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1.
Life Sci ; 260: 118388, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-32890602

RESUMO

Damage to the cholinergic system in central nervous system injuries such as traumatic brain injury (TBI) and neurodegenerative diseases leads to impaired learning and cognition. Neural stem cells (NSCs) have self-renewal capacity and multi-directional differentiation potential and considered the best source of cells for cell replacement therapy. However, how to promote the differentiation of NSCs into neurons is a major challenge in current research. Lhx8 has a specific effect on the development of the cholinergic nervous system, but its exact function is unclear. In this study, we found that Lhx8 could regulate the expression of Growth arrest-specific (GAS)5 which has been implicated in cancer but was less studied in the nervous system. Additionally, results from PCR, fluorescence in situ hybridization, and immunocytochemical analyses showed that GAS5 is mainly expressed in the cytoplasm of hippocampal neural stems cells and promotes their differentiation into neurons; the Morris water maze test demonstrated that GAS5 overexpression restored learning and memory in rats with cholinergic injury. These findings indicate that GAS5, which is regulated by Lhx8, improve brain function following cholinergic nerve injury.


Assuntos
Lesões Encefálicas/fisiopatologia , Neurônios Colinérgicos/patologia , Proteínas com Homeodomínio LIM/metabolismo , Aprendizagem/fisiologia , Memória/fisiologia , Células-Tronco Neurais/patologia , RNA Longo não Codificante/genética , Fatores de Transcrição/metabolismo , Acetilcolina/metabolismo , Animais , Colina O-Acetiltransferase/metabolismo , Neurônios Colinérgicos/metabolismo , Regulação da Expressão Gênica , Proteínas com Homeodomínio LIM/genética , Células-Tronco Neurais/metabolismo , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Fatores de Transcrição/genética
2.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 37(9): 972-975, 2020 Sep 10.
Artigo em Chinês | MEDLINE | ID: mdl-32820510

RESUMO

OBJECTIVE: To analyze variation of ISL1 gene and explore its functional characteristics in relation with congenital heart defect (CHD). METHODS: Clinical data and peripheral blood samples of 194 CHD patients and 232 healthy controls were collected for the extraction of genomic DNA. The coding exons and flanking intronic regions of the ISL1 gene were sequenced. Expression plasmid for the wild-type ISL1 gene ISL1-pcDNA3.1 was constructed, and the corresponding variants were obtained by site-specific mutagenesis. The gene expression plasmid was transfected into CHO cells with liposome, and the functional characteristics of ISL1 variant were studied by double luciferase reporter gene analysis. RESULTS: A novel variant of the ISL1 gene c.499C>T (p.Q167X) was detected in a patient with sporadic CHD. Functional study showed that the variant has lost its transcriptional activation function for the MEF2C promoter. CONCLUSION: A novel variant of the ISL1 gene related to CHD has been identified. The defect of ISL1 gene may underlay the pathogenesis for a proportion of CHD.


Assuntos
Cardiopatias Congênitas , Proteínas com Homeodomínio LIM , Fatores de Transcrição , Animais , Estudos de Casos e Controles , Cricetinae , Cricetulus , Éxons , Cardiopatias Congênitas/genética , Humanos , Proteínas com Homeodomínio LIM/genética , Fatores de Transcrição/genética
3.
Technol Cancer Res Treat ; 19: 1533033820914286, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32207384

RESUMO

Renal cell carcinoma is one of the most common kidney cancer, which accounts almost 90% of the adult renal malignancies worldwide. In recent years, a new class of endogenous noncoding RNAs, circular RNAs, exert important roles in cell function and certain types of pathological responses, especially in cancers, generally by acting as a microRNA sponge. Circular RNAs could act as sponge to regulate the microRNA and the target genes. However, the knowledge about circular RNAs in renal cell carcinoma remains unclear so far. In the research, we selected a highly expressed novel circular RNAs named circMTO1 in renal cell carcinomas. We investigated the roles of circMTO1 and found that circMTO1 overexpression could suppress cell proliferation and metastases in both A497 and 786-O renal cancer cells, while silencing of circMTO1 could promote the progression in SN12C and OS-RC-2 renal cancer cells. The study showed that circMTO1 acted as miR9 and miR223 sponge and inhibited their levels. Furthermore, silencing of circMTO1 in renal cell carcinoma could downregulate LMX1A, the target of miR-9, resulting in the promotion of renal cell carcinoma cell proliferation and invasion. In addition, LMX1A expression suppression induced by transfection of miR9 mimics confirmed that miR9 exerted its function in renal cell carcinoma by regulating LMX1A expression. What's more, miR9 inhibitor and LMX1A overexpression could block the tumor-promoting effect of circMTO1 silencing. In conclusion, circMTO1 suppresses renal cell carcinoma progression by circMTO1/miR9/ LMX1A, indicating that circMTO1 may be a potential target in renal cell carcinoma therapy.


