Exosome-Based Nucleic Acid Therapeutics

In recent years, the development of nucleic acid therapeutics has gained considerable traction, with the aim of controlling the expression of disease-related genes. In particular, numerous small interfering RNA (siRNA) drugs that act through the RNA interference (RNAi) mechanism have been developed. Growing attention has also been directed toward strategies that inhibit microRNAs (miRNAs), which endogenously mediate RNAi. Both approaches involve nucleic acid drugs that modulate the RNA-induced silencing complex (RISC)—the central effector of RNAi—to control disease-related gene expression through either gene silencing or restoration. To improve the design of nucleic acid therapeutics, artificial nucleic acids have been employed to enhance enzymatic stability and target RNA recognition. Although ribose-based artificial nucleic acids have been widely adopted, researchers have also begun exploring alternatives with chemical structures that differ substantially from those of the ribose type, especially to achieve greater enzymatic resistance. Against this backdrop, we have focused on non-ribose type acyclic artificial nucleic acids with amino-acid-based backbones, serinol nucleic acid (SNA) and acyclic L-threoninol nucleic acid (L-aTNA), and have explored their use in designing nucleic acid therapeutics. This account describes designs for both siRNA and antisense oligonucleotides against miRNA (anti-miRNA), which are referred to as AMOs. The unique properties of these acyclic nucleic acids are highlighted, along with an introduction to research on siRNA designs aimed at regulating strand selection during RISC assembly—a key strategy for minimizing off-target effects. This account also includes the design and mechanism of action for our AMO. A key limitation of acyclic artificial nucleic acids is also addressed, with a chemical strategy proposed to overcome it. Ultimately, this review suggests that the unique chemical properties of acyclic artificial nucleic acids may enable the next generation of highly stable and selective nucleic acid therapeutics, paving the way for more effective and safer clinical applications.

Nucleic Acid TherapeuticsVol. 26, No. 3 Special Issue On AptamersEditorialAptamers Coming of Age at Twenty-FiveBruce A. SullengerBruce A. SullengerSearch for more papers by this authorPublished Online:3 Jun 2016https://doi.org/10.1089/nat.2016.29001.sulAboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View article"Aptamers Coming of Age at Twenty-Five." nucleic acid therapeutics, 26(3), p. 119FiguresReferencesRelatedDetailsCited bySelective Modulation of the Protease Activated Protein C Using Exosite Inhibiting Aptamers Nasim Shahidi Hamedani, Jens Müller, Fabian Tolle, Heiko Rühl, Behnaz Pezeshkpoor, Kerstin Liphardt, Johannes Oldenburg, Günter Mayer, and Bernd Pötzsch9 October 2020 | Nucleic Acid Therapeutics, Vol. 30, No. 5Achieving Selective Targeting Using Engineered Nanomaterials5 March 2019Carbon nanomaterial-enabled pesticide biosensors: Design strategy, biosensing mechanism, and practical applicationTrAC Trends in Analytical Chemistry, Vol. 106Aptamer delivery of siRNA, radiopharmaceutics and chemotherapy agents in cancerInternational Journal of Pharmaceutics, Vol. 525, No. 2G-quadruplex-based aptamers against protein targets in therapy and diagnosticsBiochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1861, No. 5 Volume 26Issue 3Jun 2016 InformationCopyright 2016, Mary Ann Liebert, Inc.To cite this article:Bruce A. Sullenger.Aptamers Coming of Age at Twenty-Five.Nucleic Acid Therapeutics.Jun 2016.119-119.http://doi.org/10.1089/nat.2016.29001.sulPublished in Volume: 26 Issue 3: June 3, 2016PDF download

Smart Nucleic Acids as Future Therapeutics

Nucleic Acid TherapeuticsVol. 21, No. 6 PerspectiveThe Development of RNA Interference TherapeuticsJohn J. RossiJohn J. RossiSearch for more papers by this authorPublished Online:23 Dec 2011https://doi.org/10.1089/nat.2011.1503AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View article"The Development of RNA Interference Therapeutics." , 21(6), pp. 371–372FiguresReferencesRelatedDetailsCited byMethods for Assembling B-Cell Lymphoma Specific and Internalizing Aptamer–siRNA Nanoparticles Via the Sticky BridgeTumor-targeted in vivo gene silencing via systemic delivery of cRGD-conjugated siRNA15 September 2014 | Nucleic Acids Research, Vol. 42, No. 18Nanodelivery Systems for Nucleic Acid Therapeutics in Drug Resistant Tumors1 April 2014 | Molecular Pharmaceutics, Vol. 11, No. 8Ion exchange liquid chromatography method for the direct determination of small ribonucleic acidsAnalytica Chimica Acta, Vol. 799Something Old, Something New Bruce Sullenger23 December 2011 | Nucleic Acid Therapeutics, Vol. 21, No. 6 Volume 21Issue 6Dec 2011 InformationCopyright 2011, Mary Ann Liebert, Inc.To cite this article:John J. Rossi.The Development of RNA Interference Therapeutics.Nucleic Acid Therapeutics.Dec 2011.371-372.http://doi.org/10.1089/nat.2011.1503Published in Volume: 21 Issue 6: December 23, 2011PDF download

Nucleic acid based therapeutics have an allure for medicinal chemists due to their potential for specific control of gene expression. Persistent efforts have resulted in the FDA approval of the nucleic acid based drugs Vitravene™, Macugen™ and recently Kynamro™, leading to a sudden leap in the number of active clinical trials involving the nucleic acid moieties. Beginning with antigene and antisense technology, nucleic acid therapeutics have recently gained more potent strategies, e.g. RNA interference, ribozymes, decoy oligonucleotides and aptamers. Nucleic acid based therapeutics which have opened up new frontiers in medicinal chemistry research, along with various chemical modifications undertaken to improve their utility and their mechanism of action, will be discussed in this review article.

Pancreatic cancer (PC), one of the most aggressive and lethal malignancies, remains an imminent threat to human health despite enormous advancements in cancer research. Traditional therapeutic approaches are unable to significantly enhance the life expectancy of PC patients due to limitations such as lower targetability, recurrence of cancerous tumors, and side effects on healthy cells. Nucleic acid therapeutics (NATs) offer a promising alternative against traditional approaches due to their ability to target and modify genetic pathways involved in PC progression, thus providing precision treatment. However, their efficient transportation to cancerous cells and tissues is still a major obstacle. Because of the high biocompatibility, tunable physicochemical characteristics, surface functionalization potential, and scalable manufacturing potential, nanomaterials have established themselves as a frontrunner as a delivery carrier against different diseases. Considering this, different nanocarriers were explored as a vehicle for anticancer therapeutics, especially to deliver therapeutic drugs, NATs, etc., and also provided a shield against enzymatic and chemical degradation in the bloodstream while promoting tumor-specific accumulation and targetability. Pertaining to this, in the current review, we have systematically discussed the development of nanomaterial-based NATs delivery systems for the delivery of nucleic acids like siRNA, miRNA, mRNA, saRNA, pDNA, CRISPR-Cas9 guide RNA, and other nucleic acids against PC, with an emphasis on their benefits, drawbacks, and translational potential. We also highlighted and compared the preclinical assessment of nanomaterial-mediated NATs delivery in PC therapy in wild-type and drug-resistant PC cells and discussed NATs delivery in two-dimensional (2D) and three-dimensional (3D) cell culture models alongside in vivo and clinical studies. Additionally, we also explored the possibilities of the 3D cell culture model's advantage over traditional 2D cell cultures and their relevance to in vivo systems, especially in facilitating NATs delivery and clinical translation possibilities, and paved the way for future research and clinical applications toward precision medicine.

Nucleic acid therapeutics has huge potential for the treatment of a wide range of diseases including respiratory diseases. Plasmid DNA (pDNA) and small interfering RNA (siRNA) are the two most widely investigated nucleic acids for therapeutic development. However, efficient and safe delivery of nucleic acids is still a major hurdle in translating nucleic acid therapy into clinical practice. For the treatment of respiratory diseases, administration via inhalation is the most direct and effective way to deliver therapeutic nucleic acids to the lungs. Although liquid aerosol formulation is investigated in most of the studies, it is not desirable in terms of maintaining the stability of nucleic acid especially during long-term storage. This problem could be circumvented by formulating the therapeutic nucleic acids into dry powder for inhalation, and should be considered as the future direction of developing inhalable nucleic acids. In this review, the three major particle engineering methods investigated for the preparation of inhalable pDNA and siRNA formulations, including spray drying (SD), spray freeze drying (SFD) and supercritical fluid (SFC) drying, are discussed and compared. Moreover, common assessment methods and the challenges of evaluating the biological activities of inhalable nucleic acid powders are also reviewed.

