Intracellular drug delivery: aiding cross presentation of subunit vaccines
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1 Intracellular drug delivery: aiding cross presentation of subunit vaccines Last Time: Today: Reading: basic vaccine concepts Using synthetic biomaterials to enhance cytosolic delivery of molecules Wang et al. Molecularly engineered poly(ortho ester) microspheres for enhanced delivery of DNA vaccines, Nat. Mater (2004) Supplementary Reading: ANNOUNCEMENTS: Lecture 21 Spring
2 Particle-stimulated cross presentation Graph removed due to copyright restrictions. Please see: Kovacs-Bankowski et al. PNAS 90 (1993): Image removed due to copyright restrictions. Please see: Lehner, and Cresswell. Curr Opin Immunol 16, 82 (2004). Lecture 21 Spring
3 INTRACELLULAR DRUG DELIVERY AND VACCINES: Image removed due to copyright restrictions. Please see: Vijayanathan, et al Lecture 21 Spring
4 ENDOSOMAL ESCAPE: Enhancing cross presentation cytosolic delivery of large macromolecules (1) proton sponge effect (2) ph-activated polymers and peptides Lecture 21 Spring
5 INTRACELLULAR VS. EXTRACELLULAR ENVIRONMENT Image removed due to copyright restrictions. Please see: Alberts, Bruce, et al. Molecular Biology of the Cell. New York, NY: Garland,
6 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PROTON PROTON SPONGES SPONGES Proton sponge effect cytosol Endosome interior Ion transporters Proton pumps polyethyleneimine Lecture 21 Spring
7 Role of additional structural features of PEI in efficient endosomal escape: Images removed due to copyright restrictions. Please see: Dubruel, et al. Biomacromolecules 5 (2004):
8 Endosomal escape by direct membrane interactions Both polycation and polyanion headgroups with pkas = 5-7 can promote endosomal escape: ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Image removed due to copyright restrictions. Please see: Mann, Stephen. Biomineralization: Principles and Concepts in Bioinorganic Materials Chemistry. New York, NY: Oxford University Press, ph: Polyanionic liposome Lecture 21 Spring
9 Endosomal escape by direct membrane interactions Both polycation and polyanion headgroups with pkas = 5-7 can promote endosomal escape: ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS ph: polycationic liposome Lecture 21 Spring
10 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS STRATEGIES FOR CUED BURST RELEASE OF CARGO COINCIDENT WITH ENDOSOMAL ESCAPE Images removed due to copyright restrictions. Please see: Lynn, Langer, et al. Angew Chem Int Ed 40 (2001): Lecture 21 Spring
11 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Images and graph removed due to copyright restrictions. Please see: Little, Langer, et al. PNAS 101 (2004): Lecture 21 Spring
12 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Approaches to endosome escape: encrypted polymers Multi-function molecular carriers: Figure removed due to copyright restrictions. Please see: Figure 1 in Murthy, N. et al. "Bioinspired ph-responsive Polymers for the Intracellular Delivery of Biomolecular Drugs." Bioconjug Chem 14 (2003): Lecture 21 Spring
13 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS (1) (2) (3) (4) endosomes Lecture 21 Spring
14 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Figure removed due to copyright restrictions. Please see: Figure 3 in Murthy, N. et al. "Bioinspired ph-responsive Polymers for the Intracellular Delivery of Biomolecular Drugs." Bioconjug Chem 14 (2003): Lecture 21 Spring
15 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Results with peptide delivery by encrypted polymers Figure removed due to copyright restrictions. Lecture 21 Spring
16 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Example vaccine results: ph-responsive gels as vaccines Images removed due to copyright restrictions. Please see: Murthy, Frechet, et al. PNAS 100 (2003): Lecture 21 Spring
17 ENDOSOMAL ENDOSOMAL ESCAPE: ESCAPE: PH-RESPONSIVE PH-RESPONSIVE POLYMERS POLYMERS Example vaccine results: ph-responsive gels as vaccines Figure removed due to copyright restrictions. Please see: Murthy, Frechet, et al. Bioconj Chem 15 (2004): Lecture 21 Spring
18 DIRECT ENTRY TO THE CYTOSOL Membrane-penetrating peptides Pore-forming peptides Fusogenic peptides Lecture 21 Spring
19 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES PEPTIDES Cell-penetrating peptides (CPPs) [aka Protein Transduction Domains (PTDs)] Image removed due to copyright restrictions. Please see: Joliot, A., and A. Prochiantz. "Transduction Peptides: from Technology to Physiology." Nat Cell Biol 6 (2004): Lecture 21 Spring
20 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES PEPTIDES Sources and sequences Table removed due to copyright restrictions. Please see: Table 1 in Joliot, A., and A. Prochiantz. "Transduction Peptides: from Technology to Physiology." Nat Cell Biol 6 (2004): Lecture 21 Spring
