-
Folate Receptor-Mediated Small Interfering RNA Delivery: Recent Developments and Future Directions for RNA Interference Therapeutics Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-15 Sumit Gangopadhyay; Rahul R. Nikam; Kiran R. Gore
RNA interference (RNAi), a gene regulatory process mediated by small interfering RNAs (siRNAs), has made remarkable progress as a potential therapeutic agent against various diseases. However, RNAi is associated with fundamental challenges such as poor systemic delivery and susceptibility to the nucleases. Targeting ligand-bound delivery vehicles has improved the accumulation of drug at the target
-
Outcomes of the European Federation of Pharmaceutical Industries and Associations Oligonucleotide Working Group Survey on Nonclinical Practices and Regulatory Expectations for Therapeutic Oligonucleotide Safety Assessment Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Yann Tessier; William Achanzar; Lauren Mihalcik; Chidozie Amuzie; Patrik Andersson; Joel D. Parry; Jonathan Moggs; Laurence O. Whiteley
The Oligonucleotide Working Group of the European Federation of Pharmaceutical Industries and Associations (EFPIA) conducted a survey of companies to understand the trends in nonclinical practices and regulatory expectations for oligonucleotide drug safety assessment. Twenty-two companies of different types, with varying oligonucleotide experience levels in the field, participated. The survey identified
-
To Conjugate or to Package? A Look at Targeted siRNA Delivery Through Folate Receptors Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Lidya Salim; Jean-Paul Desaulniers
RNA interference (RNAi) applications have evolved from experimental tools to study gene function to the development of a novel class of gene-silencing therapeutics. Despite decades of research, it was not until August 2018 that the US FDA approved the first-ever RNAi drug, marking a new era for RNAi therapeutics. Although there are many limitations associated with the inherent structure of RNA, delivery
-
Antisense Oligonucleotides as Potential Therapeutics for Type 2 Diabetes Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Suxiang Chen; Nabayet Sbuh; Rakesh N. Veedu
Type 2 diabetes (T2D) is a chronic metabolic disorder characterized by persistent hyperglycemia resulting from inefficient signaling and insufficient production of insulin. Conventional management of T2D has largely relied on small molecule-based oral hypoglycemic medicines, which do not halt the progression of the disease due to limited efficacy and induce adverse effects as well. To this end, antisense
-
Short-Chain Guide RNA for Site-Directed A-to-I RNA Editing Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Kanako Nose; Kota Hidaka; Takashi Yano; Yohei Tomita; Masatora Fukuda
Site-directed RNA editing is a promising genetic modification technology for therapeutic and pharmaceutical applications. We previously constructed adenosine deaminases acting on RNA (ADAR)-guiding RNAs (AD-gRNAs) that direct A-to-I RNA editing activity of native human ADAR2 into a programmable target site. In this study, we developed the short-chain AD-gRNA (shAD-gRNA) as a potential basic framework
-
Human Papillomavirus G-Rich Regions as Potential Antiviral Drug Targets Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Josué Carvalho; Jéssica Lopes-Nunes; Maria Paula Cabral Campello; António Paulo; Janice Milici; Craig Meyers; Jean-Louis Mergny; Gilmar F. Salgado; João A. Queiroz; Carla Cruz
Herein, we report, for the first time, the screening of several ligands in terms of their ability to bind and stabilize G-quadruplexes (G4) found in seven human Papillomavirus (HPV) genomes. Using a variety of biophysical assays, HPV G-quadruplexes were shown to possess a high degree of structural polymorphism upon ligand binding, which may have an impact on transcription, replication, and viral protein
-
The Performance of Minicircle DNA Versus Parental Plasmid in p53 Gene Delivery Into HPV-18-Infected Cervical Cancer Cells Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-11 Dalinda Eusébio; Ana Margarida Almeida; Joel Marques Alves; Cláudio Jorge Maia; João António Queiroz; Fani Sousa; Ângela Sousa
Minicircle DNA (mcDNA) has been suggested as a vanguard technology for gene therapy, consisting of a nonviral DNA vector devoid of prokaryotic sequences. Unlike conventional plasmid DNA (pDNA), this small vector is able to sustain high expression rates throughout time. Thus, this work describes the construction, production, and purification of mcDNA-p53 and its precursor parental plasmid (PP)-p53 for
-
Splice-Switching Antisense Oligonucleotides as a Targeted Intrinsic Engineering Tool for Generating Armored Redirected T Cells Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-09 Erica Ceccarello; Tommaso Tabaglio; Sarene Koh; Vincent Oei; Winnie Teo; Owen Julianto Jonathan; Andrea Pavesi; Qingfeng Chen; Antonio Bertoletti; Keng Boon Wee; Ernesto Guccione
