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López Lizárraga Paulina - One of the best experts on this subject based on the ideXlab platform.

  • Estudio de secuenciación del gen Mir-184 en pacientes con ectasias corneales primarias
    'Universitat Autonoma de Barcelona', 2020
    Co-Authors: López Lizárraga Paulina
    Abstract:

    Les ectàsies corneals primàries representen un grup d’entitats que cursen amb aprimament corneal progressiu, bilateral i asimètric. Dins d’aquestes, es troba per ordre de major a menor freqüència el queratocono, la degeneració marginal pel·lúcida i el queratoglobo. Han estat nombrosos els estudis que estableixen un probable origen en comú de les mateixes com a part de l’espectre d’una mateixa entitat amb un probable component genètic per a la seva presentació, l’anterior amb molt més freqüència en el queratocono. El gen Mir-184 és el microRNA més abundantment expressat en còrnia i cristal·lí, regulador de l’apoptosi en aquests teixits, i la seva mutació s’ha associat a queratocono en pacients caucàsics, convertint-lo en un potencial candidat, la seva mutació podria portar a el desenvolupament d’una ectàsia corneal primària. Pregunta de recerca Existeixen mutacions en el gen Mir-184 en un grup de pacients amb ectàsies corneals primàries? Justificació Les ectàsies corneals es troben dins les principals causes de trasplantament de còrnia en països desenvolupat, identificar l’origen del seu desenvolupament canviaria significativament el plantejament de la seva fisiopatologia i tractament. Hipòtesi de Recerca Mutacions en el gen Mir-184 s’associen amb la presència de ectàsies corneals primàries. Objectiu General Identificar mutacions de el gen Mir-184 en pacients amb diagnòstic de ectàsia corneal primària. Objectius Secundaris Realitzar una recerca exhaustiva a manera de revisió sistemàtica respecte a les ectàsies corneals primàries. Disseny de l’estudi Estudi descriptiu observacional en què es va buscar la presència o absència de la mutació en el gen Mir-184 en pacients amb ectàsies corneals primàries. Material i mètodes Previ consentiment informat, es va realitzar un anàlisi de genotipificació amb 200 mostres de casos esporàdics o familiars de pacients mexicans amb diagnòstic clínic i topogràfic de ectàsia corneal primària. A partir d’ una mostra de 2 ml de sang, a cada pacient es va extreure el DNA genòmic, amb posterior anàlisi del gen Mir-184 mitjançant PCR i seqüenciació nucleotídica. Les seqüències obtingudes de cada pacient es van comparar amb la seqüència silvestre de Mir-184 a Ensembl (www.ensembl.org) per així determinar la presència o absència de mutació en el mateix gen. Es va determinar realitzar l’anàlisi estadístic mitjançant la prova de xi2 d’acord a la variable predictora dictotòmica que fa a la presència o absència de la mutació en el gen Mir-184. Resultats i Conclusions No s’ha trobat aquesta mutació en cap dels casos. Hi ha forta evidència científica que orienta les ectàsies corneals primàries com diferents graus de manifestació d’un mateix origen fisiopatològic en comú, i que orienta de manera predominant a l’queratocono com una malaltia amb un component genètic significatiu el qual s’expressa posterior a l’exposició a un factor ambiental desencadenant, de manera principal, el rascat ocular. El gen Mir-184 és el micro RNA més abundantment expressat tant en l’epiteli corneal com en el cristalí, i un important regulador de l’apoptosi, la seva mala regulació s’ha associat de manera significativa a el desenvolupament principalment de queratocono i la seva mutació representa un fort candidat per explicar el desenvolupament de ectàsies corneals primàries. S’ha observat que aproximadament el 60% dels gens codificants de proteïna són regulats per micro RNAs. El no haver pogut demostrar en aquest estudi una associació causa-efecte de manera significativa entre una mutació en el gen Mir-184 i el desenvolupament de ectàsies corneals primàries, orienta a que l’heterogeneïtat genètica juga un paper important per al desenvolupament de les mateixes, així com també l’exposició a factors ambientals desencadenants.“Las ectasias corneales primarias representan un grupo de entidades que cursan con adelgazamiento corneal progresivo, bilateral y asimétrico. Dentro de éstas, se encuentra por orden de mayor a menor frecuencia el queratocono, la degeneración marginal pelúcida y el queratoglobo. Han sido numerosos los estudios que establecen un probable origen en común de las mismas como parte del espectro de una misma entidad con un probable componente genético para su presentación, lo anterior con mucho más frecuencia en el queratocono. El gen Mir-184 es el microRNA más abundantemente expresado en córnea y cristalino, regulador de la apoptosis en dichos tejidos, cuya mutación se ha asociado a queratocono en pacientes caucásicos, convirtiéndolo en un potencial candidato cuya mutación podría llevar al desarrollo de una ectasia corneal primaria. Pregunta de investigación Existen mutaciones del gen Mir-184 en un grupo de pacientes con ectasias corneales primarias? Justificación Las ectasias corneales representan una de las principales causas de trasplante de cornea en países desarrollados. El poder identificar el origen de su desarrollo cambiaría significativamente el planteamiento respecto a su fisiopatología y tratamiento. Hipótesis de la Investigación Mutaciones en el gen MIR 184 se asocian con la presencia de ectasias corneales primarias. Objetivo general Identificar mutaciones del gen Mir-184 en pacientes con diagnóstico de ectasia corneal primaria. Objetivos secundarios Realizar una búsqueda exhaustiva a modo de revisión sistemática respecto las ectasias corneales primarias. Diseño del estudio Estudio descriptivo observacional en el que se buscó la presencia o ausencia de la mutación en el gen Mir-184 en pacientes con ectasias corneales primarias. Material y métodos Previo consentimiento informado, se realizó un análisis de genotipificación en 200 muestras de casos esporádicos o familiares de pacientes mexicanos con diagnóstico clínico y topográfico de ectasia corneal primaria. De una muestra de 2 ml de sangre, a cada paciente se extrajo el DNA genómico, con posterior análisis del gen Mir-184 mediante PCR y secuenciación nucleotídica. Las secuencias obtenidas de cada paciente se compararon con la secuencia silvestre de Mir-184 en Ensembl (www.ensembl.org) para así determinar la presencia o ausencia de mutación en el mismo. Se determinó realizar el análisis estadístico mediante la prueba de xi2 de acuerdo a la variable predictora dictotómica respecto a la presencia o ausencia de la mutación en el gen Mir-184 . Resultados y Conclusiones No se encontró dicha mutación en ninguno de los casos. Hay fuerte evidencia científica que orienta a las ectasias corneales primarias tiene un mismo origen fisiopatológico en común, y que orienta de manera predominante al queratocono con un componente genético significativo el cual se expresa posterior a la exposición a un factor ambiental desencadenante. El gen Mir-184 al ser el micro RNA más abundantemente expresado tanto en el epitelio corneal como en el cristaliniano, y al ser un importante regulador de la apoptosis, mecanismo cuya mala regulación se ha asociado de manera significativa al desarrollo principalmente de queratocono, su mutación representa un fuerte candidato para explicar el desarrollo de ectasias corneales primarias. Se ha observado que aproximadamente el 60% de los genes codificantes de proteína son regulados por micro RNAs. El no haber podido demostrar en este estudio una asociación causa-efecto de manera significativa entre una mutación en el gen Mir-184 y el desarrollo de una ectasia corneal primaria, orienta a que la heterogeneidad genética juega un papel importante para el desarrollo de las mismas, así como también la exposición a factores ambientales desencadenantes, lo cual a su vez orienta a que dichas entidades se encuentran dentro del espectro de las enfermedades multifactoriales.Primary corneal ectasies represent a group of entities that have progressive, bilateral and asymmetric corneal thinning. Keratoconus, pellucid marginal degeneration and keratoglobus are englobed in the classification of primary corneal ectasies with keratoconus being the most frequent among them. There have been numerous reports that establish a probable common origin of them as part of the spectrum of the same entity with a probable genetic component for their presentation, mainly in keratoconus. Mir-184 gene is the most abundantly expressed microRNA in corneal and lens tissue and is also an important apoptosis regulator in these tissues. Its mutation has been associated with keratoconus in Caucasian patients, making its mutation a potential candidate for the development of a primary corneal ectasia. Research question Are mutations in the Mir-184 gene associated to primary corneal ectasies? Justification Corneal ectasies represent one of the main causes of corneal transplantation in developed countries. Being able to identify the origin of their development would significantly change their approach regarding its pathophysiology and treatment. Research Hypothesis Mutations in the Mir-184 gene are associated with the presence of primary corneal ectasies. General Objective To identify mutations in the gene Mir-184 in patients diagnosed with a primary corneal ectasia. Secondary Objectives To perform research in the literature in the form of a systematic review regarding primary corneal ectasies. Study Design This was an observational descriptive study where the presence or absence of mutation in the Mir-184 gene was sought in patients with primary corneal ectasies. Material and methods Prior informed consent, a genotyping analysis was performed in 200 samples of Mexican patients with clinical and topographic diagnosis of sporadic or familial primary corneal ectasia. From a 2 ml sample of blood, genomic DNA was extracted from each patient, and a subsequent analysis of the Mir-184 gene by PCR and nucleotide sequencing was performed. The sequences obtained from each patient were compared with the wild sequence of Mir-184 in Ensembl (www.ensembl.org) in order to determine the presence or absence of a mutation. The statistical analysis was performed using the xi2 test according to the dicotomic predictive variable regarding the presence or absence of the mutation. Results and conclusions No such mutation was found in any of the cases. There is strong scientific evidence that guides primary corneal ectasies as different degrees of manifestation of the same pathophysiological origin in common, and that predominantly guides keratoconus as a disease with a significant genetic component which is expressed after exposure to a environmental triggering factor, mainly, eye scratching. The Mir-184 gene, being the most abundantly expressed micro RNA in both the corneal and crystalline epithelium, and being an important regulator of apoptosis, a mechanism whose poor regulation has been significantly associated with the development mainly of keratoconus, represents a strong candidate to explain the development of primary corneal ectasies. It has been observed that approximately 60% of the protein coding genes are regulated by micro RNAs. Not having been able to demonstrate in this study a significant cause-effect association between a mutation in the Mir-184 gene and the development of primary corneal ectasies, could orient to the fact that genetic heterogeneity plays an important role in their development, as well as exposure to environmental triggering factors, which would englobe them as part of the diseases with a multifactorial origin.Universitat Autònoma de Barcelona. Programa de Doctorat en Cirurgia i Ciències Morfològique

