Cellular Therapy Using Epitope-Imprinted Composite Nanoparticles to Remove α-Synuclein from an In Vitro Model

Mei-Hwa Lee; Jeng-Shiung Jan; James L Thomas; Yuan-Pin Shih; Jin-An Li; Chien-Yu Lin; Tooru Ooya; Lilla Barna; Mária Mészáros; András Harazin; Gergő Porkoláb; Szilvia Veszelka; Maria A Deli; Hung-Yin Lin

doi:10.3390/cells11162584

Cellular Therapy Using Epitope-Imprinted Composite Nanoparticles to Remove α-Synuclein from an In Vitro Model

Cells. 2022 Aug 19;11(16):2584. doi: 10.3390/cells11162584.

Authors

Mei-Hwa Lee¹, Jeng-Shiung Jan², James L Thomas³, Yuan-Pin Shih⁴, Jin-An Li⁴, Chien-Yu Lin⁴, Tooru Ooya^{5

6}, Lilla Barna^{7

8}, Mária Mészáros⁷, András Harazin⁷, Gergő Porkoláb^{7

8}, Szilvia Veszelka⁷, Maria A Deli⁷, Hung-Yin Lin⁴

Affiliations

¹ Department of Materials Science and Engineering, I-Shou University, Kaohsiung 84001, Taiwan.
² Department of Chemical Engineering, National Cheng Kung University, Tainan 70101, Taiwan.
³ Department of Physics and Astronomy, University of New Mexico, Albuquerque, NM 87131, USA.
⁴ Department of Chemical and Materials Engineering, National University of Kaohsiung, Kaohsiung 81148, Taiwan.
⁵ Graduate School of Engineering, Department of Chemical Science and Engineering, Kobe University, Kobe 657-8501, Japan.
⁶ Center for Advanced Medical Engineering Research & Development (CAMED), Kobe University, Kobe 657-8501, Japan.
⁷ Institute of Biophysics, Biological Research Centre, H-6726 Szeged, Hungary.
⁸ Doctoral School in Biology, University of Szeged, H-6720 Szeged, Hungary.

Abstract

Several degenerative disorders of the central nervous system, including Parkinson's disease (PD), are related to the pathological aggregation of proteins. Antibodies against toxic disease proteins, such as α-synuclein (SNCA), are therefore being developed as possible therapeutics. In this work, one peptide (YVGSKTKEGVVHGVA) from SNCA was used as the epitope to construct magnetic molecularly imprinted composite nanoparticles (MMIPs). These composite nanoparticles were characterized by dynamic light scattering (DLS), high-performance liquid chromatography (HPLC), isothermal titration calorimetry (ITC), Brunauer-Emmett-Teller (BET) analysis, and superconducting quantum interference device (SQUID) analysis. Finally, the viability of brain endothelial cells that were treated with MMIPs was measured, and the extraction of SNCA from CRISPR/dCas9a-activated HEK293T cells from the in vitro model system was demonstrated for the therapeutic application of MMIPs.

Keywords: gene activation; magnetic nanoparticles; peptide imprinting; protein extraction; α-synuclein.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endothelial Cells / metabolism
Epitopes
HEK293 Cells
Humans
Molecular Imprinting* / methods
Nanoparticles*
alpha-Synuclein / metabolism

Substances

Epitopes
alpha-Synuclein

Grants and funding

The authors would like to give their appreciation to the Ministry of Science and Technology of ROC under Contract nos. MOST 107-2923-M-006-002-MY3, MOST 107-2923-M-390-001-MY3, MOST 108-2221-E-006-034-MY3, MOST 108-2923-B-390-001-MY3, MOST 109-2221-E-214-028–and MOST 110-2221-E-390-003-MY3. This work was supported by a research grant of the National Research, Development and Innovation Office, Budapest, Hungary (NNE29617 to M.A.D.) in the frame of the nanoPD consortium, and partially supported by the Kobe University grant for promoting international joint research (to TO). M.M. was supported by the research grant of the National Research, Development and Innovation Office, Budapest, Hungary (PD138930) and the Gedeon Richter Plc Centenarial Foundation (H-1103 Budapest, Gyömrői str. 19-21. Hungary). S.V. was supported by the Premium Postdoctoral Research Program (Premium-2019-469) of the Hungarian Academy of Sciences. G.P. was supported by the Szeged Scientists Academy under the sponsorship of the Hungarian Ministry of Innovation and Technology (FEIF/646-4/2021-ITM_SZERZ), as well as by the ÚNKP-21-3-405 New National Excellence Program of the Ministry for Innovation and Technology from the source of the National Research, Development and Innovation Fund.