Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo
| Autor: | Stacey N. Barnaby, Chad A. Mirkin, Jasmine L. May, Anthony J. Sprangers, Nikunjkumar Savalia, Alexander H. Stegh, Serena T. Ghelfi, Lisa E. Cole, Timothy L. Sita, Thomas C. Cayton, Timothy J. Merkel, Lisa A. Hurley, Andrew Lee, Fotini M. Kouri, Alexandra Chalastanis, Charles David James |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Biodistribution Mice SCID Nanoconjugates Biology Fluorescence 03 medical and health sciences Mice 0302 clinical medicine In vivo RNA interference medicine Temozolomide Tumor Cells Cultured Gene silencing Animals Humans RNA Small Interfering neoplasms Antineoplastic Agents Alkylating DNA Modification Methylases Gene knockdown Multidisciplinary Brain Neoplasms Tumor Suppressor Proteins Biological Sciences Molecular biology Xenograft Model Antitumor Assays Dacarbazine 030104 developmental biology DNA Repair Enzymes 030220 oncology & carcinogenesis Spherical nucleic acid Female RNA Interference Glioblastoma Ex vivo medicine.drug |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 114(16) |
| ISSN: | 1091-6490 |
| Popis: | RNA interference (RNAi)-based gene regulation platforms have shown promise as a novel class of therapeutics for the precision treatment of cancer. Techniques in preclinical evaluation of RNAi-based nanoconjugates have yet to allow for optimization of their gene regulatory activity. We have developed spherical nucleic acids (SNAs) as a blood-brain barrier-/blood-tumor barrier-penetrating nanoconjugate to deliver small interfering (si) and micro (mi)RNAs to intracranial glioblastoma (GBM) tumor sites. To identify high-activity SNA conjugates and to determine optimal SNA treatment regimens, we developed a reporter xenograft model to evaluate SNA efficacy in vivo. Engrafted tumors stably coexpress optical reporters for luciferase and a near-infrared (NIR) fluorescent protein (iRFP670), with the latter fused to the DNA repair protein O6-methylguanine-DNA-methyltransferase (MGMT). Using noninvasive imaging of animal subjects bearing reporter-modified intracranial xenografts, we quantitatively assessed MGMT knockdown by SNAs composed of MGMT-targeting siRNA duplexes (siMGMT-SNAs). We show that systemic administration of siMGMT-SNAs via single tail vein injection is capable of robust intratumoral MGMT protein knockdown in vivo, with persistent and SNA dose-dependent MGMT silencing confirmed by Western blotting of tumor tissue ex vivo. Analyses of SNA biodistribution and pharmacokinetics revealed rapid intratumoral uptake and significant intratumoral retention that increased the antitumor activity of coadministered temozolomide (TMZ). Our study demonstrates that dual noninvasive bioluminescence and NIR fluorescence imaging of cancer xenograft models represents a powerful in vivo strategy to identify RNAi-based nanotherapeutics with potent gene silencing activity and will inform additional preclinical and clinical investigations of these constructs. |
| Databáze: | OpenAIRE |
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