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1.
Bioorg Chem ; 107: 104534, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33339664

RESUMEN

A novel strategy combining visible-light and enzyme catalysis in one pot for the synthesis of 4H-pyrimido[2,1-b] benzothiazole derivatives from alcohols is described for the first time. Fourteen 4H-pyrimido[2,1-b] benzothiazole derivatives were prepared with yields of up to 98% under mild reaction conditions by a simple operation. The photoorgano catalyst rose Bengal (rB) was employed to oxyfunctionalise alcohols to aldehydes. Compared with aldehydes, alcohols with more stable properties and lower cost, thus we used photocatalysis to oxidize alcohols into aldehydes. Next, the enzyme was used to further catalyze the reaction of Biginelli to produce the target product of 4H-pyrimidine [2,1-b] benzothiazole. Experimental results show that this method provides a more efficient and eco-friendly strategy for the synthesis of 4H-pyrimido[2,1-b] benzothiazole derivatives.


Asunto(s)
Benzotiazoles/química , Enzimas/metabolismo , Luz , Benzotiazoles/síntesis química , Benzotiazoles/metabolismo , Biocatálisis , Candida/enzimología , Catálisis , Enzimas/química , Proteínas Fúngicas/metabolismo , Humanos , Lipasa/metabolismo , Oxidación-Reducción
2.
Bioorg Chem ; 107: 104607, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33450543

RESUMEN

An efficient and green method, combining enzymatic and visible-light catalysis for synthesis of the widely applicable 2-substituted benzothiazoles, has been developed. This method features a relay catalysis protocol consisting of biocatalytic promiscuity and visible-light-induced subsequent oxidization of 2-phenyl benzothiazolines. The whole reaction process is very high-efficiency, achieving 99% yield in just 10 min, under an air atmosphere, nearly 100% atomic utilization, and the 2-substituted benzothiazole products were obtained in good to excellent yields with a wide range of substrates. This reaction is the other example of combining the non-natural catalytic activity of hydrolases with visible-light catalysis for organic synthesis and the catalytic system does not require additional oxidants or metals, which is good for the environment.


Asunto(s)
Benzotiazoles/síntesis química , Hidrolasas/metabolismo , Luz , Benzotiazoles/química , Benzotiazoles/metabolismo , Catálisis , Oxidación-Reducción , Fármacos Fotosensibilizantes/química , Solventes/química , Especificidad por Sustrato
3.
Lab Invest ; 100(9): 1208-1222, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32457352

RESUMEN

Aberrant activation of the hedgehog (Hh) signaling pathway is associated with the formation of medulloblastoma (MB), the most common malignant pediatric brain tumor. However, tumor cells from human and mouse MB can not be passaged or preserved after being adherently cultured. Moreover, Hh signaling in MB cells is inactivated in such culture. Here we demonstrate that MB cells are capable of forming tumoroids (tumor spheroids) in vitro under optimized conditions, which can be further passaged and cryopreserved. More importantly, MB cells maintain Hh pathway activation and cell proliferation in tumoroids. Our studies further reveal that tumoroids-forming capacity of MB cells relies on astrocytes, a major component of the MB microenvironment. Astrocytes facilitate the formation of MB tumoroids by secreting sonic hedgehog (Shh) and generating astrocyte-derived extracellular matrix. These findings demonstrate the critical role of stromal astrocytes in supporting the survival and proliferation of MB cells in vitro. This study establishes a valid model for long-term culture of primary MB cells, which could be greatly beneficial for future investigation of MB tumorigenicity and the development of improved approaches to treat MB.


