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1.
SAR QSAR Environ Res ; 32(2): 111-131, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33461329

RESUMEN

This paper is devoted to the analysis of available experimental data and preparation of predictive models for binding affinity of molecules with respect to two nuclear receptors involved in endocrine disruption (ED): the oestrogen (ER) and the androgen (AR) receptors. The ED-relevant data were retrieved from multiple sources, including the CERAPP, CoMPARA, and the Tox21 projects as well as ChEMBL and PubChem databases. Data analysis performed with the help of generative topographic mapping revealed the problem of low agreement between experimental values from different sources. Collected data were used to train both classification models for ER and AR binding activities and regression models for relative binding affinity (RBA) and median inhibition concentration (IC50). These models displayed relatively poor performance in classification (sensitivities ER = 0.34, AR = 0.49) and in regression (determination coefficient r 2 for the RBA and IC50 models in external validation varied from 0.44 to 0.76). Our analysis demonstrates that low models' performance resulted from misinterpreted experimental endpoints or wrongly reported values, thus confirming the observations reported in CERAPP and CoMPARA studies. Developed models and collected data sets included of 6215 (ER) and 3789 (AR) unique compounds, which are freely available.


Asunto(s)
Disruptores Endocrinos/química , Relación Estructura-Actividad Cuantitativa , Receptores Androgénicos/química , Receptores Estrogénicos/química , Humanos , Modelos Teóricos
2.
Cells ; 9(12)2020 12 10.
Artículo en Inglés | MEDLINE | ID: mdl-33321757

RESUMEN

Around 80-90% of prostate cancer (PCa) cases are dependent on androgens at initial diagnosis; hence, androgen ablation therapy directed toward a reduction in serum androgens and the inhibition of androgen receptor (AR) is generally the first therapy adopted. However, the patient's response to androgen ablation therapy is variable, and 20-30% of PCa cases become castration resistant (CRPCa). Several mechanisms can guide treatment resistance to anti-AR molecules. In this regard, AR-dependent and -independent resistance mechanisms can be distinguished within the AR pathway. In this article, we investigate the multitude of AR signaling aspects, encompassing the biological structure of AR, current AR-targeted therapies, mechanisms driving resistance to AR, and AR crosstalk with other pathways, in an attempt to provide a comprehensive review for the PCa research community. We also summarize the new anti-AR drugs approved in non-metastatic castration-resistant PCa, in the castration-sensitive setting, and combination therapies with other drugs.


Asunto(s)
Resistencia a Antineoplásicos , Neoplasias de la Próstata/metabolismo , Receptores Androgénicos/metabolismo , Antagonistas de Receptores Androgénicos/uso terapéutico , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Neoplasias de la Próstata/tratamiento farmacológico , Receptores Androgénicos/química , Transducción de Señal
3.
BMC Bioinformatics ; 21(Suppl 5): 245, 2020 Oct 26.
Artículo en Inglés | MEDLINE | ID: mdl-33106158

RESUMEN

BACKGROUND: Abnormal activation of human nuclear hormone receptors disrupts endocrine systems and thereby affects human health. There have been machine learning-based models to predict androgen receptor agonist activity. However, the models were constructed based on limited numerical features such as molecular descriptors and fingerprints. RESULT: In this study, instead of the numerical features, 2-D chemical structure images of compounds were used to build an androgen receptor toxicity prediction model. The images may provide unknown features that were not represented by conventional numerical features. As a result, the new strategy resulted in a construction of highly accurate prediction model: Mathews correlation coefficient (MCC) of 0.688, positive predictive value (PPV) of 0.933, sensitivity of 0.519, specificity of 0.998, and overall accuracy of 0.981 in 10-fold cross-validation. Validation on a test dataset showed MCC of 0.370, sensitivity of 0.211, specificity of 0.991, PPV of 0.882, and overall accuracy of 0.801. Our chemical image-based prediction model outperforms conventional models based on numerical features. CONCLUSION: Our constructed prediction model successfully classified molecular images into androgen receptor agonists or inactive compounds. The result indicates that 2-D molecular mimetic diagram would be used as another feature to construct molecular activity prediction models.


