RESUMEN
Inflammation is a fundamental defense mechanism to protect the body from danger, which becomes potentially harmful if it turns chronic. Therapeutic strategies aimed at specifically blocking proinflammatory signals, particularly cytokines, such as IL-4, IL-6, IL-13, IL-17A, or TNF-α, have substantially improved our ability to effectively and safely treat chronic inflammatory diseases. Much less effort has been made to better understand the role of potential anti-inflammatory mechanisms. Here we summarize the current understanding of regulatory cell populations in the context of chronic inflammation, namely macrophages, Langerhans cells, myeloid-derived suppressor cells, and regulatory T and B lymphocytes. Emphasis is given to the skin because many different immune-related diseases occur in the skin. Development, phenotype, function, and evidence for their role in animal models of inflammation, as well as in the corresponding human diseases, are described. Finally, the feasibility of using regulatory cells as targets for potentially disease-modifying therapeutic strategies is discussed.
Asunto(s)
Inflamación/inmunología , Piel/inmunología , Animales , Antiinflamatorios/farmacología , Linfocitos B/inmunología , Humanos , Inflamación/patología , Células de Langerhans/inmunología , Macrófagos/inmunología , Células Supresoras de Origen Mieloide/inmunología , Piel/patología , Linfocitos T Reguladores/inmunologíaRESUMEN
Recent work has shown that deregulation of the transcription factor Myb contributes to the development of leukemia and several other human cancers, making Myb and its cooperation partners attractive targets for drug development. By employing a myeloid Myb-reporter cell line we have identified Withaferin A (WFA), a natural compound that exhibits anti-tumor activities, as an inhibitor of Myb-dependent transcription. Analysis of the inhibitory mechanism of WFA showed that WFA is a significantly more potent inhibitor of C/EBPß, a transcription factor cooperating with Myb in myeloid cells, than of Myb itself. We show that WFA covalently modifies specific cysteine residues of C/EBPß, resulting in the disruption of the interaction of C/EBPß with the co-activator p300. Our work identifies C/EBPß as a novel direct target of WFA and highlights the role of p300 as a crucial co-activator of C/EBPß. The finding that WFA is a potent inhibitor of C/EBPß suggests that inhibition of C/EBPß might contribute to the biological activities of WFA.
Asunto(s)
Antineoplásicos/farmacología , Proteína beta Potenciadora de Unión a CCAAT/antagonistas & inhibidores , Witanólidos/farmacología , Células 3T3 , Animales , Sitios de Unión , Proteína beta Potenciadora de Unión a CCAAT/química , Línea Celular Tumoral , Humanos , Ratones , Unión Proteica , Factores de Transcripción p300-CBP/metabolismoRESUMEN
The transcription factor Myb is a key regulator of hematopoietic cell proliferation, differentiation, and survival and has been implicated in the development of leukemia and several other human cancers. Pharmacological inhibition of Myb is therefore emerging as a potential therapeutic strategy. Recently, the first low-molecular-weight compounds that show Myb inhibitory activity have been identified. Characterization of these compounds suggests disruption of the protein-protein-interaction of Myb and the coactivator p300 as a suitable strategy to inhibit Myb.