Modification of specificity of T cells for the use in adoptive transfer (CAR- or TCR-redirected T cells) has revolutionized the therapy of liquid tumors and some infectious diseases. However, several obstacles are still hampering the efficacy of such potent therapy, hence concurrent modification of the function is also required to obtain successful results. Here we show the use of splice-switching
-
Immortalized Canine Dystrophic Myoblast Cell Lines for Development of Peptide-Conjugated Splice-Switching Oligonucleotides Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-09 Yuichiro Tone; Kamel Mamchaoui; Maria K. Tsoumpra; Yasumasa Hashimoto; Reiko Terada; Rika Maruyama; Michael J. Gait; Andrey A. Arzumanov; Graham McClorey; Michihiro Imamura; Shin'ichi Takeda; Toshifumi Yokota; Matthew J.A. Wood; Vincent Mouly; Yoshitsugu Aoki
Duchenne muscular dystrophy (DMD) is a severe muscle-wasting disease caused by frameshift or nonsense mutations in the DMD gene, resulting in the loss of dystrophin from muscle membranes. Exon skipping using splice-switching oligonucleotides (SSOs) restores the reading frame of DMD pre-mRNA by generating internally truncated but functional dystrophin protein. To potentiate effective tissue-specific
-
Hsc70 Facilitates Mannose-6-Phosphate Receptor-Mediated Intracellular Trafficking and Enhances Endosomal Release of Phosphorothioate-Modified Antisense Oligonucleotides Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-09 Xue-Hai Liang; Joshua G. Nichols; Chih-Wei Hsu; Stanley T. Crooke
Phosphorothioate-modified antisense oligonucleotide (PS-ASO) drugs are commonly used to modulate gene expression through RNase H1-mediated cleavage of target RNAs. Upon internalization through endocytic pathways into cells, PS-ASOs must be released from membraned endosomal organelles to act on target RNAs, a limiting step of PS-ASO activity. Here we report that Hsc70 protein mediates productive release
-
Perspectives on the Designation of Oligonucleotide Starting Materials Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-02 William F. Kiesman; Andrew K. McPherson; Louis J. Diorazio; Leo Van den Bergh; Peter D. Smith; John M. Northall; Alec Fettes; Tiejun Wang; Martin Mehlmann; Syed Raza; Gary Held
The designation of starting materials (SMs) for pharmaceuticals has been a topic of great interest and debate since the first ICH quality guidance was published. The increase in the number and variety of commercialized oligonucleotides (antisense oligonucleotides—ASOs, small interfering RNAs—siRNAs, etc.) in recent years has reignited dialogue on this topic because of the unique complexity of the monomeric
-
Solid-Phase Separation of Toxic Phosphorothioate Antisense Oligonucleotide-Protein Nucleolar Aggregates Is Cytoprotective Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-02-02 Xue-hai Liang; Cheryl Li De Hoyos; Wen Shen; Lingdi Zhang; Michael Fazio; Stanley T. Crooke
Phosphorothioate antisense oligonucleotides (PS-ASOs) interact with proteins and can localize to or induce the formation of a variety of subcellular PS-ASO-protein or PS-ASO-ribonucleoprotein aggregates. In this study, we show that these different aggregates that form with varying compositions at various concentrations in the cytosol, nucleus, and nucleolus may undergo phase separations in cells. Some
-
Considerations of the Japanese Research Working Group for the ICH S6 & Related Issues Regarding Nonclinical Safety Assessments of Oligonucleotide Therapeutics: Comparison with Those of Biopharmaceuticals Nucleic Acid Ther. (IF 4.875) Pub Date : 2021-01-19 Yoko Hirabayashi; Kazushige Maki; Kiyoshi Kinoshita; Takahiro Nakazawa; Satoshi Obika; Misaki Naota; Kazuto Watanabe; Mutsumi Suzuki; Teruyo Arato; Aki Fujisaka; Osamu Fueki; Kosuke Ito; Hiroshi Onodera
This white paper summarizes the current consensus of the Japanese Research Working Group for the ICH S6 & Related Issues (WGS6) on strategies for the nonclinical safety assessment of oligonucleotide-based therapeutics (ONTs), specifically focused on the similarities and differences to biotechnology-derived pharmaceuticals (biopharmaceuticals). ONTs, like biopharmaceuticals, have high species and target
-
Oligonucleotide IMT504 Improves Glucose Metabolism and Controls Immune Cell Mediators in Female Diabetic NOD Mice IMT504, a Potential Therapy for Type 1 Diabetes Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-21 Stefania Bianchi; Verónica C. Martínez Allo; Milena Massimino; María del R. Lavignolle Heguy; Francisco R. Borzone; Sofía Gomez Bustillo; Norma A. Chasseing; Carlos Libertun; Alejandro D. Montaner; Gabriel A. Rabinovich; Marta A. Toscano; Victoria A. Lux-Lantos; María S. Bianchi