Assuntos
Carcinoma de Células Renais/genética , Neoplasias Renais/genética , Proteínas com Homeodomínio LIM/metabolismo , MicroRNAs/metabolismo , RNA Circular/genética , Fatores de Transcrição/metabolismo , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Progressão da Doença , Humanos , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Proteínas com Homeodomínio LIM/genética , MicroRNAs/genética , RNA Circular/metabolismo , Fatores de Transcrição/genética
4.
EBioMedicine ; 52: 102635, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32028069

RESUMO

BACKGROUND: The ovulatory dysfunction mechanisms underlying polycystic ovary syndrome (PCOS) are not completely understood. There is no effective therapy for PCOS so far. METHODS: We measured the expression of four and a half LIM domain 2 (FHL2) and other related-genes in human granulosa cells (hGCs) from patients with and without PCOS. To minimise the heterogeneity of patients with PCOS, we only included PCOS patients meeting all three criteria according to the revised Rotterdam consensus. The in vitro effects of FHL2 on ovulatory genes and the underlying mechanisms were examined in KGN cells. The role of FHL2 in ovulation was investigated in vivo by overexpressing FHL2 in rat ovaries via intrabursal lentivirus injection. FINDINGS: Increased FHL2 and androgen receptor (AR) expression and decreased CCAAT/enhancer-binding protein ß (C/EBPß) expression were observed in hGCs from patients with PCOS. FHL2 inhibited the expression of ovulation-related genes, including phosphorylated ERK1/2, C/EBPß, COX2 and HAS2 in KGN cells. It was partially by interacting with AR to act as its co-regulator to inhibit C/EBPß expression and by binding to ERK1/2 to inhibit its phosphorylation. Moreover, FHL2 abundance in hGCs was positively correlated with the basal serum testosterone concentration of patients with PCOS, and dihydrotestosterone (DHT)-induced FHL2 upregulation was mediated by AR signalling in KGN cells. Additionally, lentiviral-mediated functional FHL2 overexpression in rat ovaries for 1 week contributed to an impaired superovulatory response, displaying decreased numbers of retrieved oocytes and a lower MII oocyte rate. 3-week FHL2 overexpression rat models without superovulation led to acyclicity and polycystic ovary morphology. INTERPRETATION: Our findings provide novel insights into the mechanisms underlying the pathogenesis of PCOS, suggesting that FHL2 could be a potential treatment target for ovulatory obstacles in PCOS. FUND: National Key Research and Development Program of China, National Natural Science Foundation, National Institutes of Health project and Shanghai Commission of Science and Technology.


Assuntos
Regulação da Expressão Gênica , Proteínas com Homeodomínio LIM/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteínas Musculares/genética , Ovulação/genética , Síndrome do Ovário Policístico/etiologia , Síndrome do Ovário Policístico/metabolismo , Receptores Androgênicos/metabolismo , Fatores de Transcrição/genética , Animais , Biomarcadores , Proteína beta Intensificadora de Ligação a CCAAT/genética , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Modelos Animais de Doenças , Feminino , Imunofluorescência , Humanos , Proteínas com Homeodomínio LIM/metabolismo , Sistema de Sinalização das MAP Quinases , Proteínas Musculares/metabolismo , Ligação Proteica , Ratos , Receptores Androgênicos/genética , Fatores de Transcrição/metabolismo
5.
Nucleic Acids Res ; 48(7): 3525-3541, 2020 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-32086523

RESUMO

Germ-cell transcription factors control gene networks that regulate oocyte differentiation and primordial follicle formation during early, postnatal mouse oogenesis. Taking advantage of gene-edited mice lacking transcription factors expressed in female germ cells, we analyzed global gene expression profiles in perinatal ovaries from wildtype, FiglaNull, Lhx8Null and Sohlh1Null mice. Figla deficiency dysregulates expression of meiosis-related genes (e.g. Sycp3, Rad51, Ybx2) and a variety of genes (e.g. Nobox, Lhx8, Taf4b, Sohlh1, Sohlh2, Gdf9) associated with oocyte growth and differentiation. The absence of FIGLA significantly impedes meiotic progression, causes DNA damage and results in oocyte apoptosis. Moreover, we find that FIGLA and other transcriptional regulator proteins (e.g. NOBOX, LHX8, SOHLH1, SOHLH2) are co-expressed in the same subset of germ cells in perinatal ovaries and Figla ablation dramatically disrupts KIT, NOBOX, LHX8, SOHLH1 and SOHLH2 abundance. In addition, not only do FIGLA, LHX8 and SOHLH1 cross-regulate each other, they also cooperate by direct interaction with each during early oocyte development and share downstream gene targets. Thus, our findings substantiate a major role for FIGLA, LHX8 and SOHLH1 as multifunctional regulators of networks necessary for oocyte maintenance and differentiation during early folliculogenesis.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Redes Reguladoras de Genes , Proteínas com Homeodomínio LIM/metabolismo , Oócitos/metabolismo , Oogênese/genética , Fatores de Transcrição/metabolismo , Animais , Apoptose , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Proliferação de Células/genética , Dano ao DNA , Feminino , Regulação da Expressão Gênica , Células HEK293 , Humanos , Proteínas com Homeodomínio LIM/genética , Meiose/genética , Camundongos , Oócitos/citologia , Ovário/metabolismo , Fatores de Transcrição/genética
6.
Nat Commun ; 11(1): 44, 2020 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-31896750

