Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation

Abstract Clinical success of effective gene therapy is mainly hampered by the insufficiency of safe and efficient internalization of a transgene to the targeted cellular site. Therefore, the development of a safe and efficient nanocarrier system is one of the fundamental challenges to transfer the t...

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Autores principales:	Imran Tariq, Muhammad Yasir Ali, Muhammad Farhan Sohail, Muhammad Umair Amin, Sajid Ali, Nadeem Irfan Bukhari, Abida Raza, Shashank Reddy Pinnapireddy, Jens Schäfer, Udo Bakowsky
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dccf07d5182c4bb7a2141c37785af9f8
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spelling	oai:doaj.org-article:dccf07d5182c4bb7a2141c37785af9f82021-12-02T11:43:58ZLipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation10.1038/s41598-020-78123-62045-2322https://doaj.org/article/dccf07d5182c4bb7a2141c37785af9f82020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78123-6https://doaj.org/toc/2045-2322Abstract Clinical success of effective gene therapy is mainly hampered by the insufficiency of safe and efficient internalization of a transgene to the targeted cellular site. Therefore, the development of a safe and efficient nanocarrier system is one of the fundamental challenges to transfer the therapeutic genes to the diseased cells. Polyamidoamine (PAMAM) dendrimer has been used as an efficient non-viral gene vector (dendriplexes) but the toxicity and unusual biodistribution induced by the terminal amino groups (–NH2) limit its in vivo applications. Hence, a state of the art lipid modification with PAMAM based gene carrier (lipodendriplexes) was planned to investigate theirs in vitro (2D and 3D cell culture) and in vivo behaviour. In vitro pDNA transfection, lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) generation, cellular protein contents, live/dead staining and apoptosis were studied in 2D cell culture of HEK-293 cells while GFP transfection, 3D cell viability and live/dead staining of spheroids were performed in its 3D cell culture. Acute toxicity studies including organ to body index ratio, hematological parameters, serum biochemistry, histopathological profiles and in vivo transgene expression were assessed in female BALB/c mice. The results suggested that, in comparison to dendriplexes the lipodendriplexes exhibited significant improvement of pDNA transfection (p < 0.001) with lower LDH release (p < 0.01) and ROS generation (p < 0.05). A substantially higher cellular protein content (p < 0.01) and cell viability were also observed in 2D culture. A strong GFP expression with an improved cell viability profile (p < 0.05) was indicated in lipodendriplexes treated 3D spheroids. In vivo archives showed the superiority of lipid-modified nanocarrier system, depicted a significant increase in green fluorescent protein (GFP) expression in the lungs (p < 0.01), heart (p < 0.001), liver (p < 0.001) and kidneys (p < 0.001) with improved serum biochemistry and hematological profile as compared to unmodified dendriplexes. No tissue necrosis was evident in the animal groups treated with lipid-shielded molecules. Therefore, a non-covalent conjugation of lipids with PAMAM based carrier system could be considered as a promising approach for an efficient and biocompatible gene delivery system.Imran TariqMuhammad Yasir AliMuhammad Farhan SohailMuhammad Umair AminSajid AliNadeem Irfan BukhariAbida RazaShashank Reddy PinnapireddyJens SchäferUdo BakowskyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-15 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Imran Tariq Muhammad Yasir Ali Muhammad Farhan Sohail Muhammad Umair Amin Sajid Ali Nadeem Irfan Bukhari Abida Raza Shashank Reddy Pinnapireddy Jens Schäfer Udo Bakowsky Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
description	Abstract Clinical success of effective gene therapy is mainly hampered by the insufficiency of safe and efficient internalization of a transgene to the targeted cellular site. Therefore, the development of a safe and efficient nanocarrier system is one of the fundamental challenges to transfer the therapeutic genes to the diseased cells. Polyamidoamine (PAMAM) dendrimer has been used as an efficient non-viral gene vector (dendriplexes) but the toxicity and unusual biodistribution induced by the terminal amino groups (–NH2) limit its in vivo applications. Hence, a state of the art lipid modification with PAMAM based gene carrier (lipodendriplexes) was planned to investigate theirs in vitro (2D and 3D cell culture) and in vivo behaviour. In vitro pDNA transfection, lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) generation, cellular protein contents, live/dead staining and apoptosis were studied in 2D cell culture of HEK-293 cells while GFP transfection, 3D cell viability and live/dead staining of spheroids were performed in its 3D cell culture. Acute toxicity studies including organ to body index ratio, hematological parameters, serum biochemistry, histopathological profiles and in vivo transgene expression were assessed in female BALB/c mice. The results suggested that, in comparison to dendriplexes the lipodendriplexes exhibited significant improvement of pDNA transfection (p < 0.001) with lower LDH release (p < 0.01) and ROS generation (p < 0.05). A substantially higher cellular protein content (p < 0.01) and cell viability were also observed in 2D culture. A strong GFP expression with an improved cell viability profile (p < 0.05) was indicated in lipodendriplexes treated 3D spheroids. In vivo archives showed the superiority of lipid-modified nanocarrier system, depicted a significant increase in green fluorescent protein (GFP) expression in the lungs (p < 0.01), heart (p < 0.001), liver (p < 0.001) and kidneys (p < 0.001) with improved serum biochemistry and hematological profile as compared to unmodified dendriplexes. No tissue necrosis was evident in the animal groups treated with lipid-shielded molecules. Therefore, a non-covalent conjugation of lipids with PAMAM based carrier system could be considered as a promising approach for an efficient and biocompatible gene delivery system.
format	article
author	Imran Tariq Muhammad Yasir Ali Muhammad Farhan Sohail Muhammad Umair Amin Sajid Ali Nadeem Irfan Bukhari Abida Raza Shashank Reddy Pinnapireddy Jens Schäfer Udo Bakowsky
author_facet	Imran Tariq Muhammad Yasir Ali Muhammad Farhan Sohail Muhammad Umair Amin Sajid Ali Nadeem Irfan Bukhari Abida Raza Shashank Reddy Pinnapireddy Jens Schäfer Udo Bakowsky
author_sort	Imran Tariq
title	Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
title_short	Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
title_full	Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
title_fullStr	Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
title_full_unstemmed	Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
title_sort	lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/dccf07d5182c4bb7a2141c37785af9f8
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Lipodendriplexes mediated enhanced gene delivery: a cellular to pre-clinical investigation

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