Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 5 de 5
Filtrar
Mais filtros

Base de dados
Tipo de documento
País de afiliação
Intervalo de ano de publicação
1.
ACS Chem Biol ; 17(3): 556-566, 2022 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-35188729

RESUMO

Hematopoietic progenitor kinase 1 (HPK1) is an MAP4K family member within the Ste20-like serine/threonine branch of the kinome. HPK1 expression is limited to hematopoietic cells and has a predominant role as a negative regulator of T cell function. Because of the central/dominant role in negatively regulating T cell function, HPK1 has long been in the center of interest as a potential pharmacological target for immune therapy. The development of a small molecule HPK1 inhibitor remains challenging because of the need for high specificity relative to other kinases, including additional MAP4K family members, that are required for efficient immune cell activation. Here, we report the identification of the selective and potent HPK1 chemical probe, A-745. In unbiased cellular kinase-binding assays, A-745 demonstrates an excellent cellular selectivity binding profile within pharmacologically relevant concentrations. This HPK1 selectivity translates to an in vitro immune cell activation phenotype reminiscent of Hpk1-deficient and Hpk1-kinase-dead T cells, including augmented proliferation and cytokine production. The results from this work give a path forward for further developmental efforts to generate additional selective and potent small molecule HPK1 inhibitors with the pharmacological properties for immunotherapy.


Assuntos
Proteínas Serina-Treonina Quinases , Linfócitos T , Fatores Imunológicos , Imunoterapia , Transdução de Sinais
2.
J Cell Biochem ; 112(10): 2759-72, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21748788

RESUMO

Heterotopic ossification (HO) is a disabling condition associated with neurologic injury, inflammation, and overactive bone morphogenetic protein (BMP) signaling. The inductive factors involved in lesion formation are unknown. We found that the expression of the neuro-inflammatory factor Substance P (SP) is dramatically increased in early lesional tissue in patients who have either fibrodysplasia ossificans progressiva (FOP) or acquired HO, and in three independent mouse models of HO. In Nse-BMP4, a mouse model of HO, robust HO forms in response to tissue injury; however, null mutations of the preprotachykinin (PPT) gene encoding SP prevent HO. Importantly, ablation of SP(+) sensory neurons, treatment with an antagonist of SP receptor NK1r, deletion of NK1r gene, or genetic down-regulation of NK1r-expressing mast cells also profoundly inhibit injury-induced HO. These observations establish a potent neuro-inflammatory induction and amplification circuit for BMP-dependent HO lesion formation, and identify novel molecular targets for prevention of HO.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Ossificação Heterotópica/metabolismo , Substância P/metabolismo , Animais , Proteína Morfogenética Óssea 4/genética , Proteína Morfogenética Óssea 4/metabolismo , Proteínas Morfogenéticas Ósseas/genética , Feminino , Humanos , Imuno-Histoquímica , Isoindóis/farmacologia , Masculino , Camundongos , Camundongos Transgênicos , Miosite Ossificante/genética , Miosite Ossificante/metabolismo , Antagonistas dos Receptores de Neurocinina-1 , Ossificação Heterotópica/genética , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Receptores da Neurocinina-1/metabolismo , Células Receptoras Sensoriais/metabolismo , Taquicininas/genética , Taquicininas/metabolismo
3.
Sci Rep ; 11(1): 2879, 2021 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-33536571

RESUMO

Alzheimer's disease (AD) is a common neurodegenerative disease with poor prognosis. New options for drug discovery targets are needed. We developed an imaging based arrayed CRISPR method to interrogate the human genome for modulation of in vitro correlates of AD features, and used this to assess 1525 human genes related to tau aggregation, autophagy and mitochondria. This work revealed (I) a network of tau aggregation modulators including the NF-κB pathway and inflammatory signaling, (II) a correlation between mitochondrial morphology, respiratory function and transcriptomics, (III) machine learning predicted novel roles of genes and pathways in autophagic processes and (IV) individual gene function inferences and interactions among biological processes via multi-feature clustering. These studies provide a platform to interrogate underexplored aspects of AD biology and offer several specific hypotheses for future drug discovery efforts.


Assuntos
Doença de Alzheimer/genética , Autofagia/genética , Modelos Genéticos , Agregação Patológica de Proteínas/genética , Proteínas tau/metabolismo , Doença de Alzheimer/patologia , Encéfalo/patologia , Sistemas CRISPR-Cas/genética , Linhagem Celular Tumoral , Regulação da Expressão Gênica , Engenharia Genética , Humanos , Aprendizado de Máquina , Mitocôndrias/genética , Mitocôndrias/patologia , Neurônios , Agregação Patológica de Proteínas/patologia , Transdução de Sinais/genética
4.
Stem Cells Transl Med ; 4(5): 437-47, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25834120

RESUMO

Studies of human cerebral cortex development are limited by difficulties in accessing and manipulating human neural tissue at specific development stages. We have derived human radial glia (hRG), which are responsible for most cerebral cortex neurogenesis, from human pluripotent stem cells. These hRG display the hallmark morphological, cellular, and molecular features of radial glia in vitro. They can be passaged and generate layer-specific subtypes of cortical neurons in a temporal and passage-dependent fashion. In later passages, they adopt a distinct progenitor phenotype that gives rise to cortical astrocytes and GABAergic interneurons. These hRG are also capable of following developmental cues to engraft, differentiate, migrate, and integrate into the embryonic mouse cortex when injected into E14 lateral ventricles. Moreover, hRG-derived cells can be cryopreserved at specific stages and retain their stage-specific phenotypes and competence when revived. Our study demonstrates that cultured hRG maintain a cell-intrinsic clock that regulates the progressive generation of stage-specific neuronal and glial subtypes. It also describes an easily accessible cell source for studying hRG lineage specification and progression and an on-demand supply of specific cortical neuron subtypes and astrocytes.


Assuntos
Diferenciação Celular/genética , Córtex Cerebral/citologia , Células Ependimogliais/citologia , Células-Tronco Pluripotentes/citologia , Animais , Astrócitos/citologia , Humanos , Camundongos , Neurogênese , Neuroglia/citologia , Neurônios/citologia , Células-Tronco Pluripotentes/metabolismo
5.
Mol Neurodegener ; 9: 3, 2014 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-24401693

RESUMO

An early substantial loss of basal forebrain cholinergic neurons (BFCNs) is a constant feature of Alzheimer's disease (AD) and is associated with deficits in spatial learning and memory. Induced pluripotent stem cells (iPSCs) derived from patients with AD as well as from normal controls could be efficiently differentiated into neurons with characteristics of BFCNs. We used BFCNs derived from iPSCs to model sporadic AD with a focus on patients with ApoE3/E4 genotypes (AD-E3/E4). BFCNs derived from AD-E3/E4 patients showed typical AD biochemical features evidenced by increased Aß42/Aß40 ratios. AD-E3/E4 neurons also exhibited altered responses to treatment with γ-secretase inhibitors compared to control BFCNs or neurons derived from patients with familial AD. BFCNs from patients with AD-E3/E4 also exhibited increased vulnerability to glutamate-mediated cell death which correlated with increased intracellular free calcium upon glutamate exposure. The ability to generate BFCNs with an AD phenotype is a significant step both for understanding disease mechanisms and for facilitating screening for agents that promote synaptic integrity and neuronal survival.


Assuntos
Doença de Alzheimer , Técnicas de Cultura de Células/métodos , Neurônios Colinérgicos , Células-Tronco Pluripotentes Induzidas/citologia , Adulto , Idoso , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Apolipoproteína E3/genética , Apolipoproteína E4/genética , Western Blotting , Morte Celular , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Masculino , Células-Tronco Neurais/citologia , Técnicas de Patch-Clamp , Reação em Cadeia da Polimerase , Prosencéfalo/citologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA