Visible Light-Guided Gene Delivery with Nonviral Supramolecular Block Copolymer Vectors.

To achieve efficient gene delivery in vitro or in vivo, nonviral vectors should have excellent biostability across cellular and tissue barriers and also smart stimuli responsiveness toward controlled release of therapeutic genes into the cell nucleus. However, it remains a key challenge to effectively combine the biostability of covalent polymers with the stimuli responsiveness of noncovalent polymers into one nonviral vehicle. In this work, we report the construction of a kind of cationic supramolecular block copolymers (SBCs) through noncovalent polymerization of ß-cyclodextrin/azobenzene-terminated pentaethylenehexamine (DMA-Azo-PEHA-ß-CD) in aqueous media using ß-CD-monosubstituted poly(ethylene glycol) (PEG-ß-CD) as a supramolecular initiator. The resultant SBC exhibits superior biostability, biocompatibility, and light/pH dual-responsive characteristics, and it also demonstrates efficient plasmid DNA condensation capacity and the ability to rapidly release plasmid DNA into cells driven by visible light (450 nm). Eventually, this SBC-based delivery system demonstrates visible light-induced enhancement of gene delivery in both COS-7 and HeLa cells. We anticipate that this work provides a facile and robust strategy to enhance gene delivery in vitro or in vivo via visible light-guided manipulation of genes, further achieving safe, highly efficient, targeting gene therapy for cancer.

A novel docetaxel derivative exhibiting potent anti-tumor activity and high safety in preclinical animal models.

As a taxoid agent, docetaxel (DTX) exhibits potent antitumor activity. However, severe toxic side effects and acquired multidrug resistance represent its clinical challenges. Herein, a novel docetaxel derivative (DTX-AI) is synthesized via the nucleophilic addition reaction of 4-acetylphenyl carbamate at the C10 position of the DTX framework. DTX-AI exhibits superior cytotoxicity and a higher apoptotic ratio in vitro against DTX-sensitive tumor cells (MCF-7, HeLa and A549 cells) and even DTX-resistant ones (HeLa/PTX cells), but displays less toxicity against normal cells (MRC-5 and L929 cells) compared with DTX. DTX-AI can effectively suppress the growth of HeLa-tumor xenografts in vivo and even induce complete tumor regression. Furthermore, DTX-AI shows sustained effects on the inhibition of A549-tumor xenograft growth and no obvious recurrence, even after the drug administration was stopped for 30 d. More importantly, DTX-AI has significantly reduced long-term and short-term animal toxicity and extended the survival of mice (100%) compared with DTX (0%). DTX-AI is expected to be a promising 'me-better' anti-tumor drug with higher efficiency and lower toxicity for improved chemotherapy in the clinic.

In Situ Preparation of Amphibious ZnO Quantum Dots with Blue Fluorescence Based on Hyperbranched Polymers and their Application in Bio-Imaging.

A new strategy for preparing amphibious ZnO quantum dots (QDs) with blue fluorescence within hyper-branched poly(ethylenimine)s (HPEI) was proposed in this paper. By changing [Zn2+]/[OH-] molar ratio and heating time, ZnO QDs with a quantum yields (QY) of 30% in ethanol were obtained. Benefiting from the amphibious property of HPEI, the ZnO/HPEI nanocomposites in ethanol could be dissolved in chloroform and water, acquiring a QY of 53%, chloroform and 11% in water. By this strategy, the ZnO/HPEI nano-composites could be applied in not only in optoelectronics, but also biomedical fields (such as bio-imaging and gene transfection). The bio-imaging application of water-soluble ZnO/HPEI nanocomposites was investigated and it was found that they could easily be endocytosed by the COS-7 cells, without transfection reagent, and they exhibited excellent biological imaging behavior.

Nanocrystal Encapsulation, Release and Application Based on pH-Sensitive Covalent Dynamic Hyperbranched Polymers.

A new strategy for nanocrystal encapsulation, release and application based on pH-sensitive covalent dynamic hyperbranched polymers is described. The covalent dynamic hyperbranched polymers, with multi-arm hydrophobic chains and a hydrophilic hyperbranched poly(amidoamine) (HPAMAM) core connected with pH-sensitive imine bonds (HPAMAM-DA), could encapsulate CdTe quantum dots (QDs) and Au nanoparticles (NPs). Benefiting from its pH response property, CdTe QDs and Au NPs encapsulated by HPAMAM-DA could be released to aqueous phase after imine hydrolysis. The released CdTe/HPAMAM and Au/HPAMAM nanocomposites exhibited excellent biological imaging behavior and high catalytic activities on p-nitrophenol hydrogenation, respectively.

Site-dependent fluorescence enhanced polymers with a self-restricted GFP chromophore for living cell imaging.

The ß-barrel structure of green fluorescent protein (GFP) provides a confined environment to enhance its fluorescence efficiency. Inspired by the unique structure of GFP, we reported a self-restricted GFP chromophore analogue which was rationally grafted onto the middle or the terminal of poly(ethylene glycol)-block-poly(N-isopropyl acrylamide) (PEG-b-PNIPAM) via click chemistry to obtain PEG-GA-PNIPAM and PEG-PNIPAM-GA (GA: MeOBDPI). These structures were characterized through NMR, GPC, and FT-IR. By varying the length of PNIPAM and the location of the GFP chromophore, self-assembly behaviour and fluorescence intensity were correspondingly changed. PEG-GA-PNIPAM and PEG-PNIPAM-GA were assembled into nano-sized spherical micelles above the low critical solution temperature (LCST). The size of the micelles increased with the length of the PNIPAM block. These optical properties were carefully evaluated by UV-Vis and fluorescence spectroscopy. The results indicated that increasing the length of the PNIPAM block enhanced the fluorescence in water, and PEG-PNIPAM74-GA has more remarkable fluorescence intensity than PEG-GA-PNIPAM106 in living cells such as MCF-7 cells. Furthermore, the fluorescence behaviour of PEG-PNIPAM74-GA was studied in MCF-7 cells and L929 cells. The result showed that PEG-PNIPAM74-GA was mostly located in the cytoplasm. Compared with the CellTracker™ Red CMTPX dye, it could enter into MCF-7 cells and L929 cells more easily in DMEM with 10% FBS. Therefore, PEG-PNIPAM74-GA has potential application prospects for living cell imaging.

"Bottom-Up" Fabrication of BODIPY-Functionalized Fluorescent Hyperbranched Glycopolymers for Hepatoma-Targeted Imaging.

A novel type of multivalent and highly specific fluorescent hyperbranched glycopolymers h-P(GalEA-co-VBPT-co-BYMA) (hPGVB) is designed and prepared successfully via a facile "bottom-up" strategy. The acetylated hPGVB is prepared by one-pot reversible addition-fragmentation chain transfer (RAFT) copolymerization of acrylate-type galactose monomers AcGalEA and methacrylate-type fluorescent monomers BYMA in presence of an inimer-type RAFT chain transfer agent. After deacetylation, the resulting amphiphilic hPGVB can self-assemble into stable nanoparticles in aqueous media, showing strong green fluorescence with relative high quantum yields and good photostability. The cell viability study indicates the excellent biocompatibility of the hPGVB fluorescent nanoparticles (FNPs) against HepG2 and NIH3T3 cells. More importantly, comparing with the galactose-free fluorescent hyperbranched polymers h-P(OEGMA-co-VBPT-co-BYMA), hPEVB FNPs can be selectively internalized by asialoglycoprotein (ASGP) receptor-rich HepG2 cells, indicating their potential application in the bioimaging fields.

Supramolecular block copolymers for gene delivery: enhancement of transfection efficiency by charge regulation.

A class of cationic supramolecular block copolymers with readily controlled charges has been exploited. Upon post-synthetic structural optimization, this copolymer exhibits comparable biocompatibility, greatly improved pDNA condensation capability and biostability, and further enhanced transfection efficiency in vitro. This work provides valuable insight into the creation of advanced nonviral vectors for gene delivery.

Biopolymer-Drug Conjugate Nanotheranostics for Multimodal Imaging-Guided Synergistic Cancer Photothermal-Chemotherapy.