RESUMO

Sclerosing stromal tumor (SST) of the ovary is a rare type of sex cord-stromal tumor (SCST), whose genetic underpinning is currently unknown. Here, using whole-exome, targeted capture and RNA-sequencing, we report recurrent FHL2-GLI2 fusion genes in 65% (17/26) of SSTs and other GLI2 rearrangements in additional 15% (4/26) SSTs, none of which are detected in other types of SCSTs (n = 48) or common cancer types (n = 9,950). The FHL2-GLI2 fusions result in transcriptomic activation of the Sonic Hedgehog (SHH) pathway in SSTs. Expression of the FHL2-GLI2 fusion in vitro leads to the acquisition of phenotypic characteristics of SSTs, increased proliferation, migration and colony formation, and SHH pathway activation. Targeted inhibition of the SHH pathway results in reversal of these oncogenic properties, indicating its role in the pathogenesis of SSTs. Our results demonstrate that the FHL2-GLI2 fusion is likely the oncogenic driver of SSTs, defining a genotypic-phenotypic correlation in ovarian neoplasms.


Assuntos
Proteínas com Homeodomínio LIM/genética , Proteínas Musculares/genética , Proteínas Nucleares/genética , Proteínas de Fusão Oncogênica/genética , Neoplasias Ovarianas/genética , Tumores do Estroma Gonadal e dos Cordões Sexuais/genética , Fatores de Transcrição/genética , Proteína Gli2 com Dedos de Zinco/genética , Adolescente , Adulto , Feminino , Dosagem de Genes , Regulação Neoplásica da Expressão Gênica , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Humanos , Pessoa de Meia-Idade , Mutação , Proteínas de Fusão Oncogênica/metabolismo , Neoplasias Ovarianas/patologia , Esclerose , Tumores do Estroma Gonadal e dos Cordões Sexuais/patologia , Células Estromais/patologia , Sequenciamento Completo do Exoma , Adulto Jovem
7.
Medicine (Baltimore) ; 99(2): e18715, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31914083

RESUMO

BACKGROUND: ISL1 promotes cardiomyocyte differentiation and plays important roles in heart development. However, whether ISL1 rs1017 polymorphism is associated with the congenital heart disease (CHD) risk remains controversial. METHODS: Five database including PubMed, Cochrane Library, ISI Web of Science, CNKI, and Wan Fang were searched by using key words "Insulin Gene Enhancer Protein ISL1" and "Single Nucleotide Polymorphism," and "Congenital Heart Disease." Five relative articles including 6 independent studies containing 2132 cases and 3812 controls were finally recruited to our study. Meta-analyses were performed by pooling odds ratios (ORs) and 95% confidence interval (CI) from included studies using STATA 12.0 software. RESULTS: The associations between ISL1 rs1017 polymorphism and the risk of CHD were statistically significant under the allele model (T vs A; OR: 1.421; 95% CI: 1.072-1.882), heterozygous model (AT vs AA; OR: 1.342; 95% CI: 1.019-1.767), and dominant model (AT+ TT vs AA; OR: 1.466; 95% CI: 1.059-2.028). Sensitivity analysis indicated that the results were not stable. Subgroup analysis demonstrated that associations were found in Caucasians under the allele model and the heterozygous model (P < .05), but not the dominant model (P > .05). CONCLUSION: In summary, our meta-analysis results suggest that the T allele of ISL1 rs1017 is a risk factor for CHD. However, further studies based on large sample size and multi-ethnic population should be conducted to further prove this correlation.


