Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 16 de 16
Filtrar
1.
Cell ; 136(6): 1017-31, 2009 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-19303846

RESUMEN

The Disrupted in Schizophrenia 1 (DISC1) gene is disrupted by a balanced chromosomal translocation (1; 11) (q42; q14.3) in a Scottish family with a high incidence of major depression, schizophrenia, and bipolar disorder. Subsequent studies provided indications that DISC1 plays a role in brain development. Here, we demonstrate that suppression of DISC1 expression reduces neural progenitor proliferation, leading to premature cell cycle exit and differentiation. Several lines of evidence suggest that DISC1 mediates this function by regulating GSK3beta. First, DISC1 inhibits GSK3beta activity through direct physical interaction, which reduces beta-catenin phosphorylation and stabilizes beta-catenin. Importantly, expression of stabilized beta-catenin overrides the impairment of progenitor proliferation caused by DISC1 loss of function. Furthermore, GSK3 inhibitors normalize progenitor proliferation and behavioral defects caused by DISC1 loss of function. Together, these results implicate DISC1 in GSK3beta/beta-catenin signaling pathways and provide a framework for understanding how alterations in this pathway may contribute to the etiology of psychiatric disorders.


Asunto(s)
Glucógeno Sintasa Quinasa 3/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Neurogénesis , Transducción de Señal , beta Catenina/metabolismo , Células Madre Adultas/citología , Células Madre Adultas/metabolismo , Animales , Encéfalo/citología , Encéfalo/embriología , Embrión de Mamíferos/metabolismo , Técnicas de Silenciamiento del Gen , Glucógeno Sintasa Quinasa 3 beta , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Neuronas/citología , Neuronas/metabolismo , Células Madre/citología , Células Madre/metabolismo
2.
J Lipid Res ; 56(2): 266-76, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25528754

RESUMEN

Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. Previous studies have linked it to atherosclerosis. Perlecan contains a core protein and three HS side chains. Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). The functional significance of this domain has been unknown. Here, we show that perlecan DII interacts with LDL. Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. We further demonstrate an additive effect of HS and DII on LDL binding. Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. Thus, perlecan glycosylation may play a role in the early LDL retention during the development of atherosclerosis.


Asunto(s)
Aterosclerosis/metabolismo , Lipoproteínas LDL/metabolismo , Receptores de LDL/metabolismo , Animales , Células COS , Línea Celular , Chlorocebus aethiops , Glicosilación , Células HeLa , Proteoglicanos de Heparán Sulfato/metabolismo , Humanos , Inmunohistoquímica , Microscopía Confocal , Mutagénesis Sitio-Dirigida , Ácido N-Acetilneuramínico/metabolismo , Ratas
3.
Small ; 9(9-10): 1842-52, 2013 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-23423856

RESUMEN

The development of classification nano-structure-activity Relationships (nano-SARs) of nanoparticle (NP) bioactivity is presented with the aim of demonstrating the integration of multiparametric toxicity/bioactivity assays to arrive at statistically meaningful class definitions (i.e., bioactivity/inactivity endpoints), as well as the implications of nano-SAR applicability domains and decision boundaries. Nano-SARs are constructed based on a dataset of 44 iron oxide core nanoparticles (NPs), used in molecular imaging and nano-sensing, containing bioactivity profiles for four cell types and four different assays. Class definitions are developed on the basis of 'hit' (i.e., significant bioactivity) identification analysis and self-organizing map based consensus clustering; these class definitions enable construction of nano-SARs of a high classification accuracy (>78%) with different NP descriptor combinations that include primary size, spin-lattice and spin-spin relaxivities, and zeta potentials. Analysis of the nano-SAR performance for different class definitions suggests that H4 (i.e., class with at least four hits) is a reasonable endpoint (from a 'regulatory' viewpoint) for keeping the level of false negatives (i.e., incorrect labeling of bioactive NPs as inactive) low. The establishment of a quantitative nano-SAR applicability domain is demonstrated, making use of a probability density with the H4 class definition and naive Bayesian classifier (NBC) model (with spin-lattice relaxivity and zeta potential as descriptors). Decision boundaries are determined for the above H4/NBC nano-SAR for different acceptance levels of false negative to false positive predictions, illustrating a practical approach that may assist in regulatory decision making with a consideration of reducing the likelihood of identifying bioactive NPs as being inactive.


Asunto(s)
Nanopartículas , Teorema de Bayes , Nanopartículas/química , Relación Estructura-Actividad
4.
Nano Lett ; 12(11): 5808-12, 2012 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-23039907

RESUMEN

Products are increasingly incorporating nanomaterials, but we have a poor understanding of their adverse effects. To assess risk, regulatory authorities need more experimental testing of nanoparticles. Computational models play a complementary role in allowing rapid prediction of potential toxicities of new and modified nanomaterials. We generated quantitative, predictive models of cellular uptake and apoptosis induced by nanoparticles for several cell types. We illustrate the potential of computational methods to make a contribution to nanosafety.


