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Circulating Tumor Cells: Diagnostic and Therapeutic Applications.
Lin E, Cao T, Nagrath S, King MR
(2018) Annu Rev Biomed Eng 20: 329-352
MeSH Terms: Animals, Cell Separation, Electrophoresis, Epithelial Cells, Filtration, Humans, Lab-On-A-Chip Devices, Lymph Nodes, Lymphatic Metastasis, Lymphatic System, Neoplasm Metastasis, Neoplasms, Neoplastic Cells, Circulating, Prognosis, TNF-Related Apoptosis-Inducing Ligand
Show Abstract · Added April 15, 2019
Metastasis contributes to poor prognosis in many types of cancer and is the leading cause of cancer-related deaths. Tumor cells metastasize to distant sites via the circulatory and lymphatic systems. In this review, we discuss the potential of circulating tumor cells for diagnosis and describe the experimental therapeutics that aim to target these disseminating cancer cells. We discuss the advantages and limitations of such strategies and how they may lead to the development of the next generation of antimetastasis treatments.
0 Communities
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15 MeSH Terms
Colistin-Functionalized Nanoparticles for the Rapid Capture of Acinetobacter baumannii.
Miller SE, Bell CS, Mejias R, McClain MS, Cover TL, Giorgio TD
(2016) J Biomed Nanotechnol 12: 1806-19
MeSH Terms: Acinetobacter baumannii, Bacteriological Techniques, Cell Separation, Colistin, Gold, Metal Nanoparticles
Show Abstract · Added March 21, 2018
Gold nanoparticles (AuNPs) were functionalized for rapid binding of Acinetobacter baumannii (A. baumannii), a Gram-negative bacterium. AuNPs were functionalized with colistin (Col), a polycationic antibiotic, using a two-step self-assembly process, in which heterobifunctional polyethylene glycol (PEG) was used as a linker. Colistin was successfully conjugated to the AuNPs (Col-PEG-AuNP), as validated by dynamic light scattering (DLS) and proton nuclear magnetic resonance (H1 NMR). High angle annular dark field scanning transmission electron microscopy (HAADF-STEM) images, acquired simultaneously with X-ray energy dispersive spectroscopy (EDS) data, confirmed binding of Col-PEG-AuNPs to the cell envelope of A. baumannii. Results generated from a binding assay indicated that Col-PEG-AuNP complexation with A. baumannii occurred rapidly and reached half-maximum saturation in approximately 7 minutes, on average, for all A. baumannii strains evaluated. Quantitative measurement of the kinetics of Col-PEG-AuNP binding to A. baumannii is essential to inform the design of colistin-functionalized magnetic nanoparticles for magnetic separation of nanoparticle-bound A. baumannii.
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6 MeSH Terms
Differentiation of Human Pluripotent Stem Cells into Functional Lung Alveolar Epithelial Cells.
Jacob A, Morley M, Hawkins F, McCauley KB, Jean JC, Heins H, Na CL, Weaver TE, Vedaie M, Hurley K, Hinds A, Russo SJ, Kook S, Zacharias W, Ochs M, Traber K, Quinton LJ, Crane A, Davis BR, White FV, Wambach J, Whitsett JA, Cole FS, Morrisey EE, Guttentag SH, Beers MF, Kotton DN
(2017) Cell Stem Cell 21: 472-488.e10
MeSH Terms: Base Sequence, Cell Differentiation, Cell Line, Cell Proliferation, Cell Self Renewal, Cell Separation, Epithelial Cells, Gene Expression Profiling, Genes, Reporter, Humans, Lung Diseases, Models, Biological, Pluripotent Stem Cells, Pulmonary Alveoli, Pulmonary Surfactants, Thyroid Nuclear Factor 1, Time Factors, Wnt Proteins, Wnt Signaling Pathway
Show Abstract · Added April 1, 2019
Lung alveoli, which are unique to air-breathing organisms, have been challenging to generate from pluripotent stem cells (PSCs) in part because there are limited model systems available to provide the necessary developmental roadmaps for in vitro differentiation. Here we report the generation of alveolar epithelial type 2 cells (AEC2s), the facultative progenitors of lung alveoli, from human PSCs. Using multicolored fluorescent reporter lines, we track and purify human SFTPC+ alveolar progenitors as they emerge from endodermal precursors in response to stimulation of Wnt and FGF signaling. Purified PSC-derived SFTPC+ cells form monolayered epithelial "alveolospheres" in 3D cultures without the need for mesenchymal support, exhibit self-renewal capacity, and display additional AEC2 functional capacities. Footprint-free CRISPR-based gene correction of PSCs derived from patients carrying a homozygous surfactant mutation (SFTPB) restores surfactant processing in AEC2s. Thus, PSC-derived AEC2s provide a platform for disease modeling and future functional regeneration of the distal lung.
