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  1. Article: Senescence of endplate osteoclasts induces sensory innervation and spinal pain.

    Pan, Dayu / Benkato, Kheiria Gamal / Han, Xuequan / Zheng, Jinjian / Kumar, Vijay / Wan, Mei / Zheng, Junying / Cao, Xu

    bioRxiv : the preprint server for biology

    2024  

    Abstract: Spinal pain affects individuals of all ages and is the most common musculoskeletal problem globally. Its clinical management remains a challenge as the underlying mechanisms leading to it are still unclear. Here, we report that significantly increased ... ...

    Abstract Spinal pain affects individuals of all ages and is the most common musculoskeletal problem globally. Its clinical management remains a challenge as the underlying mechanisms leading to it are still unclear. Here, we report that significantly increased numbers of senescent osteoclasts (SnOCs) are observed in mouse models of spinal hypersensitivity, like lumbar spine instability (LSI) or aging, compared to controls. The larger population of SnOCs is associated with induced sensory nerve innervation, as well as the growth of H-type vessels, in the porous endplate. We show that deletion of senescent cells by administration of the senolytic drug Navitoclax (ABT263) results in significantly less spinal hypersensitivity, spinal degeneration, porosity of the endplate, sensory nerve innervation and H-type vessel growth in the endplate. We also show that there is significantly increased SnOC-mediated secretion of Netrin-1 and NGF, two well-established sensory nerve growth factors, compared to non-senescent OCs. These findings suggest that pharmacological elimination of SnOCs may be a potent therapy to treat spinal pain.
    Language English
    Publishing date 2024-05-07
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.10.26.564218
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Toehold-Mediated Shape Transition of Nucleic Acid Nanoparticles.

    Hartung, Jordan / McCann, Nathan / Doe, Erwin / Hayth, Hannah / Benkato, Kheiria / Johnson, M Brittany / Viard, Mathias / Afonin, Kirill A / Khisamutdinov, Emil F

    ACS applied materials & interfaces

    2023  Volume 15, Issue 21, Page(s) 25300–25312

    Abstract: We introduce a toehold-mediated strand displacement strategy for regulated shape-switching of nucleic acid nanoparticles (NANPs) enabling their sequential transformation from triangular to hexagonal architectures at isothermal conditions. The successful ... ...

    Abstract We introduce a toehold-mediated strand displacement strategy for regulated shape-switching of nucleic acid nanoparticles (NANPs) enabling their sequential transformation from triangular to hexagonal architectures at isothermal conditions. The successful shape transitions were confirmed by electrophoretic mobility shift assays, atomic force microscopy, and dynamic light scattering. Furthermore, implementation of split fluorogenic aptamers allowed for monitoring the individual transitions in real time. Three distinct RNA aptamers─malachite green (MG), broccoli, and mango─were embedded within NANPs as reporter domains to confirm shape transitions. While MG "lights up" within the square, pentagonal, and hexagonal constructs, the broccoli is activated only upon formation of pentagon and hexagon NANPs, and mango reports only the presence of hexagons. Moreover, the designed RNA fluorogenic platform can be employed to construct a logic gate that performs an AND operation with three single-stranded RNA inputs by implementing a non-sequential polygon transformation approach. Importantly, the polygonal scaffolds displayed promising potential as drug delivery agents and biosensors. All polygons exhibited effective cellular internalization followed by specific gene silencing when decorated with fluorophores and RNAi inducers. This work offers a new perspective for the design of toehold-mediated shape-switching nanodevices to activate different light-up aptamers for the development of biosensors, logic gates, and therapeutic devices in the nucleic acid nanotechnology.
    MeSH term(s) Nucleic Acids ; RNA/genetics ; Nanoparticles ; Nanotechnology ; Microscopy, Atomic Force ; Oligonucleotides
    Chemical Substances Nucleic Acids ; RNA (63231-63-0) ; Oligonucleotides
    Language English
    Publishing date 2023-05-19
    Publishing country United States
    Document type Journal Article
    ISSN 1944-8252
    ISSN (online) 1944-8252
    DOI 10.1021/acsami.3c01604
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Evaluation of Thermal Stability of RNA Nanoparticles by Temperature Gradient Gel Electrophoresis (TGGE) in Native Condition.

    Benkato, Kheiria / O'Brien, Benjamin / Bui, My N / Jasinski, Daniel L / Guo, Peixuan / Khisamutdinov, Emil F

    Methods in molecular biology (Clifton, N.J.)

