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OBJECTIVES: The purpose of the study is to investigate feasibility of early activation after cochlear implantation by evaluating long-term impedance change and speech perception. DESIGN: Case-control study SETTING: Between July 2015 and December 2016, we prospectively enrolled 20 subjects for early activation (within 24 hours after cochlear implantation). On the other hand, from November 2013 to July 2015, 20 age- and sex-matched control subjects from the database of cochlear implantees treated with conventional activation schedule (4 weeks after surgery) were retrospectively enrolled. PARTICIPANT: Forty patients who underwent cochlear implantation surgeries. MAIN OUTCOME MEASURES: The series impedance and speech perception score of both groups were compared. RESULTS: No statistical difference in long-term follow-up between the two groups was found using GEEs and multivariate analysis. In the early activation group, impedance reached a steady level by the 2nd postoperative week, and the hearing perception ability significantly improved by the 4th postoperative week. CONCLUSION: This comparative study illustrated sequential impedance data during early activation (24 hours) and conventional activation (4 weeks) after CI surgery. There were no major complications in either group, and the safety of early activation with respect to impedance changes, postoperative residual hearing preservation and speech perception scores were non-inferior to that of the conventional group. Therefore, in this study, we established the feasibility of early activation 24 hours after cochlear implantation.
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Implantación Coclear , Implantes Cocleares , Trastornos de la Audición/terapia , Percepción del Habla/fisiología , Pruebas de Impedancia Acústica , Adolescente , Adulto , Factores de Edad , Anciano , Estudios de Casos y Controles , Niño , Preescolar , Estudios de Factibilidad , Femenino , Trastornos de la Audición/fisiopatología , Trastornos de la Audición/psicología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
Complex genetic networks control hematopoietic stem cell differentiation into progenitors that give rise to billions of erythrocytes daily. Previously, we described a role for the master regulator of erythropoiesis, GATA-1, in inducing genes encoding components of the autophagy machinery. In this context, the Forkhead transcription factor, Foxo3, amplified GATA-1-mediated transcriptional activation. To determine the scope of the GATA-1/Foxo3 cooperativity, and to develop functional insights, we analyzed the GATA-1/Foxo3-dependent transcriptome in erythroid cells. GATA-1/Foxo3 repressed expression of Exosc8, a pivotal component of the exosome complex, which mediates RNA surveillance and epigenetic regulation. Strikingly, downregulating Exosc8, or additional exosome complex components, in primary erythroid precursor cells induced erythroid cell maturation. Our results demonstrate a new mode of controlling erythropoiesis in which multiple components of the exosome complex are endogenous suppressors of the erythroid developmental program.
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Eritrocitos/citología , Exosomas/fisiología , Factores de Transcripción Forkhead/metabolismo , Factor de Transcripción GATA1/metabolismo , Animales , Autofagia , Diferenciación Celular , Epigénesis Genética , Eritroblastos/citología , Células Eritroides/metabolismo , Eritropoyesis/genética , Proteína Forkhead Box O3 , Regulación de la Expresión Génica , Ratones , ARN/metabolismo , Activación TranscripcionalRESUMEN
Induced pluripotent stem (iPS) cell technology holds vast promises for a cure to the hemoglobinopathies. Constructs and methods to safely insert therapeutic genes to correct the genetic defect need to be developed. Site-specific insertion is a very attractive method for gene therapy because the risks of insertional mutagenesis are eliminated provided that a "safe harbor" is identified, and because a single set of validated constructs can be used to correct a large variety of mutations simplifying eventual clinical use. We report here the correction of α-thalassemia major hydrops fetalis in transgene-free iPS cells using zinc finger-mediated insertion of a globin transgene in the AAVS1 site on human chromosome 19. Homozygous insertion of the best of the 4 constructs tested led to complete correction of globin chain imbalance in erythroid cells differentiated from the corrected iPS cells.
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Endonucleasas/metabolismo , Células Madre Pluripotentes Inducidas/metabolismo , Dedos de Zinc , Talasemia alfa/metabolismo , Diferenciación Celular , Línea Celular , Eritroblastos/citología , Eritroblastos/metabolismo , Índices de Eritrocitos , Fibroblastos/citología , Fibroblastos/metabolismo , Regulación de la Expresión Génica , Orden Génico , Homocigoto , Humanos , Células Madre Pluripotentes Inducidas/citología , Recombinación Genética , Transgenes , Globinas alfa/genética , Globinas alfa/metabolismo , Talasemia alfa/genética , Talasemia alfa/terapiaRESUMEN
The reprogramming of human acute myeloid leukemia (AML) cells into induced pluripotent stem cell (iPSC) lines could provide new faithful genetic models of AML, but is currently hindered by low success rates and uncertainty about whether iPSC-derived cells resemble their primary counterparts. Here we developed a reprogramming method tailored to cancer cells, with which we generated iPSCs from 15 patients representing all major genetic groups of AML. These AML-iPSCs retain genetic fidelity and produce transplantable hematopoietic cells with hallmark phenotypic leukemic features. Critically, single-cell transcriptomics reveal that, upon xenotransplantation, iPSC-derived leukemias faithfully mimic the primary patient-matched xenografts. Transplantation of iPSC-derived leukemias capturing a clone and subclone from the same patient allowed us to isolate the contribution of a FLT3-ITD mutation to the AML phenotype. The results and resources reported here can transform basic and preclinical cancer research of AML and other human cancers. SIGNIFICANCE: We report the generation of patient-derived iPSC models of all major genetic groups of human AML. These exhibit phenotypic hallmarks of AML in vitro and in vivo, inform the clonal hierarchy and clonal dynamics of human AML, and exhibit striking similarity to patient-matched primary leukemias upon xenotransplantation. See related commentary by Doulatov, p. 252. This article is highlighted in the In This Issue feature, p. 247.
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Células Madre Pluripotentes Inducidas , Leucemia Mieloide Aguda , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Leucemia Mieloide Aguda/genética , Fenotipo , Perfilación de la Expresión Génica , Variación Genética/genéticaRESUMEN
BACKGROUND: Cochlear implants (CIs) are viable treatment options in patients with severe to profound hearing loss. Speech recognition difficulties were reported in some CI recipients even with a good-aided hearing threshold. The aim of this study was to report a mapping strategy based on different target-aided hearing thresholds to achieve optimal speech recognition and maximize functional outcomes. The safety and efficacy of the mapping strategy were also inspected in the article. METHODS: This prospective repeated measures study enrolled 20 adult CI recipients with postlingual deafness using the MED-EL CI system. Word and sentence discrimination assessment and administration of a questionnaire pertaining to comfort level were conducted at the end of each session. The electrophysiological features of the CI mapping were recorded. RESULTS: The correlation between audiometry results and word and sentence recognition was not high. CIs performed best at an audiometry threshold between 25 and 35 dB. CONCLUSION: CI performance with the best perception relies on a balance between minimizing the hearing threshold and maximizing the dynamic range while maintaining an appropriate comfort level, which was achieved when the target hearing threshold was set at 25-35 dB in this study.
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Implantación Coclear , Implantes Cocleares , Pérdida Auditiva Sensorineural , Percepción del Habla , Adulto , Implantación Coclear/métodos , Humanos , Estudios Prospectivos , Percepción del Habla/fisiologíaRESUMEN
The derivation of mesenchymal progenitors from human embryonic stem cells (hESCs) has recently been reported. We studied the immune characteristics of these hESC-derived mesenchymal progenitors (EMPs) and their interactions with T lymphocytes and natural killer cells (NKs), two populations of lymphocytes with important roles in transplantation immunology. EMPs express a number of bone marrow mesenchymal stromal cell (BMMSC) markers, as well as the hESC marker SSEA-4. Immunologically, EMPs do not express HLA-DR or costimulatory molecules. On the other hand, HLA-G, a nonclassic MHC I protein involved in mediating maternal-fetal tolerance, can be found on the surface of EMPs, and its expression is increased after interferon-gamma stimulation. EMPs can suppress CD4(+) or CD8(+) lymphocyte proliferation, similar to BMMSCs. However, EMPs are more resistant to NK-mediated lysis than BMMSCs and can suppress the cytotoxic effects of activated NKs, as well as downregulating the NK-activating receptors NKp30 and NKp46. With their broad immunosuppressive properties, EMPs may represent a new potential cell source for therapeutic use.
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Células Madre Embrionarias/citología , Células Asesinas Naturales/inmunología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/patología , Linfocitos T/inmunología , Apoptosis/efectos de los fármacos , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Antígeno CD56/metabolismo , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Línea Celular , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Inmunofenotipificación , Interferón gamma/farmacología , Interleucina-15/farmacología , Interleucina-2/farmacología , Linfocitos T/citologíaRESUMEN
OBJECTIVE: To further elucidate the role of balloon Eustachian tuboplasty (BET) in tympanoplasty, we conducted a study to compare the outcomes of tympanoplasty with and without BET for the treatment of chronic suppurative otitis media (CSOM) with obstructive Eustachian tube dysfunction (OETD). STUDY DESIGN: Case control study. SETTING: Tertiary referral center. PATIENTS: A total of 70 ears diagnosed with CSOM (tubotympanic type) and OETD were included in this study. Thirty-five patients were prospectively enrolled for BET and tympanomastoidectomy between February 2018 and June 2019. Thirty-five control subjects were matched by sex and age and retrospectively enrolled for tympanomastoidectomy between July 2016 and January 2018. INTERVENTIONS: BET, tympanomastoidectomy. MAIN OUTCOME MEASURES: The graft take rate, hearing levels, and Eustachian tube function test results. RESULTS: The graft take success rate was higher in the BET group (80.0%; 28/35) than in the control group (68.6%; 24/35). However, the difference was not statistically significant. The average air-bone gap (ABG) improvement was 10.93â±â7.70âdB in the BET group and 7.11â±â8.08âdB in the control group, with a statistically significant between-group difference (pâ=â0.033). CONCLUSIONS: Our findings suggest that BET can objectively and subjectively improve the Eustachian tube function, with a slight but significant improvement in ABG despite the lack of a clinically significant improvement overall. However, it does not affect the graft take rate. In summary, BET could be used as an adjunctive procedure in the treatment of CSOM with OETD.
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Trompa Auditiva , Otitis Media Supurativa , Estudios de Casos y Controles , Trompa Auditiva/cirugía , Humanos , Otitis Media Supurativa/cirugía , Estudios Retrospectivos , Resultado del Tratamiento , TimpanoplastiaRESUMEN
OBJECTIVE: Whereas the nature of otosclerosis has been extensively investigated, treatment modalities in advanced otosclerosis with the sensorineural hearing loss (SNHL) are relatively unexplored. MATERIALS AND METHODS: This article presents a retrospective case series study of nine patients who received a one-stage piston coupled with Vibrant Soundbridge® vibroplasty in treating otosclerosis with moderate-to-severe SNHL. RESULTS: The findings suggest that hearing loss could be restored across frequencies and no significant change in the bone-conduction threshold were measured. CONCLUSION: One-stage piston surgery coupled with incus vibroplasty is a safe procedure and has sufficient efficacy to restore hearing loss in patients with otosclerosis with moderate-to-severe SNHL.
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A glomangiopericytoma, or sinonasal type hemangiopericytoma, is a rare lesion which accounts for <0.5% of all sinonasal tumors. The mainstay treatment is wide excision. Instead of traditional open surgical approaches, such as midfacial degloving or lateral rhinotomy, we offer a case of 21-year-old male with diagnosis of glomangiopericytoma with skull base and intraorbital invasion and received navigation-assisted endoscopic excision of a glomangiopericytoma.
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TET enzymes oxidize 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), which can lead to DNA demethylation. However, direct connections between TET-mediated DNA demethylation and transcriptional output are difficult to establish owing to challenges in distinguishing global versus locus-specific effects. Here we show that TET1, TET2 and TET3 triple-knockout (TKO) human embryonic stem cells (hESCs) exhibit prominent bivalent promoter hypermethylation without an overall corresponding decrease in gene expression in the undifferentiated state. Focusing on the bivalent PAX6 locus, we find that increased DNMT3B binding is associated with promoter hypermethylation, which precipitates a neural differentiation defect and failure of PAX6 induction during differentiation. dCas9-mediated locus-specific demethylation and global inactivation of DNMT3B in TKO hESCs partially reverses the hypermethylation at the PAX6 promoter and improves differentiation to neuroectoderm. Taking these findings together with further genome-wide methylation and TET1 and DNMT3B ChIP-seq analyses, we conclude that TET proteins safeguard bivalent promoters from de novo methylation to ensure robust lineage-specific transcription upon differentiation.
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Metilación de ADN , Proteínas de Unión al ADN/fisiología , Células Madre Embrionarias/metabolismo , Oxigenasas de Función Mixta/fisiología , Regiones Promotoras Genéticas , Animales , Diferenciación Celular/genética , Células Cultivadas , Proteínas de Unión al ADN/genética , Dioxigenasas/genética , Dioxigenasas/fisiología , Células Madre Embrionarias/citología , Humanos , Ratones , Oxigenasas de Función Mixta/genética , Oxigenasas de Función Mixta/metabolismo , Mutación , Placa Neural/citología , Factor de Transcripción PAX6/biosíntesis , Factor de Transcripción PAX6/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas/fisiologíaRESUMEN
Connecting specific cancer genotypes with phenotypes and drug responses constitutes the central premise of precision oncology but is hindered by the genetic complexity and heterogeneity of primary cancer cells. Here, we use patient-derived induced pluripotent stem cells (iPSCs) and CRISPR/Cas9 genome editing to dissect the individual contributions of two recurrent genetic lesions, the splicing factor SRSF2 P95L mutation and the chromosome 7q deletion, to the development of myeloid malignancy. Using a comprehensive panel of isogenic iPSCs-with none, one, or both genetic lesions-we characterize their relative phenotypic contributions and identify drug sensitivities specific to each one through a candidate drug approach and an unbiased large-scale small-molecule screen. To facilitate drug testing and discovery, we also derive SRSF2-mutant and isogenic normal expandable hematopoietic progenitor cells. We thus describe here an approach to dissect the individual effects of two cooperating mutations to clinically relevant features of malignant diseases.
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Antineoplásicos/uso terapéutico , Células Madre Pluripotentes Inducidas/patología , Mutación/genética , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Empalme Alternativo/genética , Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/patología , Neoplasias/patología , Fenotipo , Factores de Empalme Serina-Arginina/genética , Factores de Empalme Serina-Arginina/metabolismo , Bibliotecas de Moléculas Pequeñas/farmacologíaRESUMEN
In the version of this article initially published, in the Methods, the Gene Expression Omnibus accession code for H3K36me3 ChIP-seq data was incorrectly given as GSM1003585 instead of GSM733725. The error has been corrected in the HTML, PDF and print versions of the article.
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The version of the Supplementary Text and Figures file initially posted was missing Supplementary Tables 1-6 and the Supplementary Note and used incorrect versions of the supplementary figures.
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Myeloid malignancy is increasingly viewed as a disease spectrum, comprising hematopoietic disorders that extend across a phenotypic continuum ranging from clonal hematopoiesis to myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). In this study, we derived a collection of induced pluripotent stem cell (iPSC) lines capturing a range of disease stages encompassing preleukemia, low-risk MDS, high-risk MDS, and secondary AML. Upon their differentiation, we found hematopoietic phenotypes of graded severity and/or stage specificity that together delineate a phenotypic roadmap of disease progression culminating in serially transplantable leukemia. We also show that disease stage transitions, both reversal and progression, can be modeled in this system using genetic correction or introduction of mutations via CRISPR/Cas9 and that this iPSC-based approach can be used to uncover disease-stage-specific responses to drugs. Our study therefore provides insight into the cellular events demarcating the initiation and progression of myeloid transformation and a new platform for testing genetic and pharmacological interventions.
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Transformación Celular Neoplásica/patología , Progresión de la Enfermedad , Células Madre Pluripotentes Inducidas/citología , Leucemia Mieloide Aguda/patología , Animales , Antineoplásicos/farmacología , Transformación Celular Neoplásica/efectos de los fármacos , Reprogramación Celular/efectos de los fármacos , Reprogramación Celular/genética , Análisis Mutacional de ADN , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Células Madre Pluripotentes Inducidas/metabolismo , Leucemia Mieloide Aguda/genética , Ratones , Modelos Biológicos , Síndromes Mielodisplásicos/patología , Trasplante de Neoplasias , Fenotipo , Transcriptoma/efectos de los fármacos , Transcriptoma/genéticaRESUMEN
To combat hostile viruses, bacteria and archaea have evolved a unique antiviral defense system composed of clustered regularly interspaced short palindromic repeats (CRISPRs), together with CRISPR-associated genes (Cas). The CRISPR/Cas9 system develops an adaptive immune resistance to foreign plasmids and viruses by creating site-specific DNA double-stranded breaks (DSBs). Here we adapt the CRISPR/Cas9 system to human cells for intracellular defense against foreign DNA and viruses. Using HIV-1 infection as a model, our results demonstrate that the CRISPR/Cas9 system disrupts latently integrated viral genome and provides long-term adaptive defense against new viral infection, expression and replication in human cells. We show that engineered human-induced pluripotent stem cells stably expressing HIV-targeted CRISPR/Cas9 can be efficiently differentiated into HIV reservoir cell types and maintain their resistance to HIV-1 challenge. These results unveil the potential of the CRISPR/Cas9 system as a new therapeutic strategy against viral infections.
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Linfocitos T CD4-Positivos/inmunología , Sistemas CRISPR-Cas/inmunología , Infecciones por VIH/prevención & control , Células Madre Pluripotentes/inmunología , Secuencia de Bases , Western Blotting , Cartilla de ADN/genética , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Dosificación de Gen/genética , Células HEK293 , Humanos , Mutación INDEL/genética , Microscopía Fluorescente , Datos de Secuencia Molecular , Plásmidos/genética , Células Madre Pluripotentes/metabolismo , Provirus/genética , Provirus/metabolismoRESUMEN
Chromosomal deletions associated with human diseases, such as cancer, are common, but synteny issues complicate modeling of these deletions in mice. We use cellular reprogramming and genome engineering to functionally dissect the loss of chromosome 7q (del(7q)), a somatic cytogenetic abnormality present in myelodysplastic syndromes (MDS). We derive del(7q)- and isogenic karyotypically normal induced pluripotent stem cells (iPSCs) from hematopoietic cells of MDS patients and show that the del(7q) iPSCs recapitulate disease-associated phenotypes, including impaired hematopoietic differentiation. These disease phenotypes are rescued by spontaneous dosage correction and can be reproduced in karyotypically normal cells by engineering hemizygosity of defined chr7q segments in a 20-Mb region. We use a phenotype-rescue screen to identify candidate haploinsufficient genes that might mediate the del(7q)- hematopoietic defect. Our approach highlights the utility of human iPSCs both for functional mapping of disease-associated large-scale chromosomal deletions and for discovery of haploinsufficient genes.
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Deleción Cromosómica , Ingeniería Genética , Células Madre Pluripotentes Inducidas/citología , Síndromes Mielodisplásicos/genética , Animales , Cromosomas Humanos Par 7/genética , Humanos , Cariotipificación , Ratones , Síndromes Mielodisplásicos/terapiaRESUMEN
Hepatocyte transplantation has the potential to cure inherited liver diseases, but its application is impeded by a scarcity of donor livers. Therefore, we explored whether transplantation of hepatocyte-like cells (iHeps) differentiated from human induced pluripotent stem cells (iPSCs) could ameliorate inherited liver diseases. iPSCs reprogrammed from human skin fibroblasts were differentiated to iHeps, which were transplanted into livers of uridinediphosphoglucuronate glucuronosyltransferase-1 (UGT1A1)-deficient Gunn rats, a model of Crigler-Najjar syndrome 1 (CN1), where elevated unconjugated bilirubin causes brain injury and death. To promote iHep proliferation, 30% of the recipient liver was X-irradiated before transplantation, and hepatocyte growth factor was expressed. After transplantation, UGT1A1+ iHep clusters constituted 2.5%-7.5% of the preconditioned liver lobe. A decline of serum bilirubin by 30%-60% and biliary excretion of bilirubin glucuronides indicated that transplanted iHeps expressed UGT1A1 activity, a postnatal function of hepatocytes. Therefore, iHeps warrant further exploration as a renewable source of hepatocytes for treating inherited liver diseases.
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Tratamiento Basado en Trasplante de Células y Tejidos , Síndrome de Crigler-Najjar/terapia , Hepatocitos/trasplante , Hiperbilirrubinemia/terapia , Células Madre Pluripotentes Inducidas/trasplante , Animales , Bilirrubina/sangre , Síndrome de Crigler-Najjar/sangre , Síndrome de Crigler-Najjar/patología , Glucuronosiltransferasa/deficiencia , Glucuronosiltransferasa/genética , Humanos , Hiperbilirrubinemia/sangre , Hiperbilirrubinemia/genética , Hígado/patología , Hígado/cirugía , Ratas , Ratas GunnRESUMEN
Developmental and homeostatic remodeling of cellular organelles is mediated by a complex process termed autophagy. The cohort of proteins that constitute the autophagy machinery functions in a multistep biochemical pathway. Though components of the autophagy machinery are broadly expressed, autophagy can occur in specialized cellular contexts, and mechanisms underlying cell-type-specific autophagy are poorly understood. We demonstrate that the master regulator of hematopoiesis, GATA-1, directly activates transcription of genes encoding the essential autophagy component microtubule-associated protein 1 light chain 3B (LC3B) and its homologs (MAP1LC3A, GABARAP, GABARAPL1, and GATE-16). In addition, GATA-1 directly activates genes involved in the biogenesis/function of lysosomes, which mediate autophagic protein turnover. We demonstrate that GATA-1 utilizes the forkhead protein FoxO3 to activate select autophagy genes. GATA-1-dependent LC3B induction is tightly coupled to accumulation of the active form of LC3B and autophagosomes, which mediate mitochondrial clearance as a critical step in erythropoiesis. These results illustrate a novel mechanism by which a master regulator of development establishes a genetic network to instigate cell-type-specific autophagy.
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Autofagia , Factor de Transcripción GATA1/genética , Regulación de la Expresión Génica , Hematopoyesis , Animales , Células Cultivadas , Cromatina/genética , Eritroblastos/citología , Eritroblastos/metabolismo , Factor de Transcripción GATA1/metabolismo , Estudio de Asociación del Genoma Completo , Humanos , Lisosomas/metabolismo , RatonesRESUMEN
We have previously shown that human embryonic stem cells can be differentiated into embryonic and fetal type of red blood cells that sequentially express three types of hemoglobins recapitulating early human erythropoiesis. We report here that we have produced iPS from three somatic cell types: adult skin fibroblasts as well as embryonic and fetal mesenchymal stem cells. We show that regardless of the age of the donor cells, the iPS produced are fully reprogrammed into a pluripotent state that is undistinguishable from that of hESCs by low and high-throughput expression and detailed analysis of globin expression patterns by HPLC. This suggests that reprogramming with the four original Yamanaka pluripotency factors leads to complete erasure of all functionally important epigenetic marks associated with erythroid differentiation regardless of the age or the tissue type of the donor cells, at least as detected in these assays. The ability to produce large number of erythroid cells with embryonic and fetal-like characteristics is likely to have many translational applications.
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Técnicas de Cultivo de Célula/métodos , Embrión de Mamíferos/citología , Eritrocitos/citología , Feto/citología , Células Madre Pluripotentes Inducidas/citología , Adulto , Diferenciación Celular/genética , Línea Celular , Eritrocitos/metabolismo , Regulación de la Expresión Génica , Hematopoyesis/genética , Humanos , Células Madre Pluripotentes Inducidas/metabolismoRESUMEN
Interactions between maternal natural killer lymphocytes (NKs) and fetal tissues are important in mediating maternal-fetal tolerance. We therefore investigated the interactions of NKs to placenta-derived multipotent cells (PDMCs) isolated from the term human placenta. PDMCs have similar cell surface marker expression as bone marrow mesenchymal stem cells (BMMSCs) and additionally express human embryonic stem cell markers SSEA-4 and CD-9. Differentiation into the tri-mesodermal lineages of osteoblastic, adipocytic, and chondrogenic phenotypes can be readily achieved under the appropriate conditions. We found that PDMCs are more resistant to NK-mediated lysis than the major histocompatibility complex (MHC) class-I null target cell K562, and can suppress NK secretion of interferon-γ (IFN-γ). Moreover, as third-party cells, PDMCs suppressed the cytotoxic effects of cytokine-stimulated NKs on K562. Pretreatment of PDMCs with IFN-γ, a proinflammatory cytokine, surprisingly enhanced such immunosuppressive effects. Cell-cell contact between NKs and PDMCs is required for suppressive effects, which are partially mediated by slight upregulation of the NK inhibitory receptor killer inhibitory receptor and downregulation of the activating receptor NKp30. Moreover, enhancement of PDMC suppressive effects is also mediated by IFN-γ-induced surface expression of HLA-G--an immunomodulatory nonclassical MHC class I molecule--on PDMCs, as seen by partial reversibility with HLA-G neutralizing antibodies. With its broad immunosuppressive properties, PDMCs may represent a potential cell source for therapeutic use.