55 results on '"Gwenola Manic"'
Search Results
2. Correction: Tumor microenvironment modulation enhances immunologic benefit of chemoradiotherapy
- Author
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Dmitriy Zamarin, Howard L Kaufman, Dobrin Draganov, Sofia R Gameiro, Ilio Vitale, Sandy Adjemian, Aitziber Buqué Martinez, Timothy A Chan, Richard L Edelson, Lucia Gabriele, Encouse Golden, Kevin J Harrington, Dewan Md Sakib Hossain, Michael Karin, Juan Jose Lasarte, Sherene Loi, Gwenola Manic, Alan A Melcher, Karen L Mossman, Maria Rescigno, Chiara Riganti, Antonella Sistigu, Bryan E Strauss, Kazuki Tatsuno, Stefaan W van Gool, and Peter Vandenabeele
- Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Published
- 2020
- Full Text
- View/download PDF
3. Consensus guidelines for the definition, detection and interpretation of immunogenic cell death
- Author
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Dmitriy Zamarin, Howard L Kaufman, Dobrin Draganov, Sofia R Gameiro, Ilio Vitale, Sandy Adjemian, Aitziber Buqué Martinez, Timothy A Chan, Richard L Edelson, Lucia Gabriele, Encouse Golden, Kevin J Harrington, Dewan Md Sakib Hossain, Michael Karin, Juan Jose Lasarte, Sherene Loi, Gwenola Manic, Alan A Melcher, Karen L Mossman, Maria Rescigno, Chiara Riganti, Antonella Sistigu, Bryan E Strauss, Kazuki Tatsuno, Stefaan W van Gool, and Peter Vandenabeele
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Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Cells succumbing to stress via regulated cell death (RCD) can initiate an adaptive immune response associated with immunological memory, provided they display sufficient antigenicity and adjuvanticity. Moreover, multiple intracellular and microenvironmental features determine the propensity of RCD to drive adaptive immunity. Here, we provide an updated operational definition of immunogenic cell death (ICD), discuss the key factors that dictate the ability of dying cells to drive an adaptive immune response, summarize experimental assays that are currently available for the assessment of ICD in vitro and in vivo, and formulate guidelines for their interpretation.
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- 2020
- Full Text
- View/download PDF
4. Caspase 2 in mitotic catastrophe: The terminator of aneuploid and tetraploid cells
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Ilio Vitale, Gwenola Manic, Maria Castedo, and Guido Kroemer
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chromosome instability ,mitotic slippage ,polyploidy ,replication stress ,spindle assembly checkpoint ,targeted cancer therapy ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Mitotic catastrophe is an oncosuppressive mechanism that targets cells experiencing defective mitoses via the activation of specific cell cycle checkpoints, regulated cell death pathways and/or cell senescence. This prevents the accumulation of karyotypic aberrations, which otherwise may drive oncogenesis and tumor progression. Here, we summarize experimental evidence confirming the role of caspase 2 (CASP2) as the main executor of mitotic catastrophe, and we discuss the signals that activate CASP2 in the presence of mitotic aberrations. In addition, we summarize the main p53-dependent and -independent effector pathways through which CASP2 limits chromosomal instability and non-diploidy, hence mediating robust oncosuppressive functions.
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- 2017
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- View/download PDF
5. Type-I-interferons in infection and cancer: Unanticipated dynamics with therapeutic implications
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Martina Musella, Gwenola Manic, Ruggero De Maria, Ilio Vitale, and Antonella Sistigu
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anticancer therapy ,cancer stem cells ,ifns ,immunotherapy ,tumor immunity ,Immunologic diseases. Allergy ,RC581-607 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
If there is a great new hope in the treatment of cancer, the immune system is it. Innate and adaptive immunity either promote or attenuate tumorigenesis and so can have opposing effects on the therapeutic outcome. Originally described as potent antivirals, Type-I interferons (IFNs) were quickly recognized as central coordinators of tumor-immune system interactions. Type-I-IFNs are produced by, and act on, both tumor and immune cells being either host-protecting or tumor-promoting. Here, we discuss Type-I-IFNs in infectious and cancer diseases highlighting their dichotomous role and raising the importance to deeply understand the underlying mechanisms so to reshape the way we can exploit Type-I-IFNs therapeutically.
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- 2017
- Full Text
- View/download PDF
6. Impact of the Ku complex on HIV-1 expression and latency.
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Gwenola Manic, Aurélie Maurin-Marlin, Fanny Laurent, Ilio Vitale, Sylvain Thierry, Olivier Delelis, Philippe Dessen, Michelle Vincendeau, Christine Leib-Mösch, Uriel Hazan, Jean-François Mouscadet, and Stéphanie Bury-Moné
- Subjects
Medicine ,Science - Abstract
Ku, a cellular complex required for human cell survival and involved in double strand break DNA repair and multiple other cellular processes, may modulate retroviral multiplication, although the precise mechanism through which it acts is still controversial. Recently, Ku was identified as a possible anti-human immunodeficiency virus type 1 (HIV-1) target in human cells, in two global approaches. Here we investigated the role of Ku on the HIV-1 replication cycle by analyzing the expression level of a panel of non-replicative lentiviral vectors expressing the green fluorescent protein in human colorectal carcinoma HCT 116 cells, stably or transiently depleted of Ku. We found that in this cellular model the depletion of Ku did not affect the efficiency of (pre-)integrative steps but decreased the early HIV-1 expression by acting at the transcriptional level. This negative effect was specific of the HIV-1 promoter, required the obligatory step of viral DNA integration and was reversed by transient depletion of p53. We also provided evidence on a direct binding of Ku to HIV-1 LTR in transduced cells. Ku not only promotes the early transcription from the HIV-1 promoter, but also limits the constitution of viral latency. Moreover, in the presence of a normal level of Ku, HIV-1 expression was gradually lost over time, likely due to the counter-selection of HIV-1-expressing cells. On the contrary, the reactivation of transgene expression from HIV-1 by means of trichostatin A- or tumor necrosis factor α-administration was enhanced under condition of Ku haplodepletion, suggesting a phenomenon of provirus latency. These observations plead in favor of the hypothesis that Ku has an impact on HIV-1 expression and latency at early- and mid-time after integration.
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- 2013
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- View/download PDF
7. Critical involvement of the ATM-dependent DNA damage response in the apoptotic demise of HIV-1-elicited syncytia.
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Jean-Luc Perfettini, Roberta Nardacci, Mehdi Bourouba, Frédéric Subra, Laurent Gros, Claire Séror, Gwenola Manic, Filippo Rosselli, Alessandra Amendola, Peggy Masdehors, Luciana Chessa, Giuseppe Novelli, David M Ojcius, Jan Konrad Siwicki, Magdalena Chechlinska, Christian Auclair, José R Regueiro, Hugues de Thé, Marie-Lise Gougeon, Mauro Piacentini, and Guido Kroemer
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Medicine ,Science - Abstract
DNA damage can activate the oncosuppressor protein ataxia telangiectasia mutated (ATM), which phosphorylates the histone H2AX within characteristic DNA damage foci. Here, we show that ATM undergoes an activating phosphorylation in syncytia elicited by the envelope glycoprotein complex (Env) of human immunodeficiency virus-1 (HIV-1) in vitro. This was accompanied by aggregation of ATM in discrete nuclear foci that also contained phospho-histone H2AX. DNA damage foci containing phosphorylated ATM and H2AX were detectable in syncytia present in the brain or lymph nodes from patients with HIV-1 infection, as well as in a fraction of blood leukocytes, correlating with viral status. Knockdown of ATM or of its obligate activating factor NBS1 (Nijmegen breakage syndrome 1 protein), as well as pharmacological inhibition of ATM with KU-55933, inhibited H2AX phosphorylation and prevented Env-elicited syncytia from undergoing apoptosis. ATM was found indispensable for the activation of MAP kinase p38, which catalyzes the activating phosphorylation of p53 on serine 46, thereby causing p53 dependent apoptosis. Both wild type HIV-1 and an HIV-1 mutant lacking integrase activity induced syncytial apoptosis, which could be suppressed by inhibiting ATM. HIV-1-infected T lymphoblasts from patients with inactivating ATM or NBS1 mutations also exhibited reduced syncytial apoptosis. Altogether these results indicate that apoptosis induced by a fusogenic HIV-1 Env follows a pro-apoptotic pathway involving the sequential activation of ATM, p38MAPK and p53.
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- 2008
- Full Text
- View/download PDF
8. Type I IFNs promote cancer cell stemness by triggering the epigenetic regulator KDM1B
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Martina Musella, Andrea Guarracino, Nicoletta Manduca, Claudia Galassi, Eliana Ruggiero, Alessia Potenza, Ester Maccafeo, Gwenola Manic, Luca Mattiello, Sara Soliman Abdel Rehim, Michele Signore, Marco Pietrosanto, Manuela Helmer-Citterich, Matteo Pallocca, Maurizio Fanciulli, Tiziana Bruno, Francesca De Nicola, Giacomo Corleone, Anna Di Benedetto, Cristiana Ercolani, Edoardo Pescarmona, Laura Pizzuti, Francesco Guidi, Francesca Sperati, Sara Vitale, Daniele Macchia, Massimo Spada, Giovanna Schiavoni, Fabrizio Mattei, Adele De Ninno, Luca Businaro, Valeria Lucarini, Laura Bracci, Eleonora Aricò, Giovanna Ziccheddu, Francesco Facchiano, Stefania Rossi, Massimo Sanchez, Alessandra Boe, Mauro Biffoni, Ruggero De Maria, Ilio Vitale, and Antonella Sistigu
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Histone Demethylases ,Settore BIO/11 ,Immunology ,Breast Neoplasms ,Epigenesis, Genetic ,Genetic ,Settore MED/04 - PATOLOGIA GENERALE ,Interferon Type I ,Neoplastic Stem Cells ,Humans ,Immunology and Allergy ,NA ,Anthracyclines ,Female ,Epigenesis - Abstract
Cancer stem cells (CSCs) are a subpopulation of cancer cells endowed with high tumorigenic, chemoresistant and metastatic potential. Nongenetic mechanisms of acquired resistance are increasingly being discovered, but molecular insights into the evolutionary process of CSCs are limited. Here, we show that type I interferons (IFNs-I) function as molecular hubs of resistance during immunogenic chemotherapy, triggering the epigenetic regulator demethylase 1B (KDM1B) to promote an adaptive, yet reversible, transcriptional rewiring of cancer cells towards stemness and immune escape. Accordingly, KDM1B inhibition prevents the appearance of IFN-I-induced CSCs, both in vitro and in vivo. Notably, IFN-I-induced CSCs are heterogeneous in terms of multidrug resistance, plasticity, invasiveness and immunogenicity. Moreover, in breast cancer (BC) patients receiving anthracycline-based chemotherapy, KDM1B positively correlated with CSC signatures. Our study identifies an IFN-I → KDM1B axis as a potent engine of cancer cell reprogramming, supporting KDM1B targeting as an attractive adjunctive to immunogenic drugs to prevent CSC expansion and increase the long-term benefit of therapy.
