Inhibition of Phosphatidylinositol 3-Kinase by Pictilisib Blocks Influenza Virus Propagation in Cells and in Lungs of Infected Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Treatment with Pictilisib
2.2. Cells and Viruses
2.3. In Vitro Infection
2.4. In Vivo Infection
2.5. Plaque Titrations
2.6. Cell Growth Analysis
2.7. Western Blotting
2.8. Reporter Gene Assay
2.9. Human Chip Model
2.10. Immunofluorescence Microscopy
2.11. qRT-PCR
2.12. Legendplex Assay
2.13. Statistical Analysis
3. Results
3.1. Pictilisib Treatment Leads to an Efficient Inhibition of Influenza Virus Replication in Mono-Cell-Culture
3.2. Pictilisib Treatment Does Not Affect Viability of Epithelial Cells
3.3. An Early Stage in IV-Infection Is Inhibited by Pictilisib
3.4. Pictilisib Treatment Leads to an Efficient Inhibition of IV Replication in the Human Chip Model
3.5. Pictilisib Blocks Virus Titers and The Pro-Inflammatory Response of IV-Infected Mice
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
- Chow, E.J.; Rolfes, M.A.; O’Halloran, A.; Alden, N.B.; Anderson, E.J.; Bennett, N.M.; Billing, L.; Dufort, E.; Kirley, P.D.; George, A.; et al. Respiratory and Nonrespiratory Diagnoses Associated with Influenza in Hospitalized Adults. JAMA Netw. Open 2020, 3, e201323. [Google Scholar] [CrossRef] [Green Version]
- Putri, W.C.; Muscatello, D.J.; Stockwell, M.S.; Newall, A.T. Economic burden of seasonal influenza in the United States. Vaccine 2018, 36, 3960–3966. [Google Scholar] [CrossRef] [PubMed]
- Vogel, O.A.; Manicassamy, B. Broadly Protective Strategies against Influenza Viruses: Universal Vaccines and Therapeutics. Front. Microbiol. 2020, 11, 135. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jorgensen, P.; Mereckiene, J.; Cotter, S.; Johansen, K.; Tsolova, S.; Brown, C. How close are countries of the WHO European Region to achieving the goal of vaccinating 75% of key risk groups against influenza? Results from national surveys on seasonal influenza vaccination programmes, 2008/2009 to 2014/2015. Vaccine 2018, 36, 442–452. [Google Scholar] [CrossRef]
- Duwe, S. Influenza viruses—Antiviral therapy and resistance. GMS Infect. Dis. 2017, 5. [Google Scholar] [CrossRef]
- Fukao, K.; Ando, Y.; Noshi, T.; Kitano, M.; Noda, T.; Kawai, M.; Yoshida, R.; Sato, A.; Shishido, T.; Naito, A. Baloxavir marboxil, a novel cap-dependent endonuclease inhibitor potently suppresses influenza virus replication and represents therapeutic effects in both immunocompetent and immunocompromised mouse models. PLoS ONE 2019, 14, e0217307. [Google Scholar] [CrossRef]
- Gubareva, L.V.; Mishin, V.P.; Patel, M.C.; Chesnokov, A.; Nguyen, H.T.; De La Cruz, J.; Spencer, S.; Campbell, A.P.; Sinner, M.; Reid, H.; et al. Assessing baloxavir susceptibility of influenza viruses circulating in the United States during the 2016/17 and 2017/18 seasons. Eurosurveillance 2019, 24, 1800666. [Google Scholar] [CrossRef]
- Ludwig, S.; Hrincius, E.R.; Boergeling, Y. The Two Sides of the Same Coin-Influenza Virus and Intracellular Signal Trans-duction. Cold Spring Harb. Perspect. Med. 2019, 11, a038513. [Google Scholar] [CrossRef] [Green Version]
