Efficient Therapeutic Delivery by a Novel Cell-Penetrating Peptide Derived from Acinus
Abstract
:1. Introduction
2. Results
2.1. Acinus Contains a CPP-Like Sequence
2.2. Cellular Uptake of hAP10 and hAP10DR
2.3. Cellular Uptake Mechanism of hAP10 and hAP10DR
2.4. Analysis of Cellular Toxicity, Hemolytic Activity and Immunogenicity of hAP10 and hAP10DR
2.5. Intracellular Delivery of hAP10- and hAP10DR-GFP Fusion Protein
2.6. Anti-Tumoral Effect of AAC-11 Heptad Leucine Repeat-Derived Peptides
2.7. RT33 and RT33DR Induce Targeted Killing of Circulating Malignant T Cells in Sézary Patients’ Primary PBMC
2.8. RT33 and RT33DR Induce Tumor Growth Reduction in a Xenograft Murine Model of Sézary Syndrome
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Patients and Cells
4.3. Peptides Characterization
4.4. Cellular Uptake Quantification
4.5. Live Cell Microscopy
4.6. Cell Viability and Lactate Dehydrogenase (LDH) Release Assays
4.7. Hemolysis Assay
4.8. Immunogenicity Assay
4.9. Recombinant Protein Purification
4.10. Flow Cytometry Analysis of Sézary Patients’ Cells
4.11. Xenograft Tumor Model
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Ye, J.; Liu, E.; Yu, Z.; Pei, X.; Chen, S.; Zhang, P.; Shin, M.C.; Gong, J.; He, H.; Yang, V.C. CPP-Assisted Intracellular Drug Delivery, What is Next? Int. J. Mol. Sci. 2016, 17, 1892. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Garnacho, C. Intracellular Drug Delivery: Mechanisms for Cell Entry. Curr. Pharm. Des. 2016, 22, 1210–1226. [Google Scholar] [CrossRef] [PubMed]
- Cardoso, A.M.; Trabulo, S.; Cardoso, A.L.; Lorents, A.; Morais, C.M.; Gomes, P.; Nunes, C.; Lucio, M.; Reis, S.; Padari, K.; et al. S4(13)-PV cell-penetrating peptide induces physical and morphological changes in membrane-mimetic lipid systems and cell membranes: Implications for cell internalization. Biochim. Biophys. Acta 2012, 1818, 877–888. [Google Scholar] [CrossRef] [PubMed]
- Alves, I.D.; Goasdoue, N.; Correia, I.; Aubry, S.; Galanth, C.; Sagan, S.; Lavielle, S.; Chassaing, G. Membrane interaction and perturbation mechanisms induced by two cationic cell penetrating peptides with distinct charge distribution. Biochim. Biophys. Acta 2008, 1780, 948–959. [Google Scholar] [CrossRef]
- Richard, J.P.; Melikov, K.; Vives, E.; Ramos, C.; Verbeure, B.; Gait, M.J.; Chernomordik, L.V.; Lebleu, B. Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake. J. Biol. Chem. 2003, 278, 585–590. [Google Scholar] [CrossRef] [Green Version]
- Maiolo, J.R.; Ferrer, M.; Ottinger, E.A. Effects of cargo molecules on the cellular uptake of arginine-rich cell-penetrating peptides. Biochim. Biophys. Acta 2005, 1712, 161–172. [Google Scholar] [CrossRef] [Green Version]
- Guidotti, G.; Brambilla, L.; Rossi, D. Cell-Penetrating Peptides: From Basic Research to Clinics. Trends Pharm. Sci. 2017, 38, 406–424. [Google Scholar] [CrossRef] [PubMed]
- Bechara, C.; Sagan, S. Cell-penetrating peptides: 20 years later, where do we stand? FEBS Lett. 2013, 587, 1693–1702. [Google Scholar] [CrossRef]
- Johnson, R.M.; Harrison, S.D.; Maclean, D. Therapeutic applications of cell-penetrating peptides. Methods Mol. Biol. 2011, 683, 535–551. [Google Scholar] [CrossRef] [PubMed]
- de la Fuente, J.M.; Berry, C.C. Tat peptide as an efficient molecule to translocate gold nanoparticles into the cell nucleus. Bioconjug. Chem. 2005, 16, 1176–1180. [Google Scholar] [CrossRef]
- Zatsepin, T.S.; Turner, J.J.; Oretskaya, T.S.; Gait, M.J. Conjugates of oligonucleotides and analogues with cell penetrating peptides as gene silencing agents. Curr. Pharm. Des. 2005, 11, 3639–3654. [Google Scholar] [CrossRef] [PubMed]
- Liu, H.; Zeng, F.; Zhang, M.; Huang, F.; Wang, J.; Guo, J.; Liu, C.; Wang, H. Emerging landscape of cell penetrating peptide in reprogramming and gene editing. J. Control. Release. 2016, 226, 124–137. [Google Scholar] [CrossRef] [PubMed]
- Suresh, B.; Ramakrishna, S.; Kim, H. Cell-Penetrating Peptide-Mediated Delivery of Cas9 Protein and Guide RNA for Genome Editing. Methods Mol. Biol. 2017, 1507, 81–94. [Google Scholar] [CrossRef] [PubMed]
- Bullok, K.E.; Dyszlewski, M.; Prior, J.L.; Pica, C.M.; Sharma, V.; Piwnica-Worms, D. Characterization of novel histidine-tagged Tat-peptide complexes dual-labeled with (99m)Tc-tricarbonyl and fluorescein for scintigraphy and fluorescence microscopy. Bioconjug. Chem. 2002, 13, 1226–1237. [Google Scholar] [CrossRef] [PubMed]
- Nguyen, Q.T.; Olson, E.S.; Aguilera, T.A.; Jiang, T.; Scadeng, M.; Ellies, L.G.; Tsien, R.Y. Surgery with molecular fluorescence imaging using activatable cell-penetrating peptides decreases residual cancer and improves survival. Proc. Natl. Acad. Sci. USA 2010, 107, 4317–4322. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Olson, E.S.; Jiang, T.; Aguilera, T.A.; Nguyen, Q.T.; Ellies, L.G.; Scadeng, M.; Tsien, R.Y. Activatable cell penetrating peptides linked to nanoparticles as dual probes for in vivo fluorescence and MR imaging of proteases. Proc. Natl. Acad. Sci. USA 2010, 107, 4311–4316. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Habault, J.; Poyet, J.L. Recent Advances in Cell Penetrating Peptide-Based Anticancer Therapies. Molecules 2019, 24, 927. [Google Scholar] [CrossRef] [Green Version]
- Vucetic, Z.; Zhang, Z.; Zhao, J.; Wang, F.; Soprano, K.J.; Soprano, D.R. Acinus-S’ represses retinoic acid receptor (RAR)-regulated gene expression through interaction with the B domains of RARs. Mol. Cell. Biol. 2008, 28, 2549–2558. [Google Scholar] [CrossRef] [Green Version]
- Rodor, J.; Pan, Q.; Blencowe, B.J.; Eyras, E.; Caceres, J.F. The RNA-binding profile of Acinus, a peripheral component of the exon junction complex, reveals its role in splicing regulation. RNA 2016, 22, 1411–1426. [Google Scholar] [CrossRef] [Green Version]
- Tange, T.O.; Shibuya, T.; Jurica, M.S.; Moore, M.J. Biochemical analysis of the EJC reveals two new factors and a stable tetrameric protein core. RNA 2005, 11, 1869–1883. [Google Scholar] [CrossRef] [Green Version]