Nucleic acid therapeutics can be used to control virtually every aspect of cell behavior and therefore have significant potential to treat genetic disorders, infectious diseases, and cancer. However, while clinically approved to treat a small number of diseases, the full potential of nucleic acid therapeutics is hampered by inefficient delivery. Nucleic acids are large, highly charged biomolecules that are sensitive to degradation and so the approaches to deliver these molecules differ significantly from traditional small molecule drugs. Current studies suggest less than 1% of the injected nucleic acid dose is delivered to the target cell in an active form. This inefficient delivery increases costs and limits their use to applications where a small amount of nucleic acid is sufficient. In this review, we focus on two of the major barriers to efficient nucleic acid delivery: (1) delivery to the target cell and (2) transport to the subcellular compartment where the nucleic acids are therapeutically active. We explore how nanoparticles can be modified with targeting ligands to increase accumulation in specific cells, and how the composition of the nanoparticle can be engineered to manipulate or disrupt cellular membranes and facilitate delivery to the optimal subcellular compartments. Finally, we highlight how with intelligent material design, nanoparticle delivery systems have been developed to deliver nucleic acids that silence aberrant genes, correct genetic mutations, and act as both therapeutic and prophylactic vaccines.This article is categorized under:Nanotechnology Approaches to Biology > Cells at the NanoscaleTherapeutic Approaches and Drug Discovery > Nanomedicine for Infectious DiseaseBiology‐Inspired Nanomaterials > Lipid‐Based Structures

Technology for Nucleic Acid Delivery in the Treatment of Hematological Malignancies.

Effective process development towards intensified processing for gene delivery applications using Hepatitis B core Antigen (HBcAg) virus-like particles (VLPs) relies on analytical methods for the absolute quantification of HBcAg VLP proteins and bound nucleic acids. We investigated a silica spin column (SC)-based extraction procedure, including proteinase K lysis and silica chromatography, for the absolute quantification of different species of nucleic acids bound to HBcAg VLPs analyzed by dye-based fluorescence assays. This revealed load-dependent nucleic acid recoveries of the silica-SC-based extraction. We also developed a reversed-phase high-performance liquid chromatography (RP-HPLC) method to separate and quantify the HBcAg proteins and the bound nucleic acids simultaneously without prior sample treatment by dissociation reagents. The method demonstrated sufficient linearity, accuracy, and precision coefficients and is suited for determining absolute protein and nucleic acid concentrations and HBcAg protein purities at various purification stages. Both the silica-SC-based extraction and the RP-based extraction presented overcome the limitations of analytical techniques, which are restricted to relative or qualitative analyses for HBcAg VLPs with bound nucleic acids. In combination with existing analytics, the methods for an absolute quantification of HBcAg VLPs and bound nucleic acids presented here are required to evaluate downstream purification steps, such as the removal of host cell-derived nucleic acids, concurrent protein loss, and efficient loading with therapeutic nucleic acids. Hence, the methods are key for effective process development when using HBcAg VLP as potential gene delivery vehicles.

Nucleic acid therapeutics have demonstrated tremendous potential for treating diseases by targeting the genetic underpinnings at the transcriptomic level. However, their efficacy hinges on robust strategies to protect nucleic acids from degradation during circulation and to facilitate precise delivery to diseased tissues and cells. Here the critical roles of chemical modification and bioconjugation in advancing nucleic acid therapeutics for improved binding affinity, enhanced stability, and targeted delivery are reviewed. Commencing diverse applications, the significance of different chemical modifications is discussed based on recent literature and clinical products, on oligonucleotides. These modifications encompass backbone, ribose, base alterations and bioconjugation techniques such as N‐acetylgalactosamine (GAlNac), aptamers, antibodies, and cell‐penetrating peptides (CPPs). Supported by a clinical perspective, diverse applications and ongoing developments are highlighted. Furthermore, the current landscape of nucleic acid therapeutics and their potential in addressing genetic disorders with multiple cellular/organelle targeting is discussed. Here the promising prospect of combining chemical innovation and bioconjugation strategies is underscored to propel the development of more effective nucleic acid therapeutics.

Nucleic acid therapeutics: Past, present, and future.

Nanoparticles for nucleic acid delivery: Applications in cancer immunotherapy.

Delivery approaches of immunomodulatory nucleic acids for cancer therapy

Jokūbas Leikauskas, Commissioning Editor, Nucleic Acid Insights With a background in science communication and digital publishing, Jokūbas focuses on advancing the nucleic acid therapeutics field by commissioning and shaping high-impact, open access content for Nucleic Acid Insights. As Commissioning Editor, he leads the development of interviews, expert articles, and industry perspectives that highlight emerging advances across mRNA, DNA, oligonucleotide, and drug delivery modalities. Jokūbas is driven to translate complex scientific topics into engaging, accessible content while maintaining strong connections across the nucleic acids community.