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1.
Natl Vital Stat Rep ; 70(13): 1-23, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34878381

RESUMO

Objectives-This study evaluated the quality of the causeof-death information on death certificates for injury deaths, by determining the percentage of deaths for which the underlying cause was a nonspecific injury mechanism.

3.
J Mol Endocrinol ; 68(1): R1-R9, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34546964

RESUMO

Summary: After the discovery of ERß, a novel role for dihydrotestosterone (DHT) in estrogen signaling was revealed. Instead of just being a better androgen, DHT was found to be a precursor of the ERß agonist 5α-androstane-3ß, 17ß-diol (3ßAdiol), an estrogen which does not require aromatase for its synthesis. ERß was found to oppose androgen signaling and thus is a potential target for treatment of prostate cancer. ERß was also found to have effects that were independent of androgen signaling, particularly in the CNS. Although in rodent models of neurodegenerative diseases (Parkinson's disease, multiple sclerosis, and Alzheimer's disease), ERß agonists are very effective in relieving symptoms and improving pathologies, this has not proven to be the case in humans. In this review we will focus on the main differences in ERß signaling between rodents and humans and will make the point that a very important difference between the two species is in the splice variants which are expressed in humans and not rodents. The main conclusion at this point is that before we think of using ERß agonists clinically, much more work on ERß signaling in the human or in primates needs to be done.

4.
Histopathology ; 79(6): 1072-1086, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34333806

RESUMO

AIMS: In-situ follicular neoplasia (ISFN) occurs in approximately 2-3% of reactive lymph nodes, and is currently set apart from 'partial involvement by follicular lymphoma' (PFL). ISFN can progress to overt lymphoma, but precise parameters with which to assess this risk and its association with related diseases remain incompletely understood. The aim of this study was to explore these parameters. METHODS AND RESULTS: We reviewed 11 cases of ISFN and one of PFL between 2003 and 2018. Ten patients had ISFN in the lymph nodes, and one had ISFN in the spleen. Haematoxylin and eosin and immunohistochemical stains were reviewed. Involvement of follicles by ISFN was scored with a three-tier scheme. Of five patients with low ISFN scores, one had chronic myelomonocytic leukaemia, one had mycosis fungoides, and three were free of haematopoietic disease. Among them, four are alive and one was lost to follow-up. Of the six ISFN patients with high scores, two had concurrent marginal zone lymphomas, one had concurrent diffuse large B-cell lymphoma (DLBCL), one had Castleman-like disease, one had progressive transformation of germinal centres with IgG4-related disease, and one had no haematopoietic disease; all are alive except for one who died of concurrent DLBCL. The patient with PFL developed DLBCL 7 years after diagnosis. CONCLUSIONS: On the basis of this limited series, we conclude that only cases with high scores are associated with an overt lymphoma or an abnormal lymphoid process, and that scoring may be a useful parameter with which to assess the risk of associated lymphoma, and deserves further study. We also performed a comprehensive review of the literature.

5.
Proc Natl Acad Sci U S A ; 118(33)2021 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-34389675

RESUMO

To identify regulators of triple-negative breast cancer (TNBC), gene expression profiles of malignant parts of TNBC (mTNBC) and normal adjacent (nadj) parts of the same breasts have been compared. We are interested in the roles of estrogen receptor ß (ERß) and the cytochrome P450 family (CYPs) as drivers of TNBC. We examined by RNA sequencing the mTNBC and nadj parts of five women. We found more than a fivefold elevation in mTNBC of genes already known to be expressed in TNBC: BIRC5/survivin, Wnt-10A and -7B, matrix metalloproteinases (MMPs), chemokines, anterior gradient proteins, and lysophosphatidic acid receptor and the known basal characteristics of TNBC, sox10, ROPN1B, and Col9a3. There were two unexpected findings: 1) a strong induction of CYPs involved in activation of fatty acids (CYP4), and in inactivation of calcitriol (CYP24A1) and retinoic acid (CYP26A1); and 2) a marked down-regulation of FOS, FRA1, and JUN, known tethering partners of ERß. ERß is expressed in 20 to 30% of TNBCs and is being evaluated as a target for treating TNBC. We used ERß+ TNBC patient-derived xenografts in mice and found that the ERß agonist LY500703 had no effect on growth or proliferation. Expression of CYPs was confirmed by immunohistochemistry in formalin-fixed and paraffin-embedded (FFPE) TNBC. In TNBC cell lines, the CYP4Z1-catalyzed fatty acid metabolite 20-hydroxyeicosatetraenoic acid (20-HETE) increased proliferation, while calcitriol decreased proliferation but only after inhibition of CYP24A1. We conclude that CYP-mediated pathways can be drivers of TNBC but that ERß is unlikely to be a tumor suppressor because the absence of its main tethering partners renders ERß functionless on genes involved in proliferation and inflammation.

6.
NCHS Data Brief ; (413): 1-8, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34310274

RESUMO

Drowning deaths are the second leading cause of unintentional injury deaths for children aged 0-17 years and the leading cause for those aged 1-4 (1). Previous studies using national data have shown that unintentional drowning deaths can differ by sex, age, race and ethnicity, and urban-rural category (2,3). This report uses the latest mortality data from the National Vital Statistics System (NVSS) to present national trends in unintentional drowning death rates from 1999 through 2019 for children aged 0-17.


Assuntos
Lesões Acidentais , Afogamento , Criança , Humanos , População Rural , Estados Unidos/epidemiologia
7.
Mol Psychiatry ; 2021 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-33963286

RESUMO

Astrocytes are integral components of synaptic transmission, and their dysfunction leads to neuropsychiatric disorders such as anxiety and depression. Liver X receptor ß (LXRß) is expressed in astrocytes, and LXRß global knockout mice shows impaired synaptic formation. In order to define the role of LXRß in astrocytes, we used a conditional Cre-loxP system to specifically remove LXRß from astrocytes. We found that this deletion caused anxiety-like but not depressive-like behaviors in adult male mice. This behavioral phenotype could be completely reproduced by selective deletion of LXRß in astrocytes in the medial prefrontal cortex (mPFC). Pyramidal neurons in layer V of mPFC are involved in mood behaviors. We found that there was an increased spontaneous excitatory synaptic transmission in layer V pyramidal neurons of the mPFC of these mice. This was concurrent with increased dendritic complexity, despite normal appearance and number of dendritic spines. In addition, gene ontology analysis of RNA sequencing revealed that deletion of astrocytic LXRß led to the enrichment of the process of synaptic transmission in mPFC. Finally, we also confirmed that renormalized excitatory synaptic transmission in layer V pyramidal neurons alleviated the anxiety in mice with astrocytic LXRß deletion in mPFC. Together, our findings reveal that astrocytic LXRß in mPFC is critical in the regulation of synaptic transmission, and this provides a potential new target for treatment of anxiety-like behavior.

8.
NCHS Data Brief ; (406): 1-8, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33814035

RESUMO

Deaths from drug overdose continue to contribute to the public health burden in the United States (1). The increase in the rate of drug overdose deaths involving cocaine and psychostimulants has been well-documented in recent years (1-4). This NCHS Data Brief provides additional information on drug overdose deaths involving cocaine and other psychostimulants (drugs such as methamphetamine, amphetamine, and methylphenidate) by examining the concurrent involvement of opioids. Trends from 2009 through 2019 and differences by census region in 2019 are presented.


Assuntos
Analgésicos Opioides , Cocaína , Overdose de Drogas/mortalidade , Demografia , Humanos , Mortalidade/tendências , Estados Unidos/epidemiologia
9.
NCHS Data Brief ; (398): 1-8, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33663651

RESUMO

In 2019, suicide was the 10th leading cause of death for all ages in the United States (1). As the second leading cause of death for ages 10-34 and the fourth leading cause for ages 35-54, suicide is a major contributor to premature mortality (2). Recent reports have documented a steady increase in suicide rates over the past two decades (3-6). This Data Brief uses final mortality data from the National Vital Statistics System to update trends in suicide rates from 1999 through 2019 and to describe differences by sex, age group, and means of suicide.


Assuntos
Suicídio Consumado/tendências , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mortalidade/tendências , Fatores de Risco , Fatores Sexuais , Estados Unidos/epidemiologia , Estatísticas Vitais , Adulto Jovem
10.
Proc Natl Acad Sci U S A ; 118(13)2021 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-33771918

RESUMO

Knockout of ERß in the mouse leads to nuclear expression of epidermal growth factor receptor (EGFR) in the prostate. To examine whether ERß plays a similar role in the human prostate, we used four cohorts of men: 1) a Swedish cohort of normal prostates and PCa (prostate cancer) of different Gleason grades; 2) men with benign prostatic hyperplasia (BPH) treated with the 5α-reductase inhibitor, finasteride, and finasteride together with the ERß agonists, soy isoflavones; 3) men with PCa above Gleason grade 4 (GG4), treated with ADT (androgen deprivation therapy) and abiraterone (AA), the blocker of androgen synthesis for different durations; and 4) men with GG4 PCa on ADT or ADT with the AR (androgen receptor) blocker, enzalutamide, for 4 mo to 6 mo. In men with BPH, finasteride treatment induced EGFR nuclear expression, but, when finasteride was combined with isoflavones, EGFR remained on the cell membrane. In GG4 patients, blocking of AR for 4 mo to 6 mo resulted in loss of ERß and PTEN expression and increase in patients with nuclear EGFR from 10 to 40%. In the men with GG4 PCa, blocking of adrenal synthesis of testosterone for 2 mo to 7 mo had the beneficial effect of increasing ERß expression, but, on treatment longer than 8 mo, ERß was lost and EGFR moved to the nucleus. Since nuclear EGFR is a predictor of poor outcome in PCa, addition of ERß agonists together with abiraterone should be considered as a treatment that might sustain expression of ERß and offer some benefit to patients.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Receptor beta de Estrogênio/agonistas , Hiperplasia Prostática/tratamento farmacológico , Neoplasias da Próstata/tratamento farmacológico , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Antagonistas de Androgênios/farmacologia , Antagonistas de Androgênios/uso terapêutico , Androstenos/farmacologia , Androstenos/uso terapêutico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Benzamidas/farmacologia , Benzamidas/uso terapêutico , Biópsia , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Estudos de Coortes , Receptores ErbB/metabolismo , Receptor beta de Estrogênio/metabolismo , Finasterida/farmacologia , Finasterida/uso terapêutico , Humanos , Masculino , Camundongos , Camundongos Knockout , Gradação de Tumores , Nitrilas/farmacologia , Nitrilas/uso terapêutico , PTEN Fosfo-Hidrolase/metabolismo , Feniltioidantoína/farmacologia , Feniltioidantoína/uso terapêutico , Fitoestrógenos/farmacologia , Fitoestrógenos/uso terapêutico , Próstata/citologia , Próstata/efeitos dos fármacos , Próstata/patologia , Hiperplasia Prostática/patologia , Neoplasias da Próstata/patologia , Receptores Androgênicos/metabolismo , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo
12.
NCHS Data Brief ; (426): 1-8, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34978529

RESUMO

Deaths from drug overdose continue to contribute to overall mortality and the lowering of life expectancy in the United States (1-4). This report uses the most recent data from the National Vital Statistics System (NVSS) to update statistics on deaths from drug overdose in the United States, showing rates by demographic group and by specific types of drugs involved (such as opioids or stimulants), with a focus on changes from 2019 to 2020.


Assuntos
Estimulantes do Sistema Nervoso Central , Overdose de Drogas , Estatísticas Vitais , Analgésicos Opioides , Humanos , Expectativa de Vida , Estados Unidos/epidemiologia
13.
Proc Natl Acad Sci U S A ; 117(42): 26347-26355, 2020 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-33020300

RESUMO

Loss of the tumor suppressor, PTEN, is one of the most common findings in prostate cancer (PCa). This loss leads to overactive Akt signaling, which is correlated with increased metastasis and androgen independence. However, another tumor suppressor, inositol-polyphosphate 4-phosphatase type II (INPP4B), can partially compensate for the loss of PTEN. INPP4B is up-regulated by androgens, and this suggests that androgen-deprivation therapy (ADT) would lead to hyperactivity of AKT. However, in the present study, we found that in PCa, samples from men treated with ADT, ERß, and INPP4B expression were maintained in some samples. To investigate the role of ERß1 in regulation of INPPB, we engineered the highly metastatic PCa cell line, PC3, to express ERß1. In these cells, INPP4B was induced by ERß ligands, and this induction was accompanied by inhibition of Akt activity and reduction in cell migration. These findings reveal that, in the absence of androgens, ERß1 induces INPP4B to dampen AKT signaling. Since the endogenous ERß ligand, 3ß-Adiol, is lost upon long-term ADT, to obtain the beneficial effects of ERß1 on AKT signaling, an ERß agonist should be added along with ADT.


Assuntos
Receptor beta de Estrogênio/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Neoplasias da Próstata/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Antagonistas de Androgênios/metabolismo , Androgênios/farmacologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Humanos , Masculino , Células PC-3 , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias da Próstata/patologia , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Transdução de Sinais
14.
Lancet Haematol ; 7(9): e640-e648, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32853584

RESUMO

BACKGROUND: Patients with immune thrombocytopenia are at risk of bleeding during surgery, and intravenous immunoglobulin is commonly used to increase the platelet count. We aimed to establish whether perioperative eltrombopag was non-inferior to intravenous immunoglobulin. METHODS: We did a randomised, open-label trial in eight academic hospitals in Canada. Patients were aged at least 18 years, with primary or secondary immune thrombocytopenia and platelet counts less than 100 × 109 cells per L before major surgery or less than 50 × 109 cells per L before minor surgery. Previous intravenous immunoglobulin within 2 weeks or thrombopoietin receptor agonists within 4 weeks before randomisation were not permitted. Patients were randomly assigned to receive oral daily eltrombopag 50 mg from 21 days preoperatively to postoperative day 7 or intravenous immunoglobulin 1 g/kg or 2 g/kg 7 days before surgery. Eltrombopag dose adjustments were allowed weekly based on platelet counts. The randomisation sequence was generated by a computerised random number generator, concealed and stratified by centre and surgery type (major or minor). The central study statistician was masked to treatment allocation. The primary outcome was achievement of perioperative platelet count targets (90 × 109 cells per L before major surgery or 45 × 109 cells per L before minor surgery) without rescue treatment. We did intention-to-treat and per-protocol analyses using an absolute non-inferiority margin of -10%. This trial is registered with ClinicalTrials.gov, NCT01621204. FINDINGS: Between June 5, 2013, and March 7, 2019, 92 patients with immune thrombocytopenia were screened, of whom 74 (80%) were randomly assigned: 38 to eltrombopag and 36 to intravenous immunoglobulin. Median follow-up was 50 days (IQR 49-55). By intention-to-treat analysis, perioperative platelet targets were achieved for 30 (79%) of 38 patients assigned to eltrombopag and 22 (61%) of 36 patients assigned to intravenous immunoglobulin (absolute risk difference 17·8%, one-sided lower limit of the 95% CI 0·4%; pnon-inferiority=0·005). In the per-protocol analysis, perioperative platelet targets were achieved for 29 (78%) of 37 patients in the eltrombopag group and 20 (63%) of 32 in the intravenous immunoglobulin group (absolute risk difference 15·9%, one-sided lower limit of the 95% CI -2·1%; pnon-inferiority=0·009). Two serious adverse events occurred in the eltrombopag group: one treatment-related pulmonary embolism and one vertigo. Five serious adverse events occurred in the intravenous immunoglobulin group (atrial fibrillation, pancreatitis, vulvar pain, chest tube malfunction and conversion to open splenectomy); all were related to complications of surgery. No treatment-related deaths occurred. INTERPRETATION: Eltrombopag is an effective alternative to intravenous immunoglobulin for perioperative treatment of immune thrombocytopenia. However, treatment with eltrombopag might increase risk of thrombosis. The decision to choose one treatment over the other will depend on patient preference, resource limitations, cost, and individual risk profiles. FUNDING: GlaxoSmithKline and Novartis.


Assuntos
Benzoatos/uso terapêutico , Hidrazinas/uso terapêutico , Imunoglobulinas Intravenosas/uso terapêutico , Pirazóis/uso terapêutico , Trombocitopenia/tratamento farmacológico , Administração Oral , Adulto , Idoso , Fibrilação Atrial/etiologia , Benzoatos/efeitos adversos , Feminino , Humanos , Hidrazinas/efeitos adversos , Imunoglobulinas Intravenosas/efeitos adversos , Masculino , Pessoa de Meia-Idade , Pancreatite/etiologia , Assistência Perioperatória , Contagem de Plaquetas , Embolia Pulmonar/etiologia , Pirazóis/efeitos adversos , Resultado do Tratamento , Vertigem/etiologia
15.
ACS Pharmacol Transl Sci ; 3(4): 759-772, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32832875

RESUMO

The calcitonin receptor-like class B G protein-coupled receptor (CLR) mediates adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) functions including vasodilation, cardioprotection, and nociception. Receptor activity-modifying proteins (RAMP1-3) form heterodimers with CLR and determine its peptide ligand selectivity through an unresolved mechanism. The CGRP (RAMP1:CLR) and AM (RAMP2/3:CLR) receptors are proven or promising drug targets, but short AM and CGRP plasma half-lives limit their therapeutic utility. Here, we used synthetic peptide combinatorial library and rational design approaches to probe the ligand selectivity determinants and develop truncated AM and CGRP antagonist variants with receptor extracellular domain binding affinities that were enhanced ∼1000-fold into the low nanomolar range. Receptor binding studies and a high-resolution crystal structure of a novel library-identified AM variant bound to the RAMP2-CLR extracellular domain complex explained the increased affinities and defined roles for AM Lys46 and RAMP modulation of CLR conformation in the ligand selectivity mechanism. In longer AM and CGRP scaffolds that also bind the CLR transmembrane domain, the variants generated picomolar affinity antagonists, one with an estimated 12.5 h CGRP receptor residence time, and sustained signaling agonists "ss-AM" and "ss-CGRP" that exhibited persistent cAMP signaling after ligand washout. Sustained signaling was demonstrated in primary human umbilical vein endothelial cells and the SK-N-MC cell line, which endogenously express AM and CGRP receptors, respectively. This work clarifies the RAMP-modulated CLR ligand selectivity mechanism and provides AM and CGRP variants that are valuable pharmacological tools and may have potential as long-acting therapeutics.

16.
NCHS Data Brief ; (356): 1-8, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32487285

RESUMO

Deaths from drug overdose continue to contribute to mortality in the United States (1-5). This report uses the most recent data from the National Vital Statistics System (NVSS) to update trends in drug overdose deaths for all drugs and for specific drugs and drug types, and to identify changes in rates by state from 2017 to 2018.


Assuntos
Causas de Morte/tendências , Overdose de Drogas/mortalidade , Feminino , Humanos , Masculino , Estados Unidos/epidemiologia , Estatísticas Vitais
17.
NCHS Data Brief ; (362): 1-8, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32487287

RESUMO

Suicide is the 10th leading cause of death for all ages in the United States (1). Suicide is a major contributor to premature mortality as it ranks as the second leading cause of death for ages 10-34 and the fourth leading cause for ages 35-54 (1). Despite national goals to lower the suicide rate (2), several recent reports have documented a steady increase in suicide rates in recent years (3-6). This data brief uses final mortality data from the National Vital Statistics System (NVSS) to update trends in suicide rates from 1999 through 2018 and to describe differences by sex, age group, and urbanicity of county of residence.


Assuntos
Causas de Morte/tendências , Suicídio/tendências , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Estados Unidos/epidemiologia , Adulto Jovem
18.
J Biol Chem ; 295(28): 9736-9751, 2020 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-32487746

RESUMO

Calcitonin gene-related peptide (CGRP), adrenomedullin (AM), and adrenomedullin 2/intermedin (AM2/IMD) have overlapping and unique functions in the nervous and circulatory systems including vasodilation, cardioprotection, and pain transmission. Their actions are mediated by the class B calcitonin-like G protein-coupled receptor (CLR), which heterodimerizes with three receptor activity-modifying proteins (RAMP1-3) that determine its peptide ligand selectivity. How the three agonists and RAMPs modulate CLR binding to transducer proteins remains poorly understood. Here, we biochemically characterized agonist-promoted G protein coupling to each CLR·RAMP complex. We adapted a native PAGE method to assess the formation and thermostabilities of detergent-solubilized fluorescent protein-tagged CLR·RAMP complexes expressed in mammalian cells. Addition of agonist and the purified Gs protein surrogate mini-Gs (mGs) yielded a mobility-shifted agonist·CLR·RAMP·mGs quaternary complex gel band that was sensitive to antagonists. Measuring the apparent affinities of the agonists for the mGs-coupled receptors and of mGs for the agonist-occupied receptors revealed that both ligand and RAMP control mGs coupling and defined how agonist engagement of the CLR extracellular and transmembrane domains affects transducer recruitment. Using mini-Gsq and -Gsi chimeras, we observed a coupling rank order of mGs > mGsq > mGsi for each receptor. Last, we demonstrated the physiological relevance of the native gel assays by showing that they can predict the cAMP-signaling potencies of AM and AM2/IMD chimeras. These results highlight the power of the native PAGE assay for membrane protein biochemistry and provide a biochemical foundation for understanding the molecular basis of shared and distinct signaling properties of CGRP, AM, and AM2/IMD.