Assuntos
Cardiopatias Congênitas/genética , Proteínas com Homeodomínio LIM/genética , Fatores de Transcrição/genética , Alelos , Humanos , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Sensibilidade e Especificidade
8.
Vasc Endovascular Surg ; 54(2): 175-181, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31746280

RESUMO

Nail-patella syndrome (NPS) is a rare disorder characterized by abnormal development of ectodermal and mesodermal tissues. Classically, NPS presents as a triad of nail dysplasia, dysplastic patellae, and bony exostoses of the ilia known as "iliac horns." Apart from dermatological and skeletal abnormalities, patients may also have involvement of ophthalmologic and renal systems. The underlying molecular etiology in NPS is the mutation of LMX1B homeobox gene which results in loss of function of its protein also called LMX1B, a DNA-binding protein belonging to the larger LIM-homeodomain transcription factor family. Normal LMX1B gene and protein function are essential for dorsalization of the vertebrate limb bud, development of anterior eye structures, skull formation, and differentiation and migration of neurons in the central nervous system. We report a case of confirmed NPS presenting with congenital aplasia of the internal carotid artery and believe this is the first report of cerebrovascular developmental abnormality associated with NPS.


Assuntos
Artéria Carótida Interna/anormalidades , Malformações Vasculares do Sistema Nervoso Central/genética , Proteínas com Homeodomínio LIM/genética , Mutação , Síndrome da Unha-Patela/genética , Fatores de Transcrição/genética , Artéria Carótida Interna/diagnóstico por imagem , Malformações Vasculares do Sistema Nervoso Central/diagnóstico por imagem , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Síndrome da Unha-Patela/diagnóstico , Fenótipo
9.
J Forensic Sci ; 65(2): 465-470, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31490551

RESUMO

Age estimation using DNA methylation levels has been widely investigated in recent years because of its potential application in forensic genetics. The main aim of this study was to develop an age predictor model (APM) for blood samples of deceased individuals based in five age-correlated genes. Fifty-one samples were analyzed through the bisulfite polymerase chain reaction (PCR) sequencing method for DNA methylation evaluation in genes ELOVL2, FHL2, EDARADD, PDE4C, and C1orf132. Linear regression was used to analyze relationships between methylation levels and age. The model using the highest age-correlated CpG from each locus revealed a correlation coefficient of 0.888, explaining 76.3% of age variation, with a mean absolute deviation from the chronological age (MAD) of 6.08 years. The model was validated in an independent test set of 19 samples producing a MAD of 8.84 years. The developed APM seems to be informative and could have potential application in forensic analysis.


Assuntos
Envelhecimento/genética , Metilação de DNA , Análise de Sequência de DNA/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Ilhas de CpG/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/genética , Proteína de Domínio de Morte Associada a Edar/genética , Elongases de Ácidos Graxos/genética , Feminino , Genética Forense/métodos , Marcadores Genéticos , Humanos , Proteínas com Homeodomínio LIM/genética , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/genética , Reação em Cadeia da Polimerase/métodos , Sulfitos , Fatores de Transcrição/genética , Adulto Jovem
10.
Vascul Pharmacol ; 125-126: 106634, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31866461

RESUMO

Despite the advent of new-generation drug-eluting stents, in-stent restenosis remains a significant problem in patients with coronary artery disease. In- stent restenosis is defined as the gradual re-narrowing of a stented coronary artery lesion due to arterial damage with subsequent local inflammation of the vessel wall and excessive growth of the vascular smooth muscle cells (vSMCs). Four-and-a-half LIM-domain protein 2 (FHL2) is a scaffold protein involved in regulating vSMC function and inflammation. Previously we have demonstrated that FHL2 prevents vSMC proliferation in a murine carotid artery ligation model. However, the effect of FHL2 on the inflammatory response of the vSMCs is not investigated. Therefore, we studied the inflammatory response in the vessel wall of FHL2-deficient (-KO) mice after carotid artery ligation. We found that circulating cytokines and local macrophage infiltration in the ligated carotid vessels were increased in FHL2-KO mice after carotid artery ligation. Moreover, FHL2-KO vSMCs showed increased secretion of cytokines such as SDF-1α and RANTES, and enhanced activation of the NFκB pathway. Finally, we found that blocking the NFκB signalling pathway abrogated this pro-inflammatory state in FHL2-KO vSMCs. Taken together, our results demonstrate that FHL2 decreases the inflammatory response of vSMCs through inhibition of the NFkB-signalling pathway.


Assuntos
Doenças das Artérias Carótidas/metabolismo , Inflamação/metabolismo , Proteínas com Homeodomínio LIM/metabolismo , Proteínas Musculares/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , NF-kappa B/metabolismo , Fatores de Transcrição/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Artérias Carótidas/metabolismo , Artérias Carótidas/patologia , Doenças das Artérias Carótidas/genética , Doenças das Artérias Carótidas/patologia , Doenças das Artérias Carótidas/prevenção & controle , Células Cultivadas , Citocinas/sangue , Modelos Animais de Doenças , Inflamação/genética , Inflamação/patologia , Inflamação/prevenção & controle , Proteínas com Homeodomínio LIM/deficiência , Proteínas com Homeodomínio LIM/genética , Macrófagos/metabolismo , Macrófagos/patologia , Camundongos Knockout , Proteínas Musculares/deficiência , Proteínas Musculares/genética , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , Transdução de Sinais , Fatores de Transcrição/deficiência , Fatores de Transcrição/genética
11.
J Pineal Res ; 68(1): e12616, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31609018

RESUMO

Homeobox genes generally encode transcription factors involved in regulating developmental processes. In the pineal gland, a brain structure devoted to nocturnal melatonin synthesis, a number of homeobox genes are also expressed postnatally; among these is the LIM homeobox 4 gene (Lhx4). We here report that Lhx4 is specifically expressed in the postnatal pineal gland of rats and humans. Circadian analyses revealed a fourfold rhythm in Lhx4 expression in the rat pineal gland, with rhythmic expression detectable from postnatal day 10. Pineal Lhx4 expression was confirmed to be positively driven by adrenergic signaling, as evidenced by in vivo modulation of Lhx4 expression by pharmacological (isoprenaline injection) and surgical (superior cervical ganglionectomy) interventions. In cultured pinealocytes, Lhx4 expression was upregulated by cyclic AMP, a second messenger of norepinephrine. By use of RNAscope technology, Lhx4 transcripts were found to be exclusively localized in melatonin-synthesizing pinealocytes. This prompted us to investigate the possible role of Lhx4 in regulation of melatonin-producing enzymes. By use of siRNA technology, we knocked down Lhx4 by 95% in cultured pinealocytes; this caused a reduction in transcripts encoding the melatonin-producing enzyme arylalkylamine N-acetyl transferase (Aanat). Screening the transcriptome of siRNA-treated pinealocytes by RNAseq revealed a significant impact of Lhx4 on the phototransduction pathway and on transcripts involved in development of the nervous system and photoreceptors. These data suggest that rhythmic expression of Lhx4 in the pineal gland is controlled via an adrenergic-cyclic AMP mechanism and that Lhx4 acts to promote nocturnal melatonin synthesis.


Assuntos
Proteínas com Homeodomínio LIM , Melatonina/metabolismo , Glândula Pineal , Fatores de Transcrição , Transcriptoma/genética , Adulto , Animais , Arilalquilamina N-Acetiltransferase/genética , Arilalquilamina N-Acetiltransferase/metabolismo , Ritmo Circadiano/genética , AMP Cíclico/metabolismo , Feminino , Humanos , Proteínas com Homeodomínio LIM/genética , Proteínas com Homeodomínio LIM/metabolismo , Masculino , Melatonina/genética , Pessoa de Meia-Idade , Norepinefrina/metabolismo , Glândula Pineal/química , Glândula Pineal/citologia , Glândula Pineal/crescimento & desenvolvimento , Glândula Pineal/metabolismo , RNA Interferente Pequeno/genética , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Adulto Jovem
12.
PLoS Genet ; 15(12): e1008573, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31877129

RESUMO

The ability to establish spatial organization is an essential feature of any developing tissue and is achieved through well-defined rules of cell-cell communication. Maintenance of this organization requires elimination of cells with inappropriate positional identity, a poorly understood phenomenon. Here we studied mechanisms regulating cell elimination in the context of a growing tissue, the Drosophila wing disc and its dorsal determinant Apterous. Systematic analysis of apterous mutant clones along with their twin spots shows that they are eliminated from the dorsal compartment via three different mechanisms: relocation to the ventral compartment, basal extrusion, and death, depending on the position of the clone in the wing disc. We find that basal extrusion is the main elimination mechanism in the hinge, whereas apoptosis dominates in the pouch and in the notum. In the absence of apoptosis, extrusion takes over to ensure clearance in all regions. Notably, clones in the hinge grow larger than those in the pouch, emphasizing spatial differences. Mechanistically, we find that limiting cell division within the clones does not prevent their extrusion. Indeed, even clones of one or two cells can be extruded basally, demonstrating that the clone size is not the main determinant of the elimination mechanism to be used. Overall, we revealed three elimination mechanisms and their spatial biases for preserving pattern in a growing organ.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/crescimento & desenvolvimento , Proteínas com Homeodomínio LIM/genética , Fatores de Transcrição/genética , Asas de Animais/crescimento & desenvolvimento , Animais , Divisão Celular , Células Clonais/citologia , Células Clonais/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas com Homeodomínio LIM/metabolismo , Mutação , Fatores de Transcrição/metabolismo , Asas de Animais/metabolismo
13.
Nat Commun ; 10(1): 5192, 2019 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-31729356

RESUMO

The extent of neocortical gyrification is an important determinant of a species' cognitive abilities, yet the mechanisms regulating cortical gyrification are poorly understood. We uncover long-range regulation of this process originating at the telencephalic dorsal midline, where levels of secreted Bmps are maintained by factors in both the neuroepithelium and the overlying mesenchyme. In the mouse, the combined loss of transcription factors Lmx1a and Lmx1b, selectively expressed in the midline neuroepithelium and the mesenchyme respectively, causes dorsal midline Bmp signaling to drop at early neural tube stages. This alters the spatial and temporal Wnt signaling profile of the dorsal midline cortical hem, which in turn causes gyrification of the distal neocortex. Our study uncovers early mesenchymal-neuroepithelial interactions that have long-range effects on neocortical gyrification and shows that lissencephaly in mice is actively maintained via redundant genetic regulation of dorsal midline development and signaling.