Asunto(s)
Antineoplásicos/farmacología , Terapia Molecular Dirigida , Proteínas Proto-Oncogénicas c-myb/antagonistas & inhibidores , Factores de Transcripción/antagonistas & inhibidores , Animales , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Humanos , Leucemia/tratamiento farmacológico , Leucemia/etiología , Leucemia/metabolismo , Unión Proteica , Proteínas Proto-Oncogénicas c-myb/química , Proteínas Proto-Oncogénicas c-myb/genética , Proteínas Proto-Oncogénicas c-myb/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/química , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Activación TranscripcionalRESUMEN
The transcription factor c-Myb is essential for the proliferation of hematopoietic cells and has been implicated in the development of leukemia and other human cancers. Pharmacologic inhibition of Myb is therefore emerging as a potential therapeutic strategy for these diseases. By using a Myb reporter cell line, we have identified plumbagin and several naphthoquinones as potent low-molecular weight Myb inhibitors. We demonstrate that these compounds inhibit c-Myb by binding to the c-Myb transactivation domain and disrupting the cooperation of c-Myb with the coactivator p300, a major driver of Myb activity. Naphthoquinone-induced inhibition of c-Myb suppresses Myb target gene expression and induces the differentiation of the myeloid leukemia cell line HL60. We demonstrate that murine and human primary acute myeloid leukemia cells are more sensitive to naphthoquinone-induced inhibition of clonogenic proliferation than normal hematopoietic progenitor cells. Overall, our work demonstrates for the first time the potential of naphthoquinones as small-molecule Myb inhibitors that may have therapeutic potential for the treatment of leukemia and other tumors driven by deregulated Myb. Mol Cancer Ther; 15(12); 2905-15. ©2016 AACR.
Asunto(s)
Antineoplásicos/farmacología , Proteína p300 Asociada a E1A/metabolismo , Leucemia Mieloide Aguda/metabolismo , Proteínas Proto-Oncogénicas c-myb/metabolismo , Antineoplásicos/química , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/patología , Naftoquinonas/química , Naftoquinonas/farmacología , Unión Proteica/efectos de los fármacos , Dominios y Motivos de Interacción de Proteínas , Proteínas Proto-Oncogénicas c-myb/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-myb/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The transcription factor Myb plays a key role in the hematopoietic system and has been implicated in the development of leukemia and other human cancers. Inhibition of Myb is therefore emerging as a potential therapeutic strategy for these diseases. However, because of a lack of suitable inhibitors, the feasibility of therapeutic approaches based on Myb inhibition has not been explored. We have identified the triterpenoid Celastrol as a potent low-molecular-weight inhibitor of the interaction of Myb with its cooperation partner p300. We demonstrate that Celastrol suppresses the proliferative potential of acute myeloid leukemia (AML) cells while not affecting normal hematopoietic progenitor cells. Furthermore, Celastrol prolongs the survival of mice in a model of an aggressive AML. Overall, our work demonstrates the therapeutic potential of a small molecule inhibitor of the Myb/p300 interaction for the treatment of AML and provides a starting point for the further development of Myb-inhibitory compounds for the treatment of leukemia and, possibly, other tumors driven by deregulated Myb.
Asunto(s)
Proteína p300 Asociada a E1A/metabolismo , Leucemia Mieloide Aguda/tratamiento farmacológico , Terapia Molecular Dirigida , Proteínas Proto-Oncogénicas c-myb/metabolismo , Bibliotecas de Moléculas Pequeñas/uso terapéutico , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Pollos , Modelos Animales de Enfermedad , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Células HL-60 , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Humanos , Leucemia Mieloide Aguda/patología , Ratones Endogámicos C57BL , Células Mieloides/efectos de los fármacos , Células Mieloides/patología , Triterpenos Pentacíclicos , Unión Proteica/efectos de los fármacos , Estructura Terciaria de Proteína , Bibliotecas de Moléculas Pequeñas/farmacología , Triterpenos/farmacología , Triterpenos/uso terapéuticoRESUMEN
The transcription factor c-Myb is highly expressed in hematopoietic progenitor cells and controls the transcription of genes important for lineage determination, cell proliferation, and differentiation. Deregulation of c-Myb has been implicated in the development of leukemia and certain other types of human cancer. c-Myb activity is highly dependent on the interaction of the c-Myb with the KIX domain of the coactivator p300, making the disruption of this interaction a reasonable strategy for the development of Myb inhibitors. Here, we have used bacterial Autodisplay to develop an in vitro binding assay that mimics the interaction of Myb and the KIX domain of p300. We have used this binding assay to investigate the potential of Naphthol AS-E phosphate, a compound known to bind to the KIX domain, to disrupt the interaction between Myb and p300. Our data show that Naphthol AS-E phosphate interferes with the Myb-KIX interaction in vitro and inhibits Myb activity in vivo. By using several human leukemia cell lines, we demonstrate that Naphthol AS-E phosphate suppresses the expression of Myb target genes and induces myeloid differentiation and apoptosis. Our work identifies Naphthol AS-E phosphate as the first low molecular weight compound that inhibits Myb activity by disrupting its interaction with p300, and suggests that inhibition of the Myb-KIX interaction might be a useful strategy for the treatment of leukemia and other tumors caused by deregulated c-Myb.