Type 1 diabetes occurs as a consequence of progressive autoimmune destruction of beta cells. A potential treatment for this disease should address the immune attack on beta cells and their preservation/regeneration. The objective of this study was to elucidate whether the immunomodulatory synthetic oligonucleotide IMT504 was able to ameliorate diabetes in NOD mice and to provide further understanding
-
Efficiency and Target Derepression of Anti-miR-92a: Results of a First in Human Study. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Wesley Tyler Abplanalp,Ariane Fischer,David John,Andreas M Zeiher,Willy Gosgnach,Helene Darville,Rusty Montgomery,Linda Pestano,Guillaume Allée,Isabelle Paty,Francoise Fougerousse,Stefanie Dimmeler
MicroRNA (miRNA) inhibition is a promising therapeutic strategy in several disease indications. MRG-110 is a locked nucleic acid-based antisense oligonucleotide that targets miR-92a-3p and experimentally was shown to have documented therapeutic effects on cardiovascular disease and wound healing. To gain first insights into the activity of anti-miR-92a in humans, we investigated miR-92a-3p expression
-
Development of Prodrug Type Circular siRNA for In Vivo Knockdown by Systemic Administration Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Kenji Hagiwara; Masakazu Honma; Toshimasa Harumoto; Kenji Harada; Takashi Sawada; Junichiro Yamamoto; Fumikazu Shinohara
siRNAs are being developed as a novel therapeutic modality; however, problems impeding their application in extrahepatic tissues persist, including inadequate stability in biological environments and inefficient drug delivery system to target tissues. Thus, technological improvements that enable robust silencing of target messenger RNA (mRNA) in extrahepatic tissues are necessary. We developed prodrug
-
Development of Human CBF1-Targeting Single-Stranded DNA Aptamers with Antiangiogenic Activity In Vitro. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Mari Tezuka-Kagajo,Masashi Maekawa,Atsushi Ogawa,Yoshiko Hatta,Eiichi Ishii,Mariko Eguchi,Shigeki Higashiyama
C promoter binding factor 1 (CBF1) (alias RBPJ) is a critical transcription factor involved in Notch signaling. The activation of Notch signaling through CBF1 maintains the angiostatic state of endothelial cells suppressing angiogenesis, that is, the formation of new blood vessels. Vascular endothelial growth factor (VEGF) induces angiogenesis by promoting the proteasomal degradation of CBF1, in addition
-
DNA-Encoded Glutamine Synthetase Enzyme as Ammonia-Lowering Therapeutic for Hyperammonemia. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Makan Khoshnejad,Yaya Dia,Ami Patel,Ziyang Xu,Xizhou Zhu,Kun Yun,Krzysztof Wojtak,Rehman Qureshi,Laurent Humeau,Kar Muthumani,David B Weiner
Hyperammonemia is a dangerous life-threatening metabolic complication characterized by markedly elevated ammonia levels that can lead to irreversible brain damage if not carefully monitored. Current pharmacological treatment strategies available for hyperammonemia patients are suboptimal and associated with major side effects. In this study, we focus on developing and evaluating the in vivo delivery
-
Delivery of Antisense DNA into Pathogenic Parasite Trypanosoma cruzi Using Virus-Like Protein-Based Nanoparticles. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Rosa E Cárdenas-Guerra,David S Moreno-Gutierrez,Oscar de J Vargas-Dorantes,Bertha Espinoza,Armando Hernandez-Garcia
Trypanosoma cruzi, which causes Chagas disease, is one of the most lacerating parasites in terms of health and social impacts. New approaches for its study and treatment are urgently needed since in more than 50 years only two drugs have been approved. Genetic approaches based on antisense oligonucleotides (AONs) are promising; however, to harness their full potential the development of effective carriers
-
A Novel DNA Aptamer Targeting S100P Induces Antitumor Effects in Colorectal Cancer Cells Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Wenjing Sun; Lifang Luo; Daoquan Fang; Tianbin Tang; Wuhua Ni; Bichun Dai; Hongguang Sun; Lei Jiang
Colorectal cancer (CRC) is a prevalent malignancy with poor prognosis and survival. As a Ca2+ binding protein, S100P plays a role in calcium-dependent signal transduction pathways that involve in diverse biological processes. Our previous studies have shown that S100P is overexpressed in CRC tissues and regulates cell growth, invasion, and metastasis in CRC. Therefore, S100P is expected to be an effective
-
Improved Anti-Prion Nucleic Acid Aptamers by Incorporation of Chemical Modifications Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-12-04 Jussara Amato; Tsukasa Mashima; Yuji O. Kamatari; Kazuo Kuwata; Ettore Novellino; Antonio Randazzo; Concetta Giancola; Masato Katahira; Bruno Pagano
Nucleic acid aptamers are innovative and promising candidates to block the hallmark event in the prion diseases, that is the conversion of prion protein (PrP) into an abnormal form; however, they need chemical modifications for effective therapeutic activity. This communication reports on the development and biophysical characterization of a small library of chemically modified G-quadruplex-forming