Asunto(s)
Apoptosis , Biofisica/métodos , Nanopartículas/química , Nanoestructuras/química , Nanotecnología/métodos , Animales , Teorema de Bayes , Línea Celular Tumoral , Células Endoteliales de la Vena Umbilical Humana , Humanos , Ratones , Modelos Estadísticos , Análisis de Regresión , Riesgo , Relación Estructura-Actividad
5.
Angew Chem Int Ed Engl ; 52(40): 10593-7, 2013 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-23960025

RESUMEN

A proteomics method to pull down secondary drug targets from live cells is described. The drug of interest is modified with trans-cyclooctene (TCO) and incubated with live cells. Upon cell lysis, the modified drug bound to the protein is pulled down using magnetic beads decorated with a cleavable tetrazine-modified linker. Samples are then run on an SDS-PAGE gel and isolated bands are submitted for mass spectrometry analysis to identify drug targets.


Asunto(s)
Terapia Molecular Dirigida/métodos , Proteómica/métodos , Humanos
6.
Acc Chem Res ; 44(10): 842-52, 2011 Oct 18.
Artículo en Inglés | MEDLINE | ID: mdl-21661727

RESUMEN

Advances in our understanding of the genetic basis of disease susceptibility coupled with prominent successes for molecular targeted therapies have resulted in an emerging strategy of personalized medicine. This approach envisions risk stratification and therapeutic selection based on an individual's genetic makeup and physiologic state (the latter assessed through cellular or molecular phenotypes). Molecularly targeted nanoparticles can play a key role in this vision through noninvasive assessments of molecular processes and specific cell populations in vivo, sensitive molecular diagnostics, and targeted delivery of therapeutics. A superparamagnetic iron oxide nanoparticle with a cross-linked dextran coating, or CLIO, is a powerful and illustrative nanoparticle platform for these applications. These structures and their derivatives support diagnostic imaging by magnetic resonance (MRI), optical, and positron emission tomography (PET) modalities and constitute a versatile platform for conjugation to targeting ligands. A variety of conjugation methods exist to couple the dextran surface to different functional groups; in addition, a robust bioorthogonal [4 + 2] cycloaddition reaction between 1,2,4,5-tetrazene (Tz) and trans-cyclooctene (TCO) can conjugate nanoparticles to targeting ligands or label pretargeted cells. The ready availability of conjugation methods has given rise to the synthesis of libraries of small molecule modified nanoparticles, which can then be screened for nanoparticles with specificity for a specific cell type. Since most nanoparticles display their targeting ligands in a multivalent manner, a detailed understanding of the kinetics and affinity of a nanoparticle's interaction with its target (as determined by surface plasmon resonance) can yield functionally important insights into nanoparticle design. In this Account, we review applications of the CLIO platform in several areas relevant to the mission of personalized medicine. We demonstrate rapid and highly sensitive molecular profiling of cancer markers ex vivo, as part of detailed, individualized molecular phenotyping. The CLIO platform also facilitates targeted magnetic resonance and combined modality imaging (such as MR/PET/fluorescence/CT) to enable multiplexed measurement of molecular phenotypes in vivo for early diagnosis and disease classification. Finally, the targeted delivery of a photodynamic therapy agent as part of a theranostic nanoparticle successfully increased local cell toxicity and minimized systemic side effects.


Asunto(s)
Dextranos/química , Compuestos Férricos/química , Imagen Molecular/métodos , Terapia Molecular Dirigida/métodos , Nanopartículas/uso terapéutico , Animales , Humanos , Nanomedicina , Nanopartículas/química
7.
Angew Chem Int Ed Engl ; 51(2): 450-4, 2012 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-22113923

RESUMEN

Be my guest: A supramolecular host-guest interaction is utilized for highly efficient bioorthogonal labeling of cellular targets. Antibodies labeled with a cyclodextrin host molecule bind to adamantane-labeled magnetofluorescent nanoparticles (see picture) and provide an amplifiable strategy for biomarker detection that can be adapted to different diagnostic techniques such as molecular profiling or magnetic cell sorting.