    2017  Volume 1632, Page(s) 123–133

    Abstract: Temperature gradient gel electrophoresis (TGGE) is a powerful tool used to analyze the thermal stabilities of nucleic acids. While TGGE is a decades-old technique, it has recently gained favor in the field of RNA nanotechnology, notably in assessing the ... ...

    Abstract Temperature gradient gel electrophoresis (TGGE) is a powerful tool used to analyze the thermal stabilities of nucleic acids. While TGGE is a decades-old technique, it has recently gained favor in the field of RNA nanotechnology, notably in assessing the thermal stabilities of RNA nanoparticles (NPs). With TGGE, an electrical current and a linear temperature gradient are applied simultaneously to NP-loaded polyacrylamide gel, separating the negatively charged NPs based on their thermal behavior (a more stable RNA complex will remain intact through higher temperature ranges). The linear temperature gradient can be set either perpendicular or parallel to the electrical current, as either will make the NPs undergo a transition from native to denatured conformations. Often, the melting transition is influenced by sequence variations, secondary/tertiary structures, concentrations, and external factors such as the presence of a denaturing agent (e.g., urea), the presence of monovalent or divalent metal ions, and the pH of the solvent. In this chapter, we describe the experimental setup and the analysis of the thermal stability of RNA NPs in native conditions using a modified version of a commercially available TGGE system.
    MeSH term(s) Denaturing Gradient Gel Electrophoresis/instrumentation ; Denaturing Gradient Gel Electrophoresis/methods ; Nucleic Acid Conformation ; RNA/chemistry ; Temperature ; Thermodynamics ; Transition Temperature
    Chemical Substances RNA (63231-63-0)
    Language English
    Publishing date 2017
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-7138-1_8
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Programmable Nucleic Acid Based Polygons with Controlled Neuroimmunomodulatory Properties for Predictive QSAR Modeling.

    Johnson, Morgan Brittany / Halman, Justin R / Satterwhite, Emily / Zakharov, Alexey V / Bui, My N / Benkato, Kheiria / Goldsworthy, Victoria / Kim, Taejin / Hong, Enping / Dobrovolskaia, Marina A / Khisamutdinov, Emil F / Marriott, Ian / Afonin, Kirill A

    Small (Weinheim an der Bergstrasse, Germany)

    2017  Volume 13, Issue 42

    Abstract: In the past few years, the study of therapeutic RNA nanotechnology has expanded tremendously to encompass a large group of interdisciplinary sciences. It is now evident that rationally designed programmable RNA nanostructures offer unique advantages in ... ...

    Abstract In the past few years, the study of therapeutic RNA nanotechnology has expanded tremendously to encompass a large group of interdisciplinary sciences. It is now evident that rationally designed programmable RNA nanostructures offer unique advantages in addressing contemporary therapeutic challenges such as distinguishing target cell types and ameliorating disease. However, to maximize the therapeutic benefit of these nanostructures, it is essential to understand the immunostimulatory aptitude of such tools and identify potential complications. This paper presents a set of 16 nanoparticle platforms that are highly configurable. These novel nucleic acid based polygonal platforms are programmed for controllable self-assembly from RNA and/or DNA strands via canonical Watson-Crick interactions. It is demonstrated that the immunostimulatory properties of these particular designs can be tuned to elicit the desired immune response or lack thereof. To advance the current understanding of the nanoparticle properties that contribute to the observed immunomodulatory activity and establish corresponding designing principles, quantitative structure-activity relationship modeling is conducted. The results demonstrate that molecular weight, together with melting temperature and half-life, strongly predicts the observed immunomodulatory activity. This framework provides the fundamental guidelines necessary for the development of a new library of nanoparticles with predictable immunomodulatory activity.
    MeSH term(s) Cell Line, Tumor ; DNA/chemistry ; Humans ; Immunomodulation ; Microglia/cytology ; Nucleic Acids/chemistry ; Quantitative Structure-Activity Relationship ; RNA/chemistry ; Reproducibility of Results
    Chemical Substances Nucleic Acids ; RNA (63231-63-0) ; DNA (9007-49-2)
    Language English
    Publishing date 2017-09-18
    Publishing country Germany
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2168935-0
    ISSN 1613-6829 ; 1613-6810
    ISSN (online) 1613-6829
    ISSN 1613-6810
    DOI 10.1002/smll.201701255
    Database MEDical Literature Analysis and Retrieval System OnLINE

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