-
Strategies for Identity Testing of Therapeutic Oligonucleotide Drug Substances and Drug Products. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Daniel Capaldi,Nadim Akhtar,Tom Atherton,David Benstead,Ayman Charaf,Thomas De Vijlder,Carl Heatherington,Joerg Hoernschemeyer,Hong Jiang,Ulrike Rieder,Francis Ring,Robert Peter,Jessica A Stolee,Rainer Wechselberger
A risk-based approach for routine identity testing of therapeutic oligonucleotide drug substances and drug products is described. Risk analysis of solid-phase oligonucleotide synthesis indicates that intact mass measurement is a powerful technique for confirming synthesis of the intended oligonucleotide. Further risk assessment suggests that the addition of a second, sequence-sensitive identity test
-
Immunogenicity Assessment of Inotersen, a 2'-O-(2-Methoxyethyl) Antisense Oligonucleotide in Animals and Humans: Effect on Pharmacokinetics, Pharmacodynamics, and Safety. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Rosie Z Yu,Yanfeng Wang,Dan A Norris,Tae-Won Kim,Padma Narayanan,Richard S Geary,Brett P Monia,Scott P Henry
Inotersen (TEGSEDI™) is a 2′-O-(2-methoxyethyl)-modified antisense oligonucleotide, intended for treating hereditary transthyretin (TTR) amyloidosis with polyneuropathy. The potential immunogenicity (IM) response to inotersen was evaluated in chronic nonclinical safety studies and the pivotal phase 2/3 clinical study. The evaluation was designed to assess the characteristics of antidrug antibodies
-
Selective Modulation of the Protease Activated Protein C Using Exosite Inhibiting Aptamers. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Nasim Shahidi Hamedani,Jens Müller,Fabian Tolle,Heiko Rühl,Behnaz Pezeshkpoor,Kerstin Liphardt,Johannes Oldenburg,Günter Mayer,Bernd Pötzsch
Activated protein C (APC) is a serine protease with anticoagulant and cytoprotective activities. Nonanticoagulant APC mutants show beneficial effects as cytoprotective agents. To study, if such biased APC signaling can be achieved by APC-binding ligands, the aptamer technology has been used. A G-quadruplex-containing aptamer, G-NB3, has been selected that binds to the basic exosite of APC with a KD
-
Anti-CD44 DNA Aptamers Selectively Target Cancer Cells. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Aleksandra Pęcak,Łukasz Skalniak,Katarzyna Pels,Mirosław Książek,Mariusz Madej,Dobrosława Krzemień,Stanisław Malicki,Benedykt Władyka,Adam Dubin,Tad A Holak,Grzegorz Dubin
CD44 is a type I transmembrane glycoprotein interacting with a number of extracellular components, including hyaluronic acid (HA). CD44-HA axis is involved in a variety of processes, including adhesion, migration, differentiation, trafficking, and others. CD44 is overexpressed in several cancers where binding of HA induces signal transduction leading to activation of antiapoptotic proteins and factors
-
Construction of Monomeric and Dimeric G-Quadruplex-Structured Cytosine-Phosphate-Guanine Oligodeoxynucleotides for Enhanced Uptake and Activation in Toll-Like Receptor 9-Positive Macrophages. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Wenqing Liao,Mengmeng Tan,Kosuke Kusamori,Yoshinobu Takakura,Makiya Nishikawa
The G-quadruplex (GQ) structure has potential applications in nucleic acid drug delivery because of its superior stability. In this study, we added one G-tract (five guanines) to an unmethylated phosphodiester-linked cytosine-phosphate-guanine oligodeoxynucleotide (CpG ODN), a potential immune adjuvant, to construct a GQ-structured CpG ODN with precise structural properties, increased biological stability
-
Gapmer Antisense Oligonucleotides Targeting 5S Ribosomal RNA Can Reduce Mature 5S Ribosomal RNA by Two Mechanisms. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Adam J Pollak,Justin H Hickman,Xue-Hai Liang,Stanley T Crooke
In this study, we demonstrate that 5S ribosomal RNA (rRNA), a highly structured and protein-bound RNA, is quite difficult to reduce with antisense oligonucleotides (ASOs). However, we found a single accessible site that was targetable with a high-affinity complementary ASO. The ASO appeared to bind to the site, recruit RNaseH1, and cause degradation of the 5S RNA. Intriguingly, we also observed that
-
Novel MicroRNA Sponges to Specifically Modulate Gene Expression in Colon Cancer Cells. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-10-09 Ana R Rama,Gloria Perazzoli,Laura Cabeza,Cristina Mesas,Francisco Quiñonero,Beatriz García-Pinel,Celia Vélez
MicroRNA (miRNA) sponges allow the selective blockade of a complete family of associated miRNAs, which induce post-transcriptional gene silencing in their target through binding to 3′UTR mRNA. miRNA-365 and miRNA-145 are downregulated in colorectal cancer (CRC) but not in healthy tissues. Based on this, we constructed two vectors by inserting miRNA sponges (one for miRNA-365 and other for miRNA-145)
-
Technical Considerations for Use of Oligonucleotide Solution API. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Jale Muslehiddinoglu,Robert Simler,Malcolm L Hill,Claudia Mueller,John H A Amery,Leigh Dixon,Anna Watson,Kirsten Storch,Cinzia Gazziola,Frank Gielen,Stefan Andreas Lange,Jeremy D Prail,Doug P Nesta
The most common approach for the manufacture of oligonucleotides includes isolation of the active pharmaceutical ingredient (API) via lyophilization to provide a solid product, which is then dissolved to provide an aqueous formulation. It is well known from the development and manufacture of large molecules (“biologics”) that API production does not always require isolation of solid API before drug
-
Treatment with Volanesorsen, a 2'-O-Methoxyethyl-Modified Antisense Oligonucleotide Targeting APOC3 mRNA, Does Not Affect the QTc Interval in Healthy Volunteers. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Lynnetta M Watts,Ewa Karwatowska-Prokopczuk,Eunju Hurh,Veronica J Alexander,Kristin Balogh,Louis O'Dea,Richard S Geary,Sotirios Tsimikas
The aim of this study was to assess the effect of volanesorsen on the corrected QT (QTc) interval. This thorough QT study enrolled 52 healthy male and female subjects who were randomized at a single site in a four-way crossover study. Subjects were randomly assigned to 1 of 12 treatment sequences and crossed over into four treatment periods over the course of which each subject was to receive a single
-
Delivery of Antisense Oligonucleotides to the Cornea. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Viet Q Chau,Jiaxin Hu,Xin Gong,John D Hulleman,Rafael L Ufret-Vincenty,Frank Rigo,Thahza P Prakash,David R Corey,V Vinod Mootha
Antisense oligonucleotides (ASOs) are synthetic nucleic acids that recognize complementary RNA sequences inside cells and modulate gene expression. In this study, we explore the feasibility of ASO delivery to the cornea. We used quantitative polymerase chain reaction to test the efficacy of a benchmark ASO targeting a noncoding nuclear RNA, Metastasis-Associated Lung Adenocarcinoma Transcript 1 (MALAT1)
-
Likelihood of Nonspecific Activity of Gapmer Antisense Oligonucleotides Is Associated with Relative Hybridization Free Energy. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Andrew T Watt,Grant Swayze,Eric E Swayze,Susan M Freier
Reduction of matched and nearly complementary unintended transcripts was evaluated for 96 antisense oligonucleotides (ASOs) and 832 nearly matched unintended transcripts. The ASOs were 16–20 nucleotide “gapmers” with a gap of 8–10 DNA residues and 2′-O-methoxy-ethyl or constrained-ethyl substitutions in the wings. Most unintended transcripts were not reduced or were reduced with a potency more than
-
Effects of Chemical Modifications on siRNA Strand Selection in Mammalian Cells. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Andrew J Varley,Matthew L Hammill,Lidya Salim,Jean-Paul Desaulniers
Small interfering RNAs (siRNAs) enable efficient gene silencing through RNA interference (RNAi) mechanisms. The RNAi machinery relies on an RNA-guided nuclease, Argonaute-2 (Ago2), which preferentially selects a single strand from an siRNA duplex. Complementarity between the selected strand and an RNA target strand leads to silencing through cleavage. The U.S. Food and Drug Administration's recent
-
MYCN Silencing by RNAi Induces Neurogenesis and Suppresses Proliferation in Models of Neuroblastoma with Resistance to Retinoic Acid. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-08-06 Ruhina Maeshima,Dale Moulding,Andrew W Stoker,Stephen L Hart
Neuroblastoma (NB) is the most common solid tumor in childhood. Twenty percent of patients display MYCN amplification, which indicates a very poor prognosis. MYCN is a highly specific target for an NB tumor therapy as MYCN expression is absent or very low in most normal cells, while, as a transcription factor, it regulates many essential cell activities in tumor cells. We aim to develop a therapy for
-
Modulating Polymer-siRNA Binding Does Not Promote Polyplex-Mediated Silencing. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-07-29 R Chauncey Splichal,Joseph A Gredell,Erin B Vogel,Amanda Malefyt,Georgina Comiskey,Milton R Smith,Christina Chan,S Patrick Walton
The development of delivery vehicles for small interfering RNAs (siRNAs) remains a bottleneck to widespread clinical use. Cationic polymers represent an important class of potential delivery vehicles. In this study, we used alkyne-azide click chemistry to synthesize a variety of cationic poly(propargyl glycolide) backbone polymers to bind and deliver siRNAs. We demonstrated control over the binding
-