Asunto(s)
Astrocitos/metabolismo , Neoplasias Cerebelosas/genética , Matriz Extracelular/metabolismo , Proteínas Hedgehog/genética , Meduloblastoma/genética , Transducción de Señal/genética , Animales , Astrocitos/patología , Línea Celular Tumoral , Neoplasias Cerebelosas/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteínas Hedgehog/metabolismo , Humanos , Meduloblastoma/metabolismo , Meduloblastoma/patología , Ratones Noqueados , Ratones SCID , Ratones Transgénicos , Receptor Patched-2/genética , Receptor Patched-2/metabolismo , Microambiente Tumoral/genética , Proteína con Dedos de Zinc GLI1/genética , Proteína con Dedos de Zinc GLI1/metabolismo
4.
Genes Dev ; 23(2): 157-70, 2009 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-19171780

RESUMEN

Studying the early stages of cancer can provide important insight into the molecular basis of the disease. We identified a preneoplastic stage in the patched (ptc) mutant mouse, a model for the brain tumor medulloblastoma. Preneoplastic cells (PNCs) are found in most ptc mutants during early adulthood, but only 15% of these animals develop tumors. Although PNCs are found in mice that develop tumors, the ability of PNCs to give rise to tumors has never been demonstrated directly, and the fate of cells that do not form tumors remains unknown. Using genetic fate mapping and orthotopic transplantation, we provide definitive evidence that PNCs give rise to tumors, and show that the predominant fate of PNCs that do not form tumors is differentiation. Moreover, we show that N-myc, a gene commonly amplified in medulloblastoma, can dramatically alter the fate of PNCs, preventing differentiation and driving progression to tumors. Importantly, N-myc allows PNCs to grow independently of hedgehog signaling, making the resulting tumors resistant to hedgehog antagonists. These studies provide the first direct evidence that PNCs can give rise to tumors, and demonstrate that identification of genetic changes that promote tumor progression is critical for designing effective therapies for cancer.


Asunto(s)
Diferenciación Celular/fisiología , Meduloblastoma/patología , Lesiones Precancerosas/patología , Proteínas Proto-Oncogénicas c-myc/metabolismo , Animales , Movimiento Celular , Proliferación Celular , Cerebelo/citología , Modelos Animales de Enfermedad , Expresión Génica , Genes Reporteros , Proteínas Hedgehog/antagonistas & inhibidores , Ratones , Ratones SCID , Ratones Transgénicos , Células Madre/citología , Células Madre/efectos de los fármacos , Alcaloides de Veratrum/farmacología
5.
Nature ; 468(7327): 1095-9, 2010 Dec 23.
Artículo en Inglés | MEDLINE | ID: mdl-21150899

RESUMEN

Medulloblastoma encompasses a collection of clinically and molecularly diverse tumour subtypes that together comprise the most common malignant childhood brain tumour. These tumours are thought to arise within the cerebellum, with approximately 25% originating from granule neuron precursor cells (GNPCs) after aberrant activation of the Sonic Hedgehog pathway (hereafter, SHH subtype). The pathological processes that drive heterogeneity among the other medulloblastoma subtypes are not known, hindering the development of much needed new therapies. Here we provide evidence that a discrete subtype of medulloblastoma that contains activating mutations in the WNT pathway effector CTNNB1 (hereafter, WNT subtype) arises outside the cerebellum from cells of the dorsal brainstem. We found that genes marking human WNT-subtype medulloblastomas are more frequently expressed in the lower rhombic lip (LRL) and embryonic dorsal brainstem than in the upper rhombic lip (URL) and developing cerebellum. Magnetic resonance imaging (MRI) and intra-operative reports showed that human WNT-subtype tumours infiltrate the dorsal brainstem, whereas SHH-subtype tumours are located within the cerebellar hemispheres. Activating mutations in Ctnnb1 had little impact on progenitor cell populations in the cerebellum, but caused the abnormal accumulation of cells on the embryonic dorsal brainstem which included aberrantly proliferating Zic1(+) precursor cells. These lesions persisted in all mutant adult mice; moreover, in 15% of cases in which Tp53 was concurrently deleted, they progressed to form medulloblastomas that recapitulated the anatomy and gene expression profiles of human WNT-subtype medulloblastoma. We provide the first evidence, to our knowledge, that subtypes of medulloblastoma have distinct cellular origins. Our data provide an explanation for the marked molecular and clinical differences between SHH- and WNT-subtype medulloblastomas and have profound implications for future research and treatment of this important childhood cancer.


Asunto(s)
Tronco Encefálico/patología , Neoplasias Cerebelosas/patología , Meduloblastoma/patología , Animales , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , Ratones Transgénicos , Mutación , beta Catenina/genética
6.
Cancer Cell ; 11(1): 3-5, 2007 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-17222787

RESUMEN

Cancer stem cells (CSCs) are thought to be critical for initiation and propagation of many types of cancer. Because these cells are resistant to conventional therapies, they have been very difficult to eliminate. A study in this issue of Cancer Cell suggests that brain tumor CSCs live in a "vascular niche" that promotes their long-term growth and self-renewal. Disrupting this niche impairs CSC self-renewal and thereby significantly inhibits the growth of tumors. Targeting the unique microenvironment of CSCs may be the key to effective cancer therapy.


Asunto(s)
Neoplasias Encefálicas/irrigación sanguínea , Células Madre Neoplásicas , Neovascularización Patológica , Animales , Neoplasias Encefálicas/patología , Humanos , Neuronas/patología
7.
bioRxiv ; 2024 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-38405864

RESUMEN

Hypothyroidism is commonly detected in patients with medulloblastoma (MB). A possible link between thyroid hormone (TH) signaling and MB pathogenicity has not been reported. Here, we find that TH plays a critical role in promoting tumor cell differentiation. Reduction in TH levels frees the TH receptor, TRα1, to bind to EZH2 and repress expression of NeuroD1, a transcription factor that drives tumor cell differentiation. Increased TH reverses EZH2-mediated repression of NeuroD1 by abrogating the binding of EZH2 and TRα1, thereby stimulating tumor cell differentiation and reducing MB growth. Importantly, TH-induced differentiation of tumor cells is not restricted by the molecular subgroup of MB. These findings establish an unprecedented association between TH signaling and MB pathogenicity, providing solid evidence for TH as a promising modality for MB treatment.

8.
STAR Protoc ; 4(2): 102234, 2023 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-37074909

RESUMEN

Tumor relapse is the leading adverse prognostic factor in medulloblastoma (MB). However, there is still no established mouse model for MB relapse, impeding our efforts to develop strategies to treat relapsed MB. We present a protocol for generating a mouse model for relapsed MB using irradiation by optimizing mouse breeding and age, as well as irradiation dosage and timing. We then detail procedures for determining tumor relapse based on tumor cell trans-differentiation in MB tissue, immunohistochemistry, and tumor cell isolation. For complete details on the use and execution of this protocol, please refer to Guo et al. (2021).1.

9.
Cell Death Differ ; 30(5): 1198-1210, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36813922

RESUMEN

Elevated levels of PDLIM3 expression are frequently detected in sonic hedgehog (SHH) group of medulloblastoma (MB). However, the possible role of PDLIM3 in MB tumorigenesis is still unknown. Here, we found that PDLIM3 expression is necessary for hedgehog (Hh) pathway activation in MB cells. PDLIM3 is present in primary cilia of MB cells and fibroblasts, and such cilia localization is mediated by the PDZ domain of PDLIM3 protein. Deletion of PDLIM3 significantly compromised cilia formation and interfered the Hh signaling transduction in MB cells, suggesting that PDLIM3 promotes the Hh signaling through supporting the ciliogenesis. PDLIM3 protein physically interacts with cholesterol, a critical molecule for cilia formation and hedgehog signaling. The disruption of cilia formation and Hh signaling in PDLIM3 null MB cells or fibroblasts, was significantly rescued by treatment with exogenous cholesterol, demonstrating that PDLIM3 facilitates the ciliogenesis through cholesterol provision. Finally, deletion of PDLIM3 in MB cells significantly inhibited their proliferation and repressed tumor growth, suggesting that PDLIM3 is necessary for MB tumorigenesis. Our studies elucidate the critical functions of PDLIM3 in the ciliogenesis and Hh signaling transduction in SHH-MB cells, supporting to utilize PDLIM3 as a molecular marker for defining SHH group of MB in clinics.


Asunto(s)
Neoplasias Cerebelosas , Meduloblastoma , Humanos , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Meduloblastoma/genética , Meduloblastoma/patología , Cilios/metabolismo , Colesterol/metabolismo , Neoplasias Cerebelosas/genética , Neoplasias Cerebelosas/patología , Carcinogénesis/patología , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Proteínas con Dominio LIM/genética , Proteínas con Dominio LIM/metabolismo
10.
STAR Protoc ; 2(2): 100409, 2021 06 18.
Artículo en Inglés | MEDLINE | ID: mdl-33870218

RESUMEN

This protocol provides the procedures for isolating differentiated tumor cells from medulloblastoma (MB) in mice. Procedures for transplantation into cerebella are also included to examine the tumorigenesis of differentiated MB cells. This protocol outlines the detailed steps required for (1) isolation of tumor cells from mouse MB, (2) purification of differentiated tumor cells by fluorescence-activated cell sorting, and (3) transplantation of tumor cells into cerebella. This protocol is useful to purify differentiated tumor cells for investigating mechanisms underlying MB progression. For complete details on the use and execution of this protocol, please refer to Cheng et. al. (2020).


Asunto(s)
Separación Celular/métodos , Neoplasias Cerebelosas/patología , Cerebelo/cirugía , Meduloblastoma/patología , Células Tumorales Cultivadas , Animales , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , Trasplante de Células , Ratones , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/patología , Células Tumorales Cultivadas/trasplante
11.
Oncogene ; 40(12): 2258-2272, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33649536

RESUMEN

The Hedgehog (Hh) pathway plays an indispensable role in bone development and genetic activation of the pathway results in medulloblastoma (MB), the most common malignant brain tumor in children. Inhibitors of Hh pathway (such as vismodegib and sonedigib), which are used to treat MB, cause irreversible defects in bone growth in young children. Cholesterol is required for the activation of the Hh pathway, and statins, inhibitors of cholesterol biosynthesis, suppress MB growth by repressing Hh signaling in tumor cells. Here, we investigate the role of cholesterol biosynthesis in the proliferation and Hh signaling in chondrocytes, and examine the bone development in mice after statin treatment. Statins significantly inhibited MB growth in young mice, but caused no defects in bone development. Conditional deletion of NADP steroid dehydrogenase-like (NSDHL), an enzyme necessary for cholesterol biosynthesis, suppressed cholesterol synthesis in chondrocytes, and disrupted the growth plate in mouse femur and tibia, indicating the important function of intracellular cholesterol in bone development. Hh pathway activation and the proliferation of chondrocytes were inhibited by statin treatment in vitro; however, statins did not impair bone growth in vivo due to insufficient penetration into the bone. Our studies reveal a critical role of cholesterol in bone development, and support the utilization of statins for treatment of MB as well as other Hh pathway-associated malignancies.


Asunto(s)
3-Hidroxiesteroide Deshidrogenasas/genética , Colesterol/biosíntesis , Proteínas Hedgehog/genética , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Meduloblastoma/tratamiento farmacológico , Anilidas/efectos adversos , Animales , Desarrollo Óseo/efectos de los fármacos , Desarrollo Óseo/genética , Proliferación Celular/efectos de los fármacos , Colesterol/genética , Condrocitos/efectos de los fármacos , Proteínas Hedgehog/antagonistas & inhibidores , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/efectos adversos , Lipogénesis/efectos de los fármacos , Meduloblastoma/genética , Meduloblastoma/patología , Ratones , Ratones Noqueados , Piridinas/efectos adversos , Transducción de Señal/efectos de los fármacos
12.
Mol Cancer Res ; 19(10): 1651-1665, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34158391

RESUMEN

Nestin, a class IV intermediate filament protein, is generally considered as a putative marker of neural stem and progenitor cells in the central nervous system. Glioma is a common type of adult brain tumors, and glioblastoma (GBM) represents the most aggressive form of glioma. Here, we report that Nestin expression is significantly upregulated in human GBM, compared with other types of glioma. Nestin knockdown or deletion in U251 cells and tumor cells from GBM patients derived xenografts resulted in G2-M arrest, finally leading to apoptosis in tumor cells. Using proximity-dependent biotin identification method, we identified ßII-tubulin as an interacting protein of Nestin in U251 cells. Nestin stabilized ßII-tubulin in U251 cells through physical interaction. Knockdown of Nestin or ßII-tubulin disrupted spindle morphology in tumor cells. Our studies further revealed that Nestin deficiency in U251 cells and GBM PDX cells repressed tumor growth upon transplantation. Finally, we found that Nestin deficiency sensitized GBM cells to microtubule-destabilizing drugs such as vinblastine and vincristine. Our studies demonstrate the essential functions and underlying mechanisms of Nestin in the growth and drug response of GBM cells. IMPLICATIONS: Through interaction with ßII-tubulin, Nestin facilitates cell-cycle progression and spindle assembly of tumor cells in glioblastoma.


Asunto(s)
Neoplasias Encefálicas/metabolismo , Ciclo Celular/fisiología , Glioblastoma/metabolismo , Nestina/metabolismo , Huso Acromático/metabolismo , Animales , Apoptosis/fisiología , Línea Celular , Línea Celular Tumoral , Glioma/metabolismo , Células HEK293 , Humanos , Ratones Desnudos , Ratones SCID , Tubulina (Proteína)/metabolismo
13.
J Exp Med ; 218(9)2021 09 06.
Artículo en Inglés | MEDLINE | ID: mdl-34254999

RESUMEN

Astrocytes, a major glial cell type in the brain, play a critical role in supporting the progression of medulloblastoma (MB), the most common malignant pediatric brain tumor. Through lineage tracing analyses and single-cell RNA sequencing, we demonstrate that astrocytes are predominantly derived from the transdifferentiation of tumor cells in relapsed MB (but not in primary MB), although MB cells are generally believed to be neuronal-lineage committed. Such transdifferentiation of MB cells relies on Sox9, a transcription factor critical for gliogenesis. Our studies further reveal that bone morphogenetic proteins (BMPs) stimulate the transdifferentiation of MB cells by inducing the phosphorylation of Sox9. Pharmacological inhibition of BMP signaling represses MB cell transdifferentiation into astrocytes and suppresses tumor relapse. Our studies establish the distinct cellular sources of astrocytes in primary and relapsed MB and provide an avenue to prevent and treat MB relapse by targeting tumor cell transdifferentiation.


Asunto(s)
Astrocitos/patología , Neoplasias Cerebelosas/patología , Meduloblastoma/patología , Animales , Proteínas Morfogenéticas Óseas/metabolismo , Proteínas Morfogenéticas Óseas/farmacología , Transdiferenciación Celular/efectos de los fármacos , Neoplasias Cerebelosas/genética , Neoplasias Cerebelosas/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteínas Hedgehog/metabolismo , Humanos , Meduloblastoma/genética , Meduloblastoma/metabolismo , Ratones Transgénicos , Receptor Patched-1/genética , Receptor Patched-1/metabolismo , Fosforilación , Pirazoles/farmacología , Pirimidinas/farmacología , Factor de Transcripción SOX9/metabolismo , Análisis de la Célula Individual , Ensayos Antitumor por Modelo de Xenoinjerto
14.
Mol Cell Oncol ; 7(6): 1810514, 2020 Sep 05.
Artículo en Inglés | MEDLINE | ID: mdl-33235914

RESUMEN

Medulloblastoma (MB) often originate from cerebellar granule neuron precursors (GNPs). We recently found that medulloblastoma cells undergo differentiation as GNPs. Differentiated MB cells have permanently lost their proliferative capacity and tumorigenicity. The differentiation of MB cells is driven by the transcription factor NeuroD1 (Neurogenic differentiation 1), and NeuroD1 expression in MB cells is repressed by EZH2-mediated H3K27me3.

15.
J Biotechnol ; 324: 91-98, 2020 Dec 20.
Artículo en Inglés | MEDLINE | ID: mdl-33010308

RESUMEN

Enzymes, which provide more efficient and eco-friendly strategies for various functional molecules' construction than traditional chemo-catalysts, were utilized for the synthesis of 4H-pyrimido[2,1-b] benzothiazole derivatives. Reported herein is a trypsin-catalysed three- component Biginelli reaction of aldehyde, ß-ketoester and 2-amino benzothiazole in one pot, affording a streamlined pathway to diverse ring-fused pyrimidines. In addition to using commercially available aromatic aldehydes as substrates, acetaldehyde, the chemical liquid with rather low boiling point and difficult to handle above room temperature, is utilized to further extend the range of substrates. It was verified that most of the tested substrates exhibited satisfactory reactivity. In addition, several substrates indicated AIE (Aggregation-Induced Emission) property and have been investigated as potential biomarkers.


Asunto(s)
Aldehídos , Benzotiazoles , Acetaldehído , Catálisis , Pirimidinas
16.
ACS Appl Bio Mater ; 3(8): 5193-5201, 2020 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-35021694

RESUMEN

A series of aggregation-induced emission (AIE) fluorescence probes, coined 4H-pyrimido[2,1-b]benzothiazole derivatives, has been synthesized by Biginelli reactions. Utilizing spectroscopic techniques, their photophysical properties have been comprehensively investigated in working environment both in vitro and in vivo. Density functional theory (DFT) calculations were performed to better understand the mechanism of these probes. The interactions between 4H-pyrimido[2,1-b]benzothiazoles with different substituents and bovine serum albumin (BSA) were analyzed using UV-vis and fluorescence spectroscopy, synchronous fluorescence, and docking analysis. Studies found that 4H-pyrimido[2,1-b]benzothiazole could bind to bovine serum albumin (BSA) through a hydrogen bond and hydrophobic interactions, resulting in enhancement of fluorescence emission of probes 1a-6f and fluorescence quenching of BSA. Combined with cell imaging experiments, these provide information on potential effects of benzothiazoles on disease treatment.

17.
Cell Rep ; 31(12): 107782, 2020 06 23.
Artículo en Inglés | MEDLINE | ID: mdl-32579914

RESUMEN

Tumor cells are characterized by unlimited proliferation and perturbed differentiation. Using single-cell RNA sequencing, we demonstrate that tumor cells in medulloblastoma (MB) retain their capacity to differentiate in a similar way as their normal originating cells, cerebellar granule neuron precursors. Once they differentiate, MB cells permanently lose their proliferative capacity and tumorigenic potential. Differentiated MB cells highly express NeuroD1, a helix-loop-helix transcription factor, and forced expression of NeuroD1 promotes the differentiation of MB cells. The expression of NeuroD1 in bulk MB cells is repressed by trimethylation of histone 3 lysine-27 (H3K27me3). Inhibition of the histone lysine methyltransferase EZH2 prevents H3K27 trimethylation, resulting in increased NeuroD1 expression and enhanced differentiation in MB cells, which consequently reduces tumor growth. These studies reveal the mechanisms underlying MB cell differentiation and provide rationales to treat MB (potentially other malignancies) by stimulating tumor cell differentiation.


Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diferenciación Celular , Neoplasias Cerebelosas/patología , Meduloblastoma/patología , Proteínas del Tejido Nervioso/metabolismo , Animales , Carcinogénesis/patología , Línea Celular Tumoral , Proliferación Celular , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Proteínas Hedgehog/metabolismo , Histonas/metabolismo , Humanos , Lisina/metabolismo , Metilación , Ratones , Neuronas/metabolismo , Neuronas/patología , Receptor Patched-1/metabolismo , Transducción de Señal , Análisis de la Célula Individual
18.
Eur J Med Chem ; 172: 1-15, 2019 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-30939349

RESUMEN

The Hedgehog (Hh) pathway plays a critical role during embryonic development by controlling cell patterning, growth and migration. In adults, the function of Hh pathway is curtailed to tissue repair and maintenance. Aberrant reactivation of Hh signaling has been linked to tumorigenesis in various cancers, such as basal cell carcinoma (BCC) and medulloblastoma. The Smoothened (Smo) receptor, a key component of the Hh pathway which is central to the signaling transduction, has emerged as an attractive therapeutic target for the treatment of human cancers. Taking advantage of the availability of several crystal structures of Smo in complex with different antagonists, we have previously conducted a molecular docking-based virtual screening to identify several compounds which exhibited significant inhibitory activity against the Hh pathway activation (IC50 < 10 µM) in a Gli-responsive element (GRE) reporter gene assay. The most potent compound (ChemDiv ID C794-1677: 47 nM) showed comparable Hh signaling inhibition to the marketed drug vismodegib (46 nM). Herein, we report our structural optimization based on the virtual screening hit C794-1677. Our efforts are aimed to improve potency, decrease cLogP, and remove potentially metabolic labile/toxic pyrrole and aniline functionalities presented in C794-1677. The optimization led to the identification of numerous potent compounds exemplified by 25 (7.1 nM), which was 7 folds more potent compared with vismodegib. In addition, 25 was much less lipophilic compared with C794-1677 and devoid of the potentially metabolic labile/toxic pyrrole and aniline functional groups. Furthermore, 25 exhibited promising efficacy in inhibiting Gli1 mRNA expression in NIH3T3 cells with either wildtype Smo or D473H Smo mutant. These results represented significant improvement over the virtual screening hit C794-1677 and suggested that compound 25 can be used as a good starting point to support lead optimization.