  • Estudio de secuenciación del gen Mir-184 en pacientes con ectasias corneales primarias
    2020
    Co-Authors: López Lizárraga Paulina
    Abstract:

    Les ectàsies corneals primàries representen un grup d'entitats que cursen amb aprimament corneal progressiu, bilateral i asimètric. Dins d'aquestes, es troba per ordre de major a menor freqüència el queratocono, la degeneració marginal pel·lúcida i el queratoglobo. Han estat nombrosos els estudis que estableixen un probable origen en comú de les mateixes com a part de l'espectre d'una mateixa entitat amb un probable component genètic per a la seva presentació, l'anterior amb molt més freqüència en el queratocono. El gen Mir-184 és el microRNA més abundantment expressat en còrnia i cristal·lí, regulador de l'apoptosi en aquests teixits, i la seva mutació s'ha associat a queratocono en pacients caucàsics, convertint-lo en un potencial candidat, la seva mutació podria portar a el desenvolupament d'una ectàsia corneal primària. Pregunta de recerca Existeixen mutacions en el gen Mir-184 en un grup de pacients amb ectàsies corneals primàries? Justificació Les ectàsies corneals es troben dins les principals causes de trasplantament de còrnia en països desenvolupat, identificar l'origen del seu desenvolupament canviaria significativament el plantejament de la seva fisiopatologia i tractament. Hipòtesi de Recerca Mutacions en el gen Mir-184 s'associen amb la presència de ectàsies corneals primàries. Objectiu General Identificar mutacions de el gen Mir-184 en pacients amb diagnòstic de ectàsia corneal primària. Objectius Secundaris Realitzar una recerca exhaustiva a manera de revisió sistemàtica respecte a les ectàsies corneals primàries. Disseny de l'estudi Estudi descriptiu observacional en què es va buscar la presència o absència de la mutació en el gen Mir-184 en pacients amb ectàsies corneals primàries. Material i mètodes Previ consentiment informat, es va realitzar un anàlisi de genotipificació amb 200 mostres de casos esporàdics o familiars de pacients mexicans amb diagnòstic clínic i topogràfic de ectàsia corneal primària. A partir d' una mostra de 2 ml de sang, a cada pacient es va extreure el DNA genòmic, amb posterior anàlisi del gen Mir-184 mitjançant PCR i seqüenciació nucleotídica. Les seqüències obtingudes de cada pacient es van comparar amb la seqüència silvestre de Mir-184 a Ensembl (www.ensembl.org) per així determinar la presència o absència de mutació en el mateix gen. Es va determinar realitzar l'anàlisi estadístic mitjançant la prova de xi2 d'acord a la variable predictora dictotòmica que fa a la presència o absència de la mutació en el gen Mir-184. Resultats i Conclusions No s'ha trobat aquesta mutació en cap dels casos. Hi ha forta evidència científica que orienta les ectàsies corneals primàries com diferents graus de manifestació d'un mateix origen fisiopatològic en comú, i que orienta de manera predominant a l'queratocono com una malaltia amb un component genètic significatiu el qual s'expressa posterior a l'exposició a un factor ambiental desencadenant, de manera principal, el rascat ocular. El gen Mir-184 és el micro RNA més abundantment expressat tant en l'epiteli corneal com en el cristalí, i un important regulador de l'apoptosi, la seva mala regulació s'ha associat de manera significativa a el desenvolupament principalment de queratocono i la seva mutació representa un fort candidat per explicar el desenvolupament de ectàsies corneals primàries. S'ha observat que aproximadament el 60% dels gens codificants de proteïna són regulats per micro RNAs. El no haver pogut demostrar en aquest estudi una associació causa-efecte de manera significativa entre una mutació en el gen Mir-184 i el desenvolupament de ectàsies corneals primàries, orienta a que l'heterogeneïtat genètica juga un paper important per al desenvolupament de les mateixes, així com també l'exposició a factors ambientals desencadenants."Las ectasias corneales primarias representan un grupo de entidades que cursan con adelgazamiento corneal progresivo, bilateral y asimétrico. Dentro de éstas, se encuentra por orden de mayor a menor frecuencia el queratocono, la degeneración marginal pelúcida y el queratoglobo. Han sido numerosos los estudios que establecen un probable origen en común de las mismas como parte del espectro de una misma entidad con un probable componente genético para su presentación, lo anterior con mucho más frecuencia en el queratocono. El gen Mir-184 es el microRNA más abundantemente expresado en córnea y cristalino, regulador de la apoptosis en dichos tejidos, cuya mutación se ha asociado a queratocono en pacientes caucásicos, convirtiéndolo en un potencial candidato cuya mutación podría llevar al desarrollo de una ectasia corneal primaria. Pregunta de investigación Existen mutaciones del gen Mir-184 en un grupo de pacientes con ectasias corneales primarias? Justificación Las ectasias corneales representan una de las principales causas de trasplante de cornea en países desarrollados. El poder identificar el origen de su desarrollo cambiaría significativamente el planteamiento respecto a su fisiopatología y tratamiento. Hipótesis de la Investigación Mutaciones en el gen MIR 184 se asocian con la presencia de ectasias corneales primarias. Objetivo general Identificar mutaciones del gen Mir-184 en pacientes con diagnóstico de ectasia corneal primaria. Objetivos secundarios Realizar una búsqueda exhaustiva a modo de revisión sistemática respecto las ectasias corneales primarias. Diseño del estudio Estudio descriptivo observacional en el que se buscó la presencia o ausencia de la mutación en el gen Mir-184 en pacientes con ectasias corneales primarias. Material y métodos Previo consentimiento informado, se