Researcher's Perceptions on Publishing "Negative" Results and Open Access. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-07-27 Lucía Echevarría,Alberto Malerba,Virginia Arechavala-Gomeza
Scientific advance is based on reproducibility, corroboration, and availability of research results. However, large numbers of experimental results that contradict previous work do not get published and many research results are not freely available as they are hidden behind paywalls. As part of COST Action “DARTER”, a network of researchers in the field of RNA therapeutics, we have performed a small
-
Approaches for Systemic Delivery of Dystrophin Antisense Peptide Nucleic Acid in the mdx Mouse Model. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-07-15 Camilla Brolin,Ernest Wee Kiat Lim,Sylvestre Grizot,Caroline Holkmann Olsen,Niloofar Yavari,Thomas O Krag,Peter E Nielsen
Antisense-mediated exon skipping constitutes a promising new modality for treatment of Duchenne Muscular Dystrophy (DMD), which is caused by gene mutations that typically introduce a translation stop codon in the dystrophin gene, thereby abolishing production of functional dystrophin protein. The exon removal can restore translation to produce a shortened, but still partially functional dystrophin
-
Chitosan-Shelled Nanobubbles Irreversibly Encapsulate Morpholino Conjugate Antisense Oligonucleotides and Are Ineffective for Phosphorodiamidate Morpholino-Mediated Gene Silencing of DUX4. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-07-13 Maria Sofia Falzarano,Monica Argenziano,Anne Chalotte Marsollier,Virginie Mariot,Davide Rossi,Rita Selvatici,Julie Dumonceaux,Roberta Cavalli,Alessandra Ferlini
Orphan drugs, including antisense oligonucleotides (AONs), siRNAs/miRNAs, Cas9 nuclease, and recombinant genes, have recently been made available for rare diseases. However, the main bottleneck for these new therapies is delivery. Drugs/synthetic genes need to reach the affected tissues with minimal off-target effects and immune reactions. AON molecules are currently delivered as backboned naked compounds
-
Population Pharmacokinetic-Pharmacodynamic Modeling of Inotersen, an Antisense Oligonucleotide for Treatment of Patients with Hereditary Transthyretin Amyloidosis. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 Rosie Z Yu,Jon W Collins,Shannon Hall,Elizabeth J Ackermann,Richard S Geary,Brett P Monia,Scott P Henry,Yanfeng Wang
A population pharmacokinetic (PK) and pharmacodynamic (PD) model was developed for inotersen to evaluate exposure–response relationships and to optimize therapeutic dosing regimen in patients with hereditary transthyretin (TTR) amyloidosis polyneuropathy (hATTR-PN). Inotersen PK and TTR level (PD) data were composed of one Phase 1 study in healthy subjects, one Phase 2/3 study in hATTR patients, and
-
RT-qPCR Methods to Support Pharmacokinetics and Drug Mechanism of Action to Advance Development of RNAi Therapeutics. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 Elena Castellanos-Rizaldos,Christopher R Brown,Sean Dennin,Joohwan Kim,Swati Gupta,Diana Najarian,Yongli Gu,Krishna Aluri,Jennifer Enders,Kirk Brown,Yuanxin Xu
The goal of this study was to develop a reverse transcription quantitative polymerase chain reaction (RT-qPCR) method for the accurate quantification of chemically modified small interfering RNA (siRNA) including but not restricted to thermally destabilizing modifications such as glycol nucleic acid (GNA). RT-qPCR was found to be superior to mass spectrometry-based siRNA detection in terms of sensitivity
-
Population Pharmacokinetic-Pharmacodynamic Model of Serum Transthyretin Following Patisiran Administration. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 Varun Goel,Nathalie H Gosselin,Claudia Jomphe,Xiaoping Zhang,Jean-Francois Marier,Gabriel J Robbie
Hereditary transthyretin-mediated amyloidosis is an inherited, rapidly progressive, life-threatening disease caused by mutated transthyretin (TTR) protein. Patisiran is a small interfering RNA (siRNA) formulated in a lipid nanoparticle that inhibits hepatic TTR protein synthesis by RNA interference. We have developed an indirect-response pharmacokinetic–pharmacodynamic model relating plasma siRNA (ALN-18328)
-
Oligonucleotides and the COVID-19 Pandemic: A Perspective. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 John J Rossi,Daniel Rossi
The present global health emergency involving the emergence and rapid spread of a novel coronavirus has prompted the world scientific community to consider how it can help to fight this growing viral pandemic. With few safe and effective drugs available to combat this threat to humanity and the normal functioning of our society, the oligonucleotide research community is uniquely positioned to apply
-
High Levels of Apoptosis Are Induced in the Human Colon Cancer HT-29 Cell Line by Co-Administration of Sulforaphane and a Peptide Nucleic Acid Targeting miR-15b-5p. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 Jessica Gasparello,Laura Gambari,Chiara Papi,Andrea Rozzi,Alex Manicardi,Roberto Corradini,Roberto Gambari,Alessia Finotti
