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Libo Zhao,
Yi-Tsung Lu,
Fuqiang Li,
Kui Wu,
Shuang Hou,
Juehua Yu,
Qinglin Shen,
Dongxia Wu,
Min Song,
Wei-Han Ouyang,
Zheng Luo,
Tom Lee,
Xiaohong Fang,
Chen Shao,
Xun Xu,
Mitch A Garcia,
Leland W K Chung,
Matthew Rettig, Hsian-Rong Tseng,
Edwin M Posadas
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ABSTRACT: Handpick single cancer cells: A modified NanoVelcro Chip is coupled with ArcturusXT laser capture microdissection (LCM) technology to enable the detection and isolation of single circulating tumor cells (CTCs) from patients with prostate cancer (PC). This new approach paves the way for conducting next-generation sequencing (NGS) on single CTCs.
Advanced Materials 03/2013; · 13.88 Impact Factor
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Jae-Hyun Lee,
Kuan-Ju Chen,
Seung-Hyun Noh,
Mitch André Garcia,
Hao Wang,
Wei-Yu Lin,
Heeyeong Jeong,
Brian Junoh Kong,
David B Stout,
Jinwoo Cheon, Hsian-Rong Tseng
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ABSTRACT: On-demand drug release: Magnetothermally responsive drug-encapsulated supramolecular nanoparticles for on-demand drug release in vivo have been developed. The remote application of an alternative magnetic field heats the magnetic particles that effectively trigger the release of the drug. An acute drug concentration can be delivered to the tumor in vivo, resulting in an improved therapeutic outcome.
Angewandte Chemie International Edition 03/2013; · 13.45 Impact Factor
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Qinglin Shen,
Li Xu,
Libo Zhao,
Dongxia Wu,
Yunshan Fan,
Yiliang Zhou,
Wei-Han Ouyang,
Xiaochun Xu,
Zhen Zhang,
Min Song,
Tom Lee,
Mitch A Garcia,
Bin Xiong,
Shuang Hou, Hsian-Rong Tseng,
Xiaohong Fang
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ABSTRACT: A platform for capture and release of circulating tumor cells (CTCs) is demonstrated by utilizing aptamer grafted silicon nanowires. Here, single-stranded DNA-aptamers are generated via the Cell-SELEX process to serve as capture agents, allowing specific capture and release of non-small cell lung cancer (NSCLC) CTCs from whole-blood samples with minimum contamination and negligible disruption to CTC viability and functions.
Advanced Materials 03/2013; · 13.88 Impact Factor
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Shuang Hou,
Libo Zhao,
Qinglin Shen,
Juehua Yu,
Charles Ng,
Xiangju Kong,
Dongxia Wu,
Min Song,
Xiaohong Shi,
Xiaochun Xu,
Wei-Han Ouyang,
Rongxian He,
Xing-Zhong Zhao,
Tom Lee,
F Charles Brunicardi,
Mitch André Garcia,
Antoni Ribas,
Roger S Lo, Hsian-Rong Tseng
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ABSTRACT: Confined to one cell: A method to detect and isolate single circulating melanoma cells (CMCs; see figure) has been produced by integrating a polymer-nanofiber-embedded nanovelcro cell-affinity assay with a laser microdissection (LMD) technique. This method is able to separate CMCs from normal white blood cells (WBCs) and sequence individual cells for a specific mutation related to cancer progression, allowing for more personalized cancer therapy.
Angewandte Chemie International Edition 02/2013; · 13.45 Impact Factor
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Shuang Hou,
Haichao Zhao,
Libo Zhao,
Qinglin Shen,
Kevin S Wei,
Daniel Y Suh,
Aiko Nakao,
Mitch A Garcia,
Min Song,
Tom Lee,
Bin Xiong,
Shyh-Chyang Luo, Hsian-Rong Tseng,
Hsiao-Hua Yu
[show abstract]
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ABSTRACT: A platform for capture and release of circulating tumor cells is demonstrated by utilizing polymer grafted silicon nanowires. In this platform, integration of ligand-receptor recognition, nanostructure amplification, and thermal responsive polymers enables a highly efficient and selective capture of cancer cells. Subsequently, these captured cells are released upon a physical stimulation with outstanding cell viability.
Advanced Materials 12/2012; · 13.88 Impact Factor
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Jing Sun,
Michael D Masterman-Smith,
Nicholas A Graham,
Jing Jiao,
Jack Mottahedeh,
Dan R Laks,
Minori Ohashi,
Jason DeJesus,
Ken-ichiro Kamei,
Ki-Bum Lee, [......],
R Michael van Dam,
Michael E Phelps,
Hong Wu,
Linda M Liau,
Paul S Mischel,
Jorge A Lazareff,
Harley I Kornblum,
William H Yong,
Thomas G Graeber, Hsian-Rong Tseng
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Jing Sun,
Michael D Masterman-Smith,
Nicholas A Graham,
Jing Jiao,
Jack Mottahedeh,
Dan R Laks,
Minori Ohashi,
Jason DeJesus,
Ken-ichiro Kamei,
Ki-Bum Lee, [......],
R Michael van Dam,
Michael E Phelps,
Hong Wu,
Linda M Liau,
Paul S Mischel,
Jorge A Lazareff,
Harley I Kornblum,
William H Yong,
Thomas G Graeber, Hsian-Rong Tseng
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Kuan-Ju Chen,
Li Tang,
Mitch André Garcia,
Hao Wang,
Hua Lu,
Wei-Yu Lin,
Shuang Hou,
Qian Yin,
Clifton K-F Shen,
Jianjun Cheng, Hsian-Rong Tseng
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ABSTRACT: Nanomaterials have been increasingly employed as drug(s)-incorporated vectors for drug delivery due to their potential of maximizing therapeutic efficacy while minimizing systemic side effects. However, there have been two main challenges for these vectors: (i) the existing synthetic approaches are cumbersome and incapable of achieving precise control of their structural properties, which will affect their biodistribution and therapeutic efficacies, and (ii) lack of an early checkpoint to quickly predict which drug(s)-incorporated vectors exhibit optimal therapeutic outcomes. In this work, we utilized a new rational developmental approach to rapidly screen nanoparticle (NP)-based cancer therapeutic agents containing a built-in companion diagnostic utility for optimal therapeutic efficacy. The approach leverages the advantages of a self-assembly synthetic method for preparation of two different sizes of drug-incorporated supramolecular nanoparticles (SNPs), and a positron emission tomography (PET) imaging-based biodistribution study to quickly evaluate the accumulation of SNPs at a tumor site in vivo and select the favorable SNPs for in vivo therapeutic study. Finally, the enhanced in vivo anti-tumor efficacy of the selected SNPs was validated by tumor reduction/inhibition studies. We foresee our rational developmental approach providing a general strategy in the search of optimal therapeutic agents among the diversity of NP-based therapeutic agents.
Biomaterials 11/2011; 33(4):1162-9. · 7.40 Impact Factor
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Kan Liu,
Eric J Lepin,
Ming-Wei Wang,
Feng Guo,
Wei-Yu Lin,
Yi-Chun Chen,
Shannon J Sirk,
Sebastian Olma,
Michael E Phelps,
Xing-Zhong Zhao, Hsian-Rong Tseng,
R Michael van Dam,
Anna M Wu,
Clifton K-F Shen
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ABSTRACT: Methods for tagging biomolecules with fluorine 18 as immuno-positron emission tomography (immunoPET) tracers require tedious optimization of radiolabeling conditions and can consume large amounts of scarce biomolecules. We describe an improved method using a digital microfluidic droplet generation (DMDG) chip, which provides computer-controlled metering and mixing of 18F tag, biomolecule, and buffer in defined ratios, allowing rapid scouting of reaction conditions in nanoliter volumes. The identified optimized conditions were then translated to bench-scale 18F labeling of a cancer-specific engineered antibody fragments, enabling microPET imaging of tumors in xenografted mice at 0.5 to 4 hours postinjection.
Molecular Imaging 06/2011; 10(3):168-76, 1-7. · 3.18 Impact Factor
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ABSTRACT: An integrated β-camera and microfluidic chip was developed that is capable of quantitative imaging of glycolysis radioassays using (18)F-FDG in small cell populations down to a single cell. This paper demonstrates that the integrated system enables digital control and quantitative measurements of glycolysis in B-Raf(V600E)-mutated melanoma cell lines in response to specific B-Raf inhibition.
The β-camera uses a position-sensitive avalanche photodiode to detect charged particle-emitting probes within a microfluidic chip. The integrated β-camera and microfluidic chip system was calibrated, and the linearity was measured using 4 different melanoma cell lines (M257, M202, M233, and M229). Microfluidic radioassays were performed with cell populations ranging from hundreds of cells down to a single cell. The M229 cell line has a homozygous B-Raf(V600E) mutation and is highly sensitive to a B-Raf inhibitor, PLX4032. A microfluidic radioassay was performed over the course of 3 days to assess the cytotoxicity of PLX4032 on cellular (18)F-FDG uptake.
The β-camera is capable of imaging radioactive uptake of (18)F-FDG in microfluidic chips. (18)F-FDG uptake for a single cell was measured using a radioactivity concentration of 37 MBq/mL during the radiotracer incubation period. For in vitro cytotoxicity monitoring, the β-camera showed that exposure to 1 μM PLX4032 for 3 days decreased the (18)F-FDG uptake per cell in highly sensitive M229 cells, compared with vehicle controls.
The integrated β-camera and microfluidic chip can provide digital control of live cell cultures and allow in vitro quantitative radioassays for multiple samples simultaneously.
Journal of Nuclear Medicine 05/2011; 52(5):815-21. · 6.38 Impact Factor
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Shutao Wang,
Kan Liu,
Jian Liu,
Zeta T F Yu,
Xiaowen Xu,
Libo Zhao,
Tom Lee,
Eun Kyung Lee,
Jean Reiss,
Yi-Kuen Lee,
Leland W K Chung,
Jiaoti Huang,
Matthew Rettig,
David Seligson,
Kumaran N Duraiswamy,
Clifton K-F Shen, Hsian-Rong Tseng
Angewandte Chemie International Edition 03/2011; · 13.45 Impact Factor
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Angewandte Chemie International Edition 03/2011; 50(13):3058-62. · 13.45 Impact Factor
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Kuan-Ju Chen,
Stephanie M Wolahan,
Hao Wang,
Chao-Hsiung Hsu,
Hsing-Wei Chang,
Armando Durazo,
Lian-Pin Hwang,
Mitch A Garcia,
Ziyue K Jiang,
Lily Wu,
Yung-Ya Lin, Hsian-Rong Tseng
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ABSTRACT: We introduce a new category of nanoparticle-based T(1) MRI contrast agents (CAs) by encapsulating paramagnetic chelated gadolinium(III), i.e., Gd(3+)·DOTA, through supramolecular assembly of molecular building blocks that carry complementary molecular recognition motifs, including adamantane (Ad) and β-cyclodextrin (CD). A small library of Gd(3+)·DOTA-encapsulated supramolecular nanoparticles (Gd(3+)·DOTA⊂SNPs) was produced by systematically altering the molecular building block mixing ratios. A broad spectrum of relaxation rates was correlated to the resulting Gd(3+)·DOTA⊂SNP library. Consequently, an optimal synthetic formulation of Gd(3+)·DOTA⊂SNPs with an r(1) of 17.3 s(-1) mM(-1) (ca. 4-fold higher than clinical Gd(3+) chelated complexes at high field strengths) was identified. T(1)-weighted imaging of Gd(3+)·DOTA⊂SNPs exhibits an enhanced sensitivity with a contrast-to-noise ratio (C/N ratio) ca. 3.6 times greater than that observed for free Gd(3+)·DTPA. A Gd(3+)·DOTA⊂SNPs solution was injected into foot pads of mice, and MRI was employed to monitor dynamic lymphatic drainage of the Gd(3+)·DOTA⊂SNPs-based CA. We observe an increase in signal intensity of the brachial lymph node in T(1)-weighted imaging after injecting Gd(3+)·DOTA⊂SNPs but not after injecting Gd(3+)·DTPA. The MRI results are supported by ICP-MS analysis ex vivo. These results show that Gd(3+)·DOTA⊂SNPs not only exhibits enhanced relaxivity and high sensitivity but also can serve as a potential tool for diagnosis of cancer metastasis.
Biomaterials 03/2011; 32(8):2160-5. · 7.40 Impact Factor
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ABSTRACT: Cell-based arrays offer powerful tools for genomics/proteomics and drug discovery, and are widely applicable for most cell lines. However, it is challenging to apply cell-based arrays for in vitro diagnosis due to limited amount of patient samples. Here, we utilized and demonstrated microfluidic image cytometry (MIC), capable of quantitative, single-cell profiling of multiple signaling molecules using only 300-3,000 cells from clinical brain tumor specimens for in vitro molecular diagnosis. First, we characterized the PI3K/AKT/mTOR pathway, which is often over-activated in the brain tumors, in U87 brain tumor cell lines by measuring EGFR, PTEN, pAKT, and pS6 with a MIC platform, and applied this measurement to clinical brain tumor specimens. In conjunction with statistical analysis, we were able to characterize extensive inter- and intra-tumoral molecular heterogeneity.
Methods in molecular biology (Clifton, N.J.) 01/2011; 706:191-206.
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ABSTRACT: Some efficient diagnosis and therapy systems require the isolation and quantification of circulating tumor cells (CTCs), since these species are important "biomarkers" for monitoring cancer metastasis and prognosis. Existing techniques for isolating/counting CTCs include immunomagnetic-bead-based separation and microfluidic capture. However, some of these techniques have low capture efficiency and low specificity. Through the use of a three-dimensional (3D) nanostructured substrate - specifically, a silicon-nanowire (SiNW) array coated with epithelial-cell-adhesion-molecule antibodies (anti-EpCAM) - we show that CTCs can be captured efficiently and specifically. Unlike conventional methods for isolating CTCs that depend on collision frequency and contact duration, nanoscaled local topographic interactions between the CTCs and the substrate increase their binding and markedly enhance capture efficiency.
Methods in molecular biology (Clifton, N.J.) 01/2011; 726:141-50.
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ABSTRACT: Controlling the size distribution of polymer-based nanoparticles is a challenging task due to their flexible core and surface structures. To accomplish such as task requires very precise control at the molecular level. Here we demonstrate a new approach whereby uniform-sized supramolecular nanoparticles (SNPs) can be reliably generated using a digital microfluidic droplet generator (DMDG) chip. A microfluidic environment enabled precise control over the processing parameters, and therefore high batch-to-batch reproducibility and robust production of SNPs with a very narrow size distribution could be realized. Digitally adjustment of the mixing ratios of the building blocks on the DMDG chip allowed us to rapidly scan a variety of synthesis conditions without consuming significant amounts of reagents. Nearly uniform SNPs with sizes ranging from 35 to 350 nm were obtained and characterized by transmission electron microscopy and dynamic light scattering. In addition, we could fine-tune the surface chemistry of the SNPs by incorporating an additional building block functionalized with specific ligands for targeting cells. The sizes and surface properties of these SNPs correlated strongly with their cell uptake efficiencies. This study showed a feasible method for microfluidic-assisted SNP production and provided a great means for preparing size-controlled SNPs with desired surface ligand coverage.
Nanotechnology 11/2010; 21(44):445603. · 3.98 Impact Factor
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Cong Fang,
Yanju Wang,
Nam T Vu,
Wei-Yu Lin,
Yao-Te Hsieh,
Liudmilla Rubbi,
Michael E Phelps,
Markus Müschen,
Yong-Mi Kim,
Arion F Chatziioannou, Hsian-Rong Tseng,
Thomas G Graeber
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ABSTRACT: Oncogenic kinase activity and the resulting aberrant growth and survival signaling are a common driving force of cancer. Accordingly, many successful molecularly targeted anticancer therapeutics are directed at inhibiting kinase activity. To assess kinase activity in minute patient samples, we have developed an immunocapture-based in vitro kinase assay on an integrated polydimethylsiloxane microfluidics platform that can reproducibly measure kinase activity from as few as 3,000 cells. For this platform, we adopted the standard radiometric (32)P-ATP-labeled phosphate transfer assay. Implementation on a microfluidic device required us to develop methods for repeated trapping and mixing of solid-phase affinity microbeads. We also developed a solid-state beta-particle camera imbedded directly below the microfluidic device for real-time quantitative detection of the signal from this and other microfluidic radiobioassays. We show that the resulting integrated device can measure ABL kinase activity from BCR-ABL-positive leukemia patient samples. The low sample input requirement of the device creates new potential for direct kinase activity experimentation and diagnostics on patient blood, bone marrow, and needle biopsy samples.