Copyright © 2017 Elsevier Inc. All rights reserved.
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MeSH Terms
Preparing Viable Single Cells from Human Tissue and Tumors for Cytomic Analysis.
Leelatian N, Doxie DB, Greenplate AR, Sinnaeve J, Ihrie RA, Irish JM
(2017) Curr Protoc Mol Biol 118: 25C.1.1-25C.1.23
MeSH Terms: Cell Separation, Cryopreservation, Flow Cytometry, Humans, Neoplasms, Single-Cell Analysis
Show Abstract · Added April 4, 2017
Mass cytometry is a single-cell biology technique that samples >500 cells per second, measures >35 features per cell, and is sensitive across a dynamic range of >10 relative intensity units per feature. This combination of technical assets has powered a series of recent cytomic studies where investigators used mass cytometry to measure protein and phospho-protein expression in millions of cells, characterize rare cell types in healthy and diseased tissues, and reveal novel, unexpected cells. However, these advances largely occurred in studies of blood, lymphoid tissues, and bone marrow, since the cells in these tissues are readily obtained in single-cell suspensions. This unit establishes a primer for single-cell analysis of solid tumors and tissues, and has been tested with mass cytometry. The cells obtained from these protocols can be fixed for study, cryopreserved for long-term storage, or perturbed ex vivo to dissect responses to stimuli and inhibitors. © 2017 by John Wiley & Sons, Inc.
Copyright © 2017 John Wiley & Sons, Inc.
3 Communities
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6 MeSH Terms
Mimicry of an HIV broadly neutralizing antibody epitope with a synthetic glycopeptide.
Alam SM, Aussedat B, Vohra Y, Meyerhoff RR, Cale EM, Walkowicz WE, Radakovich NA, Anasti K, Armand L, Parks R, Sutherland L, Scearce R, Joyce MG, Pancera M, Druz A, Georgiev IS, Von Holle T, Eaton A, Fox C, Reed SG, Louder M, Bailer RT, Morris L, Abdool-Karim SS, Cohen M, Liao HX, Montefiori DC, Park PK, Fernández-Tejada A, Wiehe K, Santra S, Kepler TB, Saunders KO, Sodroski J, Kwong PD, Mascola JR, Bonsignori M, Moody MA, Danishefsky S, Haynes BF
(2017) Sci Transl Med 9:
MeSH Terms: Animals, Antibodies, Neutralizing, Antibody Specificity, B-Lymphocytes, Cell Lineage, Cell Separation, Clone Cells, Epitopes, Glycopeptides, HIV Antigens, HIV Envelope Protein gp120, HIV-1, Macaca mulatta, Molecular Mimicry, Protein Domains, Protein Multimerization
Show Abstract · Added May 3, 2017
A goal for an HIV-1 vaccine is to overcome virus variability by inducing broadly neutralizing antibodies (bnAbs). One key target of bnAbs is the glycan-polypeptide at the base of the envelope (Env) third variable loop (V3). We have designed and synthesized a homogeneous minimal immunogen with high-mannose glycans reflective of a native Env V3-glycan bnAb epitope (Man-V3). V3-glycan bnAbs bound to Man-V3 glycopeptide and native-like gp140 trimers with similar affinities. Fluorophore-labeled Man-V3 glycopeptides bound to bnAb memory B cells and were able to be used to isolate a V3-glycan bnAb from an HIV-1-infected individual. In rhesus macaques, immunization with Man-V3 induced V3-glycan-targeted antibodies. Thus, the Man-V3 glycopeptide closely mimics an HIV-1 V3-glycan bnAb epitope and can be used to isolate V3-glycan bnAbs.
Copyright © 2017, American Association for the Advancement of Science.
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16 MeSH Terms
Identification of a CD4-Binding-Site Antibody to HIV that Evolved Near-Pan Neutralization Breadth.