Sulforaphane (SFN) is one of most important dietary constituents of broccoli (Brassica oleracea) and other cruciferous vegetables, which have been reported to exhibit health benefits, including prevention and therapy of cancer, such as colorectal carcinoma (CRC). The objective of this study was to determine whether the anticancer effect of SFN on colon cancer HT-29 cell line could be improved by the
-
Pyrene-Modified DNA Aptamers with High Affinity to Wild-Type EGFR and EGFRvIII. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-05-22 Elena Zavyalova,Askar Turashev,Anastasia Novoseltseva,Valeriia Legatova,Olga Antipova,Ekaterina Savchenko,Sonja Balk,Andrey Golovin,Galina Pavlova,Alexey Kopylov
Nucleic acid aptamers have been proven to be a useful tool in many applications. Particularly, aptamers to epidermal growth factor receptor (EGFR) have been successfully used for the recognition of EGFR-expressing cells, the inhibition of EGFR-dependent pathways, and targeted drug delivery into EGFR-positive cells. Several aptamers are able to discriminate wild-type EGFR from its mutant form, EGFRvIII
-
Modification of KL4 Peptide Revealed the Importance of Alpha-Helical Structure for Efficient Small Interfering RNA Delivery. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-04-29 Yingshan Qiu,Jason C K Lo,Kerry C W Kwok,A James Mason,Jenny K W Lam
A safe and effective delivery system is considered a key to the success of nucleic acid therapeutics. It has been reported that pulmonary surfactants or their components could facilitate the uptake of small interfering RNA (siRNA) into the lung epithelial cells. Previously, our group investigated the use of KL4 peptide, a synthetic cationic peptide that simulates the structural properties of surfactant
-
Pharmacodynamic and Pharmacokinetic Properties of Full Phosphorothioate Small Interfering RNAs for Gene Silencing In Vivo. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-04-20 Christian Berk,Gianluca Civenni,Yuluan Wang,Christian Steuer,Carlo V Catapano,Jonathan Hall
State-of-the-art small interfering RNA (siRNA) therapeutics such as givosiran and fitusiran are constructed from three variable components: a fully-modified RNA core that conveys metabolic stability, a targeting moiety that mediates target-cell uptake, and a linker. This structural complexity poses challenges for metabolite characterization and risk assessment after long-term patient exposure. In this
-
Underlying Immune Disorder May Predispose Some Transthyretin Amyloidosis Subjects to Inotersen-Mediated Thrombocytopenia. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-02-11 PadmaKumar Narayanan,Brian R Curtis,Lijiang Shen,Eugene Schneider,Joseph A Tami,Suzanne Paz,Sebastien A Burel,Li-Jung Tai,Todd Machemer,T Jesse Kwoh,Shuting Xia,Sanford J Shattil,Joseph L Witztum,Jeffery A Engelhardt,Scott P Henry,Brett P Monia,Steven G Hughes
Inotersen, a 2'-O-methoxyethyl (2'-MOE) phosphorothioate antisense oligonucleotide, reduced disease progression and improved quality of life in patients with hereditary transthyretin amyloidosis with polyneuropathy (hATTR-PN) in the NEURO-TTR and NEURO-TTR open-label extension (OLE) trials. However, 300 mg/week inotersen treatment was associated with platelet count reductions in several patients. Mean
-
The 10th Oligonucleotide Therapy Approved: Golodirsen for Duchenne Muscular Dystrophy. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-02-11 Annemieke Aartsma-Rus,David R Corey
-
Bifunctional Aptamer-Doxorubicin Conjugate Crosses the Blood-Brain Barrier and Selectively Delivers Its Payload to EpCAM-Positive Tumor Cells. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-02-06 Joanna Macdonald,Delphine Denoyer,Justin Henri,Adelaide Jamieson,Ingrid J G Burvenich,Normand Pouliot,Sarah Shigdar
The prognosis for breast cancer patients diagnosed with brain metastases is poor, with survival time measured merely in months. This can largely be attributed to the limited treatment options capable of reaching the tumor as a result of the highly restrictive blood-brain barrier (BBB). While methods of overcoming this barrier have been developed and employed with current treatment options, the majority
-
RNA Interference for the Masses? siRNA Targeting PCSK9 Promises Prevention of Cardiovascular Disease. Nucleic Acid Ther. (IF 4.875) Pub Date : 2020-01-13 Jonathan K Watts,Ira S Ockene
-
Endocytosis Controls siRNA Efficiency: Implications for siRNA Delivery Vehicle Design and Cell-Specific Targeting. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-11-12 Daniel Vocelle,Christina Chan,S Patrick Walton
