DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells

β-amyloid (Aβ) plays an essential role in the pathogenesis of Alzheimer’s disease (AD). Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) is indispensable for Aβ production, and knockout of BACE1 has no overt phenotypes in mouse. Thus, fine modulation of BACE1 may be a safe and effective...

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Main Authors:	Jun Xiang, Wen Zhang, Xiao-Fang Cai, Min Cai, Zhong-Hai Yu, Feng Yang, Wen Zhu, Xiang-Ting Li, Ting Wu, Jing-Si Zhang, Ding-Fang Cai
Format:	Article
Language:	English
Published:	Elsevier 2019-06-01
Series:	Molecular Therapy: Nucleic Acids
Online Access:	http://www.sciencedirect.com/science/article/pii/S2162253119300563

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collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Jun Xiang Wen Zhang Xiao-Fang Cai Min Cai Zhong-Hai Yu Feng Yang Wen Zhu Xiang-Ting Li Ting Wu Jing-Si Zhang Ding-Fang Cai
spellingShingle	Jun Xiang Wen Zhang Xiao-Fang Cai Min Cai Zhong-Hai Yu Feng Yang Wen Zhu Xiang-Ting Li Ting Wu Jing-Si Zhang Ding-Fang Cai DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells Molecular Therapy: Nucleic Acids
author_facet	Jun Xiang Wen Zhang Xiao-Fang Cai Min Cai Zhong-Hai Yu Feng Yang Wen Zhu Xiang-Ting Li Ting Wu Jing-Si Zhang Ding-Fang Cai
author_sort	Jun Xiang
title	DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells
title_short	DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells
title_full	DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells
title_fullStr	DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells
title_full_unstemmed	DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells
title_sort	dna aptamers targeting bace1 reduce amyloid levels and rescue neuronal deficiency in cultured cells
publisher	Elsevier
series	Molecular Therapy: Nucleic Acids
issn	2162-2531
publishDate	2019-06-01
description	β-amyloid (Aβ) plays an essential role in the pathogenesis of Alzheimer’s disease (AD). Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) is indispensable for Aβ production, and knockout of BACE1 has no overt phenotypes in mouse. Thus, fine modulation of BACE1 may be a safe and effective treatment for AD patients. However, the large active site of BACE1 makes it challenging to target BACE1 with classical small-molecule inhibitors. DNA aptamer can have high affinity and specificity against diverse targets, and it provides an alternative strategy to target BACE1. In this study, we used a novel cell-systematic evolution of ligands by exponential enrichment (SELEX) strategy to select specific DNA aptamers optimized to target BACE1 under physiological status. After 17 rounds of selection, we identified two DNA aptamers against BACE1: BI1 and BI2. The identified aptamers interacted with BACE1 in pull-down assay, inhibited BACE1 activity in in vitro fluorescence resonance energy transfer (FRET) assay and HEK293-APP stable cell line, reduced Aβ in the culture medium of HEK293-amyloid protein precursor (APP) stable cell line and APP-PS1 primary cultured neurons, and rescued Aβ-induced neuronal deficiency in APP-PS1 primary cultured neurons. In contrast, the identified aptamers had no effect on α- or γ-secretase. In addition, cholesteryl tetraetylene glycol (TEG) modification further improved the potency of the identified aptamers. Our study suggests that it is feasible and effective to target BACE1 with DNA aptamers, and the therapeutic potential of the identified aptamers deserves further investigation. Keywords: Alzheimer’s disease, amyloid, BACE1, aptamer, SELEX
url	http://www.sciencedirect.com/science/article/pii/S2162253119300563
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spelling	doaj-ddf2458ef5e0427794009065bf25b8a62020-11-24T21:43:37ZengElsevierMolecular Therapy: Nucleic Acids2162-25312019-06-0116302312DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured CellsJun Xiang0Wen Zhang1Xiao-Fang Cai2Min Cai3Zhong-Hai Yu4Feng Yang5Wen Zhu6Xiang-Ting Li7Ting Wu8Jing-Si Zhang9Ding-Fang Cai10Department of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Stomatology, Zhongshan Hospital, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, ChinaDepartment of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, China; Corresponding author: Jing-Si Zhang, Department of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China.Department of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Laboratory of Neurology, Institute of Integrative Medicine, Fudan University, Shanghai 200032, China; Corresponding author: Ding-Fang Cai, Department of Integrative Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China.β-amyloid (Aβ) plays an essential role in the pathogenesis of Alzheimer’s disease (AD). Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) is indispensable for Aβ production, and knockout of BACE1 has no overt phenotypes in mouse. Thus, fine modulation of BACE1 may be a safe and effective treatment for AD patients. However, the large active site of BACE1 makes it challenging to target BACE1 with classical small-molecule inhibitors. DNA aptamer can have high affinity and specificity against diverse targets, and it provides an alternative strategy to target BACE1. In this study, we used a novel cell-systematic evolution of ligands by exponential enrichment (SELEX) strategy to select specific DNA aptamers optimized to target BACE1 under physiological status. After 17 rounds of selection, we identified two DNA aptamers against BACE1: BI1 and BI2. The identified aptamers interacted with BACE1 in pull-down assay, inhibited BACE1 activity in in vitro fluorescence resonance energy transfer (FRET) assay and HEK293-APP stable cell line, reduced Aβ in the culture medium of HEK293-amyloid protein precursor (APP) stable cell line and APP-PS1 primary cultured neurons, and rescued Aβ-induced neuronal deficiency in APP-PS1 primary cultured neurons. In contrast, the identified aptamers had no effect on α- or γ-secretase. In addition, cholesteryl tetraetylene glycol (TEG) modification further improved the potency of the identified aptamers. Our study suggests that it is feasible and effective to target BACE1 with DNA aptamers, and the therapeutic potential of the identified aptamers deserves further investigation. Keywords: Alzheimer’s disease, amyloid, BACE1, aptamer, SELEXhttp://www.sciencedirect.com/science/article/pii/S2162253119300563

DNA Aptamers Targeting BACE1 Reduce Amyloid Levels and Rescue Neuronal Deficiency in Cultured Cells

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