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1.
Nat Immunol ; 15(1): 98-108, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24292363

RESUMO

Dendritic cells (DCs) that orchestrate mucosal immunity have been studied in mice. Here we characterized human gut DC populations and defined their relationship to previously studied human and mouse DCs. CD103(+)Sirpα(-) DCs were related to human blood CD141(+) DCs and to mouse intestinal CD103(+)CD11b(-) DCs and expressed markers of cross-presenting DCs. CD103(+)Sirpα(+) DCs aligned with human blood CD1c(+) DCs and mouse intestinal CD103(+)CD11b(+) DCs and supported the induction of regulatory T cells. Both CD103(+) DC subsets induced the TH17 subset of helper T cells, while CD103(-)Sirpα(+) DCs induced the TH1 subset of helper T cells. Comparative analysis of transcriptomes revealed conserved transcriptional programs among CD103(+) DC subsets and identified a selective role for the transcriptional repressors Bcl-6 and Blimp-1 in the specification of CD103(+)CD11b(-) DCs and intestinal CD103(+)CD11b(+) DCs, respectively. Our results highlight evolutionarily conserved and divergent programming of intestinal DCs.


Assuntos
Diferenciação Celular/imunologia , Células Dendríticas/imunologia , Mucosa Intestinal/imunologia , Transcriptoma/imunologia , Animais , Antígenos CD/imunologia , Antígenos CD/metabolismo , Antígenos CD1/imunologia , Antígenos CD1/metabolismo , Antígeno CD11b/imunologia , Antígeno CD11b/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Análise por Conglomerados , Apresentação Cruzada/genética , Apresentação Cruzada/imunologia , Células Dendríticas/metabolismo , Citometria de Fluxo , Glicoproteínas/imunologia , Glicoproteínas/metabolismo , Humanos , Cadeias alfa de Integrinas/imunologia , Cadeias alfa de Integrinas/metabolismo , Integrinas/genética , Integrinas/imunologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Microscopia Confocal , Análise de Sequência com Séries de Oligonucleotídeos , Receptores de Quimiocinas/genética , Receptores de Quimiocinas/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Células Th17/imunologia , Células Th17/metabolismo , Transcriptoma/genética
2.
Immunity ; 47(1): 183-198.e6, 2017 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-28723550

RESUMO

Tissue macrophages arise during embryogenesis from yolk-sac (YS) progenitors that give rise to primitive YS macrophages. Until recently, it has been impossible to isolate or derive sufficient numbers of YS-derived macrophages for further study, but data now suggest that induced pluripotent stem cells (iPSCs) can be driven to undergo a process reminiscent of YS-hematopoiesis in vitro. We asked whether iPSC-derived primitive macrophages (iMacs) can terminally differentiate into specialized macrophages with the help of growth factors and organ-specific cues. Co-culturing human or murine iMacs with iPSC-derived neurons promoted differentiation into microglia-like cells in vitro. Furthermore, murine iMacs differentiated in vivo into microglia after injection into the brain and into functional alveolar macrophages after engraftment in the lung. Finally, iPSCs from a patient with familial Mediterranean fever differentiated into iMacs with pro-inflammatory characteristics, mimicking the disease phenotype. Altogether, iMacs constitute a source of tissue-resident macrophage precursors that can be used for biological, pathophysiological, and therapeutic studies.


Assuntos
Técnicas de Cultura de Células/métodos , Hematopoese , Macrófagos/fisiologia , Neurônios/fisiologia , Células-Tronco Pluripotentes/fisiologia , Animais , Diferenciação Celular , Células Cultivadas , Embrião de Mamíferos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurogênese
4.
PLoS Genet ; 10(7): e1004471, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25010009

RESUMO

Genome maintenance in germ cells is critical for fertility and the stable propagation of species. While mechanisms of meiotic DNA repair and chromosome behavior are well-characterized, the same is not true for primordial germ cells (PGCs), which arise and propagate during very early stages of mammalian development. Fanconi anemia (FA), a genomic instability syndrome that includes hypogonadism and testicular failure phenotypes, is caused by mutations in genes encoding a complex of proteins involved in repair of DNA lesions associated with DNA replication. The signaling mechanisms underlying hypogonadism and testicular failure in FA patients or mouse models are unknown. We conducted genetic studies to show that hypogonadism of Fancm mutant mice is a result of reduced proliferation, but not apoptosis, of PGCs, resulting in reduced germ cells in neonates of both sexes. Progressive loss of germ cells in adult males also occurs, overlaid with an elevated level of meiotic DNA damage. Genetic studies indicated that ATM-p53-p21 signaling is partially responsible for the germ cell deficiency.


Assuntos
Anemia de Fanconi/genética , Proteína Supressora de Tumor p53/biossíntese , Proteínas rho de Ligação ao GTP/biossíntese , Animais , Apoptose/genética , Proteínas Mutadas de Ataxia Telangiectasia/biossíntese , Proteínas Mutadas de Ataxia Telangiectasia/genética , Reparo do DNA/genética , Replicação do DNA/genética , Anemia de Fanconi/patologia , Instabilidade Genômica , Células Germinativas/metabolismo , Células Germinativas/patologia , Humanos , Hipogonadismo/genética , Hipogonadismo/patologia , Camundongos , Mutação , Transdução de Sinais/genética , Proteína Supressora de Tumor p53/genética , Proteínas rho de Ligação ao GTP/genética
5.
Stem Cell Res ; 40: 101533, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31450191

RESUMO

Rett syndrome (RTT) is a childhood neurodevelopmental disorder caused by mutations in MECP2. To study the molecular mechanisms underlying RTT, four sublines of H1 hESCs were generated, carrying a hemizygous knockout or mutant allele of MECP2. Exons 3 and 4 of MECP2 were targeted using the CRISPR/Cas9 nuclease system.


Assuntos
Células-Tronco Embrionárias/citologia , Edição de Genes , Proteína 2 de Ligação a Metil-CpG/genética , Sistemas CRISPR-Cas/genética , Diferenciação Celular , Linhagem Celular , Células-Tronco Embrionárias/metabolismo , Éxons , Humanos , Cariótipo , Síndrome de Rett/genética , Síndrome de Rett/patologia
6.
Cell Rep ; 26(9): 2494-2508.e7, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30811996

RESUMO

In Huntington disease (HD), the analysis of tissue-specific CAG repeat length effects has been challenging, given the difficulty in obtaining relevant patient tissues with a broad range of CAG repeat lengths. We used genome editing to generate an allelic panel of isogenic HD (IsoHD) human embryonic stem cell (hESC) lines carrying varying CAG repeat lengths in the first exon of HTT. Functional analyses in differentiated neural cells revealed CAG repeat length-related abnormalities in mitochondrial respiration and oxidative stress and enhanced susceptibility to DNA damage. To explore tissue-specific effects in HD, we differentiated the IsoHD panel into neural progenitor cells, neurons, hepatocytes, and muscle cells. Transcriptomic and proteomic analyses of the resultant cell types identified CAG repeat length-dependent and cell-type-specific molecular phenotypes. We anticipate that the IsoHD panel and transcriptomic and proteomic data will serve as a versatile, open-access platform to dissect the molecular factors contributing to HD pathogenesis.


Assuntos
Células-Tronco Embrionárias/citologia , Proteína Huntingtina/genética , Doença de Huntington/genética , Repetições de Trinucleotídeos , Alelos , Diferenciação Celular , Linhagem Celular , Sistema Nervoso Central/citologia , Dano ao DNA , Perfilação da Expressão Gênica , Hepatócitos/metabolismo , Humanos , Fibras Musculares Esqueléticas/metabolismo , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Células-Tronco Pluripotentes/citologia , Proteômica
7.
Elife ; 72018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-29784083

RESUMO

Calcium/calmodulin-dependent protein kinase II (CAMK2) plays fundamental roles in synaptic plasticity that underlies learning and memory. Here, we describe a new recessive neurodevelopmental syndrome with global developmental delay, seizures and intellectual disability. Using linkage analysis and exome sequencing, we found that this disease maps to chromosome 5q31.1-q34 and is caused by a biallelic germline mutation in CAMK2A. The missense mutation, p.His477Tyr is located in the CAMK2A association domain that is critical for its function and localization. Biochemically, the p.His477Tyr mutant is defective in self-oligomerization and unable to assemble into the multimeric holoenzyme.In vivo, CAMK2AH477Y failed to rescue neuronal defects in C. elegans lacking unc-43, the ortholog of human CAMK2A. In vitro, neurons derived from patient iPSCs displayed profound synaptic defects. Together, our data demonstrate that a recessive germline mutation in CAMK2A leads to neurodevelopmental defects in humans and suggest that dysfunctional CAMK2 paralogs may contribute to other neurological disorders.


Assuntos
Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/genética , Deficiências do Desenvolvimento/genética , Homozigoto , Deficiência Intelectual/genética , Mutação com Perda de Função , Convulsões/genética , Cromossomos Humanos Par 5 , Consanguinidade , Saúde da Família , Ligação Genética , Humanos , Jordânia , Mutação de Sentido Incorreto , Análise de Sequência de DNA
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