Assuntos
Mesoderma/embriologia , Neocórtex/embriologia , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas com Homeodomínio LIM/genética , Proteínas com Homeodomínio LIM/metabolismo , Masculino , Mesoderma/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neocórtex/metabolismo , Células Neuroepiteliais/metabolismo , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas Wnt/genética , Proteínas Wnt/metabolismo
14.
Mol Cell Neurosci ; 101: 103411, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31648029

RESUMO

Onecut transcription factors are required to maintain Islet1 (Isl1) expression in developing spinal motor neurons (MNs), and this process is critical for proper MN differentiation. However, the mechanisms whereby OC stimulate Isl1 expression remain unknown. CREB-binding protein (CBP) and p300 paralogs are transcriptional coactivators that interact with OC proteins in hepatic cells. In the embryonic spinal cord, CBP and p300 play key roles in neurogenesis and MN differentiation. Here, using chromatin immunoprecipitation and in ovo electroporation in chicken spinal cord, we provide evidence that CBP and p300 contribute to the regulation of Isl1 expression by the OC factors in embryonic spinal MNs. CBP and p300 are detected on the CREST2 enhancer of Isl1 where OC factors are also bound. Inhibition of CBP and p300 activity inhibits activation of the CREST2 enhancer and prevents the stimulation of Isl1 expression by the OC factors. These observations suggest that CBP and p300 coactivators cooperate with OC factors to maintain Isl1 expression in postmitotic MNs.


Assuntos
Proteína de Ligação a CREB/metabolismo , Elementos Facilitadores Genéticos , Proteínas com Homeodomínio LIM/genética , Neurônios Motores/metabolismo , Fatores de Transcrição Onecut/metabolismo , Medula Espinal/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição de p300-CBP/metabolismo , Animais , Embrião de Galinha , Proteínas com Homeodomínio LIM/metabolismo , Medula Espinal/citologia , Fatores de Transcrição/metabolismo
15.
Int J Sports Med ; 40(14): 921-930, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31614382

RESUMO

Myocardial damage due to dysfunctional myocardium has been increasing, and the prognosis of pharmacological and device-based therapies remain poor. Isl1-expressing cells were thought to be progenitor cells for cardiomyocyte proliferation after specific stimuli. However, the true origin of the proliferating myocardiac cells and the role of Isl1 in adult mammals remain unresolved. In this study, Isl1-CreERT2 knock-in mouse model was constructed using CRISPR/Cas9 technology. Using tamoxifen-inducible Isl1-CreERT/Rosa26R-LacZ system, Isl1+cells and their progeny were permanently marked by lacZ-expression. X-gal staining, immunostaining, and quantitative PCR were then used to reveal the fate of Isl1+cells under physiological and exercise conditions in mouse hearts from embryonic stage to adulthood. Isl1+cells were found to localize to the sinoatrial node, atrioventricular node, cardiac ganglia, aortic arch, and pulmonary roots in adult mice heart. However, they did not act as cardiac progenitor cells under physiological and exercise conditions. Although Isl1+cells showed progenitor cell properties in early mouse embryos (E7.5), this ability was lost by E9.5. Furthermore, although the proliferation and regeneration of heart cell was observed in response to exercise, the cells associated were not Isl1 positive.


Assuntos
Coração/fisiologia , Proteínas com Homeodomínio LIM/genética , Miocárdio/citologia , Miócitos Cardíacos/citologia , Condicionamento Físico Animal , Células-Tronco/fisiologia , Fatores de Transcrição/genética , Animais , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Proliferação de Células/fisiologia , Mapeamento Cromossômico , Coração/embriologia , Coração/crescimento & desenvolvimento , Masculino , Camundongos Endogâmicos C57BL
16.
Int Heart J ; 60(5): 1113-1122, 2019 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-31484864