realizó un análisis de genotipificación en 200 muestras de casos esporádicos o familiares de pacientes mexicanos con diagnóstico clínico y topográfico de ectasia corneal primaria. De una muestra de 2 ml de sangre, a cada paciente se extrajo el DNA genómico, con posterior análisis del gen Mir-184 mediante PCR y secuenciación nucleotídica. Las secuencias obtenidas de cada paciente se compararon con la secuencia silvestre de Mir-184 en Ensembl (www.ensembl.org) para así determinar la presencia o ausencia de mutación en el mismo. Se determinó realizar el análisis estadístico mediante la prueba de xi2 de acuerdo a la variable predictora dictotómica respecto a la presencia o ausencia de la mutación en el gen Mir-184 . Resultados y Conclusiones No se encontró dicha mutación en ninguno de los casos. Hay fuerte evidencia científica que orienta a las ectasias corneales primarias tiene un mismo origen fisiopatológico en común, y que orienta de manera predominante al queratocono con un componente genético significativo el cual se expresa posterior a la exposición a un factor ambiental desencadenante. El gen Mir-184 al ser el micro RNA más abundantemente expresado tanto en el epitelio corneal como en el cristaliniano, y al ser un importante regulador de la apoptosis, mecanismo cuya mala regulación se ha asociado de manera significativa al desarrollo principalmente de queratocono, su mutación representa un fuerte candidato para explicar el desarrollo de ectasias corneales primarias. Se ha observado que aproximadamente el 60% de los genes codificantes de proteína son regulados por micro RNAs. El no haber podido demostrar en este estudio una asociación causa-efecto de manera significativa entre una mutación en el gen Mir-184 y el desarrollo de una ectasia corneal primaria, orienta a que la heterogeneidad genética juega un papel importante para el desarrollo de las mismas, así como también la exposición a factores ambientales desencadenantes, lo cual a su vez orienta a que dichas entidades se encuentran dentro del espectro de las enfermedades multifactoriales.Primary corneal ectasies represent a group of entities that have progressive, bilateral and asymmetric corneal thinning. Keratoconus, pellucid marginal degeneration and keratoglobus are englobed in the classification of primary corneal ectasies with keratoconus being the most frequent among them. There have been numerous reports that establish a probable common origin of them as part of the spectrum of the same entity with a probable genetic component for their presentation, mainly in keratoconus. Mir-184 gene is the most abundantly expressed microRNA in corneal and lens tissue and is also an important apoptosis regulator in these tissues. Its mutation has been associated with keratoconus in Caucasian patients, making its mutation a potential candidate for the development of a primary corneal ectasia. Research question Are mutations in the Mir-184 gene associated to primary corneal ectasies? Justification Corneal ectasies represent one of the main causes of corneal transplantation in developed countries. Being able to identify the origin of their development would significantly change their approach regarding its pathophysiology and treatment. Research Hypothesis Mutations in the Mir-184 gene are associated with the presence of primary corneal ectasies. General Objective To identify mutations in the gene Mir-184 in patients diagnosed with a primary corneal ectasia. Secondary Objectives To perform research in the literature in the form of a systematic review regarding primary corneal ectasies. Study Design This was an observational descriptive study where the presence or absence of mutation in the Mir-184 gene was sought in patients with primary corneal ectasies. Material and methods Prior informed consent, a genotyping analysis was performed in 200 samples of Mexican patients with clinical and topographic diagnosis of sporadic or familial primary corneal ectasia. From a 2 ml sample of blood, genomic DNA was extracted from each patient, and a subsequent analysis of the Mir-184 gene by PCR and nucleotide sequencing was performed. The sequences obtained from each patient were compared with the wild sequence of Mir-184 in Ensembl (www.ensembl.org) in order to determine the presence or absence of a mutation. The statistical analysis was performed using the xi2 test according to the dicotomic predictive variable regarding the presence or absence of the mutation. Results and conclusions No such mutation was found in any of the cases. There is strong scientific evidence that guides primary corneal ectasies as different degrees of manifestation of the same pathophysiological origin in common, and that predominantly guides keratoconus as a disease with a significant genetic component which is expressed after exposure to a environmental triggering factor, mainly, eye scratching. The Mir-184 gene, being the most abundantly expressed micro RNA in both the corneal and crystalline epithelium, and being an important regulator of apoptosis, a mechanism whose poor regulation has been significantly associated with the development mainly of keratoconus, represents a strong candidate to explain the development of primary corneal ectasies. It has been observed that approximately 60% of the protein coding genes are regulated by micro RNAs. Not having been able to demonstrate in this study a significant cause-effect association between a mutation in the Mir-184 gene and the development of primary corneal ectasies, could orient to the fact that genetic heterogeneity plays an important role in their development, as well as exposure to environmental triggering factors, which would englobe them as part of the diseases with a multifactorial origin

Lalage M Wakefield - One of the best experts on this subject based on the ideXlab platform.

  • a p21 zeb1 complex inhibits epithelial mesenchymal transition through the microrna 183 96 182 cluster
    Molecular and Cellular Biology, 2014
    Co-Authors: Xiaoling Li, Toshifumi Hara, Youngeun Choi, Murugan Subramanian, Princy Francis, Sven Bilke, Robert L Walker, Marbin Pineda, Yuan Yang, Lalage M Wakefield
    Abstract:

    : The tumor suppressor p21 acts as a cell cycle inhibitor and has also been shown to regulate gene expression by functioning as a transcription corepressor. Here, we identified p21-regulated microRNAs (miRNAs) by sequencing small RNAs from isogenic p21(+/+) and p21(-/-) cells. Three abundant miRNA clusters, miR-200b-200a-429, miR-200c-141, and miR-183-96-182, were downregulated in p21-deficient cells. Consistent with the known function of the miR-200 family and p21 in inhibition of the epithelial-mesenchymal transition (EMT), we observed EMT upon loss of p21 in multiple model systems. To explore a role of the miR-183-96-182 cluster in EMT, we identified its genome-wide targets and found that miR-183 and miR-96 repressed common targets, including SLUG, ZEB1, ITGB1, and KLF4. Reintroduction of miR-200, miR-183, or miR-96 in p21(-/-) cells inhibited EMT, cell migration, and invasion. Conversely, antagonizing miR-200 and miR-183-96-182 cluster miRNAs in p21(+/+) cells increased invasion and elevated the levels of VIM, ZEB1, and SLUG mRNAs. Furthermore, we found that p21 forms a complex with ZEB1 at the miR-183-96-182 cluster promoter to inhibit transcriptional repression of this cluster by ZEB1, suggesting a reciprocal feedback loop.