Huang J, Kang BH, Ishida E, Zhou T, Griesman T, Sheng Z, Wu F, Doria-Rose NA, Zhang B, McKee K, O'Dell S, Chuang GY, Druz A, Georgiev IS, Schramm CA, Zheng A, Joyce MG, Asokan M, Ransier A, Darko S, Migueles SA, Bailer RT, Louder MK, Alam SM, Parks R, Kelsoe G, Von Holle T, Haynes BF, Douek DC, Hirsch V, Seaman MS, Shapiro L, Mascola JR, Kwong PD, Connors M
(2016) Immunity 45: 1108-1121
MeSH Terms: Antibodies, Neutralizing, Antibody Specificity, Binding Sites, Antibody, CD4-Positive T-Lymphocytes, Cell Separation, HIV Antibodies, HIV Envelope Protein gp120, HIV Infections, HIV-1, Humans
Show Abstract · Added May 3, 2017
Detailed studies of the broadly neutralizing antibodies (bNAbs) that underlie the best available examples of the humoral immune response to HIV are providing important information for the development of therapies and prophylaxis for HIV-1 infection. Here, we report a CD4-binding site (CD4bs) antibody, named N6, that potently neutralized 98% of HIV-1 isolates, including 16 of 20 that were resistant to other members of its class. N6 evolved a mode of recognition such that its binding was not impacted by the loss of individual contacts across the immunoglobulin heavy chain. In addition, structural analysis revealed that the orientation of N6 permitted it to avoid steric clashes with glycans, which is a common mechanism of resistance. Thus, an HIV-1-specific bNAb can achieve potent, near-pan neutralization of HIV-1, making it an attractive candidate for use in therapy and prophylaxis.
Published by Elsevier Inc.
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10 MeSH Terms
Autofluorescence flow sorting of breast cancer cell metabolism.
Shah AT, Cannon TM, Higginbotham JN, Coffey RJ, Skala MC
(2017) J Biophotonics 10: 1026-1033
MeSH Terms: Breast Neoplasms, Cell Line, Tumor, Cell Separation, Flavin-Adenine Dinucleotide, Flow Cytometry, Fluorescence, Humans, NADP
Show Abstract · Added May 3, 2017
Clinical cancer treatment aims to target all cell subpopulations within a tumor. Autofluorescence microscopy of the metabolic cofactors NAD(P)H and FAD has shown sensitivity to anti-cancer treatment response. Alternatively, flow cytometry is attractive for high throughput analysis and flow sorting. This study measures cellular autofluorescence in three flow cytometry channels and applies cellular autofluorescence to sort a heterogeneous mixture of breast cancer cells into subpopulations enriched for each phenotype. Sorted cells were grown in culture and sorting was validated by morphology, autofluorescence microscopy, and receptor expression. Ultimately, this method could be applied to improve drug development and personalized treatment planning.
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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8 MeSH Terms
Hypoxia and Reactive Oxygen Species Homeostasis in Mesenchymal Progenitor Cells Define a Molecular Mechanism for Fracture Nonunion.
Muinos-López E, Ripalda-Cemboráin P, López-Martínez T, González-Gil AB, Lamo-Espinosa JM, Valentí A, Mortlock DP, Valentí JR, Prósper F, Granero-Moltó F
(2016) Stem Cells 34: 2342-53
MeSH Terms: Animals, Bone Morphogenetic Protein 2, Cell Hypoxia, Cell Separation, Disulfides, Fracture Healing, Fractures, Ununited, Homeostasis, Humans, Hypoxia-Inducible Factor 1, alpha Subunit, Imidazoles, Male, Mesenchymal Stem Cells, Mice, Inbred C57BL, Osteogenesis, Oxidative Stress, Periosteum, Reactive Oxygen Species
Show Abstract · Added February 3, 2017
Fracture nonunion is a major complication of bone fracture regeneration and repair. The molecular mechanisms that result in fracture nonunion appearance are not fully determined. We hypothesized that fracture nonunion results from the failure of hypoxia and hematoma, the primary signals in response to bone injury, to trigger Bmp2 expression by mesenchymal progenitor cells (MSCs). Using a model of nonstabilized fracture healing in transgenic 5'Bmp2BAC mice we determined that Bmp2 expression appears in close association with hypoxic tissue and hematoma during the early phases of fracture healing. In addition, BMP2 expression is induced when human periosteum explants are exposed to hypoxia ex vivo. Transient interference of hypoxia signaling in vivo with PX-12, a thioredoxin inhibitor, results in reduced Bmp2 expression, impaired fracture callus formation and atrophic-like nonunion by a HIF-1α independent mechanism. In isolated human periosteum-derived MSCs, BMP2 expression could be induced with the addition of platelets concentrate lysate but not with hypoxia treatment, confirming HIF-1α-independent BMP2 expression. Interestingly, in isolated human periosteum-derived mesenchymal progenitor cells, inhibition of BMP2 expression by PX-12 is accomplished only under hypoxic conditions seemingly through dis-regulation of reactive oxygen species (ROS) levels. In conclusion, we provide evidence of a molecular mechanism of hypoxia-dependent BMP2 expression in MSCs where interference with ROS homeostasis specifies fracture nonunion-like appearance in vivo through inhibition of Bmp2 expression. Stem Cells 2016;34:2342-2353.
© 2016 AlphaMed Press.
1 Communities
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18 MeSH Terms
Age-Dependent Pancreatic Gene Regulation Reveals Mechanisms Governing Human β Cell Function.
Arda HE, Li L, Tsai J, Torre EA, Rosli Y, Peiris H, Spitale RC, Dai C, Gu X, Qu K, Wang P, Wang J, Grompe M, Scharfmann R, Snyder MS, Bottino R, Powers AC, Chang HY, Kim SK
(2016) Cell Metab 23: 909-20
MeSH Terms: Adult, Aging, Cell Differentiation, Cell Separation, Child, Child, Preschool, Chromatin, Chromatin Immunoprecipitation, Diabetes Mellitus, Gene Expression Regulation, Developmental, Histone Code, Homeodomain Proteins, Humans, Infant, Insulin-Secreting Cells, Middle Aged, Transcription Factors, Transcriptome, Young Adult
Show Abstract · Added July 16, 2016
Intensive efforts are focused on identifying regulators of human pancreatic islet cell growth and maturation to accelerate development of therapies for diabetes. After birth, islet cell growth and function are dynamically regulated; however, establishing these age-dependent changes in humans has been challenging. Here, we describe a multimodal strategy for isolating pancreatic endocrine and exocrine cells from children and adults to identify age-dependent gene expression and chromatin changes on a genomic scale. These profiles revealed distinct proliferative and functional states of islet α cells or β cells and histone modifications underlying age-dependent gene expression changes. Expression of SIX2 and SIX3, transcription factors without prior known functions in the pancreas and linked to fasting hyperglycemia risk, increased with age specifically in human islet β cells. SIX2 and SIX3 were sufficient to enhance insulin content or secretion in immature β cells. Our work provides a unique resource to study human-specific regulators of islet cell maturation and function.
Copyright © 2016 Elsevier Inc. All rights reserved.
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19 MeSH Terms
Low frequency of broadly neutralizing HIV antibodies during chronic infection even in quaternary epitope targeting antibodies containing large numbers of somatic mutations.
Hicar MD, Chen X, Kalams SA, Sojar H, Landucci G, Forthal DN, Spearman P, Crowe JE
(2016) Mol Immunol 70: 94-103
MeSH Terms: Antibodies, Monoclonal, Antibodies, Neutralizing, B-Lymphocytes, Cell Separation, Enzyme-Linked Immunosorbent Assay, Epitopes, Flow Cytometry, Genes, Immunoglobulin, HIV Antibodies, HIV Infections, Humans, Immunoglobulin Heavy Chains, Immunoglobulin Variable Region, Mutation, env Gene Products, Human Immunodeficiency Virus
Show Abstract · Added January 26, 2016
Neutralizing antibodies (Abs) are thought to be a critical component of an appropriate HIV vaccine response. It has been proposed that Abs recognizing conformationally dependent quaternary epitopes on the HIV envelope (Env) trimer may be necessary to neutralize diverse HIV strains. A number of recently described broadly neutralizing monoclonal Abs (mAbs) recognize complex and quaternary epitopes. Generally, many such Abs exhibit extensive numbers of somatic mutations and unique structural characteristics. We sought to characterize the native antibody (Ab) response against circulating HIV focusing on such conformational responses, without a prior selection based on neutralization. Using a capture system based on VLPs incorporating cleaved envelope protein, we identified a selection of B cells that produce quaternary epitope targeting Abs (QtAbs). Similar to a number of broadly neutralizing Abs, the Ab genes encoding these QtAbs showed extensive numbers of somatic mutations. However, when expressed as recombinant molecules, these Abs failed to neutralize virus or mediate ADCVI activity. Molecular analysis showed unusually high numbers of mutations in the Ab heavy chain framework 3 region of the variable genes. The analysis suggests that large numbers of somatic mutations occur in Ab genes encoding HIV Abs in chronically infected individuals in a non-directed, stochastic, manner.
Copyright © 2015 Elsevier Ltd. All rights reserved.
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15 MeSH Terms