While small interfering RNAs (siRNAs) are commonly used for laboratory studies, development of siRNA therapeutics has been slower than expected, due, in part, to a still limited understanding of the endocytosis and intracellular trafficking of siRNA-containing complexes. With the recent characterization of multiple clathrin-/caveolin-independent endocytic pathways, that is, those mediated by Graf1
-
Impurity Qualification Toxicology Study for a 2'-O-Methoxyethyl-Modified Antisense Inhibitor in Mice. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-11-05 Tae-Won Kim,Chris Papagiannis,Daniel Capaldi,Andrew K McPherson,Farah Mahdi,Nhuy Luu,Andrew A Rodriguez,Christine Hoffmaster,David Serota,Scott P Henry
Safety assessment of drug impurities is a routine part of the drug development process. For oligonucleotide-based drugs, impurities can arise from impurities in starting materials, as by-products of the manufacturing process or from degradation, and are generally structurally similar to the parent oligonucleotide. To study the potential impact of impurities, a representative batch of a 2'-O-methoxyethyl
-
Expression of TNRC6 (GW182) Proteins Is Not Necessary for Gene Silencing by Fully Complementary RNA Duplexes. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-10-31 Zhongtian Liu,Samantha T Johnson,Zhiying Zhang,David R Corey
The trinucleotide repeat containing 6 (TNRC6) family of proteins are core components of RNA interference (RNAi) and consist of three paralogs (TNRC6A, TNRC6B, and TNRC6C). The TNRC6 paralogs associate with argonaute (AGO) protein, the core RNAi factor, and bridge its interactions with other proteins. We obtained TNRC6A and TNRC6B single and double knockout cell lines to investigate how the TNRC6 paralogs
-
Nuclear and Cytoplasmatic Quantification of Unconjugated, Label-Free Locked Nucleic Acid Oligonucleotides. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-10-16 Hannah Pendergraff,Steffen Schmidt,Jonas Vikeså,Christian Weile,Charlotte Øverup,Marie W Lindholm,Troels Koch
Methods for the quantification of antisense oligonucleotides (AONs) provide insightful information on biodistribution and intracellular trafficking. However, the established methods have not provided information on the absolute number of molecules in subcellular compartments or about how many AONs are needed for target gene reduction for unconjugated AONs. We have developed a new method for nuclear
-
The BACE1-Specific DNA Aptamer A1 Rescues Amyloid-β Pathology and Behavioral Deficits in a Mouse Model of Alzheimer's Disease. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-09-12 Zhi-Man Liang,Yong-Hua Peng,Yue Chen,Li-Li Long,Hong-Jie Luo,Ya-Jun Chen,Yan-Ling Liang,Ying-Hong Tian,Shu-Ji Li,Yu-Sheng Shi,Xing-Mei Zhang
Amyloid-β (Aβ) plaque deposits in the brain are considered to be one of the main pathological markers of Alzheimer's disease (AD). The sequential proteolytic cleavage of amyloid precursor protein (APP) by the aspartyl proteases β-site APP-cleaving enzyme 1 (BACE1) and γ-secretase produces Aβ. Therefore, BACE1 inhibition is a very attractive target for the treatment of AD. Our previous work identified
-
The Pharmacokinetics of 2'-O-Methyl Phosphorothioate Antisense Oligonucleotides: Experiences from Developing Exon Skipping Therapies for Duchenne Muscular Dystrophy. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-08-20 Sieto Bosgra,Jessica Sipkens,Sjef de Kimpe,Cathaline den Besten,Nicole Datson,Judith van Deutekom
Delivery to the target site and adversities related to off-target exposure have made the road to clinical success and approval of antisense oligonucleotide (AON) therapies challenging. Various classes of AONs have distinct chemical features and pharmacological properties. Understanding the similarities and differences in pharmacokinetics (PKs) among AON classes is important to make future development
-
Phosphorothioate Antisense Oligonucleotides Bind P-Body Proteins and Mediate P-Body Assembly. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-08-20 Ying Wang,Wen Shen,Xue-Hai Liang,Stanley T Crooke
Antisense oligonucleotides (ASOs) regulate gene expression by binding to complementary target RNA, and ASOs can be designed to take advantage of a growing array of post RNA binding molecular mechanisms. Intracellular trafficking of ASOs influences their efficacy. We have identified a number of membrane-less structures in the nucleus, nucleolus, and cytoplasm where phosphorothioate-modified ASOs (PS-ASOs)
-
Ligand Conjugated Multimeric siRNAs Enable Enhanced Uptake and Multiplexed Gene Silencing. Nucleic Acid Ther. (IF 4.875) Pub Date : 2019-08-08 Jonathan M Brown,James E Dahlman,Kristin K Neuman,Carla A H Prata,Monika C Krampert,Philipp M Hadwiger,Hans-Peter Vornlocher
Small interfering RNAs (siRNAs) conjugated to N-acetylgalactosamine (GalNAc) ligands have been used to treat disease in patients. However, conjugates with other ligands deliver siRNA less efficiently, limiting the development of new targeted therapies. Most approaches to enhancing the poten