- Joselin, A.P.; Schulze-Osthoff, K.; Schwerk, C. Loss of Acinus inhibits oligonucleosomal DNA fragmentation but not chromatin condensation during apoptosis. J. Biol. Chem. 2006, 281, 12475–12484. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rigou, P.; Piddubnyak, V.; Faye, A.; Rain, J.C.; Michel, L.; Calvo, F.; Poyet, J.L. The antiapoptotic protein AAC-11 interacts with and regulates Acinus-mediated DNA fragmentation. Embo. J. 2009, 28, 1576–1588. [Google Scholar] [CrossRef] [Green Version]
- Schwerk, C.; Prasad, J.; Degenhardt, K.; Erdjument-Bromage, H.; White, E.; Tempst, P.; Kidd, V.J.; Manley, J.L.; Lahti, J.M.; Reinberg, D. ASAP, a novel protein complex involved in RNA processing and apoptosis. Mol. Cell. Biol. 2003, 23, 2981–2990. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Eiriksdottir, E.; Konate, K.; Langel, U.; Divita, G.; Deshayes, S. Secondary structure of cell-penetrating peptides controls membrane interaction and insertion. Biochim. Biophys. Acta 2010, 1798, 1119–1128. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Buchan, D.W.; Minneci, F.; Nugent, T.C.; Bryson, K.; Jones, D.T. Scalable web services for the PSIPRED Protein Analysis Workbench. Nucleic Acids Res. 2013, 41, W349–W357. [Google Scholar] [CrossRef] [PubMed]
- Roy, A.; Kucukural, A.; Zhang, Y. I-TASSER: A unified platform for automated protein structure and function prediction. Nat. Protoc. 2010, 5, 725–738. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jagot-Lacoussiere, L.; Kotula, E.; Villoutreix, B.O.; Bruzzoni-Giovanelli, H.; Poyet, J.L. A Cell-Penetrating Peptide Targeting AAC-11 Specifically Induces Cancer Cells Death. Cancer Res. 2016, 76, 5479–5490. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kohnken, R.; Fabbro, S.; Hastings, J.; Porcu, P.; Mishra, A. Sezary Syndrome: Clinical and Biological Aspects. Curr. Hematol. Malig. Rep. 2016, 11, 468–479. [Google Scholar] [CrossRef] [PubMed]
- Amand, H.L.; Rydberg, H.A.; Fornander, L.H.; Lincoln, P.; Norden, B.; Esbjorner, E.K. Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans. Biochim. Biophys. Acta 2012, 1818, 2669–2678. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, D.; Wang, J.; Xu, D. Cell-penetrating peptides as noninvasive transmembrane vectors for the development of novel multifunctional drug-delivery systems. J. Control. Release Off. J. Control. Release Soc. 2016, 229, 130–139. [Google Scholar] [CrossRef] [Green Version]
- Wender, P.A.; Mitchell, D.J.; Pattabiraman, K.; Pelkey, E.T.; Steinman, L.; Rothbard, J.B. The design, synthesis, and evaluation of molecules that enable or enhance cellular uptake: Peptoid molecular transporters. Proc. Natl. Acad. Sci. USA 2000, 97, 13003–13008. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Park, C.B.; Yi, K.S.; Matsuzaki, K.; Kim, M.S.; Kim, S.C. Structure-activity analysis of buforin II, a histone H2A-derived antimicrobial peptide: The proline hinge is responsible for the cell-penetrating ability of buforin II. Proc. Natl. Acad. Sci. USA 2000, 97, 8245–8250. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Krautwald, S.; Dewitz, C.; Fandrich, F.; Kunzendorf, U. Inhibition of regulated cell death by cell-penetrating peptides. Cell Mol. Life Sci. 2016, 73, 2269–2284. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pasquereau-Kotula, E.; Habault, J.; Kroemer, G.; Poyet, J.L. The anticancer peptide RT53 induces immunogenic cell death. PLoS ONE 2018, 13, e0201220. [Google Scholar] [CrossRef]