Asunto(s)
Receptores Androgénicos/química , Simulación por Computador , Humanos , Modelos Moleculares , Sensibilidad y Especificidad
4.
Mol Cell ; 79(5): 812-823.e4, 2020 09 03.
Artículo en Inglés | MEDLINE | ID: mdl-32668201

RESUMEN

Steroid receptors activate gene transcription by recruiting coactivators to initiate transcription of their target genes. For most nuclear receptors, the ligand-dependent activation function domain-2 (AF-2) is a primary contributor to the nuclear receptor (NR) transcriptional activity. In contrast to other steroid receptors, such as ERα, the activation function of androgen receptor (AR) is largely dependent on its ligand-independent AF-1 located in its N-terminal domain (NTD). It remains unclear why AR utilizes a different AF domain from other receptors despite that NRs share similar domain organizations. Here, we present cryoelectron microscopy (cryo-EM) structures of DNA-bound full-length AR and its complex structure with key coactivators, SRC-3 and p300. AR dimerization follows a unique head-to-head and tail-to-tail manner. Unlike ERα, AR directly contacts a single SRC-3 and p300. The AR NTD is the primary site for coactivator recruitment. The structures provide a basis for understanding assembly of the AR:coactivator complex and its domain contributions for coactivator assembly and transcriptional regulation.


Asunto(s)
ADN/química , Proteína p300 Asociada a E1A/metabolismo , Coactivador 3 de Receptor Nuclear/metabolismo , Receptores Androgénicos/metabolismo , Microscopía por Crioelectrón , ADN/metabolismo , Proteína p300 Asociada a E1A/química , Células HEK293 , Humanos , Coactivador 3 de Receptor Nuclear/química , Conformación de Ácido Nucleico , Conformación Proteica , Receptores Androgénicos/química , Receptores Androgénicos/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo
5.
Mol Genet Genomics ; 295(5): 1103-1111, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32435981

RESUMEN

The androgen insensitivity syndrome (AIS) is a congenital disease characterized by androgen resistance due to androgen receptor (AR) gene mutations, resulting in disorders of sex differentiation in 46,XY individuals. However, the underlying mechanisms in the majority of AR variants and the phenotype-genotype correlations are unclear. Here, we identified a p.Y764H variant of the AR gene that results in different phenotypes in a family. Structural analyses revealed that amino acid substitution affected protein properties and spatial conformation, and in vitro, functional studies showed impaired nuclear translocation ability of the mutated protein. Moreover, the extent to which this variant reduced nuclear translocation depends on the dihydrotestosterone (DHT) concentrations. Our results, for the first time, demonstrated a pathogenesis of the p.Y764H mutations in AR resulting in AIS phenotype, and indicated that AIS patients with p.Y764H mutation and preserved gonad might have residual AR activity at high androgen levels, putting patients at risk for pubertal virilization in the future. We provide an in-depth insight into the pathogenesis in AIS based on the amino acid substitution, which may help aid its precise diagnosis, personalized treatment, and organized follow-up to avoid gender dysphoria.


Asunto(s)
Sustitución de Aminoácidos , Síndrome de Resistencia Androgénica/genética , Núcleo Celular/metabolismo , Receptores Androgénicos/genética , Receptores Androgénicos/metabolismo , Secuenciación del Exoma Completo/métodos , Síndrome de Resistencia Androgénica/metabolismo , Animales , Células COS , China , Chlorocebus aethiops , Femenino , Hemicigoto , Humanos , Lactante , Masculino , Linaje , Fenotipo , Transporte de Proteínas , Receptores Androgénicos/química , Hermanos
6.
Cancer Cell ; 37(4): 584-598.e11, 2020 04 13.
Artículo en Inglés | MEDLINE | ID: mdl-32220301

RESUMEN

Metastatic prostate cancer is characterized by recurrent genomic copy number alterations that are presumed to contribute to resistance to hormone therapy. We identified CHD1 loss as a cause of antiandrogen resistance in an in vivo small hairpin RNA (shRNA) screen of 730 genes deleted in prostate cancer. ATAC-seq and RNA-seq analyses showed that CHD1 loss resulted in global changes in open and closed chromatin with associated transcriptomic changes. Integrative analysis of this data, together with CRISPR-based functional screening, identified four transcription factors (NR3C1, POU3F2, NR2F1, and TBX2) that contribute to antiandrogen resistance, with associated activation of non-luminal lineage programs. Thus, CHD1 loss results in chromatin dysregulation, thereby establishing a state of transcriptional plasticity that enables the emergence of antiandrogen resistance through heterogeneous mechanisms.


Asunto(s)
Antagonistas de Andrógenos/farmacología , Cromatina/genética , ADN Helicasas/antagonistas & inhibidores , Proteínas de Unión al ADN/antagonistas & inhibidores , Resistencia a Antineoplásicos/genética , Neoplasias de la Próstata Resistentes a la Castración/tratamiento farmacológico , ARN Interferente Pequeño/genética , Receptores Androgénicos/química , Animales , Apoptosis , Biomarcadores de Tumor/genética , Proliferación Celular , Cromatina/metabolismo , ADN Helicasas/genética , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Regulación Neoplásica de la Expresión Génica , Ensayos Analíticos de Alto Rendimiento , Humanos , Masculino , Ratones , Neoplasias de la Próstata Resistentes a la Castración/genética , Neoplasias de la Próstata Resistentes a la Castración/patología , Receptores Androgénicos/genética , Factores de Transcripción/metabolismo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de Xenoinjerto
7.
Proc Natl Acad Sci U S A ; 117(15): 8584-8592, 2020 04 14.
Artículo en Inglés | MEDLINE | ID: mdl-32220959

RESUMEN

The androgen receptor (AR) is a type I nuclear hormone receptor and the primary drug target in prostate cancer due to its role as a lineage survival factor in prostate luminal epithelium. In prostate cancer, the AR cistrome is reprogrammed relative to normal prostate epithelium and particularly in cancers driven by oncogenic ETS fusion genes. The molecular basis for this change has remained elusive. Using purified proteins, we report a minimal cell-free system that demonstrates interdomain cooperativity between the ligand (LBD) and DNA binding domains (DBD) of AR, and its autoinhibition by the N terminus of AR. Furthermore, we identify ERG as a cofactor that activates AR's ability to bind DNA in both high and lower affinity contexts through direct interaction within a newly identified AR-interacting motif (AIM) in the ETS domain, independent of ERG's own DNA binding ability. Finally, we present evidence that this interaction is conserved among ETS factors whose expression is altered in prostate cancer. Our work highlights, at a biochemical level, how tumor-initiating ETS translocations result in reprogramming of the AR cistrome.


Asunto(s)
ADN/metabolismo , Regulación Neoplásica de la Expresión Génica , Neoplasias de la Próstata/metabolismo , Proteínas Proto-Oncogénicas c-ets/metabolismo , Receptores Androgénicos/metabolismo , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , ADN/genética , Humanos , Masculino , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Proteínas Proto-Oncogénicas c-ets/genética , Receptores Androgénicos/química , Receptores Androgénicos/genética , Regulador Transcripcional ERG/química , Regulador Transcripcional ERG/genética , Regulador Transcripcional ERG/metabolismo , Células Tumorales Cultivadas
8.
Gen Comp Endocrinol ; 291: 113418, 2020 05 15.
Artículo en Inglés | MEDLINE | ID: mdl-32027878

RESUMEN

Although testosterone (T) is a key regulator in vertebrate development, physiology, and behaviour in both sexes, studies suggest that its regulation may be sex-specific. We measured circulating T levels in Baluchistan gerbils (Gerbillus nanus) in the field and in the lab all year round and found no significant sex differences. However, we observed sex differences in circulating T levels following gonadotropin-releasing hormone (GnRH) challenge and T implants in this non-model species. Whereas only males elevated T following a GnRH challenge, females had higher serum T concentrations following T implant insertion. These differences may be a result of different points of regulation along the hypothalamic-pituitary-gonadal (HPG) axis. Consequently, we examined sex differences in the mRNA expression of the androgen receptor (AR) in multiple brain regions. We identified AR and ß-actin sequences in assembled genomic sequences of members of the Gerbillinae, which were analogous to rat sequences, and designed primers for them. The distribution of the AR in G. nanus brain regions was similar to documented expression profiles in rodents. We found lower AR mRNA levels in females in the striatum. Additionally, G. nanus that experienced housing in mixed-sex pairs had higher adrenal AR expression than G. nanus that were housed alone. Regulation of the gerbil HPG axis may reflect evolutionary sex differences in life-history strategies, with males ready to reproduce when receptive females are available, while the possible reproductive costs associated with female T direct its regulation upstream.


Asunto(s)
Gerbillinae/sangre , Caracteres Sexuales , Testosterona/sangre , Secuencia de Aminoácidos , Animales , Femenino , Regulación de la Expresión Génica , Gerbillinae/genética , Hormona Liberadora de Gonadotropina/metabolismo , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores Androgénicos/química , Receptores Androgénicos/genética , Receptores Androgénicos/metabolismo
9.
Molecules ; 25(2)2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31952272

RESUMEN

The Androgen Receptor (AR) is emerging as an important factor in the pathogenesis of breast cancer (BC), which is the most common malignancy among females worldwide. The concordance of more than 70% of AR expression in primary and metastatic breast tumors implies that AR may be a new marker and a potential therapeutic target among AR-positive breast cancer patients. Biological insight into AR-positive breast cancer reveals that AR may cross-talk with several vital signaling pathways, including key molecules and receptors. AR exhibits different behavior depending on the breast cancer subtype. Preliminary clinical research using AR-targeted drugs, which have already been FDA-approved for prostate cancer (PC), has given promising results for AR-positive breast cancer patients. However, since the prognostic and predictive value of AR positivity remains uncertain, it is difficult to identify and stratify patients that would benefit from AR-targeted therapies. Herein, through a review of preclinical studies, clinical studies, and clinical trials, we summarize the biology of AR, its prognostic and predictive value, as well as its therapeutic implications by breast cancer molecular subtype.


Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Terapia Molecular Dirigida , Receptores Androgénicos/química , Animales , Neoplasias de la Mama/patología , Evaluación Preclínica de Medicamentos , Femenino , Humanos , Receptores Androgénicos/metabolismo , Transducción de Señal
10.
Eur J Cancer ; 127: 67-75, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31986451

RESUMEN

BACKGROUND: The unmet need for predictive biomarkers emerged from the unpredictable pattern of response to androgen signalling inhibition in metastatic castration-resistant prostate cancer (mCRPC). Here, we report on the testing of a previously identified candidate androgen signalling signature associated with response to androgen signalling inhibition. PATIENTS AND METHODS: We report on the outcome of the first module of a phase II trial on abiraterone acetate (AA) followed by combination with dasatinib or sunitinib. Bone marrow biopsies (BMBs) with matched bone marrow aspirate and blood samples were collected at baseline and upon progression. End-points included assessment of a prespecified molecular signature consisting of nuclear androgen receptor (AR) overexpression, cytochrome P450, family 17, subfamily A, polypeptide 1 (CYP17) expression, and AR-C-/N terminal expression ratio of ≥0.8 by immunohistochemistry (IHC) in patients with benefit versus primary resistance to AA (i.e. progression within 4 months). Tumour markers also included v-ets avian erythroblastosis virus E26 oncogene homologue (ERG), androgen receptor splice variant (ARV7) by IHC and steroids by liquid chromatography-tandem mass spectrometry. RESULTS: Of 170 patients accrued from 03/2011 to 02/2015, 44 (26%) were primary resistant to AA. Forty-eight patients had tumour infiltrated BMB at baseline. Pretreatment androgen signalling signature was linked to benefit from AA (p < 0.001). Presence of ERG was associated with benefit (p = 0.05), whereas nuclear ARV7 presence and 20 or more bone lesions at baseline with primary resistance (p = 0.04 and p = 0.0006, respectively). CONCLUSION: Testing of a prespecified androgen signalling signature was highly supportive of its predictive value in maximal androgen deprivation strategies in mCRPC. Further validation is under way. TRIAL REGISTRATION: ClinicalTrials.gov NCT01254864.


Asunto(s)
Acetato de Abiraterona/uso terapéutico , Antagonistas de Andrógenos/uso terapéutico , Biomarcadores de Tumor/sangre , Neoplasias de la Próstata Resistentes a la Castración/sangre , Receptores Androgénicos/química , Anciano , Anciano de 80 o más Años , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia , Pronóstico , Estudios Prospectivos , Neoplasias de la Próstata Resistentes a la Castración/tratamiento farmacológico , Neoplasias de la Próstata Resistentes a la Castración/patología , Curva ROC
11.
Food Chem Toxicol ; 137: 111116, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31931072

RESUMEN

Bisphenol A (BPA), a synthetic compound widely used as a building block for polycarbonate plastics, has been declared in the European Union (EU) as a substance of very high concern (SVHC). A series of BPA alternatives and derivatives (bisphenols/BPs) with similar physical-chemical properties have been produced and used by companies for substituting it. To evaluate the estrogenic and androgenic binding activity of 26 BPs, a non-statistical in silico approach has been applied. The results of molecular docking analyses applied on six different nuclear receptors (NRs) have revealed that: i) some BPA metabolites could lower the harmful effects of BPA exposure; ii) BPS is a lower interactor for all NRs, but it does not appear safer at all for androgen receptor (AR), for which its binding activity is found similar to a pharmacological anti-androgen; iii) only a BP has been found as a safer compound for all NRs considered. Moreover, molecular dynamic simulation of three BPs on ERα have revealed that the presence of negative hydrophobic interactions could induce a decrease in receptor activity. Overall, the present results demonstrate that in silico methods could be a valid approach to screen estrogenic and androgenic activity of food contact materials (FCMs).


Asunto(s)
Compuestos de Bencidrilo/química , Fenoles/química , Andrógenos/química , Estrógenos/química , Contaminación de Alimentos/análisis , Inocuidad de los Alimentos , Simulación de Dinámica Molecular , Estructura Molecular , Receptores Androgénicos/química , Receptores Estrogénicos/química
12.
Mol Cell Endocrinol ; 499: 110577, 2020 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-31525432

RESUMEN

Androgenic hormones orchestrate the development and activation of diverse reproductive phenotypes across vertebrates. Although extensive work investigates how selection for these traits modifies individual elements of this signaling system (e.g., hormone or androgen receptor [AR] levels), we know less about natural variation in the AR sequence across vertebrates. Our knowledge of AR sequence mutations is largely limited to work in human patients or cell-lines, providing a framework to contextualize single mutations at the expense of evolutionary timescale. Here we unite both perspectives in a review that explores the functional significance of AR on a domain-by-domain basis, using existing knowledge to highlight how and why each region might evolve. We then examine AR sequence variation on different timescales by examining sequence variation in clades originating in the Cambrian (vertebrates; >500 mya) and Cretaceous (birds; >65 mya). In each case, we characterize how the receptor has changed over time and discuss which regions are most likely to evolve in response to selection. Overall, domains that are required for androgenic signaling to function (e.g., DNA- and ligand-binding) tend to be conserved. Meanwhile, areas that interface with co-regulatory molecules can exhibit notable variation even between closely related species. We propose that accumulating mutations in regulatory regions is one way that AR structure might act as a substrate for selection to guide the evolution of reproductive traits. By synthesizing literature across disciplines and highlighting the evolutionary potential of specific AR regions, we hope to inspire new avenues of integrative research into endocrine system evolution.


Asunto(s)
Aves/crecimiento & desarrollo , Evolución Molecular , Receptores Androgénicos/química , Receptores Androgénicos/genética , Animales , Proteínas Aviares/química , Proteínas Aviares/genética , Aves/genética , Humanos , Mutación , Fenotipo , Dominios Proteicos , Secuencias Reguladoras de Ácidos Nucleicos
13.
Cancer Sci ; 111(2): 369-382, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31833612

RESUMEN

The androgen receptor (AR) pathway is critical for prostate cancer carcinogenesis and development; however, after 18-24 months of AR blocking therapy, patients invariably progress to castration-resistant prostate cancer (CRPC), which remains an urgent problem to be solved. Therefore, finding key molecules that interact with AR as novel strategies to treat prostate cancer and even CRPC is desperately needed. In the current study, we focused on the regulation of RNA-binding proteins (RBPs) associated with AR and determined that the mRNA and protein levels of AR were highly correlated with Musashi2 (MSI2) levels. MSI2 was upregulated in prostate cancer specimens and significantly correlated with advanced tumor grades. Downregulation of MSI2 in both androgen sensitive and insensitive prostate cancer cells inhibited tumor formation in vivo and decreased cell growth in vitro, which could be reversed by AR overexpression. Mechanistically, MSI2 directly bound to the 3'-untranslated region (UTR) of AR mRNA to increase its stability and, thus, enhanced its transcriptional activity. Our findings illustrate a previously unknown regulatory mechanism in prostate cancer cell proliferation regulated by the MSI2-AR axis and provide novel evidence towards a strategy against prostate cancer.


Asunto(s)
Neoplasias de la Próstata/patología , Proteínas de Unión al ARN/metabolismo , Receptores Androgénicos/genética , Receptores Androgénicos/metabolismo , Regiones no Traducidas 3' , Animales , Línea Celular Tumoral , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Ratones , Clasificación del Tumor , Trasplante de Neoplasias , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Estabilidad del ARN , Receptores Androgénicos/química , Regulación hacia Arriba
14.
J Med Chem ; 62(24): 11218-11231, 2019 12 26.
Artículo en Inglés | MEDLINE | ID: mdl-31804827

RESUMEN

Androgen receptor (AR) is a validated therapeutic target for the treatment of metastatic castration-resistant prostate cancer (mCRPC). We report herein our design, synthesis, and biological characterization of highly potent small-molecule proteolysis targeting chimera (PROTAC) AR degraders using a potent AR antagonist and E3 ligase ligands with weak binding affinities to VHL protein. Our study resulted in the discovery of 11 (ARD-266), which effectively induces degradation of AR protein in AR-positive (AR+) LNCaP, VCaP, and 22Rv1 prostate cancer cell lines with DC50 values of 0.2-1 nM. ARD-266 is capable of reducing the AR protein level by >95% in these AR+ prostate cancer cell lines and effectively reduces AR-regulated gene expression suppression. For the first time, we demonstrated that an E3 ligand with micromolar binding affinity to its E3 ligase complex can be successfully employed for the design of highly potent and efficient PROTAC degraders and this finding may have a significant implication for the field of PROTAC research.


Asunto(s)
Antagonistas de Receptores Androgénicos/farmacología , Descubrimiento de Drogas , Piperidinas/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Proteolisis/efectos de los fármacos , Receptores Androgénicos/metabolismo , Bibliotecas de Moléculas Pequeñas/farmacología , Proteína Supresora de Tumores del Síndrome de Von Hippel-Lindau/metabolismo , Antagonistas de Receptores Androgénicos/química , Proliferación Celular , Diseño de Fármacos , Humanos , Ligandos , Masculino , Piperidinas/química , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Complejo de la Endopetidasa Proteasomal/efectos de los fármacos , Complejo de la Endopetidasa Proteasomal/metabolismo , Receptores Androgénicos/química , Bibliotecas de Moléculas Pequeñas/química , Células Tumorales Cultivadas , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo
15.
Cells ; 8(11)2019 11 13.
Artículo en Inglés | MEDLINE | ID: mdl-31766271