21 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES MEMBRANE-PENETRATING PEPTIDES Penetratin: Short peptide sequence from drosophila transcription factor protein Antennapedia RQIKIWFQNRRMKWKK Models of membrane-penetrating peptide function Figure removed due to copyright restrictions. Please see: Figure 7 in Derossi, D., et al. "Cell Internalization of the Third Helix of the Antennapeadia Homeodomain is Receptor-Independent." J Biol Chem 271 (1996):
22 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES PEPTIDES Uptake of penetratin by primary neuronal cells: CPP function in vitro Protein delivery using HIV tat peptide: Images removed due to copyright restrictions. Please see: Derossi, D., et al. "Cell Internalization of the Third Helix of the Antennapeadia Homeodomain is Receptor-Independent." J Biol Chem 271 (1996): Images and graph removed due to copyright restrictions. Please see: Schwarze, S. R., et al. "Vivo Protein Transduction: Delivery of a Biologically Active Protein into the Mouse." In Science 285 (1999): Lecture 21 Spring
23 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES PEPTIDES CPP function in vivo Images removed due to copyright restrictions. Please see: Schwarze, S. R., et al. "In Vivo Protein Transduction: Delivery of a Biologically Active Protein into the Mouse." Science 285 (1999): Lecture 21 Spring
24 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: MEMBRANE-PENETRATING PEPTIDES PEPTIDES ACTIVATION ON ENTRY TO THE CYTOSOL Selective bond dissociation using reversible disulfide linkages Figure removed due to copyright restrictions. Please see: Falnes, P. O., and K. Sandvig. "Penetration of Protein Toxins into Cells." Curr Opin Cell Biol 12 (2000): Lecture 21 Spring
25 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: PORE-FORMING PORE-FORMING PEPTIDES PEPTIDES Pore-forming proteins/peptides as a tool for membrane-penetrating drug carriers Figure removed due to copyright restrictions. Please see: Figure 1 in Bhakdi, S., et al. "Staphylococcal Alpha-Toxin, Streptolysin-O and Escherichia Coli Hemolysin: P rototypes of Pore-Forming Bacterial Cytlysins." Arch Microbiol 165: Lecture 21 Spring
26 DIRECT DIRECT ENTRY ENTRY TO TO CYTOSOL: CYTOSOL: FUSOGENIC FUSOGENIC PEPTIDES PEPTIDES fusogenic peptides: using viral entry strategies for drug delivery Images removed due to copyright restrictions. Please see: Hawiger, J. "Noninvasive Intracellular Delivery of Functional Peptides and Proteins." Curr Opin Chem Biol 3 (1999): Lecture 21 Spring
27 Further Reading 1. Moghimi, S. M., Hunter, A. C. & Murray, J. C. Long-circulating and target-specific nanoparticles: theory to practice. Pharmacol Rev 53, (2001). 2. Hawiger, J. Noninvasive intracellular delivery of functional peptides and proteins. Curr Opin Chem Biol 3, (1999). 3. Derossi, D. et al. Cell internalization of the third helix of the Antennapedia homeodomain is receptor-independent. J Biol Chem 271, (1996). 4. Falnes, P. O. & Sandvig, K. Penetration of protein toxins into cells. Curr Opin Cell Biol 12, (2000). 5. Joliot, A. & Prochiantz, A. Transduction peptides: from technology to physiology. Nat Cell Biol 6, (2004). 6. Schwarze, S. R., Ho, A., Vocero-Akbani, A. & Dowdy, S. F. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285, (1999). 7. Snyder, E. L. & Dowdy, S. F. Cell penetrating peptides in drug delivery. Pharm Res 21, (2004). 8. Thoren, P. E. et al. Membrane binding and translocation of cell-penetrating peptides. Biochemistry 43, (2004). 9. Asokan, A. & Cho, M. J. Exploitation of intracellular ph gradients in the cellular delivery of macromolecules. J Pharm Sci 91, (2002). 10. Sandgren, S., Cheng, F. & Belting, M. Nuclear targeting of macromolecular polyanions by an HIV-Tat derived peptide. Role for cell-surface proteoglycans. J Biol Chem 277, (2002). 11. Yatvin, M. B., Kreutz, W., Horwitz, B. A. & Shinitzky, M. Ph-Sensitive Liposomes - Possible Clinical Implications. Science 210, (1980). 12. Lee, K. D., Oh, Y. K., Portnoy, D. A. & Swanson, J. A. Delivery of macromolecules into cytosol using liposomes containing hemolysin from Listeria monocytogenes. J Biol Chem 271, (1996). 13. Bhakdi, S. et al. Staphylococcal alpha-toxin, streptolysin-o, and Escherichia coli hemolysin: prototypes of pore-forming bacterial cytolysins. Arch Microbiol 165, 73-9 (1996). 14. Raychaudhuri, S. & Rock, K. L. Fully mobilizing host defense: building better vaccines. Nat Biotechnol 16, (1998). 15. Falo, L. D., Jr., Kovacsovics-Bankowski, M., Thompson, K. & Rock, K. L. Targeting antigen into the phagocytic pathway in vivo induces protective tumour immunity. Nat Med 1, (1995). 16. Murthy, N., Campbell, J., Fausto, N., Hoffman, A. S. & Stayton, P. S. Bioinspired ph- Responsive Polymers for the Intracellular Delivery of Biomolecular Drugs. Bioconjug Chem 14, (2003). 17. Shi, G., Guo, W., Stephenson, S. M. & Lee, R. J. Efficient intracellular drug and gene delivery using folate receptor-targeted ph-sensitive liposomes composed of cationic/anionic lipid combinations. J Control Release 80, (2002). Lecture 21 Spring
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