase-8-mediated apoptosis. Excitingly, MC2494 displays tumor-preventive activity by blocking 7,12-dimethylbenz(α)anthracene-induced mammary gland hyperproliferation in vivoConclusions: These preventive features might prove useful in patients who may benefit from a recurrence-preventive approach with low toxicity during follow-up phases and in cases of established cancer predisposition. Thus, targeting the newly identified RIP1 complex may represent an attractive novel paradigm in cancer treatment and prevention.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1116555
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