- Tewari, M.; Yu, M.; Ross, B.; Dean, C.; Giordano, A.; Rubin, R. AAC-11, a novel cDNA that inhibits apoptosis after growth factor withdrawal. Cancer Res. 1997, 57, 4063–4069. [Google Scholar]
- Do, T.N.; Rosal, R.V.; Drew, L.; Raffo, A.J.; Michl, J.; Pincus, M.R.; Friedman, F.K.; Petrylak, D.P.; Cassai, N.; Szmulewicz, J.; et al. Preferential induction of necrosis in human breast cancer cells by a p53 peptide derived from the MDM2 binding site. Oncogene 2003, 22, 1431–1444. [Google Scholar] [CrossRef] [Green Version]
- Kanovsky, M.; Raffo, A.; Drew, L.; Rosal, R.; Do, T.; Friedman, F.K.; Rubinstein, P.; Visser, J.; Robinson, R.; Brandt-Rauf, P.W.; et al. Peptides from the amino terminal mdm-2-binding domain of p53, designed from conformational analysis, are selectively cytotoxic to transformed cells. Proc. Natl. Acad. Sci. USA 2001, 98, 12438–12443. [Google Scholar] [CrossRef] [Green Version]
- Sarafraz-Yazdi, E.; Bowne, W.B.; Adler, V.; Sookraj, K.A.; Wu, V.; Shteyler, V.; Patel, H.; Oxbury, W.; Brandt-Rauf, P.; Zenilman, M.E.; et al. Anticancer peptide PNC-27 adopts an HDM-2-binding conformation and kills cancer cells by binding to HDM-2 in their membranes. Proc. Natl. Acad. Sci. USA 2010, 107, 1918–1923. [Google Scholar] [CrossRef] [Green Version]
- Polyansky, A.A.; Chugunov, A.O.; Vassilevski, A.A.; Grishin, E.V.; Efremov, R.G. Recent advances in computational modeling of alpha-helical membrane-active peptides. Curr. Protein Pept. Sci. 2012, 13, 644–657. [Google Scholar] [CrossRef]
- Hughes, C.F.; Khot, A.; McCormack, C.; Lade, S.; Westerman, D.A.; Twigger, R.; Buelens, O.; Newland, K.; Tam, C.; Dickinson, M.; et al. Lack of durable disease control with chemotherapy for mycosis fungoides and Sezary syndrome: A comparative study of systemic therapy. Blood 2015, 125, 71–81. [Google Scholar] [CrossRef]
- Gautam, A.; Chaudhary, K.; Kumar, R.; Sharma, A.; Kapoor, P.; Tyagi, A.; Raghava, G.P.; Open Source Drug Discovery Consortium. In silico approaches for designing highly effective cell penetrating peptides. J. Transl. Med. 2013, 11, 74. [Google Scholar] [CrossRef] [PubMed] [Green Version]
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Habault, J.; Fraser, C.; Pasquereau-Kotula, E.; Born-Bony, M.; Marie-Cardine, A.; Poyet, J.-L. Efficient Therapeutic Delivery by a Novel Cell-Penetrating Peptide Derived from Acinus. Cancers 2020, 12, 1858. https://doi.org/10.3390/cancers12071858
Habault J, Fraser C, Pasquereau-Kotula E, Born-Bony M, Marie-Cardine A, Poyet J-L. Efficient Therapeutic Delivery by a Novel Cell-Penetrating Peptide Derived from Acinus. Cancers. 2020; 12(7):1858. https://doi.org/10.3390/cancers12071858
Chicago/Turabian StyleHabault, Justine, Claire Fraser, Ewa Pasquereau-Kotula, Maëlys Born-Bony, Anne Marie-Cardine, and Jean-Luc Poyet. 2020. "Efficient Therapeutic Delivery by a Novel Cell-Penetrating Peptide Derived from Acinus" Cancers 12, no. 7: 1858. https://doi.org/10.3390/cancers12071858