Properties of Parallel Tetramolecular G-Quadruplex Carrying N-Acetylgalactosamine as Potential Enhancer for Oligonucleotide Delivery to Hepatocytes
| العنوان: | Properties of Parallel Tetramolecular G-Quadruplex Carrying N-Acetylgalactosamine as Potential Enhancer for Oligonucleotide Delivery to Hepatocytes |
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| المؤلفون: | Anna Clua, Santiago Grijalvo, Namrata Erande, Swati Gupta, Kristina Yucius, Raimundo Gargallo, Stefania Mazzini, Muthiah Manoharan, Ramon Eritja |
| المصدر: | Molecules; Volume 27; Issue 12; Pages: 3944 Dipòsit Digital de la UB Universidad de Barcelona |
| بيانات النشر: | Multidisciplinary Digital Publishing Institute, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | G-quadruplex, N-acetylgalactosamine, Organic Chemistry, ADN, Oligonucleotides, Pharmaceutical Science, Oligonucleòtids, Gapmers, DNA, Luciferase gene, Molecules, Asialoglycoprotein receptor, Analytical Chemistry, antisense, oligonucleotide conjugates, asialoglycoprotein receptor, luciferase gene, gapmers, Oligonucleotide conjugates, Chemistry (miscellaneous), Drug Discovery, Molecular Medicine, Antisense, Physical and Theoretical Chemistry, Molècules, Ensure healthy lives and promote well-being for all at all ages |
| الوصف: | The development of oligonucleotide conjugates for in vivo targeting is one of the most exciting areas for oligonucleotide therapeutics. A major breakthrough in this field was the development of multifunctional GalNAc-oligonucleotides with high affinity to asialoglycoprotein receptors (ASGPR) that directed therapeutic oligonucleotides to hepatocytes. In the present study, we explore the use of G-rich sequences functionalized with one unit of GalNAc at the 3'-end for the formation of tetrameric GalNAc nanostructures upon formation of a parallel G-quadruplex. These compounds are expected to facilitate the synthetic protocols by providing the multifunctionality needed for the binding to ASGPR. To this end, several G-rich oligonucleotides carrying a TGGGGGGT sequence at the 3'-end functionalized with one molecule of N-acetylgalactosamine (GalNAc) were synthesized together with appropriate control sequences. The formation of a self-assembled parallel G-quadruplex was confirmed through various biophysical techniques such as circular dichroism, nuclear magnetic resonance, polyacrylamide electrophoresis and denaturation curves. Binding experiments to ASGPR show that the size and the relative position of the therapeutic cargo are critical for the binding of these nanostructures. The biological properties of the resulting parallel G-quadruplex were evaluated demonstrating the absence of the toxicity in cell lines. The internalization preferences of GalNAc-quadruplexes to hepatic cells were also demonstrated as well as the enhancement of the luciferase inhibition using the luciferase assay in HepG2 cell lines versus HeLa cells. All together, we demonstrate that tetramerization of G-rich oligonucleotide is a novel and simple route to obtain the beneficial effects of multivalent N-acetylgalactosamine functionalization. This research was funded by Spanish Ministerio de Ciencia e Innovación (MICINN) (Projects CTQ2017-84415-R, PID2019-107158GB-I100, PID2020-118145RB-I100), CIBER-BBN grant number CB06/01/0019, PIANO DI SOSTEGNO ALLA RICERCA 2020—Linea 2 azione B (DEFENS) and a predoctoral contract grant (PRE2018-084056) to A.C. CIBER-BBN is an initiative funded by the VI National R + D + I Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development. The APC was funded by MICINN, project PID2020-118145RB-I100. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1420-3049 2017-8441 |
| DOI: | 10.3390/molecules27123944 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9ebebdb249dd1239fb958291817c0271 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....9ebebdb249dd1239fb958291817c0271 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14203049 20178441 |
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| DOI: | 10.3390/molecules27123944 |