Some of the biomedical polymer-drug conjugates are being translated into clinical trials; however, they intrinsically lack photothermal and multi-imaging capabilities, hindering them from imaging-guided precision cancer therapy and complete tumor regression. We introduce a new concept of all-in-one biopolymer-drug conjugate nanotheranostics and prepare a kind of intracellular pH-sensitive polydopamine-doxorubicin (DOX) conjugate nanoparticles (PDCNs) under mild conditions. Significantly, this strategy integrates polymeric prodrug-induced chemotherapy (CT), near-infrared (NIR) light-mediated photothermal therapy (PT), and triple modalities including DOX self-fluorescence, photothermal, and photoacoustic (PA) imaging into one conjugate nanoparticle. The PDCNs present excellent photothermal property, dual stimuli-triggered drug release behavior, and about 12.4-fold blood circulation time compared to free DOX. Small animal fluorescent imaging technique confirms that PDCNs have preferential tumor accumulation effect in vivo, giving a 12.8-fold DOX higher than the control at 12 h postinjection. Upon NIR laser irradiation (5 min, 808 nm, and 2 W·cm-2), the PDCN-mediated photothermal effect can quickly elevate the tumor over 50 °C, exhibiting good photothermal and PA imaging functions, of which the PA amplitude is 3.6-fold greater than the control. In vitro and in vivo assays persuasively verify that intravenous photothermal-CT of PDCNs produces synergistic antitumor activity compared to single PT or CT, achieving complete tumor ablation during the evaluation period.

Synthesis of a Cationic Supramolecular Block Copolymer with Covalent and Noncovalent Polymer Blocks for Gene Delivery.

The design and fabrication of safe and highly efficient nonviral vectors is the key scientific issue for the achievement of clinical gene therapy. Supramolecular cationic polymers have unique structures and specific functions compared to covalent cationic polymers, such as low cytotoxicity, excellent biodegradability, and smart environmental responsiveness, thereby showing great application prospect for gene therapy. However, supramolecular gene vectors are facile to be degraded under physiological conditions, leading to a significant reduction of gene transfection efficiency. In order to achieve highly efficient gene expression, it is necessary for supramolecular gene vectors being provided with appropriate biostability to overcome various cell obstacles. To this end, a novel cationic supramolecular block copolymer composed of a conventional polymer and a noncovalent polymer was constructed through robust ß-cyclodextrin/ferrocene host-guest recognition. The resultant supramolecular block copolymer perfectly combines the advantages of both conventional polymers and supramolecular polymers ranging from structures to functions. This supramolecular copolymer not only has the ability to effectively condense pDNA for enhanced cell uptake, but also releases pDNA inside cancer cells triggered by H2O2, which can be utilized as a prospective nonviral delivery vehicle for gene delivery. The block polymer exhibited low cytotoxicity, good biostability, excellent biodegradability, and intelligent responsiveness, ascribing to the dynamic/reversible nature of noncovalent linkages. In vitro studies further illustrated that the supramolecular block polymer exhibited greatly improved gene transfection efficiency in cancer cells. This work offers an alternative platform for the exploitation of smart nonviral vehicles for specific cancer gene therapy in the future.

Matrix Metalloproteinase Responsive Nanoparticles for Synergistic Treatment of Colorectal Cancer via Simultaneous Anti-Angiogenesis and Chemotherapy.

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers worldwide, especially in developed countries. Although patients' overall survival has been improved by either conventional chemotherapy or newly developed anti-angiogenesis treatment based on its highly vascularized feature, the relatively low therapeutic efficacy and severe side effects remain big problems in clinical practice. In this study, we describe an easy method to construct a novel matrix metalloproteinase-2 (MMP-2) responsive nanocarrier, which can load hydrophobic agents (camptothecin and sorafenib) with high efficiency to exert synergistic efficacy for CRC treatment. The drug-containing nanoparticles can particularly respond to the MMP-2 and realize the controlled release of payloads at the tumor site. Moreover, both in vitro and in vivo studies have demonstrated that this responsive nanoparticle exhibits much higher therapeutic efficacy than that of single antitumor agents or combined drugs coadministrated in traditional ways.

miR-223 increases gallbladder cancer cell sensitivity to docetaxel by downregulating STMN1.