RESUMO

Occurring in about 1% of all live births, congenital heart defects (CHDs) represent the most frequent type of developmental abnormality and account for remarkably increased infant morbidity and mortality. Aggregating studies demonstrate that genetic components have a key role in the occurrence of CHDs. Nevertheless, due to pronounced genetic heterogeneity, the genetic causes of CHDs remain unclear in most patients. In this research, 114 unrelated patients affected with CHDs and 218 unrelated individuals without CHDs served as controls were recruited. The coding regions and splicing donors/acceptors of the ISL1 gene, which codes for a transcription factor required for proper cardiovascular development, were screened for mutations by sequencing in all study participants. The functional characteristics of an identified ISL1 mutation were delineated with a dual-luciferase reporter assay system. As a result, a new heterozygous ISL1 mutation, NM_002202.2: c.225C>G; p. (Tyr75*), was discovered in an index patient with double outlet right ventricle and ventricular septal defect. Analysis of the proband's family unveiled that the mutation co-segregated with the CHD phenotype. The nonsense mutation was absent in the 436 control chromosomes. Biological analysis showed that the mutant ISL1 protein had no transcriptional activity. Furthermore, the mutation nullified the synergistic activation between ISL1 and TBX20, another CHD-associated transcription factor. This research for the first time links an ISL1 loss-of-function mutation to double outlet right ventricle in humans, which adds insight to the molecular pathogenesis underpinning CHDs, suggesting potential implications for timely personalized management of CHD patients.


Assuntos
Dupla Via de Saída do Ventrículo Direito/genética , Genes Reporter/genética , Predisposição Genética para Doença/epidemiologia , Proteínas com Homeodomínio LIM/genética , Mutação com Perda de Função/genética , Fatores de Transcrição/genética , Estudos de Casos e Controles , Causalidade , Pré-Escolar , China/epidemiologia , Dupla Via de Saída do Ventrículo Direito/diagnóstico por imagem , Feminino , Cardiopatias Congênitas/diagnóstico por imagem , Cardiopatias Congênitas/epidemiologia , Cardiopatias Congênitas/genética , Heterozigoto , Hospitais Universitários , Humanos , Incidência , Lactente , Masculino , Mutação , Linhagem , Prognóstico , Estudos Retrospectivos , Medição de Risco
17.
Acta Derm Venereol ; 99(12): 1110-1115, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31513274

RESUMO

Nail-patella syndrome is an autosomal dominant disorder characterized by nail dysplasia and skeletal anomaly. Some patients have been shown to have ultrastructural abnormalities of the glomerular basement membrane that result in nephrosis. However, little has been reported on the epidermal basement membrane in this condition. This paper reports 2 families with nail-patella syndrome. Direct sequencing analysis of LMX1B revealed that family 1 and family 2 were heterozygous for the mutations c.140-1G>C and c.326+1G>C, respectively. To evaluate the epidermal basement membrane zone, ultrastructural and immunohistochemical analyses were performed using skin specimens obtained from the dorsal thumb. Electron microscopy showed intact hemidesmosomes, lamina lucida, lamina densa, and anchoring fibrils. Immunofluorescence studies with antibodies against components of the epidermal basement membrane zone revealed a normal expression pattern among the components, including type IV collagen. These data suggest that nail dysplasia in patients with nail-patella syndrome is not caused by structural abnormalities of the epidermal basement membrane.


Assuntos
Membrana Basal/química , Membrana Basal/ultraestrutura , Colágeno Tipo IV/análise , Epiderme/química , Epiderme/ultraestrutura , Imunofluorescência , Microscopia Eletrônica de Transmissão , Síndrome da Unha-Patela/diagnóstico , Biomarcadores/análise , Criança , Feminino , Predisposição Genética para Doença , Heterozigoto , Humanos , Lactente , Proteínas com Homeodomínio LIM/genética , Masculino , Mutação , Síndrome da Unha-Patela/genética , Síndrome da Unha-Patela/metabolismo , Síndrome da Unha-Patela/patologia , Fenótipo , Valor Preditivo dos Testes , Fatores de Transcrição/genética
18.
PLoS One ; 14(9): e0221640, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31557193

RESUMO

Our research group has showed that the LIM homeobox transcription factor 1 alpha (LMX1A) is inactivated in gastric cancers. Overexpression of LMX1A inhibits tumor growth. However, the mechanisms remains unclear. Considering LMX1A as a transcription factor, a comparison of RNA-seq between gastric cancer cells (GCCs) and GCCs with LMX1A overexpressed was performed to identify genes transcriptionally activated by LMX1A. Among the potential LMX1A target genes, angiopoietin-like 4 (ANGPTL4) has been reported to be an important tumor suppressor and thus was selected for further validation and research. Both LMX1A and ANGPTL4 showed downregulated expression in gastric cancer samples. More importantly, the expression of LMX1A is positively correlated with ANGPTL4, without including other family members in gastric cancer cell lines. What's more, knockdown of ANGPTL4 rescued the tumor suppressive phenotype of LMX1A overexpression, which indicated that LMX1A upregulates ANGPTL4 to exert its role. Mechanistically, we found that LMX1A inhibited the expression of the oncogene C-Myc, which is alleviated by ANGPTL4 knockdown. In general, our results showed that LMX1A exerts its tumor suppressive role by activating ANGPTL4 to inhibit C-Myc.