RESUMEN

The androgen receptor (AR) is a transcription factor that plays a key role in sexual phenotype and neuromuscular development. AR can be modulated by exogenous compounds such as pharmaceuticals or chemicals present in the environment, and particularly by AR agonist compounds that mimic the action of endogenous agonist ligands and whether restore or alter the AR endocrine system functions. The activation of AR must be correctly balanced and identifying potent AR agonist compounds is of high interest to both propose treatments for certain diseases, or to predict the risk related to agonist chemicals exposure. The development of in silico approaches and the publication of structural, affinity and activity data provide a good framework to develop rational AR hits prediction models. Herein, we present a docking and a pharmacophore modeling strategy to help identifying AR agonist compounds. All models were trained on the NR-DBIND that provides high quality binding data on AR and tested on AR-agonist activity assays from the Tox21 initiative. Both methods display high performance on the NR-DBIND set and could serve as starting point for biologists and toxicologists. Yet, the pharmacophore models still need data feeding to be used as large scope undesired effect prediction models.


Asunto(s)
Andrógenos/química , Simulación por Computador , Descubrimiento de Drogas/métodos , Receptores Androgénicos/química , Andrógenos/farmacología , Evaluación Preclínica de Medicamentos , Humanos , Ligandos , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Estructura Molecular , Unión Proteica , Receptores Androgénicos/metabolismo , Sensibilidad y Especificidad , Bibliotecas de Moléculas Pequeñas , Relación Estructura-Actividad
16.
Sci Rep ; 9(1): 16734, 2019 11 13.
Artículo en Inglés | MEDLINE | ID: mdl-31723170

RESUMEN

The androgen receptor (AR) regulates male sexual development. We have now investigated AR homodimerization, hormone-dependent monomerization and nuclear translocation in PC-3 and COS-1 cells, by utilizing mutations associated with the androgen insensitivity syndrome: Pro767Ala, Phe765Leu, Met743Val and Trp742Arg. AR wild type (WT) was expressed as a homodimer in the cytoplasm, while none of these mutants formed homodimers. Unlike AR WT which responded to 1 nM dihydrotestosterone (DHT) to dissociate and translocate into the nucleus, AR Pro767Ala and Phe765Leu mutants remain as the monomer in the cytoplasm. In the crystal structure of the AR LBD homodimer, Pro767 and Phe765 reside closely on a loop that constitutes the dimer interface; their sidechains interact with the Pro767 of the other monomer and with the DHT molecule in the ligand-binding pocket. These observations place Phe765 at a position to facilitate DHT binding to Pro767 and lead to dissociation of the AR homodimer in the cytoplasm. This Pro-Phe Met relay may constitute a structural switch that mediates androgen signaling and is conserved in other steroid hormone receptors.


Asunto(s)
Núcleo Celular/metabolismo , Citoplasma/metabolismo , Dihidrotestosterona/metabolismo , Neoplasias de la Próstata/metabolismo , Multimerización de Proteína , Receptores Androgénicos/química , Receptores Androgénicos/metabolismo , Andrógenos/metabolismo , Animales , Células COS , Chlorocebus aethiops , Humanos , Ligandos , Masculino , Mutación , Neoplasias de la Próstata/patología , Conformación Proteica , Transporte de Proteínas , Receptores Androgénicos/genética , Transducción de Señal , Células Tumorales Cultivadas
17.
Int J Mol Sci ; 20(21)2019 Oct 30.
Artículo en Inglés | MEDLINE | ID: mdl-31671693

RESUMEN

We analyzed three cases of Complete Androgen Insensitivity Syndrome (CAIS) and report three hitherto undisclosed causes of the disease. RNA-Seq, Real-timePCR, Western immunoblotting, and immunohistochemistry were performed with the aim of characterizing the disease-causing variants. In case No.1, we have identified a novel androgen receptor (AR) mutation (c.840delT) within the first exon in the N-terminal transactivation domain. This thymine deletion resulted in a frameshift and thus introduced a premature stop codon at amino acid 282. In case No.2, we observed a nonsynonymous mutation in the ligand-binding domain (c.2491C>T). Case No.3 did not reveal AR mutation; however, we have found a heterozygous mutation in CYP11A1 gene, which has a role in steroid hormone biosynthesis. Comparative RNA-Seq analysis of CAIS and control revealed 4293 significantly deregulated genes. In patients with CAIS, we observed a significant increase in the expression levels of PLCXD3, TM4SF18, CFI, GPX8, and SFRP4, and a significant decrease in the expression of SPATA16, TSACC, TCP10L, and DPY19L2 genes (more than 10-fold, p < 0.05). Our findings will be helpful in molecular diagnostics of patients with CAIS, as well as the identified genes could be also potential biomarkers for the germ cells differentiation process.