Cancer Research 11/2010; 70(21):8299-308. · 7.86 Impact Factor
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ABSTRACT: Using liquid slugs as microreactors and microvessels enable precise control over the conditions of their contents on short-time scales for a wide variety of applications. Particularly for screening applications, there is a need for control of slug parameters such as size and composition. We describe a new microfluidic approach for creating slugs in air, each comprising a size and composition that can be selected individually for each slug. Two-component slugs are formed by first metering the desired volume of each reagent, merging the two volumes into an end-to-end slug, and propelling the slug to induce mixing. Volume control is achieved by a novel mechanism: two closed chambers on the chip are initially filled with air, and a valve in each is briefly opened to admit one of the reagents. The pressure of each reagent can be individually selected and determines the amount of air compression, and thus the amount of liquid that is admitted into each chamber. We describe the theory of operation, characterize the slug generation chip, and demonstrate the creation of slugs of different compositions. The use of microvalves in this approach enables robust operation with different liquids, and also enables one to work with extremely small samples, even down to a few slug volumes. The latter is important for applications involving precious reagents such as optimizing the reaction conditions for radiolabeling biological molecules as tracers for positron emission tomography. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10404-010-0617-0) contains supplementary material, which is available to authorized users.
Microfluidics and Nanofluidics 10/2010; 9(4-5):933-943. · 3.37 Impact Factor
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Hao Wang,
Kan Liu,
Kuan-Ju Chen,
Yujie Lu,
Shutao Wang,
Wei-Yu Lin,
Feng Guo,
Ken-ichiro Kamei,
Yi-Chun Chen,
Minori Ohashi,
Mingwei Wang,
Mitch André Garcia,
Xing-Zhong Zhao,
Clifton K-F Shen, Hsian-Rong Tseng
[show abstract]
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ABSTRACT: Nanoparticles are regarded as promising transfection reagents for effective and safe delivery of nucleic acids into a specific type of cells or tissues providing an alternative manipulation/therapy strategy to viral gene delivery. However, the current process of searching novel delivery materials is limited due to conventional low-throughput and time-consuming multistep synthetic approaches. Additionally, conventional approaches are frequently accompanied with unpredictability and continual optimization refinements, impeding flexible generation of material diversity creating a major obstacle to achieving high transfection performance. Here we have demonstrated a rapid developmental pathway toward highly efficient gene delivery systems by leveraging the powers of a supramolecular synthetic approach and a custom-designed digital microreactor. Using the digital microreactor, broad structural/functional diversity can be programmed into a library of DNA-encapsulated supramolecular nanoparticles (DNA⊂SNPs) by systematically altering the mixing ratios of molecular building blocks and a DNA plasmid. In vitro transfection studies with DNA⊂SNPs library identified the DNA⊂SNPs with the highest gene transfection efficiency, which can be attributed to cooperative effects of structures and surface chemistry of DNA⊂SNPs. We envision such a rapid developmental pathway can be adopted for generating nanoparticle-based vectors for delivery of a variety of loads.
ACS Nano 10/2010; 4(10):6235-43. · 10.77 Impact Factor
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Jing Sun,
Michael D Masterman-Smith,
Nicholas A Graham,
Jing Jiao,
Jack Mottahedeh,
Dan R Laks,
Minori Ohashi,
Jason DeJesus,
Ken-ichiro Kamei,
Ki-Bum Lee, [......],
R Michael van Dam,
Michael E Phelps,
Hong Wu,
Linda M Liau,
Paul S Mischel,
Jorge A Lazareff,
Harley I Kornblum,
William H Yong,
Thomas G Graeber, Hsian-Rong Tseng
[show abstract]
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ABSTRACT: The clinical practice of oncology is being transformed by molecular diagnostics that will enable predictive and personalized medicine. Current technologies for quantitation of the cancer proteome are either qualitative (e.g., immunohistochemistry) or require large sample sizes (e.g., flow cytometry). Here, we report a microfluidic platform-microfluidic image cytometry (MIC)-capable of quantitative, single-cell proteomic analysis of multiple signaling molecules using only 1,000 to 2,800 cells. Using cultured cell lines, we show simultaneous measurement of four critical signaling proteins (EGFR, PTEN, phospho-Akt, and phospho-S6) within the oncogenic phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway. To show the clinical application of the MIC platform to solid tumors, we analyzed a panel of 19 human brain tumor biopsies, including glioblastomas. Our MIC measurements were validated by clinical immunohistochemistry and confirmed the striking intertumoral and intratumoral heterogeneity characteristic of glioblastoma. To interpret the multiparameter, single-cell MIC measurements, we adapted bioinformatic methods including self-organizing maps that stratify patients into clusters that predict tumor progression and patient survival. Together with bioinformatic analysis, the MIC platform represents a robust, enabling in vitro molecular diagnostic technology for systems pathology analysis and personalized medicine.
Cancer Research 08/2010; 70(15):6128-38. · 7.86 Impact Factor