Assuntos
Proteína 4 Semelhante a Angiopoietina/genética , Proteína 4 Semelhante a Angiopoietina/metabolismo , Genes myc , Proteínas com Homeodomínio LIM/genética , Proteínas com Homeodomínio LIM/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteína 4 Semelhante a Angiopoietina/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Genes Supressores de Tumor , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Proteínas Proto-Oncogênicas c-myc/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo
19.
Biomed Pharmacother ; 117: 109174, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31387183

RESUMO

This study aimed to explore the prognostic value of LMX1B mRNA expression and the methylation of its CpG sites in patients with laryngeal squamous cell carcinoma (LSCC). An in-silicon analysis was performed using data from the cancer genome atlas (TCGA)-Head and Neck Squamous Carcinoma (HNSC). After screening, 112 LSCC and 10 adjacent normal tissues were identified as eligible samples for analysis. Results showed that LMX1B expression was significantly upregulated in the cancer tissues (p < 0.01) and was an independent prognostic indicator in terms of OS (HR: 1.233, 95%CI: 1.082-1.405, p = 0.002) and RFS (HR: 1.200, 95%CI: 1.002-1.438, p = 0.048). By examining the methylation profile of 55 CpG sites in LMX1B locus, we found that the promoter methylation status was irrelevant to LMX1B expression. In comparison, LMX1B expression was generally positively correlated with gene body methylation. Among the gene body CpG sites, cg13600622 methylation showed a better predictive value than LMX1B expression in terms of OS (HR: 12.363, 95%CI: 1.076-142.033, p = 0.043), while cg14204784 methylation was a better marker of shorter RFS (HR: 12.363, 95%CI: 1.076-142.033, p = 0.043). Among the known downstream genes of LMX1B, only NR4A2 expression showed a moderately negative correlation (Pearson's r = -0.54) with it in LSCC tissues. However, this correlation was inconsistent with previous publications those reported a positive correlation between them. Based on these findings, we infer that upregulated LMX1B mRNA expression had an independent prognostic value in LSCC patients. Increased gene body methylation might be an important mechanism of its upregulation. Among the gene body CpG sites, cg13600622 and cg14204784 methylation level might be better prognostic markers than LMX1B mRNA expression in terms of OS and RFS respectively.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Ilhas de CpG/genética , Metilação de DNA/genética , Proteínas com Homeodomínio LIM/genética , Neoplasias Laríngeas/genética , RNA Mensageiro/genética , Fatores de Transcrição/genética , Carcinoma de Células Escamosas/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Neoplasias Laríngeas/patologia , Masculino , Prognóstico , Regiões Promotoras Genéticas/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Regulação para Cima/genética
20.
Gene Expr Patterns ; 34: 119065, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31376504

RESUMO

PURPOSE: To investigate the expression patterns of LIM Homeobox 6 (Lhx6) in the adult and developing mouse retina. METHODS: The Lhx6-GFP knock-in allele was used to activate constitutive expression of a GFP reporter in Lhx6 expressing cells. Double labeling with GFP and retinal markers in the mouse retina at postnatal day 56 (P56) was performed to identify the cell types expressing Lhx6. To determine the neuronal cell types that express Lhx6, double labeling with GFP and various retinal markers was employed in the differentiating retina at P7 and P15. RESULTS: GFP + Lhx6 lineage cells were determined in Brn3a + retinal ganglion cells (RGCs), ChAT + amacrine cells (ACs), and Islet-class LIM-homeodomain 1 (Isl1+) ACs in the mouse retina at P56. In the ganglion cell layer (GCL), Lhx6 was expressed in Brn3a + RGCs but not Brn3b + RGCs at P15. Moreover, in the inner nuclear layer (INL), Lhx6 was not expressed in Bhlhb5+ ACs at P15. However, Lhx6 was weakly expressed in Glyt1+ ACs and Pax6+ ACs, and strongly expressed in Isl1+ and ChAT + ACs at P15. CONCLUSION: Lhx6 was expressed in RGCs and ACs in both the adult and developing mouse retina.


Assuntos
Proteínas com Homeodomínio LIM/genética , Proteínas do Tecido Nervoso/genética , Retina/crescimento & desenvolvimento , Retina/metabolismo , Fatores de Transcrição/genética , Fatores Etários , Células Amácrinas/metabolismo , Animais , Linhagem da Célula , Proteínas de Ligação a DNA/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas com Homeodomínio LIM/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/metabolismo , Células Ganglionares da Retina/metabolismo , Fatores de Transcrição/metabolismo , Transcriptoma/genética
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