Asunto(s)
Proteína p300 Asociada a E1A/metabolismo , Naftoles/farmacología , Organofosfatos/farmacología , Proteínas Proto-Oncogénicas c-myb/metabolismo , Apoptosis/efectos de los fármacos , Apoptosis/genética , Sitios de Unión/genética , Western Blotting , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Línea Celular Tumoral , Proteína p300 Asociada a E1A/genética , Regulación Leucémica de la Expresión Génica/genética , Células HL-60 , Humanos , Microscopía Fluorescente , Naftoles/metabolismo , Organofosfatos/metabolismo , Unión Proteica/efectos de los fármacos , Proteínas Proto-Oncogénicas c-myb/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-myb/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células U937RESUMEN
Histone-lysine acetylation is a vital chromatin post-translational modification involved in the epigenetic regulation of gene transcription. Bromodomains bind acetylated lysines, acting as readers of the histone-acetylation code. Competitive inhibitors of this interaction have antiproliferative and anti-inflammatory properties. With 57 distinct bromodomains known, the discovery of subtype-selective inhibitors of the histone-bromodomain interaction is of great importance. We have identified the 3,5-dimethylisoxazole moiety as a novel acetyl-lysine bioisostere, which displaces acetylated histone-mimicking peptides from bromodomains. Using X-ray crystallographic analysis, we have determined the interactions responsible for the activity and selectivity of 4-substituted 3,5-dimethylisoxazoles against a selection of phylogenetically diverse bromodomains. By exploiting these interactions, we have developed compound 4d, which has IC(50) values of <5 µM for the bromodomain-containing proteins BRD2(1) and BRD4(1). These compounds are promising leads for the further development of selective probes for the bromodomain and extra C-terminal domain (BET) family and CREBBP bromodomains.
Asunto(s)
Isoxazoles/síntesis química , Lisina/metabolismo , Alcohol Feniletílico/análogos & derivados , Factores de Transcripción/metabolismo , Proteína de Unión a CREB/metabolismo , Proteínas de Ciclo Celular , Cristalografía por Rayos X , Citotoxinas/síntesis química , Citotoxinas/química , Citotoxinas/farmacología , Células HeLa , Histonas/metabolismo , Humanos , Isoxazoles/química , Isoxazoles/farmacología , Ligandos , Modelos Moleculares , Imitación Molecular , Estructura Molecular , Proteínas Nucleares/metabolismo , Alcohol Feniletílico/síntesis química , Alcohol Feniletílico/química , Alcohol Feniletílico/farmacología , Unión Proteica , Proteínas Serina-Treonina Quinasas/metabolismo , Estructura Terciaria de Proteína , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Histone lysine acetylation is a key component of epigenetic regulation of gene transcription. Bromodomains, found in histone acetyl transferases and other chromatin-associated proteins, bind selectively to acetylated lysines, acting as "readers" of the histone code, and have recently been shown to contain a druggable binding pocket. Here we report the development of high-throughput assays that quantify the binding of bromodomains to acetylated histone peptides. We have used these assays to screen for histone binding partners of as yet uncharacterized bromodomains, adding to current knowledge of the histone code and expanding the repertoire of assays for chemical probe discovery. We have also demonstrated that these assays can be used to detect small molecule binding from the very weak to the nanomolar range. This assay methodology is thereby anticipated to provide the basis both for broader interactome profiling and for small molecule inhibitor discovery.