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BACKGROUND & AIMS: The ratio of liver size to body weight (hepatostat) is tightly controlled, but little is known about how the physiologic functions of the liver help determine its size. Livers of mice repopulated with human hepatocytes (humanized livers) grow to larger than normal; the human hepatocytes do not recognize the fibroblast growth factor (FGF)-15 produced by mouse intestine. This results in up-regulation of bile acid synthesis in the human hepatocytes and enlargement of the bile acid pool. We investigated whether abnormal bile acid signaling affects the hepatostat in mice. METHODS: We crossed Fah(-/-), Rag2(-/-), Il2r(-/-) mice with nonobese diabetic mice to create FRGN mice, whose livers can be fully repopulated with human hepatocytes. We inserted the gene for human FGF19 (ortholog to mouse Fgf15), including regulatory sequences, into the FRGN mice to create FRGN19(+) mice. Livers of FRGN19(+) mice and their FRGN littermates were fully repopulated with human hepatocytes. Liver tissues were collected and bile acid pool sizes and RNA sequences were analyzed and compared with those of mice without humanized livers (controls). RESULTS: Livers were larger in FRGN mice with humanized livers (13% of body weight), compared with control FRGN mice; they also had much larger bile acid pools and aberrant bile acid signaling. Livers from FRGN19(+) normalized to 7.8% of body weight, and their bile acid pool and signaling more closely resembled that of control FRGN19(+) mice. RNA sequence analysis showed activation of the Hippo pathway, and immunohistochemical and transcription analyses revealed increased hepatocyte proliferation, but not apoptosis, in the enlarged humanized livers of FRGN mice. Cell sorting experiments showed that although healthy human liver does not produce FGF19, nonparenchymal cells from cholestatic livers produce FGF19. CONCLUSIONS: In mice with humanized livers, expression of an FGF19 transgene corrects bile acid signaling defects, resulting in normalization of bile acid synthesis, the bile acid pool, and liver size. These findings indicate that liver size is, in part, regulated by the size of the bile acid pool that the liver must circulate.
Assuntos
Proliferação de Células , Fatores de Crescimento de Fibroblastos/biossíntese , Fatores de Crescimento de Fibroblastos/metabolismo , Hepatócitos/transplante , Fígado/cirurgia , Transdução de Sinais , Animais , Ácidos e Sais Biliares/metabolismo , Proteínas de Ligação a DNA/deficiência , Proteínas de Ligação a DNA/genética , Fatores de Crescimento de Fibroblastos/genética , Regulação da Expressão Gênica , Hepatócitos/metabolismo , Xenoenxertos , Humanos , Hidrolases/deficiência , Hidrolases/genética , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Camundongos Knockout , Camundongos Transgênicos , Tamanho do Órgão , Receptores de Interleucina-2/deficiência , Receptores de Interleucina-2/genética , Fatores de Tempo , Transcrição GênicaRESUMO
UNLABELLED: Proliferating ducts, termed "oval cells," have long been thought to be bipotential, that is, produce both biliary ducts and hepatocytes during chronic liver injury. The precursor to oval cells is considered to be a facultative liver stem cell (LSC). Recent lineage tracing experiments indicated that the LSC is SRY-related HMG box transcription factor 9 positive (Sox9(+) ) and can replace the bulk of hepatocyte mass in several settings. However, no clonal relationship between Sox9(+) cells and the two epithelial liver lineages was established. We labeled Sox9(+) mouse liver cells at low density with a multicolor fluorescent confetti reporter. Organoid formation validated the progenitor activity of the labeled population. Sox9(+) cells were traced in multiple oval cell injury models using both histology and fluorescence-activated cell sorting. Surprisingly, only rare clones containing both hepatocytes and oval cells were found in any experiment. Quantitative analysis showed that Sox9(+) cells contributed only minimally (<1%) to the hepatocyte pool, even in classic oval cell injury models. In contrast, clonally marked mature hepatocytes demonstrated the ability to self-renew in all classic mouse oval cell activation injuries. A hepatocyte chimera model to trace hepatocytes and nonparenchymal cells also demonstrated the prevalence of hepatocyte-driven regeneration in mouse oval cell injury models. CONCLUSION: Sox9(+) ductal progenitor cells give rise to clonal oval cell proliferation and bipotential organoids, but rarely produce hepatocytes in vivo. Hepatocytes themselves are the predominant source of new parenchyma cells in prototypical mouse models of oval cell activation.
Assuntos
Doença Hepática Terminal/genética , Doença Hepática Terminal/patologia , Hepatócitos/citologia , Fatores de Transcrição SOX9/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo , Animais , Sistema Biliar/citologia , Biomarcadores/metabolismo , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Células Cultivadas , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Doença Hepática Induzida por Substâncias e Drogas/fisiopatologia , Quimera , Deficiência de Colina/genética , Deficiência de Colina/patologia , Deficiência de Colina/fisiopatologia , Modelos Animais de Doenças , Doença Hepática Terminal/fisiopatologia , Feminino , Ducto Hepático Comum/citologia , Hepatócitos/fisiologia , Hepatócitos/transplante , Fígado/citologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fatores de Transcrição SOX9/genéticaAssuntos
Infecções por Coronavirus , Máscaras , Pandemias , Pneumonia Viral , Betacoronavirus , COVID-19 , Pessoal de Saúde , Humanos , SARS-CoV-2RESUMO
Introduction: During a pandemic, when the supply of N95 filtering facepiece respirators (FFRs) is limited, health care workers may reuse N95 FFRs. Room temperature storage of N95 FFRs-waiting before reuse-could be a simple low-cost method to reduce severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) bioburden in such a situation. The U.S. Centers for Disease Control and Prevention specify this as a strategy for reducing self-contamination risk during a time of N95 FFR shortage. Objective: To review the literature on persistence of SARS-CoV-2 on surfaces to assess room temperature waiting times for bioburden reduction on N95 FFRs. Methods: The literature was searched for studies evaluating room temperature persistence of SARS-CoV-2. A 3-log decay time was extracted from published data for quantitative comparison between different studies. Studies using surgical masks and non-peer-reviewed studies that include N95 FFRs were used to draw conclusions. Key Findings: Experimental and analytical choices vary between studies and impact the estimated 3-log decay time. There is not a clear understanding of which material properties are significant. There are no peer-reviewed studies of virus persistence on an N95 FFR. Discussion and Conclusions: SARS-COV-2 inactivation occurs spontaneously at room temperature. The precise timing depends on factors including humidity, temperature, and surface material. In reviewed studies, a 7-day waiting period encompasses the 3-log reduction in infectious titer of SARS-COV-2 on specific N95 FFRs and surgical masks. Owing to variations between studies and among N95 FFR materials and room temperature conditions, it is impossible to extrapolate from these limited data to assign a precise 3-log decay time for all used N95 FFRs.
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Most cells in the liver are polyploid, but the functional role of polyploidy is unknown. Polyploidization occurs through cytokinesis failure and endoreduplication around the time of weaning. To interrogate polyploidy while avoiding irreversible manipulations of essential cell-cycle genes, we developed orthogonal mouse models to transiently and potently alter liver ploidy. Premature weaning, as well as knockdown of E2f8 or Anln, allowed us to toggle between diploid and polyploid states. While there was no detectable impact of ploidy alterations on liver function, metabolism, or regeneration, mice with more polyploid hepatocytes suppressed tumorigenesis and mice with more diploid hepatocytes accelerated tumorigenesis in mutagen- and high-fat-induced models. Mechanistically, the diploid state was more susceptible to Cas9-mediated tumor-suppressor loss but was similarly susceptible to MYC oncogene activation, indicating that polyploidy differentially protected the liver from distinct genomic aberrations. This suggests that polyploidy evolved in part to prevent malignant outcomes of liver injury.
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Transformação Celular Neoplásica/patologia , Neoplasias Hepáticas Experimentais/patologia , Regeneração Hepática/fisiologia , Fígado/patologia , Proteínas dos Microfilamentos/fisiologia , Poliploidia , Proteínas Repressoras/fisiologia , Animais , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Citocinese/fisiologia , Feminino , Hepatócitos/metabolismo , Hepatócitos/patologia , Fígado/metabolismo , Neoplasias Hepáticas Experimentais/genética , Neoplasias Hepáticas Experimentais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos KnockoutRESUMO
Activation of intracellular mitogenic signal transduction pathways driven by the ErbB family of receptor tyrosine kinases has been implicated in the development and/or progression of a variety of cancers. Studies on ErbB receptors in osteosarcoma have focused on HER-2 and have produced conflicting results with few studies evaluating the expression of the epidermal growth factor receptor (EGFR). In this study, we determined the level of expression of EGFR and the mutational status of the EGFR receptor in a subset of osteosarcoma tumor samples as well as in a series of established bone tumor-derived cell lines. EGFR protein expression was detected in the form of strong membranous staining by immunohistochemistry in 21 (57%) of 37 cases analyzed. Six of 12 (50%) osteosarcoma cell lines revealed moderate to high expression levels of EGFR. Two somatic alterations (E829E and R831C) were identified in the cytoplasmic domain of the EGFR gene in 1 of 10 tumor samples. The significance of these findings for the pathobiology of osteosarcomas will be investigated further.
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Neoplasias Ósseas/metabolismo , DNA de Neoplasias/análise , Receptores ErbB/metabolismo , Osteossarcoma/metabolismo , Adolescente , Adulto , Western Blotting , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Criança , Análise Mutacional de DNA , Receptores ErbB/química , Receptores ErbB/genética , Feminino , Humanos , Imuno-Histoquímica , Masculino , Osteossarcoma/genética , Osteossarcoma/secundário , Reação em Cadeia da Polimerase , Estrutura Terciária de ProteínaRESUMO
Human pancreatic islets of Langerhans contain five distinct endocrine cell types, each producing a characteristic hormone. The dysfunction or loss of the insulin-producing ß cells causes diabetes mellitus, a disease that harms millions. Until now, ß cells were generally regarded as a single, homogenous cell population. Here we identify four antigenically distinct subtypes of human ß cells, which we refer to as ß1-4, and which are distinguished by differential expression of ST8SIA1 and CD9. These subpopulations are always present in normal adult islets and have diverse gene expression profiles and distinct basal and glucose-stimulated insulin secretion. Importantly, the ß cell subtype distribution is profoundly altered in type 2 diabetes. These data suggest that this antigenically defined ß cell heterogeneity is functionally and likely medically relevant.
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Diabetes Mellitus Tipo 2/metabolismo , Células Secretoras de Insulina/citologia , Sialiltransferases/metabolismo , Tetraspanina 29/metabolismo , Adulto , Idoso , Feminino , Citometria de Fluxo , Hemoglobinas Glicadas/metabolismo , Humanos , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Adult liver progenitor cells are biliary-like epithelial cells that emerge only under injury conditions in the periportal region of the liver. They exhibit phenotypes of both hepatocytes and bile ducts. However, their origin and their significance to injury repair remain unclear. Here, we used a chimeric lineage tracing system to demonstrate that hepatocytes contribute to the progenitor pool. RNA-sequencing, ultrastructural analysis, and in vitro progenitor assays revealed that hepatocyte-derived progenitors were distinct from their biliary-derived counterparts. In vivo lineage tracing and serial transplantation assays showed that hepatocyte-derived proliferative ducts retained a memory of their origin and differentiated back into hepatocytes upon cessation of injury. Similarly, human hepatocytes in chimeric mice also gave rise to biliary progenitors in vivo. We conclude that human and mouse hepatocytes can undergo reversible ductal metaplasia in response to injury, expand as ducts, and subsequently contribute to restoration of the hepatocyte mass.