Projects | Masaharu Somiya

Protein design for drug delivery applications

Wed, 03 Apr 2024 03:00:00 +0000

Viruses have evolved to use proteins to overcome cell membrane to deliver their genetic information into host cells. Inspired by the viral spike/capsid proteins, we are trying to design new proteins that can either (1) induce membrane fusion between host membrane and membrane of drug carrier (liposomes or LNPs), or (2) disrupt host cell membrane. By using current cutting-edge machine learning-based protein design tools, we have made significant progress on making new designed proteins towards efficient intracellular delivery of various therapeutic modalities (protein, RNA etc).

References

Yang EC, Divine R, Miranda MC, Borst AJ, Sheffler W, Zhang JZ, Decarreau J, Saragovi A, Abedi M, Goldbach N, Ahlrichs M, Dobbins C, Hand A, Cheng S, Lamb M, Levine PM, Chan S, Skotheim R, Fallas J, Ueda G, Lubner J, Somiya M, Khmelinskaia A, King NP, Baker D., “Computational design of non-porous, pH-responsive antibody nanoparticles,” bioRxiv, p. 2023.04.17.537263, Apr. 2023, doi: 10.1101/2023.04.17.537263.

Cargo transfer hypothesis on extracellular vesicles

Tue, 26 Jul 2022 03:00:00 +0000

Intracellular trafficking and cargo release mechanisms of extracellular vesicles (EVs) are still largely unknown. We have developed experimental tools to study the efficiency of cargo release and membrane fusion of EVs inside recipient cells. These tools enable the quantitative assessment of the intracellular cargo delivery process of EVs, and even real-time imaging of cargo delivery in living cells (see pictures, purple dots are the cytoplasmically delivered EV cargo, with a little help from virus-derived fusion protein VSV-G).

Plasmids required for these tools are available through Addgene.

Reference

M. Somiya, “Where does the cargo go?: Solutions to provide experimental support for the ‘extracellular vesicle cargo transfer hypothesis,’” Journal of Cell Communication and Signaling, vol. 14, no. 2, pp. 135–146, Jun. 2020, doi: 10.1007/s12079-020-00552-9.
M. Somiya and S. Kuroda, “Real-Time Luminescence Assay for Cytoplasmic Cargo Delivery of Extracellular Vesicles,” Anal. Chem., vol. 93, no. 13, pp. 5612–5620, Apr. 2021, doi: 10.1021/acs.analchem.1c00339.
M. Somiya and S. Kuroda, “Reporter gene assay for membrane fusion of extracellular vesicles,” J. Extracel. Vesicle, e12171, Nov. 2021, doi: 10.1002/jev2.12171.
M. Somiya and S. Kuroda, “Verification of extracellular vesicle-mediated functional mRNA delivery via RNA editing", bioRxiv, Jan. 2022, doi: 10.1101/2022.01.25.477620.

RNA delivery using bio-inspired nanoparticles

Tue, 26 Jul 2022 00:33:07 +0000

mRNA vaccines/therapeutics have been an emerging topic since the COVID-19 pandemic starting in 2019. The potential of mRNA-based vaccines and therapeutics seems to be enormous against various diseases, including emerging infections diseases, rare diseases, genetic diseases. A typical delivery technology for RNAs is known as lipid nanoparticle (LNP), which is a tiny ball of fats and RNAs. The LNP technology has been demonstrating great potential to deliver mRNA and siRNA, however, the limited capacity of cytoplasmic delivery (which means a large fraction of LNPs are taken up by cells but degraded inside cells and could not functionally deliver RNA) is the bottleneck for LNP-mediated delivery.

I’ve been working on the functional delivery of RNAs into cells. I believe extracellular vesicles (EVs) have great potential for this purpose and engineering the EVs to improve the delivery capability will lead to the development of a novel delivery platform.

In addition, we are using neutral liposomes as a delivery system for RNAs instead of cationic liposomes. Usually, cationic liposomes are frequently used for the delivery of RNAs, however, due to the positive charge of cationic lipids, cationic liposomes exert toxicity and are easily inactivated in vivo. We discovered that non-cationic liposomes modified with photosensitizer can functionally deliver the siRNA cargo into cells upon light irradiation. This light-responsive delivery system may be an ideal tool to deliver the cargo in a spatiotemporally controlled manner.