- Ehrhardt, C.; Rückle, A.; Hrincius, E.R.; Haasbach, E.; Anhlan, D.; Ahmann, K.; Banning, C.; Reiling, S.J.; Kühn, J.; Strobl, S.; et al. The NF-kappaB inhibitor SC75741 efficiently blocks influenza virus propagation and confers a high barrier for development of viral resistance. Cell. Microbiol. 2013, 15, 1198–1211. [Google Scholar] [CrossRef]
- Mazur, I.; Wurzer, W.J.; Ehrhardt, C.; Pleschka, S.; Puthavathana, P.; Silberzahn, T.; Wolff, T.; Planz, O.; Ludwig, S. Acetylsalicylic acid (ASA) blocks influenza virus propagation via its NF-kappaB-inhibiting activity. Cell. Microbiol. 2007, 9, 1683–1694. [Google Scholar] [CrossRef]
- Meineke, R.; Rimmelzwaan, G.F.; Elbahesh, H. Influenza Virus Infections and Cellular Kinases. Viruses 2019, 11, 171. [Google Scholar] [CrossRef] [Green Version]
- Ehrhardt, C.; Ludwig, S. A new player in a deadly game: Influenza viruses and the PI3K/Akt signalling pathway. Cell. Microbiol. 2009, 11, 863–871. [Google Scholar] [CrossRef] [PubMed]
- Ehrhardt, C.; Marjuki, H.; Wolff, T.; Nurnberg, B.; Planz, O.; Pleschka, S.; Ludwig, S. Bivalent role of the phosphatidylinositol-3-kinase (PI3K) during influenza virus infection and host cell defence. Cell. Microbiol. 2006, 8, 1336–1348. [Google Scholar] [CrossRef] [PubMed]
- Eierhoff, T.; Hrincius, E.R.; Rescher, U.; Ludwig, S.; Ehrhardt, C. The Epidermal Growth Factor Receptor (EGFR) Promotes Uptake of Influenza A Viruses (IAV) into Host Cells. PLoS Pathog. 2010, 6, e1001099. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fujioka, Y.; Tsuda, M.; Hattori, T.; Sasaki, J.; Sasaki, T.; Miyazaki, T.; Ohba, Y. The Ras–PI3K Signaling Pathway Is Involved in Clathrin-Independent Endocytosis and the Internalization of Influenza Viruses. PLoS ONE 2011, 6, e16324. [Google Scholar] [CrossRef] [Green Version]
- Marjuki, H.; Gornitzky, A.; Marathe, B.M.; Ilyushina, N.A.; Aldridge, J.R.; Desai, G.; Webby, R.J.; Webster, R.G. Influenza A virus-induced early activation of ERK and PI3K mediates V-ATPase-dependent intracellular pH change required for fusion. Cell. Microbiol. 2011, 13, 587–601. [Google Scholar] [CrossRef] [Green Version]
- Sarkar, S.; Peters, K.L.; Elco, C.P.; Sakamoto, S.; Pal, S.; Sen, G.C. Novel roles of TLR3 tyrosine phosphorylation and PI3 kinase in double-stranded RNA signaling. Nat. Struct. Mol. Biol. 2004, 11, 1060–1067. [Google Scholar] [CrossRef]
- Hrincius, E.R.; Dierkes, R.; Anhlan, D.; Wixler, V.; Ludwig, S.; Ehrhardt, C. Phosphatidylinositol-3-kinase (PI3K) is activated by influenza virus vRNA via the pathogen pattern receptor Rig-I to promote efficient type I interferon production. Cell. Microbiol. 2011, 13, 1907–1919. [Google Scholar] [CrossRef]
- Hale, B.G.; Jackson, D.; Chen, Y.-H.; Lamb, R.A.; Randall, R.E. Influenza A virus NS1 protein binds p85beta and activates phosphatidylinositol-3-kinase signaling. Proc. Natl. Acad. Sci. USA 2006, 103, 14194–14199. [Google Scholar] [CrossRef] [Green Version]
- Ehrhardt, C.; Wolff, T.; Pleschka, S.; Planz, O.; Beermann, W.; Bode, J.G.; Schmolke, M.; Ludwig, S. Influenza A Virus NS1 Protein Activates the PI3K/Akt Pathway to Mediate Antiapoptotic Signaling Responses. J. Virol. 2007, 81, 3058–3067. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rosário-Ferreira, N.; Preto, A.; Melo, R.; Moreira, I.S.; Brito, R.M.M. The Central Role of Non-Structural Protein 1 (NS1) in Influenza Biology and Infection. Int. J. Mol. Sci. 2020, 21, 1511. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Krop, I.E.; Mayer, I.A.; Ganju, V.; Dickler, M.; Johnston, S.; Morales, S.; Yardley, D.A.; Melichar, B.; Forero-Torres, A.; Lee, S.C.; et al. Pictilisib for oestrogen receptor-positive, aromatase inhibitor-resistant, advanced or metastatic breast cancer (FERGI): A randomised, double-blind, placebo-controlled, phase 2 trial. Lancet Oncol. 2016, 17, 811–821. [Google Scholar] [CrossRef] [Green Version]
- Sarker, D.; Ang, J.E.; Baird, R.D.; Kristeleit, R.; Shah, K.; Moreno, V.; Clarke, P.A.; Raynaud, F.; Levy, G.; Ware, J.A.; et al. First-in-Human Phase I Study of Pictilisib (GDC-0941), a Potent Pan–Class I Phosphatidylinositol-3-Kinase (PI3K) Inhibitor, in Patients with Advanced Solid Tumors. Clin. Cancer Res. 2015, 21, 77–86. [Google Scholar] [CrossRef] [Green Version]
- Leong, S.; Moss, R.A.; Bowles, D.W.; Ware, J.A.; Zhou, J.; Spoerke, J.M.; Lackner, M.R.; Shankar, G.; Schutzman, J.L.; Van Der Noll, R.; et al. A Phase I Dose-Escalation Study of the Safety and Pharmacokinetics of Pictilisib in Combination with Erlotinib in Patients with Advanced Solid Tumors. Oncologist 2017, 22, 1491–1499. [Google Scholar] [CrossRef] [Green Version]
- O’Brien, C.; Wallin, J.J.; Sampath, D.; GuhaThakurta, D.; Savage, H.; Punnoose, E.A.; Guan, J.; Berry, L.; Prior, W.W.; Amler, L.C.; et al. Predictive Biomarkers of Sensitivity to the Phosphatidylinositol 3′ Kinase Inhibitor GDC-0941 in Breast Cancer Preclinical Models. Clin. Cancer Res. 2010, 16, 3670–3683. [Google Scholar] [CrossRef] [Green Version]
- Seidel, N.; Sauerbrei, A.; Wutzler, P.; Schmidtke, M. Hemagglutinin 222D/G Polymorphism Facilitates Fast Intra-Host Evolution of Pandemic (H1N1) 2009 Influenza A Viruses. PLoS ONE 2014, 9, e104233. [Google Scholar] [CrossRef] [Green Version]
- Manchanda, H.; Seidel, N.; Krumbholz, A.; Sauerbrei, A.; Schmidtke, M.; Guthke, R. Within-host influenza dynamics: A small-scale mathematical modeling approach. Biosystems 2014, 118, 51–59. [Google Scholar] [CrossRef]
- Basler, C.F.; Mikulasova, A.; Martinez-Sobrido, L.; Paragas, J.; Mühlberger, E.; Bray, M.; Klenk, H.-D.; Palese, P.; García-Sastre, A. The Ebola Virus VP35 Protein Inhibits Activation of Interferon Regulatory Factor 3. J. Virol. 2003, 77, 7945–7956. [Google Scholar] [CrossRef] [Green Version]
- Deinhardt-Emmer, S.; Rennert, K.; Schicke, E.; Cseresnyés, Z.; Windolph, M.; Nietzsche, S.; Heller, R.; Siwczak, F.; Haupt, K.F.; Carlstedt, S.; et al. Co-infection with Staphylococcus aureus after primary influenza virus infection leads to damage of the endothelium in a human alveolus-on-a-chip model. Biofabrication 2020, 12, 025012. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Deinhardt-Emmer, S.; Böttcher, S.; Häring, C.; Giebeler, L.; Henke, A.; Zell, R.; Jungwirth, J.; Jordan, P.M.; Werz, O.; Hornung, F.; et al. SARS-CoV-2 causes severe epithelial inflammation and barrier dysfunction. J. Virol. 2021. [Google Scholar] [CrossRef]