Asunto(s)
Adamantano/química , Anticuerpos/análisis , Ciclodextrinas/química , Colorantes Fluorescentes/análisis , Nanopartículas/análisis , Animales , Anticuerpos/química , Biomarcadores/análisis , Línea Celular , Colorantes Fluorescentes/química , Humanos , Nanopartículas/química
8.
Bioconjug Chem ; 22(12): 2390-4, 2011 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-22043803

RESUMEN

The development of faster and more sensitive detection methods capable of identifying specific bacterial species and strains has remained a longstanding clinical challenge. Thus to date, the diagnosis of bacterial infections continues to rely on the performance of time-consuming microbiological cultures. Here, we demonstrate the use of bioorthogonal chemistry for magnetically labeling specific pathogens to enable their subsequent detection by nuclear magnetic resonance. Antibodies against a bacterial target of interest were first modified with trans-cyclooctene and then coupled to tetrazine-modified magnetic nanoprobes, directly on the bacteria. This labeling method was verified by surface plasmon resonance as well as by highly specific detection of Staphylococcus aureus using a miniaturized diagnostic magnetic resonance system. Compared to other copper-free bioorthogonal chemistries, the cycloaddition reaction reported here displayed faster kinetics and yielded higher labeling efficiency. Considering the short assay times and the portability of the necessary instrumentation, it is feasible that this approach could be adapted for clinical use in resource-limited settings.


Asunto(s)
Espectroscopía de Resonancia Magnética/instrumentación , Infecciones Estafilocócicas/diagnóstico , Staphylococcus aureus/aislamiento & purificación , Diseño de Equipo , Humanos , Espectroscopía de Resonancia Magnética/economía , Factores de Tiempo
9.
Bioconjug Chem ; 21(1): 14-9, 2010 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20028085

RESUMEN

Nanoparticles bearing surface-conjugated targeting ligands are increasingly being explored for a variety of biomedical applications. The multivalent conjugation of targeting ligands on the surface of nanoparticles is presumed to enhance binding to the desired target. However, given the complexities inherent in the interactions of nanoparticle surfaces with proteins, and the structural diversity of nanoparticle scaffolds and targeting ligands, our understanding of how conjugation of targeting ligands affects nanoparticle binding remains incomplete. Here, we use surface plasmon resonance (SPR) to directly and quantitatively study the affinity and binding kinetics of nanoparticles that display small molecules conjugated to their surface. We studied the interaction between a single protein target and a structurally related series of targeting ligands whose intrinsic affinity varies over a 4500-fold range and performed SPR at protein densities that reflect endogenous receptor densities. We report that even weak small molecule targeting ligands can significantly enhance target-specific avidity (by up to 4 orders of magnitude) through multivalent interactions and also observe a much broader range of kinetic effects than has been previously reported. Quantitative measurement of how the affinity and kinetics of nanoparticle binding vary as a function of different surface conjugations is a rapid, generalizable approach to nanoparticle characterization that can inform the design of nanoparticles for biomedical applications.


Asunto(s)
Técnicas de Sonda Molecular , Nanopartículas/análisis , Nanopartículas/química , Proteínas/metabolismo , Cinética , Ligandos , Unión Proteica , Proteínas/química , Receptores de Superficie Celular/metabolismo , Resonancia por Plasmón de Superficie
10.
Org Lett ; 10(13): 2837-40, 2008 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-18537250

RESUMEN

Alkyne acids 11 were prepared in an enantioselective fashion from allylic ester derivatives 18 or 20 by Ireland-Claisen rearrangement, followed by Si-assisted elimination of HBr. The title compounds are attractive ring D synthons for an ongoing synthesis of cobyric acid.


Asunto(s)
Vitamina B 12/análogos & derivados , Ácidos/química , Estructura Molecular , Pirroles/química , Estereoisomerismo , Vitamina B 12/síntesis química , Vitamina B 12/química
11.
ACS Med Chem Lett ; 8(2): 196-200, 2017 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-28197311

RESUMEN

Several monoclonal antibodies and inhibitors targeting CD38, an ectoenzyme overexpressed on malignant plasma cells, have previously been discovered. Herein, we expand structure-activity relationships of reported small-molecule thiazoloquinolinones and show that several 4-cyclohexylamino analogues have potent binding affinity for CD38 using surface plasmon resonance. Moreover, active amine analogues could be acylated and functionalized with alkyne and fluorescein groups. Fluorescein analogue 21 bound selectively to CD38 overexpressing cells, demonstrating the potential utility of thiazoloquinolinones as small-molecule conjugates for the delivery of therapeutic and imaging agents.

12.
Org Lett ; 5(25): 4879-82, 2003 Dec 11.
Artículo en Inglés | MEDLINE | ID: mdl-14653697

RESUMEN

Homochiral ring-C precursors 34 of Vitamin B(12) have been prepared by Ireland-Claisen rearrangement of allyl esters 32, followed by a novel Si-assisted elimination of HBr. [reaction: see text]


Asunto(s)
Bromuros/química , Silicio/química , Vitamina B 12/análogos & derivados , Vitamina B 12/síntesis química , Estructura Molecular , Estereoisomerismo , Vitamina B 12/química
13.
J Vis Exp ; (79): e50772, 2013 Sep 23.
Artículo en Inglés | MEDLINE | ID: mdl-24084440