BACKGROUND: MicroRNAs (miRs) are involved in cancer carcinogenesis, and certain regulatory miRs could provide promising therapeutic methods for refractory malignancies, such as gallbladder cancer (GBC). miR-223 was found to play a pivotal role in enhancing chemotherapeutic effects, therefore evoking interest in the role of miR-223 in GBC. RESULTS: miR-223 was decreased in GBC tissues and cell lines, and ectopic miR- 223 expression exhibited multiple anti-tumorigenic effects in GBC cells, including decreased proliferation, migration and invasion in vitro. However, treatment with a miR-223 inhibitor increased cell viability. We determined that STMN1 was negatively correlated with and regulated by miR-223 in GBC. miR-223 increased GBC sensitivity to docetaxel in vitro and in vivo, and the induced sensitivity to docetaxel was suppressed by the restoration of STMN1 expression. METHODS: We examined miR-223 expression in GBC tissue and GBC cell lines using qRT-PCR. The effects of modulated miR-223 expression in GBC cells were assayed using Cell Counting Kit-8 (CCK8), flow cytometry, and wound-healing and invasion assays. Susceptibility to docetaxel was evaluated in miR-223/STMN1-modulated GBC cells and xenograft tumor models. The protein expression of relevant genes was examined by Western blotting. CONCLUSIONS: These findings indicated that miR-223 might serve as an onco-suppressor that enhances susceptibility to docetaxel by downregulating STMN1 in GBC, highlighting its promising therapeutic value.

Plasmonic, Targeted, and Dual Drugs-Loaded Polypeptide Composite Nanoparticles for Synergistic Cocktail Chemotherapy with Photothermal Therapy.

To integrate cocktail chemotherapy with photothermal therapy into one biocompatible and biodegradable nanocarrier, the plasmonic, lactose-targeted, and dual anticancer drugs-loaded polypeptide composite nanoparticles were for the first time fabricated under mild conditions. The glyco-PEGylated polypeptide micelles that self-assembled from the lactose (LAC) and PEG grafted polycysteine terpolymer were used as templates to generate the plasmonic composite nanoparticles, as mainly characterized by DLS, TEM, SEM, and XPS. These composite nanoparticles showed a broad and strong near-infrared (NIR) absorption at 650-1100 nm and increased the temperature of phosphate buffer solution by 30.1 °C upon a continuous-wave laser irradiation (808 nm, 5 min, 2 W·cm(-2)), while the same dose of NIR-mediated heating completely killed HepG2 cancer cells in vitro, presenting excellent photothermal properties. Two anticancer drugs, doxorubicin (DOX) and 6-mercaptopurine (6-MP), were loaded into the composite nanoparticles through physical interactions and Au-S bond, respectively. The dual drugs-loaded composite nanoparticles exhibited reduction-sensitive and NIR-triggered cocktail drugs release profiles and trigger-enhanced cytotoxicity. As evidenced by flow cytometry, fluorescence microscopy, and MTT assay, the LAC-coated composite nanoparticles were more internalized by the HepG2 than the HeLa cell line, demonstrating a LAC-targeting enhanced cytotoxicity toward HepG2. The combination cocktail chemo-photothermal therapy produced a lower half maximal inhibitory concentration than cocktail chemotherapy or photothermal therapy alone, displaying a good synergistic antitumor effect.

Dual-responsive aggregation-induced emission-active supramolecular nanoparticles for gene delivery and bioimaging.

Dual-responsive aggregation-induced emission-active supramolecular fluorescent nanoparticles are reported, which have the ability to undergo a unique morphological transition combining with a cooperative optical variation in response to pH and light stimuli. The dynamic supramolecular nanoparticles show excellent biocompatibility and effective plasmid DNA condensation capability, further achieving efficient in vitro gene delivery and bioimaging.

Facile Approach To Construct Ternary Cocktail Nanoparticles for Cancer Combination Therapy.

Drug combinations have been widely used in cancer treatment. However, it remains a formidable challenge to deliver three or more therapeutic agents in one nanoparticle with a precise and tunable molar ratio because of differences in pharmacokinetics and biodistribution of various anticancer drugs. Herein, we reported a facile approach to construct ternary cocktail nanoparticles, which are composed of three different anticancer drugs, such as gemcitabine, chlorambucil, and irinotecan, through the molecular coassembly of two amphiphilic drug-drug conjugates. The component of these nanoparticles can be simply adjusted by changing the feed ratio of two amphiphilic drug-drug conjugates in the coassembly process. Without the help of any drug carriers, they can self-deliver, release three drugs synchronally, and obtain the optimal synergistic therapeutic effect. This facile strategy may open a new way for cancer combination therapy.

An autoreduction method to prepare plasmonic gold-embedded polypeptide micelles for synergistic chemo-photothermal therapy.

Biodegradable, biocompatible polypeptide micelles were used as a reducing agent and template in an autoreduction method for preparing plasmonic gold-embedded polypeptide micelles under mild conditions. The micelles were fully characterized by DLS, TEM, SEM, and AFM. The in situ reduced gold was embedded in the interior core of the disulfide bond-cross-linked polypeptide micelles by forming multivalent Au-S bonds. The plasmonic gold-embedded micelles showed strong near-infrared (NIR) light absorption and NIR-mediated photothermal properties including high photothermal conversion efficiency and good photostability. After continuous-wave diode laser irradiation for 5 min (808 nm, 2 W cm-2), the NIR light-induced heating of the gold-embedded micelles efficiently killed cancer cells in vitro, as observed by a double fluorescent staining technique. A standard MTT assay, flow cytometry, and fluorescence microscopy showed that the anticancer drug doxorubicin (DOX)-loaded and gold-embedded micelles quickly entered HeLa cells and gave a lower half-maximal inhibitory concentration (IC50) than for chemotherapy or photothermal therapy alone, demonstrating a good synergistic effect for the combination chemo-photothermal therapy. Consequently, this work provides a versatile strategy for fabricating plasmonic polypeptide composite nanoparticles, which are promising for synergistic chemo-photothermal cancer therapy.

Facile construction of near-monodisperse and dual responsive polycarbonate mixed micelles with the ability of pH-induced charge reversal for intracellular delivery of antitumor drugs.

A mixed strategy was used to construct size-adjustable, pH and redox dual-responsive, near-monodisperse and charge-conversional core-shell polycarbonate carriers. First, two kinds of random polycarbonates (PC(Arss-N2CH3)) with different ratios of hydrophobic disulfide (Arss) and hydrophilic dimethylamine functional groups (N2CH3) (relatively hydrophobic polycarbonate (BPC) with more disulfide groups and relatively hydrophilic polycarbonate (LPC) with more dimethylamine groups) were used to construct near-monodisperse core micelles in pH 7.4 PBS. Then, a negatively charged shell polymer, poly(ethylene glycol)-b-1,2-dicarboxylic-cyclohexene anhydride modified amino polycarbonate (PEG-PCDCA), bearing acid-labile ß-carboxylic amides, was added to the core micelle solution to get the desired mixed micelle. The assembly process, pH induced charge-reversal, and dual-response abilities of the resultant mixed micelles were thoroughly studied using zeta potential and dynamic light scattering (DLS). Two model drugs, Nile red (NR) and doxorubicin (DOX), were successfully loaded into the micelle and could be released in response to dithiothreitol (DTT) and acidic conditions. Confocal microscopy indicated that the micelle system could achieve effective cellular uptake and DOX release in Hela cells. Moreover, the blank micelle and the drug-loaded mixed micelle showed slight and great cytotoxicity against Hela cells, respectively, indicating their significant potential in cancer therapy.

miR-122 inhibits cancer cell malignancy by targeting PKM2 in gallbladder carcinoma.

Gallbladder cancer (GBC) is one of the lethal diseases of digestive system. Increasing evidence prompt that microRNAs (miRs) might provide a novel therapeutical target for malignant disease. The antitumor effect of miR-122 to GBC is worth to be investigated. miR-122 expression level in GBC tissue sample and cell lines were assayed by qRT-PCR. miR-122 mimics were transfected for upregulation of miR-122 expression. Cell function was assayed by CCK8, flow cytometry, wound healing assay, migration assay, and invasion assay. The target genes of miR-122 were predicated by TargetScan online program and verified by western blot and luciferase report gene assay. miR-122 was decreased in GBC tissue and cell lines. The exogenous introduction of miR-122 exhibits multiple antitumor effect in GBC cell proliferation, invasion, and metastasis. Further studies revealed that the PKM2 was a regulative target of miR-122 in GBC cell. miR-122 also inhibits TGF-ß-induced epithelium mesenchymal transformation of GBC cell by downregulating PKM2 expression. These findings suggest that miR-122 plays an important role in tumorigenesis of GBC through interfering PKM2, highlighting its usefulness as a potential therapeutic agent in GBC.

Synergistic Combination Chemotherapy of Camptothecin and Floxuridine through Self-Assembly of Amphiphilic Drug-Drug Conjugate.

Combination chemotherapy has been widely applied in cancer treatment; however, the cocktail administration of combination chemotherapy could cause the nonuniform biodistribution of anticancer agents, thus impairing the therapeutic efficacy. In the present study, to address this concern, we proposed a novel strategy of preparing self-assembled nanoparticles from amphiphilic drug-drug conjugate for synergistic combination chemotherapy. The conjugate was synthesized by two-step esterification of hydrophobic camptothecin (CPT) and hydrophilic floxuridine (FUDR) through a linker compound. Because of its amphiphilic nature, the CPT-FUDR conjugate self-assembled into stable nanoparticles which could simultaneously release fixed dosage of the two drugs in cancer cells. In vitro studies demonstrated synergistic anticancer efficacy of the CPT-FUDR nanoparticles including improved cell apoptosis, varied cell cycle arrest, as well as effective inhibition of cancer cell proliferation.

Self-Assembled Nanoparticles of Amphiphilic Twin Drug from Floxuridine and Bendamustine for Cancer Therapy.

We report here an amphiphilic twin drug strategy directly using small molecular hydrophilic and hydrophobic anticancer drugs to self-assemble into nanoparticles with a high and fixed drug content, which can solve problems of anticancer drug delivery including poor water solubility, low therapeutic indices, and severe side effects. The twin drug has been prepared by the esterification of the hydrophilic anticancer drug floxuridine (FdU) with the hydrophobic anticancer drug bendamustine (BdM). Due to its inherent amphiphilicity, the FdU-BdM twin drug can self-assemble into stable and well-defined nanoparticles. After FdU-BdM twin drug enters into cells, the ester linkage between hydrophilic and hydrophobic drugs is readily cleaved by hydrolysis to release free FdU and BdM. Since both FdU and BdM can kill cancer cells, the FdU-BdM twin drug nanoparticles can overcome the multidrug resistance (MDR) of tumor cells and present an excellent anticancer activity. This strategy can be extended to other hydrophilic and hydrophobic anticancer drugs to synthesize amphiphilic twin drugs which can form nanoparticles to self-deliver drugs for cancer therapy.

A sweet polydopamine nanoplatform for synergistic combination of targeted chemo-photothermal therapy.

Inspired by sweet or sugar-coated bullets that are used for medications in clinics and the structure and function of biological melanin, a novel kind of sweet polydopamine nanoparticles and their anticancer drug doxorubicin loaded counterparts are prepared, which integrate an active targeting function, photothermal therapy, and chemotherapy into one polymeric nanocarrier. The oxidative polymerization of lactosylated dopamine and/or with dopamine are performed under mild conditions and the resulting sweet nanoparticles are thoroughly characterized. When exposed to an 808 nm continuous-wave diode laser, the magnitude of temperature elevation not only increases with the concentration of nanoparticles, but can also be tuned by the laser power density. The nanoparticles possess strong near infrared light absorption, high photothermal conversion efficiency, and good photostability. The nanoparticles present tunable binding with RCA120 lectin and a targeting effect to HepG2 cells, confirmed by dynamic light scattering, turbidity analysis, MTT assay, and flow cytometry. Importantly, the sweet nanoparticles give the lowest IC50 value of 11.67 µg mL(-1) for chemo-photothermal therapy compared with 43.19 µg mL(-1) for single chemotherapy and 67.38 µg mL(-1) for photothermal therapy alone, demonstrating a good synergistic effect for the combination therapy.