Asunto(s)
Síndrome de Resistencia Androgénica/genética , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Mutación , Receptores Androgénicos/genética , Análisis de Secuencia de ADN/métodos , Adolescente , Adulto , Síndrome de Resistencia Androgénica/metabolismo , Estudios de Casos y Controles , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Exones , Femenino , Mutación del Sistema de Lectura , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Dominios Proteicos , Receptores Androgénicos/química , Receptores Androgénicos/metabolismo , Adulto Joven
18.
J Chem Theory Comput ; 15(11): 6504-6512, 2019 Nov 12.
Artículo en Inglés | MEDLINE | ID: mdl-31584802

RESUMEN

We present an explicit solvent alchemical free-energy method for optimizing the partial charges of a ligand to maximize the binding affinity with a receptor. This methodology can be applied to known ligand-protein complexes to determine an optimized set of ligand partial atomic changes. Three protein-ligand complexes have been optimized in this work: FXa, P38, and the androgen receptor. The sets of optimized charges can be used to identify design principles for chemical changes to the ligands which improve the binding affinity for all three systems. In this work, beneficial chemical mutations are generated from these principles and the resulting molecules tested using free-energy perturbation calculations. We show that three quarters of our chemical changes are predicted to improve the binding affinity, with an average improvement for the beneficial mutations of approximately 1 kcal/mol. In the cases where experimental data are available, the agreement between prediction and experiment is also good. The results demonstrate that charge optimization in explicit solvent is a useful tool for predicting beneficial chemical changes such as pyridinations, fluorinations, and oxygen to sulfur mutations.


Asunto(s)
Factor Xa/química , Ligandos , Receptores Androgénicos/química , Proteínas Quinasas p38 Activadas por Mitógenos/química , Sitios de Unión , Factor Xa/metabolismo , Simulación de Dinámica Molecular , Unión Proteica , Receptores Androgénicos/metabolismo , Electricidad Estática , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
20.
Nat Commun ; 10(1): 3562, 2019 08 08.
Artículo en Inglés | MEDLINE | ID: mdl-31395886

RESUMEN

Molecular chaperones such as Hsp40 and Hsp70 hold the androgen receptor (AR) in an inactive conformation. They are released in the presence of androgens, enabling transactivation and causing the receptor to become aggregation-prone. Here we show that these molecular chaperones recognize a region of the AR N-terminal domain (NTD), including a FQNLF motif, that interacts with the AR ligand-binding domain (LBD) upon activation. This suggests that competition between molecular chaperones and the LBD for the FQNLF motif regulates AR activation. We also show that, while the free NTD oligomerizes, binding to Hsp70 increases its solubility. Stabilizing the NTD-Hsp70 interaction with small molecules reduces AR aggregation and promotes its degradation in cellular and mouse models of the neuromuscular disorder spinal bulbar muscular atrophy. These results help resolve the mechanisms by which molecular chaperones regulate the balance between AR aggregation, activation and quality control.


Asunto(s)
Andrógenos/metabolismo , Proteínas del Choque Térmico HSP40/metabolismo , Proteínas HSP70 de Choque Térmico/metabolismo , Receptores Androgénicos/metabolismo , Animales , Técnicas de Sustitución del Gen , Células HEK293 , Humanos , Ligandos , Masculino , Ratones , Ratones Transgénicos , Resonancia Magnética Nuclear Biomolecular , Agregado de Proteínas , Dominios Proteicos , Multimerización de Proteína , Receptores Androgénicos/química , Receptores Androgénicos/genética , Solubilidad
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