References

M. Somiya, K. Sakaeda, Y. Ishii, and S. Kuroda, “Cytoplasmic delivery of small interfering RNA by photoresponsive non-cationic liposomes,” Journal of Drug Delivery Science and Technology, vol. 63, p. 102488, Mar. 2021, doi: 10.1016/j.jddst.2021.102488.
M. Somiya et al., “One-step scalable preparation method for non-cationic liposomes with high siRNA content,” International Journal of Pharmaceutics, vol. 490, no. 1–2, pp. 316–323, Jul. 2015, doi: 10.1016/j.ijpharm.2015.05.072.
M. Somiya and S. Kuroda, “Verification of extracellular vesicle-mediated functional mRNA delivery via RNA editing", bioRxiv, Jan. 2022, doi: 10.1101/2022.01.25.477620.

Hepatitis B virus: Identification of early infection mechanism and therapeutic target

Wed, 06 Oct 2021 14:13:18 +0000

We have been studying the infection mechanism of hepatitis B virus, HBV by using virus-like particles (VLPs) produced by yeasts. This tool enables us to study the receptor binding and intracellular trafficking of HBV without using infectious viral particles. Based on the infection mechanism of HBV, we’re now trying to identify a novel compound that can inhibit the infection of HBV into cells.

References

M. Somiya et al., “Intracellular trafficking of bio-nanocapsule-liposome complex: Identification of fusogenic activity in the pre-S1 region of hepatitis B virus surface antigen L protein.,” Journal of controlled release, vol. 212, pp. 10–8, 2015, doi: 10.1016/j.jconrel.2015.06.012.
Q. Liu et al., “Mutational analysis of hepatitis B virus pre-S1 (9-24) fusogenic peptide,” Biochemical and Biophysical Research Communications, vol. 474, no. 2, pp. 406–412, 2016, doi: 10.1016/j.bbrc.2016.04.125.
M. Somiya et al., “Cellular uptake of hepatitis B virus envelope L particles is independent of sodium taurocholate cotransporting polypeptide, but dependent on heparan sulfate proteoglycan,” Virology, vol. 497, pp. 23–32, 2016, doi: 10.1016/j.virol.2016.06.024.
Q. Liu, M. Somiya, M. Iijima, K. Tatematsu, and S. Kuroda, “A hepatitis B virus-derived human hepatic cell-specific heparin-binding peptide: identification and application to a drug delivery system,” Biomater. Sci., vol. 7, no. 1, pp. 322–335, 2019, doi: 10.1039/C8BM01134F.
M. Nanahara, Y.-T. Chang, M. Somiya, and S. Kuroda, “HBV Pre-S1-Derived Myristoylated Peptide (Myr47): Identification of the Inhibitory Activity on the Cellular Uptake of Lipid Nanoparticles,” Viruses, vol. 13, no. 5, 2021, doi: 10.3390/v13050929.
K. Takagi, M. Somiya, J. Jung, M. Iijima, and S. Kuroda, “Polymerized Albumin Receptor of Hepatitis B Virus for Evading the Reticuloendothelial System,” Pharmaceuticals, vol. 14, no. 5, 2021, doi: 10.3390/ph14050408.

Virus-inspired intracellular drug delivery

Wed, 06 Oct 2021 08:30:42 +0000

All drug has side effects, unfortunately. Side effects are often induced because drugs can exert biological activity against not only diseases cells but also non-target cells. So, what if we can deliver the drugs into specific cells or tissues without compromising healthy cells in the body? “Drug delivery system” may overcome the bad side of drugs.

Intracellular delivery is one of the hardest parts in the drug delivery; because the cell’s defense system, the cell membrane, has a stable structure and makes nanoparticles difficult to penetrate.

Viruses are sophisticated invaders that can penetrate the cell membrane and establish infection. The secrets of these tiny nanoparticles may be the hint to develop a novel type of drug delivery system to achieve efficient cytoplasmic drug delivery.

I have been studying the mechanisms of cellular entry of the hepatitis B virus. I’m particularly interested in membrane fusion proteins of viruses; these fascinating proteins enable viruses to penetrate the stubborn cell membrane.

Publications

M. Somiya et al., “Intracellular trafficking of bio-nanocapsule-liposome complex: Identification of fusogenic activity in the pre-S1 region of hepatitis B virus surface antigen L protein.,” Journal of controlled release, vol. 212, pp. 10–8, 2015, doi: 10.1016/j.jconrel.2015.06.012.
M. Somiya and S. Kuroda, “Development of a virus-mimicking nanocarrier for drug delivery systems: The bio-nanocapsule,” Advanced Drug Delivery Reviews, vol. 95, pp. 77–89, Dec. 2015, doi: 10.1016/j.addr.2015.10.003.
M. Somiya et al., “Cellular uptake of hepatitis B virus envelope L particles is independent of sodium taurocholate cotransporting polypeptide, but dependent on heparan sulfate proteoglycan,” Virology, vol. 497, pp. 23–32, 2016, doi: 10.1016/j.virol.2016.06.024.