  • abstract 5331 a novel function of p21 in inhibition of epithelial mesenchymal transition through micrornas
    Cancer Research, 2013
    Co-Authors: Xiao L Li, Toshifumi Hara, Youngeun Choi, Murugan Subramanian, Princy Francis, Sven Bilke, Marbin Pineda, Yuan Yang, Robert Walker, Lalage M Wakefield
    Abstract:

    The tumor suppressor p21 inhibits cell proliferation during the stress response. However, p21 can also directly regulate gene expression by repressing specific transcription factors. Here, we identified p21-regulated miRNAs by sequencing small RNAs from HCT116-p21+/+ and HCT116-p21-/- cells. Three abundant clusters, miR-200b-200a-429, miR-200c-141 and miR-183-96-182 were down-regulated in p21-depleted HCT116 and MCF10A cells. Loss of p21 induced epithelial-mesenchymal transition (EMT) and enhanced migration and invasion in multiple model systems. Identification of genome-wide targets of the miR-183-96-182 cluster indicated that miR-183 and miR-96 repressed common targets, including SLUG, ZEB1, ITGB1 and KLF4, to inhibit EMT, migration and invasion. In turn, elevated ZEB1 levels in HCT116-p21-/- cells directly repressed miR-183-96-182 cluster transcription, revealing a feedback loop. Re-introduction of miR-200, miR-183 or miR-96 in HCT116-p21-/- cells inhibited migration and invasion. These novel findings suggest that coordinated down-regulation of three miRNA clusters upon loss of p21 in unstressed cells promotes EMT, migration and invasion. Citation Format: Xiao L. Li, Toshifumi Hara, Youngeun Choi, Murugan Subramanian, Princy Francis, Sven Bilke, Robert L. Walker, Marbin Pineda, Yu-an Yang, Ji Luo, Lalage M. Wakefield, Ben H. Park, Thomas Brabletz, Dipanjan Chowdhury, Paul S. Meltzer, Ashish Lal. A novel function of p21 in inhibition of epithelial-mesenchymal transition through microRNAs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5331. doi:10.1158/1538-7445.AM2013-5331

Larisa Nonn - One of the best experts on this subject based on the ideXlab platform.

  • the mir 183 family cluster alters zinc homeostasis in benign prostate cells organoids and prostate cancer xenografts
    Scientific Reports, 2017
    Co-Authors: Shweta Dambal, Larisa Nonn, Bethany Baumann, Tara Mccray, Latanya V Williams, Zachary Richards, Ryan Deaton, Gail S Prins
    Abstract:

    The miR-183 cluster, which is comprised of paralogous miRs-183, -96 and -182, is overexpressed in many cancers, including prostate adenocarcinoma (PCa). Prior studies showed that overexpression of individual pre-miRs-182, -96 and -183 in prostate cells decreased zinc import, which is a characteristic feature of PCa tumours. Zinc is concentrated in healthy prostate 10-fold higher than any other tissue, and an >80% decrease in zinc is observed in PCa specimens. Here, we studied the effect of overexpression of the entire 4.8 kb miR-183 family cluster, including the intergenic region which contains highly conserved genomic regions, in prostate cells. This resulted in overexpression of mature miR-183 family miRs at levels that mimic cancer-related changes. Overexpression of the miR-183 cluster reduced zinc transporter and intracellular zinc levels in benign prostate cells, PCa xenografts and fresh prostate epithelial organoids. Microarray analysis of miR-183 family cluster overexpression in prostate cells showed an enrichment for cancer-related pathways including adhesion, migration and wound healing. An active secondary transcription start site was identified within the intergenic region of the miR-183 cluster, which may regulate expression of miR-182. Taken together, this study shows that physiologically relevant expression of the miR-183 family regulates zinc levels and carcinogenic pathways in prostate cells.

  • mir 183 96 182 cluster is overexpressed in prostate tissue and regulates zinc homeostasis in prostate cells
    Journal of Biological Chemistry, 2011
    Co-Authors: Brittany Mihelich, Ekaterina A Khramtsova, Nicole N Arva, Avani Vaishnav, Daniel N Johnson, Angeline Antonio Giangreco, Elena S Martensuzunova, Omar Bagasra, Andre Kajdacsyballa, Larisa Nonn
    Abstract:

    Decreased zinc levels are a hallmark of prostate cancer tumors as zinc uniquely concentrates in healthy prostate tissue. Increased dietary zinc correlates with decreased risk of advanced prostate cancer and decreased mortality from prostate cancer. The mechanisms of prostatic zinc homeostasis are not known. Lower zinc levels in the tumor are correlated directly with decreased expression of the zinc transporter hZIP1. We report identification of a microRNA cluster that regulates multiple zinc transporters, including hZIP1. Screening in laser capture microdissected prostate cancer tumors identified miR-182 as a potential regulator of hZIP1. Regulation of hZIP1 by miR-182 via two binding sites was confirmed in primary prostate cell cultures. miR-96 and miR-183 are expressed as a cluster with miR-182 and share similar sequences. Array profiling of tissue showed that miR-183, -96, and -182 are higher in prostate cancer tissue compared with normal prostate. Overexpression of the entire miR-183-96-182 cluster suppressed five additional zinc transporters. Overexpression of miR-183, -96, and -182 individually or as a cluster diminished labile zinc pools and reduced zinc uptake, demonstrating this miR cluster as a regulator of zinc homeostasis. We observed regulation of zinc homeostasis by this cluster in prostate cells and HEK-293 cells, suggesting a universal mechanism that is not prostate-specific. To our knowledge, this is the first report of a miR cluster targeting a family of metal transport proteins. Individually or as a cluster, miR-183, -96, and -182 are overexpressed in other cancers too, implicating this miR cluster in carcinogenesis.

Shunbin Xu - One of the best experts on this subject based on the ideXlab platform.

  • the microrna 183 96 182 cluster is essential for stereociliary bundle formation and function of cochlear sensory hair cells
    Scientific Reports, 2018
    Co-Authors: Ruishuang Geng, David N Furness, Chithra K Muraleedharan, Jinsheng Zhang, Alain Dabdoub, Shunbin Xu
    Abstract:

    The microRNA (miR)-183/96/182 cluster plays important roles in the development and functions of sensory organs, including the inner ear. Point-mutations in the seed sequence of miR-96 result in non-syndromic hearing loss in both mice and humans. However, the lack of a functionally null mutant has hampered the evaluation of the cluster’s physiological functions. Here we have characterized a loss-of-function mutant mouse model (miR-183CGT/GT), in which the miR-183/96/182 cluster gene is inactivated by a gene-trap (GT) construct. The homozygous mutant mice show profound congenital hearing loss with severe defects in cochlear hair cell (HC) maturation, alignment, hair bundle formation and the checkboard-like pattern of the cochlear sensory epithelia. The stereociliary bundles retain an immature appearance throughout the cochlea at postnatal day (P) 3 and degenerate soon after. The organ of Corti of mutant newborn mice has no functional mechanoelectrical transduction. Several predicted target genes of the miR-183/96/182 cluster that are known to play important roles in HC development and function, including Clic5, Rdx, Ezr, Rac1, Myo1c, Pvrl3 and Sox2, are upregulated in the cochlea. These results suggest that the miR-183/96/182 cluster is essential for stereociliary bundle formation, morphogenesis and function of the cochlear HCs.

  • microrna 183 96 182 cluster regulates inkt cell development and function irm7p 706
    Journal of Immunology, 2015
    Co-Authors: Li Zhou, Shunbin Xu, Guihua Li, Xiling Zhang, Qing Sheng Mi
    Abstract:

    MicroRNAs (miRNAs), a class of non-coding small RNAs. Invariant Natural Killer T (iNKT) cells are potent regulators of diverse immune responses. Our previous study indicated that lack of miRNAs following the deletion of Dicer, a miRNAs-processing enzyme, affect iNKT cell development and function (PNAS, 2009). However, the roles of specific miRNA in iNKT cell development and function remain unclear. The miR-183/96/182 cluster, composed of 3 miRNA genes, is significantly upregulated upon T cell activation. In this study, we investigate the role of miR-183/96/182 cluster on development and function of iNKT cells, using miR-183/96/182 knockout (KO) mice. We found that the frequency of immature iNKT cells significantly increased, while mature iNKT cells significantly decreased in thymus of KO mice compared to littermate control (P i NKT cells displayed defective cytokine production, including IL-4 and IFNγ, compared to iNKT cells from littermate control (P

Dawit Tesfaye - One of the best experts on this subject based on the ideXlab platform.

  • microrna 183 96 182 cluster regulates bovine granulosa cell proliferation and cell cycle transition by coordinately targeting foxo1
    Biology of Reproduction, 2016
    Co-Authors: Samuel Gebremedhn, Dessie Salilewwondim, M Hoelker, F Rings, Christiane Neuhoff, Ernst Tholen, K Schellander, Dawit Tesfaye
    Abstract:

    Large-scale expression profiling of micro-RNAs (miRNAs) in bovine granulosa cells from dominant and subordinate follicles on Day 19 of the estrous cycle revealed enriched micro-RNA-183-96-182 cluster miRNAs in preovulatory dominant follicles that coordinately regulate the forkhead box protein O1 (FOXO1) gene. However, little is known about the role of this cluster in bovine granulosa cell function. We used an in vitro granulosa cell culture model to investigate this role. Granulosa cells aspirated from small growing follicles (3–5 mm in diameter) were cultured in Dulbecco modified Eagle medium/F-12 medium supplemented with fetal bovine serum and transfected with locked nucleic acid-based miRNA mimics, inhibitors, and corresponding negative controls. Overexpression of the miRNA cluster resulted in suppression of FOXO1 mRNA and protein, whereas inhibition of the cluster increased expression of FOXO1 mRNA. Overexpression also increased the relative rate of cell proliferation, whereas inhibition slowed it down. Similarly, the proportion of cells under G0/G1 arrest declined, whereas the ratio of cells in S phase increased in response to miR-183-96-182 overexpression. Selective knockdown of FOXO1 mRNA using anti-FOXO1 small interfering RNA increased the rate of granulosa cell proliferation, decreased the proportion of cells under G0/G1 arrest, and increased the proportion of cells in the S phase of cell cycle. Our data suggest that miR-183-96-182 cluster miRNAs promote proliferation and G1/S transition of bovine granulosa cells by coordinately targeting FOXO1, suggesting a critical role in granulosa cell function. MicroRNA-183-96-182 cluster regulates bovine granulosa cell function by targeting FOXO1 gene.

  • 129 microrna 183 96 182 cluster promote bovine granulosa cell proliferation through coordinated regulation of foxo1
    Reproduction Fertility and Development, 2016
    Co-Authors: Samuel Gebremedhn, Dessie Salilewwondim, M Hoelker, F Rings, Christiane Neuhoff, Ernst Tholen, Christian Looft, K Schellander, Dawit Tesfaye
    Abstract:

    Among other microRNA clusters, we previously showed that the miR-183~96~182 cluster (miR-183, miR-96, and miR-182) is abundantly expressed in bovine granulosa cells (bGC) of preovulatory dominant follicles obtained at the follicular phase of the bovine oestrous cycle. Moreover, this miRNA cluster are validated to coordinately target the Fork head O1 (FOXO1), a subfamily of transcription factors that regulate genes involved in cell proliferation, apoptosis, cell cycle arrest, and metabolism. However, the functional involvement of miR-183~96~182 cluster in bGC function by regulation of FOXO1 is not yet determined. Here, we aimed to investigate the function of miR-183~96~182 cluster in bGC using in vitro cell culture model. For this, bGC were aspirated from ovarian follicles (O 3–5 mm) obtained from local abattoir. Cells were plated in 24-well plate (2.5 × 105 cells well–1) in DMEM/F-12 (Sigma, Germany) supplemented with 10% FBS (GIBCO, Grand Island, NY) and 1% penicillin/streptomycin (GIBCO) and incubated at 37°C in 5% CO2. Transfection of bGC with miRNA mimics, inhibitors, FOXO1-siRNA, and appropriate controls (Exiqon, Vedbaek, Denmark) was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). Cell proliferation was determined using Cell Counting Kit-8 (CCK-8; Dojindo Molecular Technology, Kumamoto, Japan). Cell cycle distribution was determined with flow cytometric analysis. Total RNA was isolated using miRNeasy mini kit (Qiagen, Hilden, Germany), quantification of target gene was performed using qPCR, and data were analysed using ΔΔCT method. Differences in the mean expression values between treatments were analysed with two-tailed Student’s t-test and statistical significance was defined at P ≤ 0.05. Results showed that a sponge effect was observed upon inhibition in individual miRNA of the cluster, which could be attributed to the partial sequence similarity among cluster members. Both FOXO1 mRNA and protein expression were significantly reduced upon transfection of bGC with miR-183~96~182 cluster mimics, while miR-183~96~182 cluster inhibition increased both FOXO1 mRNA and protein expression. Transfection of bGC with miR-183~96~182 mimics promoted cell proliferation, while inhibition tends to slow down proliferation. Furthermore, the proportion of bGC under G0/G1 arrest markedly declined (P < 0.05), while the S and G2/M phases increased in response to miR-183~96~182 mimicking. Selective knockdown of FOXO1 with FOXO1-siRNA significantly reduced FOXO1 mRNA and protein expression. Interestingly, knockdown of FOXO1 showed similar phenotypic effects such as that of miR-183~96~182 mimics transfection, which resulted in elevated bGC proliferation and reduction in the proportion of cells under G0/G1 arrest. In conclusion, overexpression of miR-183~96~182 cluster promote bGC proliferation and G0/G1 to S and G2/M cell cycle transition through coordinated regulation of genes in the FOXO1 signaling axis.