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- 2022
9. Catastrophic DNA replication in unscheduled tetraploid cells
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Gwenola Manic, Lorenzo Galluzzi, and Ilio Vitale
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DNA Replication ,Tetraploidy ,Genetics ,Humans ,Mitosis ,Cell Cycle Proteins ,Aneuploidy ,S Phase - Abstract
Unscheduled tetraploidy is a metastable state that rapidly evolves into aneuploidy. Recent findings reported by Gemble et al. demonstrate that freshly formed tetraploid cells fail to accumulate the required amounts of DNA replication factors during the first G
- Published
- 2022
10. Type I IFNs promote cancer cell stemness by triggering the epigenetic regulator KDM1B
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Sara Vitale, Valeria Lucarini, Eleonora Aricò, Cristiana Ercolani, Francesca Sperati, Adele De Ninno, Mauro Biffoni, Alessandra Boe, Gwenola Manic, Luca Businaro, Andrea Guarracino, Martina Musella, Laura Pizzuti, Manuela Helmer-Citterich, Maurizio Fanciulli, Francesco Facchiano, Anna Di Benedetto, Giovanna Ziccheddu, Fabrizio Mattei, Massimo Sanchez, Giovanna Schiavoni, Matteo Pallocca, Laura Bracci, Michele Signore, Ilio Vitale, Edoardo Pescarmona, Antonella Sistigu, Nicoletta Manduca, Marco Pietrosanto, Claudia Galassi, Stefania Rossi, and Ruggero De Maria
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Cancer cell ,Cancer research ,Regulator ,Epigenetics ,Biology - Abstract
Cancer stem cells (CSCs) are immature, immortal cells within tumors, adept at resisting therapeutic pressure and responsible for local and distant disease recurrence. Non-genetic mechanisms of acquired resistance are increasingly being described, however molecular insights into this evolutionary process still lack. Here, we showed that Type I interferons (IFNs-I) act as molecular hubs of resistance during immunogenic chemotherapy, as they trigger the epigenetic regulator KDM1B, responsible for an adaptive, yet reversible, transcriptional rewiring of cancer cells towards stemness and immune escape. Accordingly, KDM1B pharmacological inhibition antagonizes the appearance of IFN-I-adapted CSCs, both in vitro and in vivo. Notably, IFN-I-adapted CSCs are heterogeneous in terms of multidrug resistance, plasticity, invasiveness and immunogenicity. Moreover, in breast cancer patients receiving anthracycline-based chemotherapy, IFN-I and KDM1B signatures positively correlate with CSC and immune evasion markers. Our study identifies IFN-I→KDM1B axis as a potent engine of cancer cell reprogramming and recommends KDM1B targeting an attractive adjunctive to immunogenic drugs to prevent CSC expansion and increase the long-term benefit of therapy.
- Published
- 2021
11. Control of replication stress and mitosis in colorectal cancer stem cells through the interplay of PARP1, MRE11 and RAD51
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Sara Vitale, Mauro Biffoni, Annapaola Franchitto, Claudia Galassi, Francesca De Nicola, Matteo Pallocca, Antonella Sistigu, Fabrizio Antonangeli, Maurizio Fanciulli, Rosa Pennisi, Eva Malacaria, Sara Soliman Abdel Rehim, Martina Musella, Stefano Scalera, Frauke Goeman, Gwenola Manic, Michele Signore, Pietro Pichierri, Marcello Maugeri-Saccà, Francesca Sperati, Andrea Guarracino, Luca Mattiello, Ruggero De Maria, Marta Baiocchi, Ilio Vitale, and Francesca Corradi
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DNA Replication ,Cell death ,cancer stem cell ,Colorectal cancer ,replication stress ,RAD51 ,Poly (ADP-Ribose) Polymerase-1 ,colorectal cancer stem cells ,Mitosis ,Antineoplastic Agents ,Biology ,PARP1 ,Downregulation and upregulation ,Cancer stem cell ,Settore MED/04 - PATOLOGIA GENERALE ,Cell Line, Tumor ,medicine ,Humans ,Molecular Biology ,Mitotic catastrophe ,MRE11 Homologue Protein ,Cancer stem cells ,Comment ,Cell Biology ,medicine.disease ,Cancer research ,Neoplastic Stem Cells ,Rad51 Recombinase ,Stem cell ,Colorectal Neoplasms - Abstract
Cancer stem cells (CSCs) are tumor subpopulations driving disease development, progression, relapse and therapy resistance, and their targeting ensures tumor eradication. CSCs display heterogeneous replication stress (RS), but the functionality/relevance of the RS response (RSR) centered on the ATR-CHK1 axis is debated. Here, we show that the RSR is efficient in primary CSCs from colorectal cancer (CRC-SCs), and describe unique roles for PARP1 and MRE11/RAD51. First, we demonstrated that PARP1 is upregulated in CRC-SCs resistant to several replication poisons and RSR inhibitors (RSRi). In these cells, PARP1 modulates replication fork speed resulting in low constitutive RS. Second, we showed that MRE11 and RAD51 cooperate in the genoprotection and mitosis execution of PARP1-upregulated CRC-SCs. These roles represent therapeutic vulnerabilities for CSCs. Indeed, PARP1i sensitized CRC-SCs to ATRi/CHK1i, inducing replication catastrophe, and prevented the development of resistance to CHK1i. Also, MRE11i + RAD51i selectively killed PARP1-upregulated CRC-SCs via mitotic catastrophe. These results provide the rationale for biomarker-driven clinical trials in CRC using distinct RSRi combinations.
- Published
- 2021
12. The targeting of mre11 or rad51 sensitizes colorectal cancer stem cells to chk1 inhibition
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Antonella Sistigu, Martina Musella, Ilio Vitale, Gwenola Manic, Francesca Corradi, Ruggero De Maria, Claudia Galassi, Sara Soliman Abdel Rehim, Andrea Guarracino, Luca Mattiello, Sara Vitale, and Francesca Sperati
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0301 basic medicine ,Cancer Research ,Cell cycle checkpoint ,medicine.medical_treatment ,Context (language use) ,Tumor initiation ,Biology ,Article ,Targeted therapy ,03 medical and health sciences ,0302 clinical medicine ,Cancer stem cell ,Settore MED/04 - PATOLOGIA GENERALE ,medicine ,Mitotic catastrophe ,RC254-282 ,Tumor-initiating cells ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Colorectal cancer ,Chromosomal instability ,Prexasertib ,enzymes and coenzymes (carbohydrates) ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,DNA damage ,Stem cell ,biological phenomena, cell phenomena, and immunity - Abstract
Simple Summary The ATR-CHK1 axis of the DNA damage response is crucial for the survival of most colorectal cancer stem cells (CRC-SCs), but a significant fraction of primary CRC-SCs either is resistant to ATR or CHK1 inhibitors or survives the abrogation of the ATR-CHK1 cascade despite an initial response. Here, we demonstrate that the targeting of RAD51 or MRE11 improves the sensitivity of primary CRC-SCs to the CHK1/2 inhibitor prexasertib by sequentially inducing replication stress, the abrogation of cell cycle checkpoints, and the emergence of mitotic defects. This results in the induction of mitotic catastrophe and CRC-SC killing via a caspase-dependent apoptosis. Abstract Cancer stem cells (CSCs) drive not only tumor initiation and expansion, but also therapeutic resistance and tumor relapse. Therefore, CSC eradication is required for effective cancer therapy. In preclinical models, CSCs demonstrated high capability to tolerate even extensive genotoxic stress, including replication stress, because they are endowed with a very robust DNA damage response (DDR). This favors the survival of DNA-damaged CSCs instead of their inhibition via apoptosis or senescence. The DDR represents a unique CSC vulnerability, but the abrogation of the DDR through the inhibition of the ATR-CHK1 axis is effective only against some subtypes of CSCs, and resistance often emerges. Here, we analyzed the impact of druggable DDR players in the response of patient-derived colorectal CSCs (CRC-SCs) to CHK1/2 inhibitor prexasertib, identifying RAD51 and MRE11 as sensitizing targets enhancing prexasertib efficacy. We showed that combined inhibition of RAD51 and CHK1 (via B02+prexasertib) or MRE11 and CHK1 (via mirin+prexasertib) kills CSCs by affecting multiple genoprotective processes. In more detail, these two prexasertib-based regimens promote CSC eradication through a sequential mechanism involving the induction of elevated replication stress in a context in which cell cycle checkpoints usually activated during the replication stress response are abrogated. This leads to uncontrolled proliferation and premature entry into mitosis of replication-stressed cells, followed by the induction of mitotic catastrophe. CRC-SCs subjected to RAD51+CHK1 inhibitors or MRE11+CHK1 inhibitors are eventually eliminated, and CRC-SC tumorspheres inhibited or disaggregated, via a caspase-dependent apoptosis. These results support further clinical development of these prexasertib-based regimens in colorectal cancer patients.
- Published
- 2021
13. Cytofluorometric assessment of dendritic cell-mediated uptake of cancer cell apoptotic bodies
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Martina, Musella, Gwenola, Manic, Claudia, Galassi, Ilio, Vitale, and Antonella, Sistigu
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Extracellular Vesicles ,Mice ,Phagocytosis ,Cell Line, Tumor ,Neoplasms ,Animals ,Humans ,Immunogenic Cell Death ,Dendritic Cells ,Flow Cytometry ,Coculture Techniques - Abstract
Dendritic cells (DCs) are specialized antigen presenting cells (APCs) able to intake and crosspresent antigens (Ags) on major histocompatibility complex (MHC) class I and II molecules to T cells thus initiating primary and memory immune responses. DC-mediated Ag uptake and crosspresentation represent crucial steps toward cancer recognition and eventually elimination. Cytofluorometry is a standardized procedure to study phagocytosis. By fast and reproducible single cell measurements, flow cytometry allows for simultaneous biochemical and functional analyses of Ag intake. In this chapter, we discuss a two-color flow cytometric analysis of DC-mediated uptake of apoptotic bodies. We also show data on the adjuvanticity of Type-I-interferons (Type-I-IFNs) during Ag retention as we offer a guideline and a range of advice on sample preparation and acquisition.
- Published
- 2020
14. Assessment of IFN-γ and granzyme-B production by in 'sitro' technology
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Claudia, Galassi, Gwenola, Manic, Martina, Musella, Antonella, Sistigu, and Ilio, Vitale
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Interferon-gamma ,Mice ,Neoplasms ,Programmed Cell Death 1 Receptor ,Animals ,Enzyme-Linked Immunosorbent Assay ,Immunotherapy ,CD8-Positive T-Lymphocytes ,Calreticulin ,Cytotoxicity Tests, Immunologic ,Flow Cytometry ,Granzymes ,Recombinant Proteins - Abstract
Tumor neantigens (TNAs) and tumor-associated antigens (TAAs) are crucial triggers of anticancer immune responses. Through major histocompatibility complex, such antigens activate T cells, which, by releasing interferon gamma (IFN-γ) and granzyme B (GRZB), act as crucial effectors against tumor onset and progression. However, in response to immune pressure, cancer cells use different strategies to favor the establishment of an immunosuppressive tumor microenvironment (TME). Elucidating the dynamics of tumor-host co-evolution provides novel opportunities for personalized cancer immunotherapies. The in sitro (in vitro+in situ) technology is an experimental approach involving the preparation of heterocellular cell suspensions from fresh tumors and their use in vitro. The in sitro experimental setup offers the possibility to (1) analyze immune-related parameters (e.g., quantification of cytokines released in the TME), (2) reveal the mechanism of action of drugs, and (3) unveil crucial cell-intrinsic and cell-extrinsic processes boosting anticancer immune responses. Nonetheless, the in sitro technology does not fully recapitulate the complexity of the tissue "in situ" nor models the patterns of infiltrating immune cell localization, and hence parallel experimentation should be scheduled. In this chapter we discuss in sitro technology to analyze and quantify IFN-γ and GRZB production by T cells either co-cultured with cancer cells in the presence of exogenous adjuvant stimuli (i.e., an antibody targeting the immune checkpoint programmed cell death protein 1, and recombinant calreticulin) and boosting with TAAs (i.e., the model SIINFEKL ovalbumin antigen). Specifically, we describe IFN-γ and GRZB quantification by flow cytometry, ELISA and ELISpot technologies.
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- 2020
15. Assessment of IFN-γ and granzyme-B production by in 'sitro' technology
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Ilio Vitale, Antonella Sistigu, Claudia Galassi, Martina Musella, and Gwenola Manic
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Granzyme B production ,Granzyme B ,Tumor microenvironment ,Immune system ,Antigen ,medicine.medical_treatment ,medicine ,Cancer research ,biology.protein ,Immunotherapy ,Biology ,Major histocompatibility complex ,Immune checkpoint - Abstract
Tumor neantigens (TNAs) and tumor-associated antigens (TAAs) are crucial triggers of anticancer immune responses. Through major histocompatibility complex, such antigens activate T cells, which, by releasing interferon gamma (IFN-γ) and granzyme B (GRZB), act as crucial effectors against tumor onset and progression. However, in response to immune pressure, cancer cells use different strategies to favor the establishment of an immunosuppressive tumor microenvironment (TME). Elucidating the dynamics of tumor-host co-evolution provides novel opportunities for personalized cancer immunotherapies. The in sitro (in vitro+in situ) technology is an experimental approach involving the preparation of heterocellular cell suspensions from fresh tumors and their use in vitro. The in sitro experimental setup offers the possibility to (1) analyze immune-related parameters (e.g., quantification of cytokines released in the TME), (2) reveal the mechanism of action of drugs, and (3) unveil crucial cell-intrinsic and cell-extrinsic processes boosting anticancer immune responses. Nonetheless, the in sitro technology does not fully recapitulate the complexity of the tissue "in situ" nor models the patterns of infiltrating immune cell localization, and hence parallel experimentation should be scheduled. In this chapter we discuss in sitro technology to analyze and quantify IFN-γ and GRZB production by T cells either co-cultured with cancer cells in the presence of exogenous adjuvant stimuli (i.e., an antibody targeting the immune checkpoint programmed cell death protein 1, and recombinant calreticulin) and boosting with TAAs (i.e., the model SIINFEKL ovalbumin antigen). Specifically, we describe IFN-γ and GRZB quantification by flow cytometry, ELISA and ELISpot technologies.
- Published
- 2020
16. Consensus guidelines for the definition, detection and interpretation of immunogenic cell death
- Author
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Sarah Warren, Taha Merghoub, Michael Karin, Radek Spisek, Howard L. Kaufman, Dmitriy Zamarin, John Stagg, Daolin Tang, Timothy A. Chan, Dobrin Draganov, Kazuki Tatsuno, Aitziber Burque Martinez, Jitka Fucikova, Peter Vandenabeele, Udo S. Gaipl, Lorenzo Galluzzi, Juan José Lasarte, Karen L. Mossman, Dewan Md Sakib Hossain, George Coukos, Alessandra Cesano, Patrizia Agostinis, Silvia C. Formenti, Bryan E. Strauss, Sandy Adjemian, Encouse B. Golden, Øystein Rekdal, Guido Kroemer, Mark J. Smyth, Stefaan Van Gool, Oliver Kepp, Felipe Prosper, Sandra Demaria, Kevin J. Harrington, Ilio Vitale, Alan Melcher, Jian Han, Maria Rescigno, Richard L. Edelson, Lucia Gabriele, Sofia R. Gameiro, Gwenola Manic, Laurence Zitvogel, Takahiro Yamazaki, Sherene Loi, Eric Deutsch, James W. Hodge, Chiara Riganti, Abhishek D. Garg, Timothy M Illidge, Antonella Sistigu, Akseli Hemminki, Francesco M. Marincola, and Michael T. Lotze
- Subjects
0301 basic medicine ,Cancer Research ,Molecular biology ,NF-KAPPA-B ,ANTICANCER CHEMOTHERAPY ,Immunogenic Cell Death ,Review ,VDP::Medisinske Fag: 700::Klinisk medisinske fag: 750::Infeksjonsmedisin: 776 ,0302 clinical medicine ,Medicine and Health Sciences ,ANTITUMOR IMMUNITY ,Immunology and Allergy ,Medicine ,RC254-282 ,Manchester Cancer Research Centre ,NOD-LIKE RECEPTORS ,Pattern recognition receptor ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Acquired immune system ,3. Good health ,IMMUNE CHECKPOINT BLOCKADE ,Oncology ,030220 oncology & carcinogenesis ,Molecular Medicine ,Immunogenic cell death ,Intracellular ,Antigenicity ,Consensus ,Antigen presentation ,Immunology ,VDP::Medical disciplines: 700::Clinical medical disciplines: 750::Communicable diseases: 776 ,Guidelines as Topic ,immunogenic cell death ,cancer ,03 medical and health sciences ,Adjuvanticity ,ANTIGEN PRESENTATION ,PATTERN-RECOGNITION RECEPTORS ,APOPTOTIC CELLS ,Humans ,CANCER-CELLS ,Molecular Biology ,Pharmacology ,CALRETICULIN EXPOSURE ,Settore MED/06 - ONCOLOGIA MEDICA ,immunology ,molecular biology ,oncology ,business.industry ,ResearchInstitutes_Networks_Beacons/mcrc ,Inflammatory and immune system ,Correction ,Biology and Life Sciences ,030104 developmental biology ,Cancer cell ,business ,Neuroscience - Abstract
Cells succumbing to stress via regulated cell death (RCD) can initiate an adaptive immune response associated with immunological memory, provided they display sufficient antigenicity and adjuvanticity. Moreover, multiple intracellular and microenvironmental features determine the propensity of RCD to drive adaptive immunity. Here, we provide an updated operational definition of immunogenic cell death (ICD), discuss the key factors that dictate the ability of dying cells to drive an adaptive immune response, summarize experimental assays that are currently available for the assessment of ICD in vitro and in vivo, and formulate guidelines for their interpretation. ispartof: JOURNAL FOR IMMUNOTHERAPY OF CANCER vol:8 issue:1 ispartof: location:England status: published
- Published
- 2020
17. Cytofluorometric assessment of dendritic cell-mediated uptake of cancer cell apoptotic bodies
- Author
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Ilio Vitale, Antonella Sistigu, Claudia Galassi, Martina Musella, and Gwenola Manic
- Subjects
biology ,medicine.diagnostic_test ,Chemistry ,Dendritic cell ,Major histocompatibility complex ,Cell biology ,Flow cytometry ,Immune system ,Antigen ,Cancer cell ,biology.protein ,medicine ,Immunogenic cell death ,Antigen-presenting cell - Abstract
Dendritic cells (DCs) are specialized antigen presenting cells (APCs) able to intake and crosspresent antigens (Ags) on major histocompatibility complex (MHC) class I and II molecules to T cells thus initiating primary and memory immune responses. DC-mediated Ag uptake and crosspresentation represent crucial steps toward cancer recognition and eventually elimination. Cytofluorometry is a standardized procedure to study phagocytosis. By fast and reproducible single cell measurements, flow cytometry allows for simultaneous biochemical and functional analyses of Ag intake. In this chapter, we discuss a two-color flow cytometric analysis of DC-mediated uptake of apoptotic bodies. We also show data on the adjuvanticity of Type-I-interferons (Type-I-IFNs) during Ag retention as we offer a guideline and a range of advice on sample preparation and acquisition.
- Published
- 2020
18. Methods in Stem Cell Biology - Part B
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Ilio Vitale, Gwenola Manic, Lorenzo Galluzzi, Ilio Vitale, Gwenola Manic, and Lorenzo Galluzzi
- Abstract
Methods in Cancer Stem Cell Biology: Part B, Volume 171 in the Methods in Cell Biology series highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including Orthotopic brain tumor models derived from glioblastoma stem-like cells, RNA sequencing in hematopoietic stem cells, Generation of inducible pluripotent stem cells from human dermal fibroblasts, In vitro preparation of dental pulp stem cell grafts combined with biocompatible scaffolds for tissue engineering, Gene expression knockdown in chronic myeloid leukemia stem cells, Identification and isolation of slow-cycling GSCs, Assessment of CD133, EpCAM, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Cell Biology series - Includes the latest information on the topic of Methods in Cancer Stem Cell Biology
- Published
- 2022
19. Methods in Stem Cell Biology - Part A
- Author
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Ilio Vitale, Gwenola Manic, Lorenzo Galluzzi, Ilio Vitale, Gwenola Manic, and Lorenzo Galluzzi
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- Cytology, Stem cells
- Abstract
Methods in Cancer Stem Cell Biology: Part A, Volume 170 in the Methods in Cell Biology series highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including Orthotopic brain tumor models derived from glioblastoma stem-like cells, RNA sequencing in hematopoietic stem cells, Generation of inducible pluripotent stem cells from human dermal fibroblasts, In vitro preparation of dental pulp stem cell grafts combined with biocompatible scaffolds for tissue engineering, Gene expression knockdown in chronic myeloid leukemia stem cells, Identification and isolation of slow-cycling GSCs, Assessment of CD133, EpCAM, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Cell Biology series - Includes the latest information on the topic of Methods in Cancer Stem Cell Biology
- Published
- 2022
20. Deciphering the loop of epithelial-mesenchymal transition, inflammatory cytokines and cancer immunoediting
- Author
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Gwenola Manic, Antonella Sistigu, Paola Nisticò, and Francesca Di Modugno
- Subjects
0301 basic medicine ,Epithelial-Mesenchymal Transition ,Endocrinology, Diabetes and Metabolism ,Immunology ,Tumor-associated macrophage ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,Cancer stem cell ,Neoplasms ,Tumor Microenvironment ,Animals ,Humans ,Immunology and Allergy ,Mesenchymal–epithelial transition ,Epithelial–mesenchymal transition ,Inflammation ,Tumor microenvironment ,Extracellular Matrix ,Cell Transformation, Neoplastic ,030104 developmental biology ,Immunoediting ,Tumor progression ,Disease Progression ,Neoplastic Stem Cells ,Cancer research ,Myeloid-derived Suppressor Cell ,Cytokines ,Signal Transduction - Abstract
Tumorigenesis and tumor progression relies on the dialectics between tumor cells, the extracellular matrix and its remodelling enzymes, neighbouring cells and soluble cues. The host immune response is crucial in eliminating or promoting tumor growth and the reciprocal coevolution of tumor and immune cells, during disease progression and in response to therapy, shapes tumor fate by activating innate and adaptive mechanisms. The phenotypic plasticity is a common feature of epithelial and immune cells and epithelial-mesenchymal transition (EMT) is a dynamic process, governed by microenvironmental stimuli, critical in tumor cell shaping, increased tumor cell heterogeneity and stemness. In this review we will outline how the dysregulation of microenvironmental signaling is crucial in determining tumor plasticity and EMT, arguing how therapy resistance hinges on these dynamics.
- Published
- 2017
21. DNA Damage in Stem Cells
- Author
-
Gwenola Manic, Lorenzo Galluzzi, Ilio Vitale, Ruggero De Maria, and Guido Kroemer
- Subjects
Pluripotent Stem Cells ,0301 basic medicine ,DNA Repair ,DNA repair ,Cellular differentiation ,Biology ,Radiation Tolerance ,Immortal DNA strand hypothesis ,Genomic Instability ,03 medical and health sciences ,stem cells ,Cancer stem cell ,Neoplasms ,Animals ,Humans ,Induced pluripotent stem cell ,Molecular Biology ,Embryonic Stem Cells ,Settore MED/06 - ONCOLOGIA MEDICA ,Genetic Drift ,Cell Biology ,Molecular biology ,Embryonic stem cell ,Cell biology ,Adult Stem Cells ,030104 developmental biology ,Mutation ,Neoplastic Stem Cells ,Stem cell ,DNA Damage ,Adult stem cell - Abstract
Both embryonic and adult stem cells are endowed with a superior capacity to prevent the accumulation of genetic lesions, repair them, or avoid their propagation to daughter cells, which would be particularly detrimental to the whole organism. Inducible pluripotent stem cells also display a robust DNA damage response, but the stability of their genome is often conditioned by the mutational history of the cell population of origin, which constitutes an obstacle to clinical applications. Cancer stem cells are particularly tolerant to DNA damage and fail to undergo senescence or regulated cell death upon accumulation of genetic lesions. Such a resistance contributes to the genetic drift of evolving tumors as well as to their limited sensitivity to chemo- and radiotherapy. Here, we discuss the pathophysiological and therapeutic implications of the molecular pathways through which stem cells cope with DNA damage.
- Published
- 2017
22. ATM kinase sustains breast cancer stem-like cells by promoting ATG4C expression and autophagy
- Author
-
Ilio Vitale, Manolo Sambucci, Ivan Arisi, Daniela Barilà, Rossella Brandi, Venturina Stagni, Martina Antonelli, Flavie Strappazzon, Mara D'Onofrio, and Gwenola Manic
- Subjects
0301 basic medicine ,autophagy ,Tumor suppressor gene ,Blotting, Western ,Autophagy-Related Proteins ,Fluorescent Antibody Technique ,Breast Neoplasms ,Ataxia Telangiectasia Mutated Proteins ,Polymerase Chain Reaction ,Cell Line ,03 medical and health sciences ,Breast cancer ,Cell Line, Tumor ,medicine ,Humans ,Gene silencing ,Oligonucleotide Array Sequence Analysis ,Neoplastic ,Tumor ,Blotting ,business.industry ,Kinase ,Cancer ,mammospheres ,ATG4 ,ATM kinase ,breast cancer stem cells (BCSCs) ,Cysteine Endopeptidases ,Female ,Gene Expression Regulation, Neoplastic ,Neoplastic Stem Cells ,Autophagy ,Cell cycle ,medicine.disease ,3. Good health ,Settore BIO/18 - Genetica ,030104 developmental biology ,Gene Expression Regulation ,Oncology ,Immunology ,Cancer research ,Ectopic expression ,Stem cell ,business ,Western ,Research Paper - Abstract
// Martina Antonelli 1, 2 , Flavie Strappazzon 1 , Ivan Arisi 3 , Rossella Brandi 3 , Mara D’Onofrio 3 , Manolo Sambucci 4 , Gwenola Manic 2 , Ilio Vitale 2, 5 , Daniela Barila 1, 2 , Venturina Stagni 1 1 Instituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Fondazione Santa Lucia, Rome, Italy 2 Department of Biology, University of Rome ‘Tor Vergata’, Rome, Italy 3 Genomics Facility, European Brain Research Institute (EBRI) ‘Rita Levi-Montalcini’, Rome, Italy 4 Neuroimmunology Unit, Fondazione Santa Lucia, IRCCS, Rome, Italy 5 Regina Elena National Cancer Center Institute, Rome, Italy Correspondence to: Venturina Stagni, email: venturina.stagni@gmail.com Daniela Barila, email: daniela.barila@uniroma2.it Keywords: ATM kinase, breast cancer stem cells (BCSCs), mammospheres, autophagy, ATG4 Received: July 05, 2016 Accepted: January 23, 2017 Published: February 20, 2017 ABSTRACT The efficacy of Ataxia-Telangiectasia Mutated (ATM) kinase signalling inhibition in cancer therapy is tempered by the identification of new emerging functions of ATM, which suggests that the role of this protein in cancer progression is complex. We recently demonstrated that this tumor suppressor gene could act as tumor promoting factor in HER2 (Human Epidermal Growth Factor Receptor 2) positive breast cancer. Herein we put in evidence that ATM expression sustains the proportion of cells with a stem-like phenotype, measured as the capability to form mammospheres, independently of HER2 expression levels. Transcriptomic analyses revealed that, in mammospheres, ATM modulates the expression of cell cycle-, DNA repair- and autophagy-related genes. Among these, the silencing of the autophagic gene, autophagy related 4C cysteine peptidase (ATG4C), impairs mammosphere formation similarly to ATM depletion. Conversely, ATG4C ectopic expression in cells silenced for ATM expression, rescues mammospheres growth. Finally, tumor array analyses, performed using public data, identify a significant correlation between ATM and ATG4C expression levels in all human breast cancer subtypes, except for the basal-like one. Overall, we uncover a new connection between ATM kinase and autophagy regulation in breast cancer. We demonstrate that, in breast cancer cells, ATM and ATG4C are essential drivers of mammosphere formation, suggesting that their targeting may improve current approaches to eradicate breast cancer cells with a stem-like phenotype.
- Published
- 2017
23. Macrophages and Metabolism in the Tumor Microenvironment
- Author
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Lorenzo Galluzzi, Gwenola Manic, Ilio Vitale, Guido Kroemer, Lisa M. Coussens, Université Paris Descartes - Paris 5 (UPD5), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université de Paris (UP)
- Subjects
0301 basic medicine ,De facto ,Physiology ,medicine.medical_treatment ,[SDV]Life Sciences [q-bio] ,Population ,Cell ,Biology ,Oxidative Phosphorylation ,03 medical and health sciences ,0302 clinical medicine ,stomatognathic system ,Tumor Microenvironment ,medicine ,Animals ,Humans ,skin and connective tissue diseases ,education ,Molecular Biology ,education.field_of_study ,Tumor microenvironment ,Macrophages ,Cell Biology ,Immunotherapy ,Metabolism ,030104 developmental biology ,medicine.anatomical_structure ,Tumor progression ,Cancer research ,hormones, hormone substitutes, and hormone antagonists ,030217 neurology & neurosurgery ,Metabolic profile - Abstract
Tumor-associated macrophages (TAMs) constitute a plastic and heterogeneous cell population of the tumor microenvironment (TME) that can account for up to 50% of some solid neoplasms. Most often, TAMs support disease progression and resistance to therapy by providing malignant cells with trophic and nutritional support. However, TAMs can mediate antineoplastic effects, especially in response to pharmacological agents that boost their phagocytic and oxidative functions. Thus, TAMs and their impact on the overall metabolic profile of the TME have a major influence on tumor progression and resistance to therapy, de facto constituting promising targets for the development of novel anticancer agents. Here, we discuss the metabolic circuitries whereby TAMs condition the TME to support tumor growth and how such pathways can be therapeutically targeted.
- Published
- 2019
24. Stress responses in stromal cells and tumor homeostasis
- Author
-
Gwenola Manic, Lorenzo Galluzzi, Ilio Vitale, Claudia Galassi, Department of Physics, University of Rome Tor Vergata, Fondazione Ospedale San Camillo [Venezia] (IRCCS), Istituto di Biologia e Patologia Molecolari - CNR [Roma, Italy], Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Weill Medical College of Cornell University [New York], Sandra and Edward Meyer Cancer Center [New-York], Yale University School of Medicine, and Université Paris Descartes - Paris 5 (UPD5)
- Subjects
0301 basic medicine ,Stromal cell ,[SDV]Life Sciences [q-bio] ,medicine.medical_treatment ,Context (language use) ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Stress, Physiological ,Neoplasms ,medicine ,Tumor Microenvironment ,Animals ,Homeostasis ,Humans ,Pharmacology (medical) ,Pharmacology ,Tumor microenvironment ,Mesenchymal stem cell ,Immunotherapy ,3. Good health ,Crosstalk (biology) ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,Stromal Cells - Abstract
In most (if not all) solid tumors, malignant cells are outnumbered by their non-malignant counterparts, including immune, endothelial and stromal cells. However, while the mechanisms whereby cancer cells adapt to microenvironmental perturbations have been studied in great detail, relatively little is known on stress responses in non-malignant compartments of the tumor microenvironment. Here, we discuss the mechanisms whereby cancer-associated fibroblasts and other cellular components of the tumor stroma react to stress in the context of an intimate crosstalk with malignant, endothelial and immune cells, and how such crosstalk influences disease progression and response to treatment.
- Published
- 2019
25. Mutational and Antigenic Landscape in Tumor Progression and Cancer Immunotherapy
- Author
-
Nils Rudqvist, Ilio Vitale, Antonella Sistigu, Lorenzo Galluzzi, Gwenola Manic, and Zlatko Trajanoski
- Subjects
cancer stem cells ,tumor evolution ,medicine.medical_treatment ,Biology ,immune evasion ,tumor microenvironment ,tumor antigen ,Epitope ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Cancer immunotherapy ,Antigens, Neoplasm ,Neoplasms ,medicine ,Animals ,Humans ,030304 developmental biology ,0303 health sciences ,Tumor microenvironment ,Settore MED/06 - ONCOLOGIA MEDICA ,Cell Biology ,Immunotherapy ,Immunosurveillance ,Tumor progression ,Immunology ,Mutation ,Central tolerance ,030217 neurology & neurosurgery - Abstract
Evolving neoplasms accumulate non-synonymous mutations at a high rate, potentially enabling the expression of antigenic epitopes that can be recognized by the immune system. Since they are not covered by central tolerance, such tumor neoantigens (TNAs) should be under robust immune control as they surge. However, genetic defects that impair cancer cell eradication by the immune system coupled with the establishment of local immunosuppression can enable TNA accumulation, which is generally associated with improved clinical sensitivity to various immunotherapies. Here, we explore how tumor-intrinsic factors and immunological processes shape the mutational and antigenic landscape of evolving neoplasms to influence clinical responses to immunotherapy, and propose strategies to achieve robust immunological control of the disease despite disabled immunosurveillance.
- Published
- 2018
26. Replication stress response in cancer stem cells as a target for chemotherapy
- Author
-
Ilio Vitale, Antonella Sistigu, Martina Musella, Francesca Corradi, Ruggero De Maria, and Gwenola Manic
- Subjects
0301 basic medicine ,DNA Replication ,Cancer Research ,DNA Repair ,DNA damage ,Antineoplastic Agents ,Biology ,Aneuploidy Chromosomal instability Fork reversal Replication catastrophe Tumor-initiating cells ,Embryonic stem cell ,03 medical and health sciences ,030104 developmental biology ,PARP1 ,Cancer stem cell ,Multipotent Stem Cell ,Chromosome instability ,Neoplasms ,Cancer cell ,Cancer research ,Neoplastic Stem Cells ,Animals ,Humans ,Homologous recombination ,DNA Damage ,Signal Transduction - Abstract
Cancer stem cells (CSCs) are subpopulations of multipotent stem cells (SCs) responsible for the initiation, long-term clonal maintenance, growth and spreading of most human neoplasms. Reportedly, CSCs share a very robust DNA damage response (DDR) with embryonic and adult SCs, which allows them to survive endogenous and exogenous genotoxins. A range of experimental evidence indicates that CSCs have high but heterogeneous levels of replication stress (RS), arising from, and being boosted by, endogenous causes, such as specific genetic backgrounds (e.g., p53 deficiency) and/or aberrant karyotypes (e.g., supernumerary chromosomes). A multipronged RS response (RSR) is put in place by CSCs to limit and ensure tolerability to RS. The characteristics of such dedicated cascade have two opposite consequences, both relevant for cancer therapy. On the one hand, RSR efficiency often increases the reliance of CSCs on specific DDR components. On the other hand, the functional redundancy of pathways of the RSR can paradoxically promote the acquisition of resistance to RS- and/or DNA damage-inducing agents. Here, we provide an overview of the molecular mechanisms of the RSR in cancer cells and CSCs, focusing on the role of CHK1 and some emerging players, such as PARP1 and components of the homologous recombination repair, whose targeting can represent a long-term effective anti-CSC strategy.
- Published
- 2018
27. Karyotypic Aberrations in Oncogenesis and Cancer Therapy
- Author
-
Laura Senovilla, Ilio Vitale, Gwenola Manic, Lorenzo Galluzzi, and Guido Kroemer
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,fungi ,Cancer therapy ,food and beverages ,Cancer ,Biology ,medicine.disease ,medicine.disease_cause ,Response to treatment ,Oncology ,Tumor progression ,Cancer cell ,Cancer research ,medicine ,Malignant cells ,Carcinogenesis ,Immunological Surveillance - Abstract
The propagation of whole-chromosome (aneuploid) or whole-genome (polyploid) defects is normally prevented by robust cell-intrinsic mechanisms. Moreover, non-diploid cells are under strict immunological surveillance. Nonetheless, tumors contain a high percentage of non-diploid genomes, indicating that malignant cells acquire the ability to bypass these control mechanisms and obtain a survival/proliferation benefit from bulky karyotypic defects. The non-diploid state imposes a significant metabolic burden on cancer cells and hence can be selectively targeted for therapeutic purposes. Here we discuss the impact of abnormal karyotypes on oncogenesis, tumor progression, and response to treatment, focusing on the biochemical and metabolic liabilities of non-diploid cells that can be harnessed for the development of novel chemo(immuno)therapeutic regimens against cancer.
- Published
- 2015
28. Type-I-interferons in infection and cancer: Unanticipated dynamics with therapeutic implications
- Author
-
Antonella Sistigu, Martina Musella, Ruggero De Maria, Gwenola Manic, and Ilio Vitale
- Subjects
0301 basic medicine ,cancer stem cells ,lcsh:Immunologic diseases. Allergy ,medicine.medical_treatment ,Immunology ,Tumor immunity ,Review ,Biology ,medicine.disease_cause ,Bioinformatics ,Anticancer therapy ,IFNs ,immunotherapy ,tumor immunity ,Immunology and Allergy ,Oncology ,lcsh:RC254-282 ,03 medical and health sciences ,Immune system ,Cancer stem cell ,Settore MED/04 - PATOLOGIA GENERALE ,medicine ,anticancer therapy ,Settore MED/06 - ONCOLOGIA MEDICA ,Cancer ,ifns ,Immunotherapy ,Acquired immune system ,medicine.disease ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,interferons ,030104 developmental biology ,Carcinogenesis ,lcsh:RC581-607 - Abstract
If there is a great new hope in the treatment of cancer, the immune system is it. Innate and adaptive immunity either promote or attenuate tumorigenesis and so can have opposing effects on the therapeutic outcome. Originally described as potent antivirals, Type-I interferons (IFNs) were quickly recognized as central coordinators of tumor-immune system interactions. Type-I-IFNs are produced by, and act on, both tumor and immune cells being either host-protecting or tumor-promoting. Here, we discuss Type-I-IFNs in infectious and cancer diseases highlighting their dichotomous role and raising the importance to deeply understand the underlying mechanisms so to reshape the way we can exploit Type-I-IFNs therapeutically.
- Published
- 2017
29. Molecular Mechanisms of Immunogenic Cell Death
- Author
-
Antonella Sistigu, Gwenola Manic, Ilio Vitale, and Martina Musella
- Subjects
Immune system ,biology ,Immunogenicity ,medicine.medical_treatment ,Cancer cell ,Immunology ,Autophagy ,biology.protein ,medicine ,Immunogenic cell death ,Immunotherapy ,Cytotoxicity ,Calreticulin - Abstract
The magnitude of the tumor response to therapy crucially depends on the synergic interaction between immune cells and hitten cancer cells. The ideal cancer treatment should elicit immunogenic cell death (ICD) thus merging direct cytotoxicity with enhanced immunogenicity. The immunogenicity of the dying cancer cells relies on subtle changes in their surface proteome and the secretion of soluble molecules known as damage-associated molecular patterns (DAMPs). This concept of “dying the right way” is vital for the immune system to be instructed to control residual ill cells and maintain micrometastases in a stage of dormancy. Cancer medicine seeks winning strategies combining chemotherapy and immunotherapy. Decoding the concepts and mechanisms that underlie chemotherapy-induced ICD may help shed light on the design of new combined regimens.
- Published
- 2017
30. Synchronization and Desynchronization of Cells by Interventions on the Spindle Assembly Checkpoint
- Author
-
Mohamed, Jemaà, Gwenola, Manic, and Ilio, Vitale
- Subjects
Microscopy, Video ,Cell Cycle ,Mitosis ,Flow Cytometry ,HCT116 Cells ,Microtubules ,Histones ,Polyploidy ,Tetraploidy ,Humans ,M Phase Cell Cycle Checkpoints ,Phosphorylation ,RNA, Small Interfering ,Anaphase ,Software - Abstract
Cell cycle checkpoints are surveillance mechanisms that sequentially and continuously monitor cell cycle progression thereby contributing to the preservation of genetic stability. Among them, the spindle assembly checkpoint (SAC) prevents the occurrence of abnormal divisions by halting the metaphase to anaphase transition following the detection of erroneous microtubules-kinetochore attachment(s). Most synchronization strategies are based on the activation of cell cycle checkpoints to enrich the population of cells in a specific phase of the cell cycle. Here, we develop a two-step protocol of sequential cell synchronization and desynchronization employing antimitotic SAC-inducing agents (i.e., nocodazole or paclitaxel) in combination with the depletion of the SAC kinase MPS1. We describe cytofluorometric and time-lapse videomicroscopy methods to detect cell cycle progression, including the assessment of cell cycle distribution, quantification of mitotic cell fraction, and analysis of single cell fate profile of living cells. We applied these methods to validate the synchronization-desynchronization protocol and to qualitatively and quantitatively determine the impact of SAC inactivation on the activity of antimitotic agents.
- Published
- 2016
31. Synchronization and Desynchronization of Cells by Interventions on the Spindle Assembly Checkpoint
- Author
-
Gwenola Manic, Ilio Vitale, and Mohamed Jemaà
- Subjects
0301 basic medicine ,Cell cycle checkpoint ,Kinetochore ,Cell cycle ,Biology ,Cell biology ,03 medical and health sciences ,Spindle checkpoint ,Nocodazole ,chemistry.chemical_compound ,030104 developmental biology ,chemistry ,Cell synchronization ,Mitosis ,Metaphase - Abstract
Cell cycle checkpoints are surveillance mechanisms that sequentially and continuously monitor cell cycle progression thereby contributing to the preservation of genetic stability. Among them, the spindle assembly checkpoint (SAC) prevents the occurrence of abnormal divisions by halting the metaphase to anaphase transition following the detection of erroneous microtubules-kinetochore attachment(s). Most synchronization strategies are based on the activation of cell cycle checkpoints to enrich the population of cells in a specific phase of the cell cycle. Here, we develop a two-step protocol of sequential cell synchronization and desynchronization employing antimitotic SAC-inducing agents (i.e., nocodazole or paclitaxel) in combination with the depletion of the SAC kinase MPS1. We describe cytofluorometric and time-lapse videomicroscopy methods to detect cell cycle progression, including the assessment of cell cycle distribution, quantification of mitotic cell fraction, and analysis of single cell fate profile of living cells. We applied these methods to validate the synchronization-desynchronization protocol and to qualitatively and quantitatively determine the impact of SAC inactivation on the activity of antimitotic agents.
- Published
- 2016
32. Type I interferons in infection and cancer: Unanticipated dynamics with therapeutic implications
- Author
-
Martina, Musella, Gwenola, Manic, De Maria Marchiano, Ruggero, Ilio, Vitale, Sistigu, Antonella, De Maria Marchiano Ruggero (ORCID:0000-0003-2255-0583), Antonella Sistigu (ORCID:0000-0002-2528-1238), Martina, Musella, Gwenola, Manic, De Maria Marchiano, Ruggero, Ilio, Vitale, Sistigu, Antonella, De Maria Marchiano Ruggero (ORCID:0000-0003-2255-0583), and Antonella Sistigu (ORCID:0000-0002-2528-1238)
- Abstract
If there is a great new hope in the treatment of cancer, the immune system is it. Innate and adaptive immunity either promote or attenuate tumorigenesis and so can have opposing effects on the therapeutic outcome. Originally described as potent antivirals, Type-I interferons (IFNs) were quickly recognized as central coordinators of tumor-immune system interactions. Type-I-IFNs are produced by, and act on, both tumor and immune cells being either host-protecting or tumor-promoting. Here, we discuss Type-I-IFNs in infectious and cancer diseases highlighting their dichotomous role and raising the importance to deeply understand the underlying mechanisms so to reshape the way we can exploit Type-I-IFNs therapeutically.
- Published
- 2017
33. DNA Damage in Stem Cells
- Author
-
Ilio, Vitale, Gwenola, Manic, De Maria Marchiano, Ruggero, Guido, Kroemer, Lorenzo, Galluzzi, De Maria Marchiano Ruggero (ORCID:0000-0003-2255-0583), Ilio, Vitale, Gwenola, Manic, De Maria Marchiano, Ruggero, Guido, Kroemer, Lorenzo, Galluzzi, and De Maria Marchiano Ruggero (ORCID:0000-0003-2255-0583)
- Abstract
Both embryonic and adult stem cells are endowed with a superior capacity to prevent the accumulation of genetic lesions, repair them, or avoid their propagation to daughter cells, which would be particularly detri- mental to the whole organism. Inducible pluripotent stem cells also display a robust DNA damage response, but the stability of their genome is often conditioned by the mutational history of the cell population of origin, which constitutes an obstacle to clinical applications. Cancer stem cells are particularly tolerant to DNA dam- age and fail to undergo senescence or regulated cell death upon accumulation of genetic lesions. Such a resistance contributes to the genetic drift of evolving tumors as well as to their limited sensitivity to chemo- and radiotherapy. Here, we discuss the pathophysiological and therapeutic implications of the molecular pathways through which stem cells cope with DNA damage.
- Published
- 2017
34. CHK1-targeted therapy to deplete DNA replication-stressed, p53-deficient, hyperdiploid colorectal cancer stem cells
- Author
-
Maria Laura De Angelis, Matteo Pallocca, Marta Baiocchi, Ilio Vitale, Sabina Barresi, Francesca Corradi, Sara Vitale, Francesca Sperati, Marcella Mottolese, Francesca De Nicola, Simone Di Franco, Giorgio Stassi, Giorgio Russo, Eleonora Policicchio, Carla Azzurra Amoreo, Gabriele De Luca, Marco Tartaglia, Martina Musella, Gwenola Manic, Antonella Sistigu, Silvia Siteni, Marcello Maugeri-Saccà, Ruggero De Maria, Maurizio Fanciulli, Ann Zeuner, Michele Signore, Manic, G., Signore, M., Sistigu, A., Russo, G., Corradi, F., Siteni, S., Musella, M., Vitale, S., De Angelis, M., Pallocca, M., Amoreo, C., Sperati, F., DI FRANCO, S., Barresi, S., Policicchio, E., De Luca, G., De Nicola, F., Mottolese, M., Zeuner, A., Fanciulli, M., Stassi, G., Maugeri-saccà, M., Baiocchi, M., Tartaglia, M., Vitale, I., and De Maria, R.
- Subjects
0301 basic medicine ,p53 ,DNA Replication ,CELL CYCLE CONTROL ,DNA damage ,Colorectal cancer ,Colon ,medicine.medical_treatment ,Antineoplastic Agents ,Biology ,Bioinformatics ,medicine.disease_cause ,DNA DAMAGE ,Targeted therapy ,03 medical and health sciences ,Cancer stem cell ,Cell Line, Tumor ,medicine ,Humans ,CHEK1 ,DRUG DEVELOPMENT ,Oligonucleotide Array Sequence Analysis ,Mutation ,COLORECTAL CANCER ,Settore MED/06 - ONCOLOGIA MEDICA ,Gastroenterology ,CHEMOTHERAPY ,medicine.disease ,Immunohistochemistry ,Prexasertib ,030104 developmental biology ,Pyrazines ,Checkpoint Kinase 1 ,Cancer research ,Neoplastic Stem Cells ,Pyrazoles ,Stem cell ,Tumor Suppressor Protein p53 ,Colorectal Neoplasms - Abstract
ObjectiveCancer stem cells (CSCs) are responsible for tumour formation and spreading, and their targeting is required for tumour eradication. There are limited therapeutic options for advanced colorectal cancer (CRC), particularly for tumours carrying RAS-activating mutations. The aim of this study was to identify novel CSC-targeting strategies.DesignTo discover potential therapeutics to be clinically investigated as single agent, we performed a screening with a panel of FDA-approved or investigational drugs on primary CRC cells enriched for CSCs (CRC-SCs) isolated from 27 patients. Candidate predictive biomarkers of efficacy were identified by integrating genomic, reverse-phase protein microarray (RPPA) and cytogenetic analyses, and validated by immunostainings. DNA replication stress (RS) was increased by employing DNA replication-perturbing or polyploidising agents.ResultsThe drug-library screening led to the identification of LY2606368 as a potent anti-CSC agent acting in vitro and in vivo in tumour cells from a considerable number of patients (∼36%). By inhibiting checkpoint kinase (CHK)1, LY2606368 affected DNA replication in most CRC-SCs, including RAS-mutated ones, forcing them into premature, lethal mitoses. Parallel genomic, RPPA and cytogenetic analyses indicated that CRC-SCs sensitive to LY2606368 displayed signs of ongoing RS response, including the phosphorylation of RPA32 and ataxia telangiectasia mutated serine/threonine kinase (ATM). This was associated with mutation(s) in TP53 and hyperdiploidy, and made these CRC-SCs exquisitely dependent on CHK1 function. Accordingly, experimental increase of RS sensitised resistant CRC-SCs to LY2606368.ConclusionsLY2606368 selectively eliminates replication-stressed, p53-deficient and hyperdiploid CRC-SCs independently of RAS mutational status. These results provide a strong rationale for biomarker-driven clinical trials with LY2606368 in patients with CRC.
- Published
- 2016
35. Cytofluorometric Quantification of Cell Death Elicited by NLR Proteins
- Author
-
Valentina, Sica, Gwenola, Manic, Guido, Kroemer, Ilio, Vitale, and Lorenzo, Galluzzi
- Subjects
Cell Death ,Caspase 1 ,Humans ,NLR Proteins ,Flow Cytometry ,HCT116 Cells ,Immunity, Innate - Abstract
Nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins, also known as NOD-like receptors, are critical components of the molecular machinery that senses intracellular danger signals to initiate an innate immune response against invading pathogens or endogenous sources of hazard. The best characterized effect of NLR signaling is the secretion of various cytokines with immunostimulatory effects, including interleukin (IL)-1β and IL-18. Moreover, at least under specific circumstances, NLRs can promote regulated variants of cell death. Here, we detail two protocols for the cytofluorometric quantification of cell death-associated parameters that can be conveniently employed to assess the lethal activity of specific NLRs or their ligands.
- Published
- 2016
36. LTX-315, CAPtivating immunity with necrosis
- Author
-
Ilio Vitale, Antonella Sistigu, and Gwenola Manic
- Subjects
0301 basic medicine ,Necrosis ,Cell Biology ,Biology ,Editorials: Cell Cycle Features ,Bioinformatics ,03 medical and health sciences ,030104 developmental biology ,Immunity ,Apoptosis ,Immunology ,Cationic Antimicrobial Peptides ,medicine ,Immunogenic cell death ,medicine.symptom ,Molecular Biology ,Oligopeptides ,Developmental Biology - Published
- 2016
37. Cytofluorometric Quantification of Cell Death Elicited by NLR Proteins
- Author
-
Valentina Sica, Ilio Vitale, Gwenola Manic, Guido Kroemer, and Lorenzo Galluzzi
- Subjects
0301 basic medicine ,Programmed cell death ,Innate immune system ,fungi ,Caspase 1 ,Interleukin ,Biology ,NLR Proteins ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Secretion ,Receptor ,Intracellular - Abstract
Nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins, also known as NOD-like receptors, are critical components of the molecular machinery that senses intracellular danger signals to initiate an innate immune response against invading pathogens or endogenous sources of hazard. The best characterized effect of NLR signaling is the secretion of various cytokines with immunostimulatory effects, including interleukin (IL)-1β and IL-18. Moreover, at least under specific circumstances, NLRs can promote regulated variants of cell death. Here, we detail two protocols for the cytofluorometric quantification of cell death-associated parameters that can be conveniently employed to assess the lethal activity of specific NLRs or their ligands.
- Published
- 2016
38. Mitochondria in Cell Death Regulation
- Author
-
I. Vitale, Gwenola Manic, G. Kroemer, and L. Galluzzi
- Subjects
Programmed cell death ,Mitochondrial permeability transition pore ,Necroptosis ,Apoptosome ,Cell fate determination ,Biology ,Mitochondrion ,Signal transduction ,Intracellular ,Cell biology - Abstract
Besides producing the majority of intracellular adenosine triphosphate and participating in many anabolic circuitries that are required for the survival of eukaryotic cells, mitochondria play a central role in various signaling pathways that actively provoke cell death, including distinct forms of apoptosis and regulated necrosis. Here, we discuss the lethal mechanisms activated by mitochondria in response to adverse conditions or developmental cues, placing special emphasis on the signal transduction cascades that control the cell fate at mitochondrial membranes.
- Published
- 2016
39. 3′ Self-Inactivating Long Terminal Repeat Inserts for the Modulation of Transgene Expression from Lentiviral Vectors
- Author
-
Stéphanie Bury-Moné, Gwenola Manic, Lorenzo Galluzzi, Jean-François Mouscadet, Aurélie Maurin-Marlin, and Frédéric Subra
- Subjects
Transgene ,Genetic enhancement ,Genetic Vectors ,Biology ,Real-Time Polymerase Chain Reaction ,Applied Microbiology and Biotechnology ,Transduction (genetics) ,Genetics ,Animals ,Humans ,3' Flanking Region ,Transgenes ,Genetics (clinical) ,Pharmacology ,Lentivirus ,Gene Transfer Techniques ,Terminal Repeat Sequences ,Genetic Therapy ,Molecular biology ,Long terminal repeat ,Antisense Orientation ,Titer ,Real-time polymerase chain reaction ,Molecular Medicine ,Sense Orientation ,Chickens ,HeLa Cells - Abstract
Gene transfer for research or gene therapy requires the design of vectors that allow for adequate and safe transgene expression. Current methods to modulate the safety and expression profile of retroviral vectors can involve the insertion of insulators or scaffold/matrix-attachment regions in self-inactivating long terminal repeats (SIN-LTRs). Here, we generated a set of lentiviral vectors (with internal CMV or PGK promoter) in which we inserted (at the level of SIN-LTRs) sequences of avian (i.e., chicken hypersensitive site-4, cHS4), human (i.e., putative insulator and desert sequence), or bacterial origin. We characterized them with respect to viral titer, integration, transduction efficiency and transgene expression levels, in both integrase-proficient and -deficient contexts. We found that the cHS4 insulator enhanced transgene expression by a factor of 1.5 only when cloned in the antisense orientation. On the other hand, cHS4 in the sense orientation as well as all other inserts decreased transgene expression. This attenuation phenomenon persisted over long periods of time and did not correspond to extinction or variegation. Decreased transgene expression was associated with lower mRNA levels, yet RNA stability was not affected. Insertions within the SIN-LTRs may negatively affect transgene transcription in a direct fashion through topological rearrangements. The lentiviral vectors that we generated constitute valuable genetic tools for manipulating the level of transgene expression. Moreover, this study demonstrates that SIN-LTR inserts can decrease transgene expression, a phenomenon that might be overcome by modifying insert orientation, thereby highlighting the importance of careful vector design for gene therapy.
- Published
- 2012
40. PO-288 Replication stress response as a target for eradicating colorectal cancer stem cells
- Author
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Gwenola Manic, R. De Maria, Francesca Corradi, A. Sistigu, Michele Signore, and Ilio Vitale
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Cancer Research ,DNA damage ,DNA replication ,Context (language use) ,Biology ,Embryonic stem cell ,chemistry.chemical_compound ,Oncology ,chemistry ,In vivo ,Cancer stem cell ,Cancer research ,Stem cell ,DNA - Abstract
Introduction Cancer stem cells (CSCs) are subsets of multipotent SCs responsible for tumour development, propagation and evolution, whose targeting is required for tumour eradication. There is (pre)clinical evidence on a role of CSCs in therapeutic resistance and intra-tumour heterogeneity, which limits the efficacy of antineoplastic regimens. In this context, CSCs reportedly share with embryonic/adult SCs a very robust DNA damage response, which favours the survival and resistance to genotoxins, and can be exploited for therapeutic purposes. Material and methods We generated a panel of ~30 CRC patient-derived tumorspheres enriched for CSCs (CRC-SCs) and characterised them at the genetic level. To discover potential monotherapeutic anti-CSC agents, we performed high-throughput screenings on multiple CRC-SCs with a library of clinically-relevant drugs. Flow cytometry, fluorescence microscopy and epistatic analyses were conducted to uncover the mechanism of action of identified compound(s), while genetic, cytogenetic and phosphoproteomic studies were carried out to identify predictive biomarkers. DNA replication stress (RS) levels were evaluated by analysing phosphorylated ATM/RPA foci and by performing COMET and DNA fibre assays, and were modulated by single administration of genome destabilising agents or by prolonged exposure to increased doses of compounds targeting the replication stress response (RSR). Results and discussions We demonstrated that the CHK1 inhibitor LY2606368 is a potent anti-CSC agent able to kill more than one third of CRC-SCs, both in vitro and in vivo. Moreover, we provided evidence of high but heterogeneous RS levels in CRC-SCs, showing that, in CRC-SCs, RS is mainly boosted endogenously by p53 deficiency, supernumerary chromosomes and DNA replication abnormalities, which results in high dependency on CHK1-mediated RSR. Accordingly, formerly LY2606368-resistant CRC-SCs were sensitised by boosting DNA replication errors or inducing whole-genome doubling, while formerly LY2606368-sensitive CRC-SCs made resistant by the continuous in vitro or in vivo administration of LY2606368 displayed diminished RS levels due to RSR rewiring, and became independent on CHK1. Conclusion Our results demonstrate that RSR is efficient and rewirable in CSCs thereby constituting a prominent therapeutic target. In particular, we designed dedicated RS-modulating or RSR-targeting strategies for long-term CSC depletion in CRC.
- Published
- 2018
41. PO-300 Unveiling and exploiting cancer stem cell editing and immunogenicity for precision medicine
- Author
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A. Sistigu, C. Galassi, Gwenola Manic, Ilio Vitale, R. De Maria, and Martina Musella
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Cancer Research ,education.field_of_study ,Immunogenicity ,Population ,Cancer ,Biology ,medicine.disease ,Immune checkpoint ,Immune system ,Oncology ,Cancer stem cell ,Cancer research ,medicine ,Immunogenic cell death ,education ,Reprogramming - Abstract
Introduction Immunogenic chemotherapy (IC) induces immunogenic cell death (ICD), which, similar to viral infection, leads to a cancer-cell autonomous Type-I-Interferon (IFN-I) signalling. This immunological signature is crucial for effective antitumor responses but may paradoxically promote the emergence of a rare population of cancer stem cells (CSCs) acting as a chemoresistant niche within the tumour and roots for metastasis and relapse. In this study, we have investigated the role of IFN-I during IC in inducing a cancer editing program resulting in the appearance of poor immunogenic CSCs. Material and methods Human and murine tumour cell lines were treated in vitro with ICD-inducers or IFN-I as control and the induction of CSC were analysed by cytofluorometry, quantitative real time (qRT)-PCR, 3D culture and functional assays. Free and vesicle-mediated nucleic acid transfer during ICD has been characterised by co-culture experiments. IC-induced CSC immunogenicity has been studied through cytofluorometry, microfluidic devices and in vivo experiments. All experiments have been done in triplicate and statistical significance evaluated by two-tailed Student’s t test and two-way ANOVA. Results and discussions The transient/acute induction of IFN-I during ICD is followed by the appearance of a rare population of CSCs. Both free nucleic acids and extracellular vesicles are released during tumour ICD constituting the upstream inducers of IFN-I-mediated reprogramming of neighbouring cells. IC-induced CSCs display epithelial-to-mesenchymal transition traits, multidrug resistance and regenerative properties, and a significant tumorigenic potential when inoculated in immunodeficient and immunocompetent mice. As expected, tumour growth and size are reduced in the presence of an intact immune system. Experiments on microfluidic devices reveal a poor immunogenic potential of CSCs, further confirmed by the expression of immune checkpoint blockers. Conclusion Our results pinpoint a surprising link between ICD, IFN-I and CSCs. Elucidating the mechanisms of CSC editing together with a deep characterisation of CSC (immune) properties could be crucial to prevent tumour relapse. This could undoubtedly have dramatic implications for the clinical management of cancer in an era of terrific development of precision combined chemo-immune therapy.
- Published
- 2018
42. Multipolar mitosis of tetraploid cells: inhibition by p53 and dependency on Mos
- Author
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Gwenola Manic, Mickaël Michaud, Maria Castedo, Nicholas Joza, Alfredo Criollo, Ilio Vitale, Santiago Rello-Varona, Nicolas Tajeddine, Mohamed Jemaà, Laura Senovilla, Guido Kroemer, Oliver Kepp, Alexander Valent, Didier Métivier, Lisa Nanty, Sonia Vivet, and Lorenzo Galluzzi
- Subjects
Mice, Nude ,Mitosis ,Aneuploidy ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Polyploidy ,Mice ,Cell Line, Tumor ,Chromosomal Instability ,Chromosome instability ,Asymmetric cell division ,medicine ,Animals ,Humans ,Molecular Biology ,Mitotic catastrophe ,Centrosome ,Genes, mos ,General Immunology and Microbiology ,General Neuroscience ,Carcinoma ,fungi ,food and beverages ,Chromosome ,medicine.disease ,Molecular biology ,Gene Expression Regulation, Neoplastic ,Colonic Neoplasms ,Female ,Tumor Suppressor Protein p53 ,Ploidy ,Gene Deletion - Abstract
Tetraploidy can constitute a metastable intermediate between normal diploidy and oncogenic aneuploidy. Here, we show that the absence of p53 is not only permissive for the survival but also for multipolar asymmetric divisions of tetraploid cells, which lead to the generation of aneuploid cells with a near-to-diploid chromosome content. Multipolar mitoses (which reduce the tetraploid genome to a sub-tetraploid state) are more frequent when p53 is downregulated and the product of the Mos oncogene is upregulated. Mos inhibits the coalescence of supernumerary centrosomes that allow for normal bipolar mitoses of tetraploid cells. In the absence of p53, Mos knockdown prevents multipolar mitoses and exerts genome-stabilizing effects. These results elucidate the mechanisms through which asymmetric cell division drives chromosomal instability in tetraploid cells.
- Published
- 2010
43. Chemotherapy-induced antitumor immunity requires formyl peptide receptor 1
- Author
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Guido Kroemer, Oliver Kepp, Fabrice Andre, Patrick Günther, José Manuel Bravo-San Pedro, Thomas Ulas, Sandy Adjemian, Elisa E. Baracco, Adele De Ninno, Gwenola Manic, Luca Businaro, Ilio Vitale, Valeria Lucarini, Monica Lucattelli, Federico Pietrocola, Suzette Delaloge, Laurence Zitvogel, Yuting Ma, Joachim L. Schultze, Michaela Semeraro, Giovanna Schiavoni, Celine Lefebvre, Heng Yang, Kariman Chaba, Lionel Apetoh, Antonella Sistigu, Sylvain Ladoire, Gautier Stoll, Christophe Borg, David Enot, Valérie Boige, Michel Ducreux, Francesca Castoldi, Francesca Peschiaroli, Fabrizio Mattei, Sylvie Rusakiewicz, Erika Vacchelli, Cécile Delarasse, Michele Signore, Annamaria Gerardino, Claire Mulot, and Pierre Laurent-Puig
- Subjects
Anthracycline ,Colorectal cancer ,medicine.medical_treatment ,T-Lymphocytes ,Breast Neoplasms ,microfluidic chip ,chemotherapy ,Polymorphism, Single Nucleotide ,Formyl peptide receptor 1 ,immune response ,Mice ,Immune system ,Immunity ,Cell Line, Tumor ,Neoplasms ,medicine ,Leukocytes ,Animals ,Humans ,Anthracyclines ,Alleles ,Annexin A1 ,Chemotherapy ,Multidisciplinary ,business.industry ,Dendritic Cells ,medicine.disease ,Receptors, Formyl Peptide ,Immunity, Innate ,Chemotherapy, Adjuvant ,Cancer cell ,Immunology ,Cancer research ,Female ,business ,Colorectal Neoplasms ,Adjuvant ,FPR1 microfluidic - Abstract
How dying tumor cells get noticed Besides killing tumor cells directly, some chemotherapies, such as anthracyclines, also activate the immune system to kill tumors. Vacchelli et al. discovered that in mice, anthracycline-induced antitumor immunity requires immune cells to express the protein formyl peptide receptor 1 (FPR1). Dendritic cells (DCs) near tumors expressed especially high amounts of FPR1. DCs normally capture fragments of dying tumor cells and use them to activate nearby T cells to kill tumors, but DCs lacking FPR1 failed to do this effectively. Individuals with breast or colon cancer expressing a variant of FPR1 and treated with anthracyclines showed poor metastasis-free and overall survival. Thus, FPR1 may affect anti-tumor immunity in people, too. Science , this issue p. 972
- Published
- 2015
44. Whole-genome duplication increases tumor cell sensitivity to MPS1 inhibition
- Author
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Mohamed, Jemaà, Gwenola, Manic, Gwendaline, Lledo, Delphine, Lissa, Christelle, Reynes, Nathalie, Morin, Frédéric, Chibon, Antonella, Sistigu, Maria, Castedo, Ilio, Vitale, Guido, Kroemer, and Ariane, Abrieu
- Subjects
Paclitaxel ,Cell Survival ,Morpholines ,Immunoblotting ,Mitosis ,AZ 3146 ,Apoptosis ,Cell Cycle Proteins ,Protein Serine-Threonine Kinases ,reversine ,Time-Lapse Imaging ,Cell Line, Tumor ,Neoplasms ,Humans ,Protein Kinase Inhibitors ,polyploidy ,Nocodazole ,regulated cell death ,Protein-Tyrosine Kinases ,HCT116 Cells ,Diploidy ,Tubulin Modulators ,Tetraploidy ,Microscopy, Fluorescence ,Purines ,mitotic spindle ,M Phase Cell Cycle Checkpoints ,RNA Interference ,Research Paper - Abstract
Several lines of evidence indicate that whole-genome duplication resulting in tetraploidy facilitates carcinogenesis by providing an intermediate and metastable state more prone to generate oncogenic aneuploidy. Here, we report a novel strategy to preferentially kill tetraploid cells based on the abrogation of the spindle assembly checkpoint (SAC) via the targeting of TTK protein kinase (better known as monopolar spindle 1, MPS1). The pharmacological inhibition as well as the knockdown of MPS1 kills more efficiently tetraploid cells than their diploid counterparts. By using time-lapse videomicroscopy, we show that tetraploid cells do not survive the aborted mitosis due to SAC abrogation upon MPS1 depletion. On the contrary diploid cells are able to survive up to at least two more cell cycles upon the same treatment. This effect might reflect the enhanced difficulty of cells with whole-genome doubling to tolerate a further increase in ploidy and/or an elevated level of chromosome instability in the absence of SAC functions. We further show that MPS1-inhibited tetraploid cells promote mitotic catastrophe executed by the intrinsic pathway of apoptosis, as indicated by the loss of mitochondrial potential, the release of the pro-apoptotic cytochrome c from mitochondria, and the activation of caspases. Altogether, our results suggest that MPS1 inhibition could be used as a therapeutic strategy for targeting tetraploid cancer cells.
- Published
- 2015
45. Trial Watch: Targeting ATM-CHK2 and ATR-CHK1 pathways for anticancer therapy
- Author
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Antonella Sistigu, Ilio Vitale, Florine Obrist, and Gwenola Manic
- Subjects
Genome instability ,UCN-01 ,Cancer Research ,Cell cycle checkpoint ,animal structures ,Review ,Biology ,medicine.disease_cause ,environment and public health ,medicine ,CHEK1 ,Checkpoint Kinase 2 ,CHEK2 ,double-strand breaks ,drug resistance ,Kinase ,genomic instability ,DNA lesions ,enzymes and coenzymes (carbohydrates) ,cell death ,Tumor progression ,Cancer research ,Molecular Medicine ,cell cycle checkpoint ,biological phenomena, cell phenomena, and immunity ,Carcinogenesis - Abstract
The ataxia telangiectasia mutated serine/threonine kinase (ATM)/checkpoint kinase 2 (CHEK2, best known as CHK2) and the ATM and Rad3-related serine/threonine kinase (ATR)/CHEK1 (best known as CHK1) cascades are the 2 major signaling pathways driving the DNA damage response (DDR), a network of processes crucial for the preservation of genomic stability that act as a barrier against tumorigenesis and tumor progression. Mutations and/or deletions of ATM and/or CHK2 are frequently found in tumors and predispose to cancer development. In contrast, the ATR-CHK1 pathway is often upregulated in neoplasms and is believed to promote tumor growth, although some evidence indicates that ATR and CHK1 may also behave as haploinsufficient oncosuppressors, at least in a specific genetic background. Inactivation of the ATM-CHK2 and ATR-CHK1 pathways efficiently sensitizes malignant cells to radiotherapy and chemotherapy. Moreover, ATR and CHK1 inhibitors selectively kill tumor cells that present high levels of replication stress, have a deficiency in p53 (or other DDR players), or upregulate the ATR-CHK1 module. Despite promising preclinical results, the clinical activity of ATM, ATR, CHK1, and CHK2 inhibitors, alone or in combination with other therapeutics, has not yet been fully demonstrated. In this Trial Watch, we give an overview of the roles of the ATM-CHK2 and ATR-CHK1 pathways in cancer initiation and progression, and summarize the results of clinical studies aimed at assessing the safety and therapeutic profile of regimens based on inhibitors of ATR and CHK1, the only 2 classes of compounds that have so far entered clinics.
- Published
- 2014
46. Trial watch – inhibiting PARP enzymes for anticancer therapy
- Author
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Gwenola Manic, Florine Obrist, Ilio Vitale, and Antonella Sistigu
- Subjects
0301 basic medicine ,chemistry.chemical_classification ,Cancer Research ,DNA damage ,Poly ADP ribose polymerase ,Review ,Synthetic lethality ,Biology ,Olaparib ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,PARP1 ,Enzyme ,chemistry ,Biochemistry ,Downregulation and upregulation ,030220 oncology & carcinogenesis ,Cancer research ,Molecular Medicine ,Nuclear protein - Abstract
Poly(ADP-ribose) polymerases (PARPs) are a members of family of enzymes that catalyze poly(ADP-ribosyl)ation (PARylation) and/or mono(ADP-ribosyl)ation (MARylation), two post-translational protein modifications involved in crucial cellular processes including (but not limited to) the DNA damage response (DDR). PARP1, the most abundant family member, is a nuclear protein that is activated upon sensing distinct types of DNA damage and contributes to their resolution by PARylating multiple DDR players. Recent evidence suggests that, along with DDR, activated PARP1 mediates a series of prosurvival and proapoptotic processes aimed at preserving genomic stability. Despite this potential oncosuppressive role, upregulation and/or overactivation of PARP1 or other PARP enzymes has been reported in a variety of human neoplasms. Over the last few decades, several pharmacologic inhibitors of PARP1 and PARP2 have been assessed in preclinical and clinical studies showing potent antineoplastic activity, particularly against homologous recombination (HR)-deficient ovarian and breast cancers. In this Trial Watch, we describe the impact of PARP enzymes and PARylation in cancer, discuss the mechanism of cancer cell killing by PARP1 inactivation, and summarize the results of recent clinical studies aimed at evaluating the safety and therapeutic profile of PARP inhibitors in cancer patients.
- Published
- 2015
47. Trial Watch: Proteasomal inhibitors for anticancer therapy
- Author
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Gwenola Manic, Guido Kroemer, Lorenzo Galluzzi, Ilio Vitale, and Florine Obrist
- Subjects
chemistry.chemical_classification ,ixazomib ,Cancer Research ,Hyperactivation ,Intracellular protein ,dexamethasone ,Review ,Biology ,NHL - Non-Hodgkin's lymphoma ,rituximab ,Enzyme ,chemistry ,Ubiquitin ,immunomodulatory drugs ,oprozomib ,Oncogenic signaling ,biology.protein ,Cancer research ,Molecular Medicine ,marizomib ,Signal transduction ,Homeostasis - Abstract
The so-called "ubiquitin-proteasome system" (UPS) is a multicomponent molecular apparatus that catalyzes the covalent attachment of several copies of the small protein ubiquitin to other proteins that are generally (but not always) destined to proteasomal degradation. This enzymatic cascade is crucial for the maintenance of intracellular protein homeostasis (both in physiological conditions and in the course of adaptive stress responses), and regulates a wide array of signaling pathways. In line with this notion, defects in the UPS have been associated with aging as well as with several pathological conditions including cardiac, neurodegenerative, and neoplastic disorders. As transformed cells often experience a constant state of stress (as a result of the hyperactivation of oncogenic signaling pathways and/or adverse microenvironmental conditions), their survival and proliferation are highly dependent on the integrity of the UPS. This rationale has driven an intense wave of preclinical and clinical investigation culminating in 2003 with the approval of the proteasomal inhibitor bortezomib by the US Food and Drug Administration for use in multiple myeloma patients. Another proteasomal inhibitor, carfilzomib, is now licensed by international regulatory agencies for use in multiple myeloma patients, and the approved indications for bortezomib have been extended to mantle cell lymphoma. This said, the clinical activity of bortezomib and carfilzomib is often limited by off-target effects, innate/acquired resistance, and the absence of validated predictive biomarkers. Moreover, the antineoplastic activity of proteasome inhibitors against solid tumors is poor. In this Trial Watch we discuss the contribution of the UPS to oncogenesis and tumor progression and summarize the design and/or results of recent clinical studies evaluating the therapeutic profile of proteasome inhibitors in cancer patients.
- Published
- 2014
48. Trial Watch: Radioimmunotherapy for oncological indications
- Author
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Norma Bloy, Jonathan Pol, Gwenola Manic, Ilio Vitale, Alexander Eggermont, Jérôme Galon, Eric Tartour, Laurence Zitvogel, Guido Kroemer, and Lorenzo Galluzzi
- Subjects
Chemotherapy ,business.industry ,medicine.medical_treatment ,Immunology ,Ibritumomab tiuxetan ,Cancer ,Review ,medicine.disease ,Radiation therapy ,Immune system ,Oncology ,Treatment modality ,Radioimmunotherapy ,Cancer research ,Immunology and Allergy ,Medicine ,Immunogenic cell death ,business ,medicine.drug - Abstract
During the past two decades, it has become increasingly clear that the antineoplastic effects of radiation therapy do not simply reflect the ability of X-, β- and γ-rays to damage transformed cells and directly cause their permanent proliferative arrest or demise, but also involve cancer cell-extrinsic mechanisms. Indeed, among other activities, radiotherapy has been shown to favor the establishment of tumor-specific immune responses that operate systemically, underpinning the so-called ‘out-of-field’ or ‘abscopal’ effect. Thus, ionizing rays appear to elicit immunogenic cell death, a functionally peculiar variant of apoptosis associated with the emission of a particularly immunostimulatory combination of damage-associated molecular patterns. In line with this notion, radiation therapy fosters, and thus exacerbates, the antineoplastic effects of various treatment modalities, including surgery, chemotherapy and various immunotherapeutic agents. Here, we summarize recent advances in the use of ionizing rays as a means to induce or potentiate therapeutically relevant anticancer immune responses. In addition, we present clinical trials initiated during the past 12 months to test the actual benefit of radioimmunotherapy in cancer patients.
- Published
- 2014
49. Chloroquine and hydroxychloroquine for cancer therapy
- Author
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Guido Kroemer, Florine Obrist, Gwenola Manic, Ilio Vitale, and Lorenzo Galluzzi
- Subjects
Cancer Research ,Autophagy ,beclin 1 ,Cancer ,Context (language use) ,Hydroxychloroquine ,Review ,Biology ,Pharmacology ,medicine.disease ,medicine.disease_cause ,lysosomes ,proteasome ,Chloroquine ,Tumor progression ,bafilomycin A1 ,mTOR ,medicine ,Molecular Medicine ,autophagosomes ,Carcinogenesis ,PI3K/AKT/mTOR pathway ,medicine.drug - Abstract
Macroautophagy (herein referred to as autophagy) is a highly conserved mechanism for the lysosomal degradation of cytoplasmic components. Autophagy is critical for the maintenance of intracellular homeostasis, both in baseline conditions and in the context of adaptive responses to stress. In line with this notion, defects in the autophagic machinery have been etiologically associated with various human disorders including infectious, inflammatory and neoplastic conditions. Once tumors are established, however, autophagy sustains the survival of malignant cells, hence representing an appealing target for the design of novel anticancer regimens. Accordingly, inhibitors of autophagy including chloroquine and hydroxychloroquine have been shown to mediate substantial antineoplastic effects in preclinical models, especially when combined with chemo- or radiotherapeutic interventions. The pharmacological profile of chloroquine and hydroxychloroquine, however, appear to involve mechanisms other than autophagy inhibition. Here, we discuss the dual role of autophagy in oncogenesis and tumor progression, and summarize the results or design of clinical studies recently completed or initiated to evaluate the therapeutic activity of chloroquine derivatives in cancer patients.
- Published
- 2014
50. Impact of the Ku Complex on HIV-1 Expression and Latency
- Author
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Christine Leib-Mösch, Uriel Hazan, Michelle Vincendeau, Jean-François Mouscadet, Ilio Vitale, Stéphanie Bury-Moné, Sylvain Thierry, Olivier Delelis, Fanny Laurent, Philippe Dessen, Aurélie Maurin-Marlin, and Gwenola Manic
- Subjects
Small interfering RNA ,DNA repair ,Virus Integration ,Transgene ,DNA transcription ,Retrovirology and HIV immunopathogenesis ,lcsh:Medicine ,Viral diseases ,Biology ,Biochemistry ,Microbiology ,Vector Biology ,Molecular cell biology ,Transcription (biology) ,Virology ,DNA-binding proteins ,Humans ,lcsh:Science ,Ku Autoantigen ,Transcription factor ,Multidisciplinary ,lcsh:R ,DNA Helicases ,Proteins ,HIV ,Transfection ,Provirus ,Flow Cytometry ,HCT116 Cells ,Viral Replication ,Viral Persistence and Latency ,Virus Latency ,Cell biology ,HIV-1 ,Medicine ,Infectious diseases ,lcsh:Q ,Gene expression ,Viral Vectors ,Tumor Suppressor Protein p53 ,Cellular model ,Viral Latency ,Cytometry ,Research Article - Abstract
Ku, a cellular complex required for human cell survival and involved in double strand break DNA repair and multiple other cellular processes, may modulate retroviral multiplication, although the precise mechanism through which it acts is still controversial. Recently, Ku was identified as a possible anti-human immunodeficiency virus type 1 (HIV-1) target in human cells, in two global approaches. Here we investigated the role of Ku on the HIV-1 replication cycle by analyzing the expression level of a panel of non-replicative lentiviral vectors expressing the green fluorescent protein in human colorectal carcinoma HCT 116 cells, stably or transiently depleted of Ku. We found that in this cellular model the depletion of Ku did not affect the efficiency of (pre-)integrative steps but decreased the early HIV-1 expression by acting at the transcriptional level. This negative effect was specific of the HIV-1 promoter, required the obligatory step of viral DNA integration and was reversed by transient depletion of p53. We also provided evidence on a direct binding of Ku to HIV-1 LTR in transduced cells. Ku not only promotes the early transcription from the HIV-1 promoter, but also limits the constitution of viral latency. Moreover, in the presence of a normal level of Ku, HIV-1 expression was gradually lost over time, likely due to the counter-selection of HIV-1-expressing cells. On the contrary, the reactivation of transgene expression from HIV-1 by means of trichostatin A- or tumor necrosis factor α-administration was enhanced under condition of Ku haplodepletion, suggesting a phenomenon of provirus latency. These observations plead in favor of the hypothesis that Ku has an impact on HIV-1 expression and latency at early- and mid-time after integration.
- Published
- 2013
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