- Alessi, D.R.; Andjelkovic, M.; Caudwell, B.; Cron, P.; Morrice, N.; Cohen, P.; Hemmings, B.A. Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J. 1996, 15, 6541–6551. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sarbassov, D.D.; Guertin, D.A.; Ali, S.M.; Sabatini, D.M. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 2005, 307, 1098–1101. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sieczkarski, S.B.; Brown, H.A.; Whittaker, G.R. Whittaker, Role of protein kinase C βII in influenza virus entry via late endosomes. J. Virol. 2003, 77, 460–469. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Paget, J.; Spreeuwenberg, P.; Charu, V.; Taylor, R.J.; Iuliano, A.D.; Bresee, J.; Simonsen, L.; Viboud, C. Global Seasonal Influenza-Associated Mortality Collaborator Network and GLaMOR Collaborating Teams. Global mortality associated with seasonal influenza epidemics: New burden estimates and predictors from the GLaMOR Project. J. Glob. Health 2019, 9, 020421. [Google Scholar] [CrossRef] [PubMed]
- Zeng, S.-X.; Zhu, Y.; Ma, A.-H.; Yu, W.; Zhang, H.; Lin, T.-Y.; Shi, W.; Tepper, C.G.; Henderson, P.T.; Airhart, S.; et al. The Phosphatidylinositol 3-Kinase Pathway as a Potential Therapeutic Target in Bladder Cancer. Clin. Cancer Res. 2017, 23, 6580–6591. [Google Scholar] [CrossRef] [Green Version]
- Schöffski, P.; Cresta, S.; Mayer, I.A.; Wildiers, H.; Damian, S.; Gendreau, S.; Rooney, I.; Morrissey, K.M.; Spoerke, J.M.; Ng, V.W.; et al. A phase Ib study of pictilisib (GDC-0941) in combination with paclitaxel, with and without bevacizumab or trastuzumab, and with letrozole in advanced breast cancer. Breast Cancer Res. 2018, 20, 109. [Google Scholar] [CrossRef]
- Elmenier, F.M.; Lasheen, D.S.; Abouzid, K.A. Phosphatidylinositol 3 kinase (PI3K) inhibitors as new weapon to combat cancer. Eur. J. Med. Chem. 2019, 183, 111718. [Google Scholar] [CrossRef]
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Deinhardt-Emmer, S.; Jäckel, L.; Häring, C.; Böttcher, S.; Wilden, J.J.; Glück, B.; Heller, R.; Schmidtke, M.; Koch, M.; Löffler, B.; et al. Inhibition of Phosphatidylinositol 3-Kinase by Pictilisib Blocks Influenza Virus Propagation in Cells and in Lungs of Infected Mice. Biomolecules 2021, 11, 808. https://doi.org/10.3390/biom11060808
Deinhardt-Emmer S, Jäckel L, Häring C, Böttcher S, Wilden JJ, Glück B, Heller R, Schmidtke M, Koch M, Löffler B, et al. Inhibition of Phosphatidylinositol 3-Kinase by Pictilisib Blocks Influenza Virus Propagation in Cells and in Lungs of Infected Mice. Biomolecules. 2021; 11(6):808. https://doi.org/10.3390/biom11060808
Chicago/Turabian StyleDeinhardt-Emmer, Stefanie, Laura Jäckel, Clio Häring, Sarah Böttcher, Janine J. Wilden, Brigitte Glück, Regine Heller, Michaela Schmidtke, Mirijam Koch, Bettina Löffler, and et al. 2021. "Inhibition of Phosphatidylinositol 3-Kinase by Pictilisib Blocks Influenza Virus Propagation in Cells and in Lungs of Infected Mice" Biomolecules 11, no. 6: 808. https://doi.org/10.3390/biom11060808