Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing

RNA interference (RNAi)-mediated gene silencing is a promising therapeutic approach to treat various diseases, but safe and efficient delivery remains a major challenge to its clinical application. Non-viral gene vectors, such as poly(β-amino esters) (pBAEs), have emerged as a potential candidate du...

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Main Authors: Ooi, Ying Jie, Huang, Chongquan, Lau, Kieran, Chew, Sing Yian, Park, Jong Gu, Chan-Park, Mary B.
Other Authors: School of Chemistry, Chemical Engineering and Biotechnology
Format: Article
Language:English
Published: 2024
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Online Access:https://hdl.handle.net/10356/176048
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-1760482024-05-13T02:45:45Z Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing Ooi, Ying Jie Huang, Chongquan Lau, Kieran Chew, Sing Yian Park, Jong Gu Chan-Park, Mary B. School of Chemistry, Chemical Engineering and Biotechnology Lee Kong Chian School of Medicine (LKCMedicine) School of Materials Science and Engineering Interdisciplinary Graduate School (IGS) Medicine, Health and Life Sciences Gene silencing Primary neural cells RNA interference (RNAi)-mediated gene silencing is a promising therapeutic approach to treat various diseases, but safe and efficient delivery remains a major challenge to its clinical application. Non-viral gene vectors, such as poly(β-amino esters) (pBAEs), have emerged as a potential candidate due to their biodegradability, low toxicity profile, ease of synthesis, and high gene transfection efficiency for both DNA and siRNA delivery. However, achieving significant gene silencing using pBAEs often requires a large amount of polymer carrier (with polymer/siRNA weight ratio >100) or high siRNA dose (>100 nM), which might potentially exacerbate toxicity concerns during delivery. To overcome these barriers, we designed and optimized a series of hyperbranched pBAEs capable of efficiently condensing siRNA and achieving excellent silencing efficiency at a lower polymer/siRNA weight ratio (w/w) and siRNA dose. Through modulation of monomer combinations and branching density, we identified the top-performing hyperbranched pBAEs, named as h(A2B3)-1, which possess good siRNA condensation ability, low cytotoxicity, and high cellular uptake efficiency. Compared with Lipofectamine 2000, h(A2B3)-1 achieved lower cytotoxicity and higher siRNA silencing efficiency in HeLa cells at a polymer/siRNA weight ratio of 30 and 30 nM siRNA dose. Notably, h(A2B3)-1 enhanced the gene uptake in primary neural cells and effectively silenced the target gene in hard-to-transfect primary cortical neurons and oligodendrocyte progenitor cells, with gene knockdown efficiencies of 34.8 and 53.4% respectively. By incorporating a bioreducible disulfide compartment into the polymer backbone, the cytocompatibility of the h(A2B3)-1 was greatly enhanced while maintaining their good transfection efficiency. Together, the low cytotoxicity and high siRNA transfection efficiency of hyperbranched h(A2B3)-1 in this study demonstrated their great potential as a non-viral gene vector for efficient siRNA delivery and RNAi-mediated gene silencing. This provides valuable insight into the future development of safe and efficient non-viral siRNA delivery systems as well as their translation into clinical applications. Ministry of Education (MOE) This work was financially supported by Welgene and Clearlab (REQ0149704). Partial funding support from MOE Tier 1Grant (RG37/20) is also acknowledged. 2024-05-13T02:45:45Z 2024-05-13T02:45:45Z 2024 Journal Article Ooi, Y. J., Huang, C., Lau, K., Chew, S. Y., Park, J. G. & Chan-Park, M. B. (2024). Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing. ACS Applied Materials and Interfaces, 16(11), 14093-14112. https://dx.doi.org/10.1021/acsami.3c10620 1944-8244 https://hdl.handle.net/10356/176048 10.1021/acsami.3c10620 38449351 2-s2.0-85187156401 11 16 14093 14112 en RG37/20 ACS Applied Materials and Interfaces © 2024 American Chemical Society. All rights reserved.
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Medicine, Health and Life Sciences
Gene silencing
Primary neural cells
spellingShingle Medicine, Health and Life Sciences
Gene silencing
Primary neural cells
Ooi, Ying Jie
Huang, Chongquan
Lau, Kieran
Chew, Sing Yian
Park, Jong Gu
Chan-Park, Mary B.
Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
description RNA interference (RNAi)-mediated gene silencing is a promising therapeutic approach to treat various diseases, but safe and efficient delivery remains a major challenge to its clinical application. Non-viral gene vectors, such as poly(β-amino esters) (pBAEs), have emerged as a potential candidate due to their biodegradability, low toxicity profile, ease of synthesis, and high gene transfection efficiency for both DNA and siRNA delivery. However, achieving significant gene silencing using pBAEs often requires a large amount of polymer carrier (with polymer/siRNA weight ratio >100) or high siRNA dose (>100 nM), which might potentially exacerbate toxicity concerns during delivery. To overcome these barriers, we designed and optimized a series of hyperbranched pBAEs capable of efficiently condensing siRNA and achieving excellent silencing efficiency at a lower polymer/siRNA weight ratio (w/w) and siRNA dose. Through modulation of monomer combinations and branching density, we identified the top-performing hyperbranched pBAEs, named as h(A2B3)-1, which possess good siRNA condensation ability, low cytotoxicity, and high cellular uptake efficiency. Compared with Lipofectamine 2000, h(A2B3)-1 achieved lower cytotoxicity and higher siRNA silencing efficiency in HeLa cells at a polymer/siRNA weight ratio of 30 and 30 nM siRNA dose. Notably, h(A2B3)-1 enhanced the gene uptake in primary neural cells and effectively silenced the target gene in hard-to-transfect primary cortical neurons and oligodendrocyte progenitor cells, with gene knockdown efficiencies of 34.8 and 53.4% respectively. By incorporating a bioreducible disulfide compartment into the polymer backbone, the cytocompatibility of the h(A2B3)-1 was greatly enhanced while maintaining their good transfection efficiency. Together, the low cytotoxicity and high siRNA transfection efficiency of hyperbranched h(A2B3)-1 in this study demonstrated their great potential as a non-viral gene vector for efficient siRNA delivery and RNAi-mediated gene silencing. This provides valuable insight into the future development of safe and efficient non-viral siRNA delivery systems as well as their translation into clinical applications.
author2 School of Chemistry, Chemical Engineering and Biotechnology
author_facet School of Chemistry, Chemical Engineering and Biotechnology
Ooi, Ying Jie
Huang, Chongquan
Lau, Kieran
Chew, Sing Yian
Park, Jong Gu
Chan-Park, Mary B.
format Article
author Ooi, Ying Jie
Huang, Chongquan
Lau, Kieran
Chew, Sing Yian
Park, Jong Gu
Chan-Park, Mary B.
author_sort Ooi, Ying Jie
title Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
title_short Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
title_full Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
title_fullStr Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
title_full_unstemmed Nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient siRNA delivery and gene silencing
title_sort nontoxic, biodegradable hyperbranched poly(β-amino ester)s for efficient sirna delivery and gene silencing
publishDate 2024
url https://hdl.handle.net/10356/176048
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