Asunto(s)
Anilidas/farmacología , Simulación por Computador , Evaluación Preclínica de Medicamentos , Piridinas/farmacología , Receptor Smoothened/antagonistas & inhibidores , Anilidas/química , Animales , Relación Dosis-Respuesta a Droga , Ratones , Simulación del Acoplamiento Molecular , Estructura Molecular , Células 3T3 NIH , Piridinas/química , Relación Estructura-Actividad
19.
Acta Neuropathol Commun ; 7(1): 179, 2019 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-31727166

RESUMEN

In the original version of this article [1], there was 1 error in the affiliation of the European Institute of Oncology (affiliation 3). In this correction article the updated affiliation is shown for clarification.

20.
Clin Cancer Res ; 25(21): 6475-6486, 2019 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-31300449

RESUMEN

PURPOSE: Here, we examined the role of leukotrienes, well-known inflammatory mediators, in the tumorigenesis of hedgehog pathway-associated medulloblastoma, and tested the efficacies of antagonists of leukotriene biosynthesis in medulloblastoma treatment.Experimental Design: We examined the leukotriene levels in medulloblastoma cells by ELISA. We next tested whether leukotriene synthesis in medulloblastoma cells relied on activation of hedgehog pathway, or the presence of hedgehog ligand secreted by astrocytes. We then investigated whether leukotriene mediated hedgehog-induced Nestin expression in tumor cells. The functions of leukotriene in tumor cell proliferation and tumor growth in medulloblastoma were determined through knocking down 5-lipoxygenase (a critical enzyme for leukotriene synthesis) by shRNAs, or using 5-lipoxygenase-deficient mice. Finally, the efficacies of antagonists of leukotriene synthesis in medulloblastoma treatment were tested in vivo and in vitro. RESULTS: Leukotriene was significantly upregulated in medulloblastoma cells. Increased leukotriene synthesis relied on hedgehog ligand secreted by astrocytes, a major component of medulloblastoma microenvironment. Leukotriene stimulated tumor cells to express Nestin, a cytoskeletal protein essential for medulloblastoma growth. Genetic blockage of leukotriene synthesis dramatically suppressed medulloblastoma cell proliferation and tumor growth in vivo. Pharmaceutical inhibition of leukotriene synthesis markedly repressed medulloblastoma cell proliferation, but had no effect on proliferation of normal neuronal progenitors. Moreover, antagonists of leukotriene synthesis exhibited promising tumor inhibitory efficacies on drug-resistant medulloblastoma. CONCLUSIONS: Our findings reveal a novel signaling pathway that is critical for medulloblastoma cell proliferation and tumor progression, and that leukotriene biosynthesis represents a promising therapeutic target for medulloblastoma treatment.


Asunto(s)
Araquidonato 5-Lipooxigenasa/genética , Carcinogénesis/genética , Leucotrienos/genética , Meduloblastoma/genética , Animales , Araquidonato 5-Lipooxigenasa/deficiencia , Astrocitos/metabolismo , Astrocitos/patología , Línea Celular Tumoral , Proliferación Celular/genética , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica/genética , Proteínas Hedgehog/genética , Humanos , Leucotrienos/biosíntesis , Meduloblastoma/patología , Ratones , Ratones Noqueados , ARN Interferente Pequeño/genética , Transducción de Señal/genética
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