With the development of synthesis technology, modified messenger RNA (mRNA) has emerged as a novel category of therapeutic agents for a broad of diseases. However, effective intracellular delivery of mRNA remains challenging, especially for its sensitivity to enzymatic degradation. Here, we propose a polyphenol-assisted handy delivery strategy for efficient in vivo delivery of IL-10 mRNA. IL-10 mRNA binds to polyphenol ellagic acid through supramolecular binding to yield a negatively charged core, followed by complexing with linear polyetherimide and coating with bilirubin-modified hyaluronic acid to obtain a layer-by-layer nanostructure. The nanostructure specifically up-regulated the level of IL-10, effectively inhibited the expression of inflammatory factors, promoted mucosal repair, protected colonic epithelial cells against apoptosis, and exerted potent therapeutic efficacy in dextran sulfate sodium salt-induced acute and chronic murine models of colitis. The designed delivery system without systemic toxicity has the potential to facilitate the development of a promising platform for mRNA delivery in ulcerative colitis treatment.
<div>AbstractPurpose:<p>Exploration of novel strategies to extend the benefit of PARP inhibitors beyond <i>BRCA</i>-mutant cancers is of great interest in personalized medicine. Here, we identified <i>EGFR</i> amplification as a potential biomarker to predict sensitivity to PARP inhibition, providing selection for the glioblastoma (GBM) patient population who will benefit from PARP inhibition therapy.</p>Experimental Design:<p>Selective sensitivity to the PARP inhibitor talazoparib was screened and validated in two sets [test set (<i>n</i> = 14) and validation set (<i>n</i> = 13)] of well-characterized patient-derived glioma sphere-forming cells (GSC). FISH was used to detect <i>EGFR</i> copy number. DNA damage response following talazoparib treatment was evaluated by γH2AX and 53BP1 staining and neutral comet assay. PARP–DNA trapping was analyzed by subcellular fractionation. The selective monotherapy of talazoparib was confirmed using <i>in vivo</i> glioma models.</p>Results:<p><i>EGFR</i>-amplified GSCs showed remarkable sensitivity to talazoparib treatment. <i>EGFR</i> amplification was associated with increased reactive oxygen species (ROS) and subsequent increased basal expression of DNA-repair pathways to counterelevated oxidative stress, and thus rendered vulnerability to PARP inhibition. Following talazoparib treatment, <i>EGFR</i>-amplified GSCs showed enhanced DNA damage and increased PARP–DNA trapping, which augmented the cytotoxicity. <i>EGFR</i> amplification–associated selective sensitivity was further supported by the <i>in vivo</i> experimental results showing that talazoparib significantly suppressed tumor growth in <i>EGFR</i>-amplified subcutaneous models but not in nonamplified models.</p>Conclusions:<p><i>EGFR</i>-amplified cells are highly sensitive to talazoparib. Our data provide insight into the potential of using <i>EGFR</i> amplification as a selection biomarker for the development of personalized therapy.</p></div>
The potential therapeutic effect of nitric oxide (NO) for cancers has received considerable attention as a "killer" that causes damage to mitochondria and DNA by oxidation or nitrosation. However, the fabrication of an intelligent and controllable NO release system has remained elusive in the desired location to realize selective cancer therapy. Herein, an intelligent endogenous esterase-triggered nitric oxide (NO) generator for synergetic cancer therapy is fabricated by integrating NO prodrug and doxorubicin (DOX) into a single glutathione (GSH)-responsive mesoporous silica nanoparticle (MPND). When the MPND is internalized into the cancer cell, the rupture of -S-S- bridges and the degradation of MPND occur in the tumor microenvironment with a high level of GSH, inducing the on-demand release of DOX. Importantly, the high endogenic esterase concentration can activate the prodrug to generate abundant NO, which further enhances the release performance of DOX. In vitro results verify that the release profiles of NO and DOX show the stimuli-responsive dependence of endogenic esterase and GSH, respectively, demonstrating the potential for on-demand release in the cancer cells. Consequently, MPND shows a high antitumor efficiency in MCF-7 cancer cells. Furthermore, using multicellular tumor spheroids to mimic in vivo experiment, MPND can enhance the tumor penetration and therapeutic effect for killing the deep tumor tissue at the central location. Therefore, the endogenous esterase-triggered NO nanogenerators may provide a potential alternative strategy to develop NO-relevant platforms for synergistic cancer therapy.
Glioblastoma (GBM) is the most common and lethal primary intracranial tumor. Actin cytoskeleton regulator ARP2/3 complex modulates glioma cell invasion and migration. Besides, inside the tumor entity contains brain tumor initiating cells (BTICs), which are resistant to chemical and radiation therapies and form recurrent tumor. It has been shown that NOTCH signaling is essential for BTICs to maintain the stem cell phenotype. But little is known regarding the role of actin cytoskeleton in NOTCH signaling and the stem cell phenotype maintenance. Here, we show ARP2/3 complex is required for NOTCH ligand DLL1 to activate NOTCH1 signaling, thus maintaining the stem cell phenotype. CD133-positive glioma cells co-expressed NESTIN and DLL1, and exhibited high NOTCH1 activity both in specimens and in vitro U87-MG and U251-MG glioma neurospheres. Silencing DLL1 decreased NOTCH1 activity as well as BTICs markers CD133 and NESTIN expression in both glioma neurospheres. Self-renewal ability was also impaired in DLL1 knockdown cells detected through single-cell neurosphere formation assay. Both ARP2/3 complex specific inhibitor CK636 and silencing subunit ARP2 induced the same effect with DLL1 knockdown. Subcellular fractionation experiments showed that membrane protein DLL1 was less expressed on the cell membrane of ARP2/3 inhibited and knockdown cells than untreated cells. An original method was introduced to study the expression pattern among cells inside the sphere. It showed that DLL1 was accumulated in the cytoplasm of the treated cells detected by confocal immunofluorescence microscope, which suggests that ARP2/3 complex regulates the transportation of DLL1 from cytoplasm to membrane to activate NOTCH1 receptor through cell contact. In vivo, silencing ARP2 also impaired the tumorigenicity of U87-MG neurosphere cells. These findings indicate that actin cytoskeleton protein participates in the stem cell phenotype maintenance, which provides evidence that anti-invasive targeted therapies may help eliminate BTICs.
Developing two-dimensional (2D) hybrid nanosheet arrays integrating inorganic and organic components is highly significant for third-order nonlinear optical (NLO) applications. Herein, an oriented 2D porphyrin-based MOF (ZnTPyP(Co)) thin film composed of vertically stacked ultrathin nanosheets was fabricated via the liquid-phase epitaxial (LPE) layer-by-layer (LBL) method. The prepared ZnTPyP(Co) thin film exhibits an outstanding third-order NLO response with a high third-order nonlinear susceptibility of ∼2.63 × 10–7 esu, which is ascribed to the hybrid nanosheet array structure. Additionally, experimental Z-scan measurement and theoretical calculations also demonstrate that the substitution of Co metal ions in the porphyrinic core can increase the level of delocalization of the porphyrinic group and contribute to the material's enhanced NLO properties. These findings not only provide new film candidates for NLO application but also highlight the potential of 2D MOF nanosheets in advanced optical devices.
Targeted protein degradation is a powerful tool for determining the function of specific proteins nowadays. Survivin is the smallest member of the inhibitor of the apoptosis protein (IAP) family. It exists in the cytoplasm and nucleus of cells, but the exact function of survivin in different subcellular locations retained unclear updates due to the lack of effective and simple technical means. In this study, we created a novel nanoantibody-based molecular toolkit, namely, the ubiquitin-proteasome system (Nb4A-Fc-T2A-TRIM21), that can target to degrade survivin localized in cytoplasmic and cell nuclear by ubiquitinating, and by which to verify the potential roles of survivin subcellular localization. Also, the results showed that the cytoplasmic survivin mainly plays an anti-apoptotic function by directly or indirectly inhibiting the caspase pathway, and the nuclear survivin mainly promotes cell proliferation and participates in the regulation of the cell cycle. In addition, the Nb4A-Fc-T2A-TRIM21 system can degrade the endogenous survivin protein in a large amount by the ubiquitin-proteasome pathway, and the system can provide theoretical support for ubiquitination degradation targeting other endogenous proteins.
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