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Scientists have studied intensively the gene delivery carriers for treating genetic diseases. However, there are challenges that impede the application of naked gene-based therapy at the clinical level, such as quick elimination of the circulation, lack of membrane penetrability, and poor endosome trapping. Herein, we develop graphene quantum dots (GQDs)-derivative nanocarriers and introduce polyethylenimine (PEI) to equip the system with enhanced biocompatibility and abundant functional groups for modification. In addition to carrying green fluorescent protein (GFP) as an example of gene delivery, this system covalently binds colon cancer cells targeted antibody and epidermal growth factor receptor (EGFR) to enhance cell membrane penetrability and cell uptake of nanocarriers. To achieve multistrategy cancer therapy, the anticancer drug doxorubicin (Dox) is noncovalently encapsulated to achieve pH-induced drug release at tumor sites and leaves space for further functional gene modification. This nanoparticle serves as a multifunctional gene delivery system, which facilitates improved cytotoxicity and longer-sustained inhibition capacity compared to free Dox treatments in colon cancer cells. Moreover, our GQD composites display compatible tumor suppression ability compared with the free Dox treatment group in xenograft mice experiment with significantly less toxicity. This GQD nanoplatform was demonstrated as a multifunctional gene delivery system that could contribute to treating other genetic diseases in the future.

In this work, we demonstrated that building different linking groups between nanodiamond (ND) and TiO2 (P25) could provide more effective protection under oxidative stress and ultraviolet (UV) light irradiation compared with the use of TiO2 alone. The establishment of ester (-C-O-O-R), amide (-CONH-), and epoxide-amine adduct (-NHCCO-) groups between ND-TiO2 composites was found to be critical in the generation of reactive oxygen species (ROS) by controlling their charge transfer behaviors. We hypothesized that linking groups between the composites dictate the performance of ROS generation from nano-TiO2 under UV-light irradiation due to the differences in linking groups. The results showed that hydroxyl radicals were attenuated by the incorporation of ND. An MTT cell proliferation assay was performed in human cells under the treatment of ND-TiO2 composites to investigate the impacts of composites on cell viability. The results from the luciferase reporter assay suggested they have anti-inflammatory activity and can reduce cellular DNA damage under ROS stimulation. A zebrafish model was also applied with the ND-TiO2 composite treatment to demonstrate the safety aspects of the composites in vivo and their biomedical application potential. Studies exploring ROS generation behaviors in different linking groups suggested that interactive functionalization between nanoparticles might be an ideal antioxidant and anti-inflammatory strategy.

As a two-dimensional nanomaterial, bismuth oxybromide (BiOBr) have attracted tremendous interest in the area of visible-light photocatalysis since it can provide the internal electric field (IEF) through z-axis through its unique electronic band structure. However, the insufficient active sites and rapid recombination rate of charged carriers hamper the efficiency of the photocatalysis. To address these two major obstacles, an enticing strategy of constructing heterojunction was established by introducing Bi

There has been great improvement in nanomaterial fields, and the biomedical potential of nanomaterials as drug delivery system is under intensive studies. Among them, graphene quantum dots (GQDs) are considered to be the next generation of carbon-based nanomaterial for biomedical applications. In this study, we utilized green fluorescent protein (GFP) nucleic acid, DNA-targeting chemoreagent doxorubicin (DOX), and branched polyethyleneimine (PEI) conjugated GQDs, to form pH-responsive nanostar drug carrier for colon cancer suppression investigation. GFP expression plasmid was applied to examine the delivery capacity of our drug carrier. DOX was encapsulated by intrinsic π-π interaction to form GQDs-polymer-DOX conjugates (GECD) as drug carrier. We proposed that once GECD enters the tumor lesion, the acidic tumor microenvironment protonated the tertiary amine of GECD from neutral to mild positive, possessed higher affinity to negatively charged cell membrane, triggered the drug release, and then cancer cells would be inhibited. The anti-cancer ability of our drug carrier system, GECD, was demonstrated in cancer cells by cell proliferation assay. Moreover, GECD exhibited its powerful anti-tumor capacity in the mice xenograft model. The in vivo and in vitro results indicate that our GECD with high cancer suppression capacity could be adopted for future cancer therapy.

Giving patients right dosage is an essential concept of precision medicine. Most of nanocarriers lack of flexible drug capacity and structural stability to be customized for specific treatment, resulting in low therapeutic efficacy and unexpected side effects. Thus, a growing need emerges for fast and rigorous approaches to develop nanoparticles with properties of adjustable dosage and controllable particle size. Poly- l -Lysine is known for its enhanced bioadhesivity and pH-triggered structural swelling effect, which is utilized as the main agent to activate the multistage drug releasing. Inspired by natural bio-assembly system, we report a simple method to self-assemble Poly- l -Lysine-based nanoparticles via supramolecular recognitions of cross-linked pyrenes, which provides noncovalent force to flexibly encapsulate Doxorubincin and to construct robust nanostructures. Pyrene-modified polypeptide self-assemblies are able to adjust drug payload from 1: 10 to 2:1 (drug: polypeptide) without changing its uniform nano-spherical morphology. This nanostructure remained the as-made morphology even after experiencing the long-term (~ 10 weeks) storage at room temperature. Also, the nanoparticles displayed multi-step drug release behaviours and exhibited great in vitro and in vivo cytotoxicity towards colon cancer cells. The as-mentioned nanoparticles provide a novel perspective to compensate the clinical needs of specific drug feedings and scalable synthesis with advantages of simple-synthesis, size-adaptivity, and morphology reversibility.

In this article, we have synthesized Co2+-doped BiOBrxCl1-x hierarchical nanostructured microspheres, featuring different degrees of Co2+ doping, displaying excellent photocatalytic performance. X-ray diffraction and Raman spectroscopy indicated that the Co2+ ions were successfully doped into the BiOBrxCl1-x nanocrystals. The photodegradation rate of rhodamine B mediated by a doped BiOBrxCl1-x was 150 % greater than that of the non-doped BiOBr. We ascribe the improved photocatalytic capability of the Co2+-doped BiOBrxCl1-x to a combination of its superior degree of light absorption, more efficient carrier separation, and faster interfacial charge migration. The major active species involved in the photodegradation of RhB also has been investigated. Moreover, the doped BiOBrxCl1-x possessed excellent cellular biocompatibility and displayed remarkable performance in the photocatalytic bacterial inactivation.

As a two-dimensional nanomaterial, bismuth oxybromide (BiOBr) have attracted tremendous interest in the area of visible-light photocatalysis since it can provide the internal electric field (IEF) through z-axis through its unique electronic band structure. However, the insufficient active sites and rapid recombination rate of charged carriers hamper the efficiency of the photocatalysis. To address these two major obstacles, an enticing strategy of constructing heterojunction was established by introducing Bi2O2(OH)(NO3) (BiON) in BiOBr with the same precursor. Through a facile one-pot hydrothermal synthesis, two Sillen-type layered photocatalysts, with intimately constructed ultrathin heterostructure, was synthesized by the co-precipitation method. In this work, the formation of Bismuth-based heterojunction for charge separation is established by the excessive bismuth nitrate, which subsequently participates with the in situ growth of ultrathin hierarchical microspheres. By attenuating the thickness of BiOBr from 20 nm to 8 nm with the aid of BiON, the photogenerated charges could migrate to the active sites through shorter charge diffusion pathway. Also, the BiOBr and BiON act as an active bridge to promote the separation of electron-hole pairs, which also brings out more active sites due to its increased specific surface area. BiON/BiOBr ultrathin hierarchical microspheres exhibited enhanced visible-light photocatalytic activity for decontaminating several types of pollutants. Besides, the activity of as-prepared BiON/BiOBr was further evaluated by inhibiting the growth of kanamycin-resistant bacteria strains. This study presents a novel strategy to incorporate the crystalline bismuth hydrate nitrate into BiOBr to form ultrathin hierarchical microspheres with high surface area for environmental remediation.

Carbon nanomaterials, include carbon nanotubes and graphene nanosheets, have drawn an increasing amount of attention because of their potential applications in daily life or in providing novel therapeutic possibilities for treating diseases. However, the overall biocompatibility, the potential toxic effects of carbon nanomaterials toward human cells, and their modulations in cellular mechanism, are not fully understood. Herein, four types of carbon nanomaterials, include long and short carbon nanotubes and graphene nanosheets, at low and high concentrations, were functionalized and dispersed in the biocompatible buffer for assessment. The surface structure, the morphology, and chemical composition of carbon nanomaterials were characterized. Also, biological assays investigating cellular viability, vitality, cell cycle, and apoptotic cell death were applied on cells co-incubated with nanomaterials, to evaluate the biocompatibility of these nanomaterials in human cells. Our data suggested that even though co-incubation of nanomaterials did not seem to affect the viability of cells notably, high concentrations (50 ug/ml) of SW could lead to unhealthy cells, and we observed dramatic G2 arrest effect mediated by p21 induction in high SW incubated cells. Other nanomaterials at high concentration may also alter cell cycle profile of the cells. In summary, our data demonstrated that these nanomaterials could regulate cell cycle and lead to apoptosis at high concentrations, and the underling molecular mechanisms have been addressed. Caution should be taken on their concentration when nanomaterials are in used in future medical applications.

Hematogenous metastasis accounts for the majority of cancer-related deaths, yet the mechanism remains unclear. Circulating tumor cells (CTCs) in blood may employ different pathways to cross blood endothelial barrier and establish a metastatic niche. Several studies provide evidence that prostate cancer (PCa) cell tethering and rolling on microvascular endothelium via E-selectin/E-selectin ligand interactions under shear flow theoretically promote extravasation and contribute to the development of metastases. However, it is unknown if CTCs from PCa patients interact with E-selectin expressed on endothelium, initiating a route for tumor metastases. Here we report that CTCs derived from PCa patients showed interactions with E-selectin and E-selectin expressing endothelial cells. To examine E-selectin-mediated interactions of PCa cell lines and CTCs derived from metastatic PCa patients, we used fluorescently-labeled anti-prostate specific membrane antigen (PSMA) monoclonal antibody J591-488 which is internalized following cell-surface binding. We employed a microscale flow device consisting of E-selectin-coated microtubes and human umbilical vein endothelial cells (HUVECs) on parallel-plate flow chamber simulating vascular endothelium. We observed that J591-488 did not significantly alter the rolling behavior in PCa cells at shear stresses below 3 dyn/cm(2). CTCs obtained from 31 PCa patient samples showed that CTCs tether and stably interact with E-selectin and E-selectin expressing HUVECs at physiological shear stress. Interestingly, samples collected during disease progression demonstrated significantly more CTC/E-selectin interactions than samples during times of therapeutic response (p=0.016). Analysis of the expression of sialyl Lewis X (sLe(x)) in patient samples showed that a small subset comprising 1.9-18.8% of CTCs possess high sLe(x) expression. Furthermore, E-selectin-mediated interactions between prostate CTCs and HUVECs were diminished in the presence of anti-E-selectin neutralizing antibody. CTC-Endothelial interactions provide a novel insight into potential adhesive mechanisms of prostate CTCs as a means to initiate metastasis.

Circulating tumor cells (CTCs) are tumor cells found in the peripheral blood that putatively originate from established sites of malignancy and likely have metastatic potential. Analysis of CTCs has demonstrated promise as a prognostic marker as well as a source of identifying potential targets for novel therapeutics. Isolation and characterization of these cells for study, however, remain challenging owing to their rarity in comparison with other cellular components of the peripheral blood. Several techniques that exploit the unique biochemical properties of CTCs have been developed to facilitate their isolation. Positive selection of CTCs has been achieved using microfluidic surfaces coated with antibodies against epithelial cell markers or tumor-specific antigens such as EpCAM or prostate-specific membrane antigen (PSMA). Following isolation, characterization of CTCs may help guide clinical decision making. For instance, molecular and genetic characterization may shed light on the development of chemotherapy resistance and mechanisms of metastasis without the need for a tissue biopsy. This paper will review novel isolation techniques to capture CTCs from patients with advanced prostate cancer, as well as efforts to characterize the CTCs. We will also review how these analyzes can assist in clinical decision making. Conclusion: The study of CTCs provides insight into the molecular biology of tumors of prostate origin that will eventually guide the development of tailored therapeutics. These advances are predicated on high yield and accurate isolation techniques that exploit the unique biochemical features of these cells.

Prostate cancer progression into castration-resistant prostate cancer (CRPC) is driven by continued androgen receptor (AR) signaling despite surgical and chemical androgen ablation. The taxanes represent the only class of chemotherapy that improves overall survival in CRPC patients. Despite their success, CRPC patients do progress on taxane treatment rendering taxane-resistant tumors. The molecular mechanisms underlying clinical taxane resistance in CRPC have not been well elucidated due to the lack of available tumor tissue to study. Circulating tumor cells (CTCs) represent a liquid biopsy of the original tumor and isolation of them can lead to their molecular characterization potentially revealing predictive biomarkers for taxane sensitivity or resistance. Here, we use a geometrically enhanced differential immunocapture (GEDI) microfluidic device that couples an anti-prostate specific membrane antigen (PSMA) antibody with optimized 3D geometry to capture and isolate live CTCs from whole blood of CRPC patients. The GEDI-microfluidic device was shown to have a 2-400 fold higher sensitivity for CTC capture than the FDA-approved CellSearch® system. We have previously shown that CRPC patient CTCs can be used to derive functional information that correlates to clinical response to taxane chemotherapy, namely AR subcellular localization status. We have developed a suite of other functional assays that can be performed on live GEDI-captured CTCs that enable their molecular characterization and allow us to test specific mechanistic hypotheses based on our extensive preclinical data. Included, and herein described, are the determination of AR subcellular localization, extent of effective drug-target engagement assessed by microtubule bundling, identification of RNA species relevant to the mechanism of taxane resistance and computer vision algorithms that will allow for enriched and automated analysis of high-volume image sets of GEDI-captured CTCs. In addition, we will be testing the hypothesis that distinct AR splice variants may affect patient sensitivity to taxane-based chemotherapy. This suite of assays are being rigorously applied in a phase II clinical trial in which chemotherapy-naïve CRPC patients will be initially treated with either docetaxel or cabazitaxel and clinically evaluated for an early switch to the other taxane following disease progression. This prospective, randomized, multi-site clinical trial will enroll 100 CRPC patients within one year. Patients will be followed until relapse and each patient will have 15 independent GEDI assays performed across five time points from baseline to chemotherapy crossover to relapse. The depth of coverage this suite of assays provides will offer unique insights for potential mechanisms of clinical taxane resistance and predictive biomarkers for taxane sensitivity in CRPC patient CTCs. Citation Format: Matthew S. Sung, Ada Gjyrezi, Guang Yu Lee, Alexandre Matov, Giuseppe Galletti, Matthew Loftus, Yusef Syed, Timothy Lannin, Atanas Hristov, Christopher Mason, Scott Tagawa, Brian Kirby, David Nanus, Paraskevi Giannakakou. Using CTCs to interrogate mechanisms of taxane resistance in the prospective TAXYNERGY clinical trial in prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3492. doi:10.1158/1538-7445.AM2013-3492

Androgen receptor (AR) signaling drives castration resistant prostate cancer (CRPC) disease progression. The taxanes (TXN) represent the only class of chemotherapy agents that improve survival in CRPC. Our recent work revealed that TXNs work by blocking AR nuclear accumulation and signaling downstream of microtubule (MT) stabilization. Moreover, we showed that AR cytoplasmic sequestration in CRPC patients’ circulating tumor cells (CTCs) significantly correlated with clinical response to TXN therapy. Our data, suggest that TXN-induced blockade of AR nuclear accumulation partly underlies the clinical activity of TXNs in CRPC. Recently several AR splice variants, lacking the ligand binding domain, have been identified in CRPC. These variants are constitutively active in the nucleus and likely to be insensitive to current therapies targeting either ligand biosynthesis or ligand-receptor interactions. Currently, the role of AR splice variants on TXN chemotherapy response in CRPC is unknown. Our preliminary data in preclinical PC models suggest that certain AR variants depend on MTs for nuclear accumulation and are TXN sensitive while others are MT-independent and TXN-insensitive. In this study we are prospectively evaluating whether the presence of AR splice variants and the integrity of the MT-AR axis in CTCs can predict clinical response to TXN chemotherapy in CRPC. CTCs are captured by the geometrically enhanced differential immunocapture (GEDI) microfluidic device that uses PSMA-based immunocapture and size-selective cell transport for maximum specificity of CRPC-derived CTCs. We captured viable CTCs from CRPC patients (median 54 CTC/ml) and performed molecular and functional analyses. Comparison of CTCs captured by CellSearch or GEDI, using same day blood draw from 30 CRPC patients, revealed a 2-400 fold increase in CTC numbers with the GEDI. GEDI-capture of live CTCs allowed us to evaluate ex-vivo TXN sensitivity and determine drug-target-engagement (DTE) by MT bundling. CTCs isolated from docetaxel (DTX)-resistant CRPC patients did not show any evidence of DTX activity in this assay suggesting that it can be used to monitor patient's response in real time. In addition, we have miniaturized RNA sequencing (RNA-Seq) to enable molecular analyses of GEDI-captured CTCs. RNA-Seq of 50 GEDI-captured LNCap 86.2 cells spiked into 1 ml healthy donor blood, detected both the T868A AR point mutation and ARv567 splice variant. In an ongoing prospective clinical study, we are capturing CTCs from CRPC patients treated with TXNs and are evaluating the integrity of the MT-AR signaling axis, in relation to clinical response to therapy. Our CTC analyses include RNA-Seq for the detection of AR splice variants, alterations in tubulin isotypes and MT related proteins, and multiplex confocal microscopy of DTE and AR localization. These analyses are performed longitudinally and may identify biomarkers predictive of clinical TXN efficacy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3634. doi:1538-7445.AM2012-3634