Assuntos
Peptídeo Relacionado com Gene de Calcitonina , Eletroforese em Gel de Poliacrilamida Nativa , Receptores de Adrenomedulina , Animais , Células COS , Peptídeo Relacionado com Gene de Calcitonina/química , Peptídeo Relacionado com Gene de Calcitonina/genética , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Chlorocebus aethiops , AMP Cíclico/metabolismo , Células HEK293 , Humanos , Domínios Proteicos , Receptores de Adrenomedulina/química , Receptores de Adrenomedulina/genética , Receptores de Adrenomedulina/metabolismo , Sistemas do Segundo Mensageiro
19.
Proc Natl Acad Sci U S A ; 117(9): 4902-4909, 2020 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-32075916

RESUMO

Disagreements about the phenotype of estrogen receptor ß (ERß) knockout mouse, created by removing the DNA-binding domain of the ERß gene or interruption of the gene with a neocassette (Oliver Smithies ERß knockout mice [ERßOS-/-]), prompted us to create an ERß knockout mouse by deleting the ERß gene with the use of CRISPR/Cas9 technology. We confirmed that the ERß gene was eliminated from the mouse genome and that no ERß mRNA or protein was detectable in tissues of this mouse. Overall the phenotype of the ventral prostate (VP) and mammary gland (MG) in ERßcrispr-/- mice was similar to, but more severe than, that in the ERßOS-/-mice. In the VP of 6-mo-old ERßcrispr-/- mice there was epithelial hyperplasia, fibroplasia, inflammation, stromal overgrowth, and intraductal cancer-like lesions. This was accompanied by an increase in Ki67 and P63 and loss in DACH1 and PURα, two androgen receptor (AR) repressors. In the MG there was overexpression of estrogen receptor α and progesterone receptor, loss of collagen, increase in proliferation and expression of metalloproteases, and invasive epithelium. Surprisingly, by 18 mo of age, the number of hyperplastic foci was reduced, the ducts of the VP and MG became atrophic, and, in the VP, there was massive immune infiltration and massive desquamation of the luminal epithelial cells. These changes were coincident with reduced levels of androgens in males and estrogens in females. We conclude that ERß is a tumor suppressor gene in the VP and MG where its loss increases the activity AR and ERα, respectively.


Assuntos
Receptor beta de Estrogênio/genética , Glândulas Mamárias Humanas/metabolismo , Fenótipo , Próstata/metabolismo , Receptores Androgênicos/metabolismo , Deleção de Sequência , Androgênios/metabolismo , Animais , Sistemas CRISPR-Cas , Quimiocinas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Epitélio/metabolismo , Receptor alfa de Estrogênio/metabolismo , Estrogênios/metabolismo , Proteínas do Olho , Feminino , Humanos , Hiperplasia/patologia , Inflamação , Antígeno Ki-67/metabolismo , Masculino , Camundongos , Camundongos Knockout , Proteínas do Tecido Nervoso/metabolismo , Próstata/patologia , RNA Mensageiro/metabolismo , Transdução de Sinais , Células Estromais , Transativadores , Fatores de Transcrição/metabolismo , Transcriptoma , Proteínas Supressoras de Tumor/metabolismo
20.
J Mol Biol ; 432(7): 1996-2014, 2020 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-32035902

RESUMO

The class B G protein-coupled receptor (GPCR) calcitonin receptor (CTR) is a drug target for osteoporosis and diabetes. N-glycosylation of asparagine 130 in its extracellular domain (ECD) enhances calcitonin hormone affinity with the proximal GlcNAc residue mediating this effect through an unknown mechanism. Here, we present two crystal structures of salmon calcitonin-bound, GlcNAc-bearing CTR ECD at 1.78 and 2.85 Å resolutions and analyze the mechanism of the glycan effect. The N130 GlcNAc does not contact the hormone. Surprisingly, the structures are nearly identical to a structure of hormone-bound, N-glycan-free ECD, which suggested that the GlcNAc might affect CTR dynamics not observed in the static crystallographic snapshots. Hydrogen-deuterium exchange mass spectrometry and molecular dynamics simulations revealed that glycosylation stabilized a ß-sheet adjacent to the N130 GlcNAc and the N-terminal α-helix near the peptide-binding site while increasing flexibility of the peptide-binding site turret loop. These changes due to N-glycosylation increased the ligand on-rate and decreased its off-rate. The glycan effect extended to RAMP-CTR amylin receptor complexes and was also conserved in the related CGRP receptor. These results reveal that N-glycosylation can modulate GPCR function by altering receptor dynamics.


Assuntos
Conformação Proteica , Receptores da Calcitonina/química , Receptores da Calcitonina/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Glicosilação , Humanos , Ligantes , Modelos Moleculares , Ligação Proteica , Domínios Proteicos , Transdução de Sinais
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