RESUMEN

Methods for rapid surface immobilization of bioactive small molecules with control over orientation and immobilization density are highly desirable for biosensor and microarray applications. In this Study, we use a highly efficient covalent bioorthogonal [4+2] cycloaddition reaction between trans-cyclooctene (TCO) and 1,2,4,5-tetrazine (Tz) to enable the microfluidic immobilization of TCO/Tz-derivatized molecules. We monitor the process in real-time under continuous flow conditions using surface plasmon resonance (SPR). To enable reversible immobilization and extend the experimental range of the sensor surface, we combine a non-covalent antigen-antibody capture component with the cycloaddition reaction. By alternately presenting TCO or Tz moieties to the sensor surface, multiple capture-cycloaddition processes are now possible on one sensor surface for on-chip assembly and interaction studies of a variety of multi-component structures. We illustrate this method with two different immobilization experiments on a biosensor chip; a small molecule, AP1497 that binds FK506-binding protein 12 (FKBP12); and the same small molecule as part of an immobilized and in situ-functionalized nanoparticle.


Asunto(s)
Técnicas Biosensibles/métodos , Ciclooctanos/química , Técnicas Analíticas Microfluídicas/métodos , Tetrazoles/química , Técnicas Biosensibles/instrumentación , Ciclización , Técnicas Analíticas Microfluídicas/instrumentación , Nanopartículas/química , Resonancia por Plasmón de Superficie
14.
Neoplasia ; 15(7): 684-93, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23814481

RESUMEN

When resectable, invasive pancreatic ductal adenocarcinoma (PDAC) is most commonly treated with surgery and radiochemotherapy. Given the intricate local anatomy and locoregional mode of dissemination, achieving clean surgical margins can be a significant challenge. On the basis of observations that cathepsin E (CTSE) is overexpressed in PDAC and that an United States Food and Drug Administration (FDA)-approved protease inhibitor has high affinity for CTSE, we have developed a CTSE optical imaging agent [ritonavir tetramethyl-BODIPY (RIT-TMB)] for potential intraoperative use. We show nanomolar affinity [half maximal inhibitory concentration (IC50) of 39.9 ± 1.2 nM] against CTSE of the RIT-TMB in biochemical assays and intracellular accumulation and target-to-background ratios that allow specific delineation of individual cancer cells. This approach should be useful for more refined surgical staging, planning, and resection with curative intent.


Asunto(s)
Compuestos de Boro , Carcinoma Ductal Pancreático/metabolismo , Catepsina E/metabolismo , Colorantes Fluorescentes , Inhibidores de la Proteasa del VIH , Neoplasias Pancreáticas/metabolismo , Ritonavir , Animales , Biopsia , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/patología , Catepsina E/antagonistas & inhibidores , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Inhibidores de la Proteasa del VIH/farmacología , Humanos , Concentración 50 Inhibidora , Ratones , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patología , Ritonavir/análogos & derivados , Ritonavir/farmacología
15.
ACS Nano ; 4(10): 5703-12, 2010 Oct 26.
Artículo en Inglés | MEDLINE | ID: mdl-20857979

RESUMEN

Evaluation of biological effects, both desired and undesired, caused by manufactured nanoparticles (MNPs) is of critical importance for nanotechnology. Experimental studies, especially toxicological, are time-consuming, costly, and often impractical, calling for the development of efficient computational approaches capable of predicting biological effects of MNPs. To this end, we have investigated the potential of cheminformatics methods such as quantitative structure-activity relationship (QSAR) modeling to establish statistically significant relationships between measured biological activity profiles of MNPs and their physical, chemical, and geometrical properties, either measured experimentally or computed from the structure of MNPs. To reflect the context of the study, we termed our approach quantitative nanostructure-activity relationship (QNAR) modeling. We have employed two representative sets of MNPs studied recently using in vitro cell-based assays: (i) 51 various MNPs with diverse metal cores (Proc. Natl. Acad. Sci. 2008, 105, 7387-7392) and (ii) 109 MNPs with similar core but diverse surface modifiers (Nat. Biotechnol. 2005, 23, 1418-1423). We have generated QNAR models using machine learning approaches such as support vector machine (SVM)-based classification and k nearest neighbors (kNN)-based regression; their external prediction power was shown to be as high as 73% for classification modeling and having an R(2) of 0.72 for regression modeling. Our results suggest that QNAR models can be employed for: (i) predicting biological activity profiles of novel nanomaterials, and (ii) prioritizing the design and manufacturing of nanomaterials toward better and safer products.


Asunto(s)
Nanopartículas del Metal/química , Nanotecnología/métodos , Animales , Línea Celular Tumoral , Humanos , Microscopía de Fuerza Atómica/métodos , Modelos Estadísticos , Modelos Teóricos , Óxidos/química , Relación Estructura-Actividad Cuantitativa , Ratas , Análisis de Regresión , Propiedades de Superficie , Células U937
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA