RESUMEN
Aldehyde dehydrogenase 1A1 (ALDH1A1), a retinoic acid (RA) synthase, is selectively expressed by the nigrostriatal dopaminergic (nDA) neurons that preferentially degenerate in Parkinson's disease (PD). ALDH1A1-positive axons mainly project to the dorsal striatum. However, whether ALDH1A1 and its products regulate the activity of postsynaptic striatal neurons is unclear. Here we show that µ-type opioid receptor (MOR1) levels were severely decreased in the dorsal striatum of postnatal and adult Aldh1a1 knockout mice, whereas dietary supplement of RA restores its expression. Furthermore, RA treatment also upregulates striatal MOR1 levels and signaling and alleviates L-DOPA-induced dyskinetic movements in pituitary homeobox 3 (Pitx3)-deficient mice that lack of ALDH1A1-expressing nDA neurons. Therefore, our findings demonstrate that ALDH1A1-synthesized RA is required for postsynaptic MOR1 expression in the postnatal and adult dorsal striatum, supporting potential therapeutic benefits of RA supplementation in moderating L-DOPA-induced dyskinesia.
Asunto(s)
Familia de Aldehído Deshidrogenasa 1/fisiología , Cuerpo Estriado/efectos de los fármacos , Neuronas Dopaminérgicas/patología , Discinesias/prevención & control , Proteínas de Homeodominio/fisiología , Receptores Opioides mu/metabolismo , Retinal-Deshidrogenasa/fisiología , Factores de Transcripción/fisiología , Tretinoina/farmacología , Animales , Cuerpo Estriado/patología , Dopamina/metabolismo , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/metabolismo , Discinesias/etiología , Discinesias/metabolismo , Discinesias/patología , Femenino , Masculino , Ratones , Ratones Noqueados , Receptores Opioides mu/genéticaRESUMEN
BACKGROUND: CD44 and aldehyde dehydrogenase 1 (ALDH1) has been reputed to be cancer stem cell (CSC) markers in breast cancer. Yet, the clinicopathologic and prognostic significance of these markers remain unclear. In this study, we have investigated the expression of these markers and their relation with conventional clinicopathologic tumor characteristic including molecular subtype. METHODS: CD44 and ALDH1 expression were investigated by immunohistochemistry in a series of 157 formalin-fixed paraffin-embedded breast cancer tissues. RESULTS: Overall, CD44 and ALDH1 are, respectively, detected in 33% (52 of 157) and 7% (10 of 157) of breast cancer cases. We also observed that CD44 expression was associated with histological grade (p = 0.005). For ALDH1, we found that its expression is more frequent with elderly women (> 50 years, p = 0.03). The investigation of relationship between the stem cell phenotype and breast cancer molecular subtype, revealed that CD44 and ALDH1 expression was more frequent in basal-like tumors (p = 0.005). Among the two cancer stem cell markers tested, ALDH1 showed a strong association with the basal marker EGFR (p = 0.05). CONCLUSIONS: These findings suggest that CD44 and ALDH1 play a role in the clinical behavior in breast cancer and might be interesting biomarkers and therapeutic targets.
Asunto(s)
Neoplasias de la Mama/patología , Receptores de Hialuranos/análisis , Isoenzimas/análisis , Células Madre Neoplásicas/química , Retinal-Deshidrogenasa/análisis , Adulto , Anciano , Anciano de 80 o más Años , Familia de Aldehído Deshidrogenasa 1 , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/química , Femenino , Humanos , Receptores de Hialuranos/fisiología , Isoenzimas/fisiología , Persona de Mediana Edad , Retinal-Deshidrogenasa/fisiologíaRESUMEN
All-trans-retinoic acid (RA) inhibits adipogenesis in established preadipocyte cell lines. Dosing pharmacological amounts of RA reduces weight gain in mice fed a high-fat diet, i.e. counteracts diet-induced obesity (DIO). The aldehyde dehydrogenase Raldh1 (Aldh1a1) functions as one of three enzymes that converts the retinol metabolite retinal into RA, and one of many proteins that contribute to RA homeostasis. Female Raldh1-ablated mice resist DIO. This phenotype contrasts with ablations of other enzymes and binding-proteins that maintain RA homeostasis, which gain adiposity. The phenotype observed prompted the conclusion that loss of Raldh1 causes an increase in adipose tissue retinal, and therefore, retinal functions independently of RA to prevent DIO. A second deduction proposed that low nM concentrations of RA stimulate adipogenesis, in contrast to higher concentrations. Using peer-reviewed LC/MS/MS assays developed and validated for quantifying tissue RA and retinal, we show that endogenous retinal and RA concentrations in adipose tissues from Raldh1-null mice do not correlate with the phenotype. Moreover, male Raldh1-null mice resist weight gain regardless of dietary fat content. Resistance to weight gain occurs during adolescence in both sexes. We show that RA concentrations as low as 1 nM, i.e. in the sub-physiological range, impair adipogenesis of embryonic fibroblasts from wild-type mice. Embryonic fibroblasts from Raldh1-null mice resist differentiating into adipocytes, but retain ability to generate RA. These fibroblasts remain sensitive to an RA receptor pan-agonist, and are not affected by an RA receptor pan-antagonist. Thus, the data do not support the hypothesis that retinal itself represses weight gain and adipogenesis independently of RA. Instead, the data indicate that Raldh1 functions as a retinal and atRA-independent promoter of adiposity during adolescence, and enhances adiposity through pre-adipocyte cell autonomous actions.
Asunto(s)
Adiposidad , Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Retinaldehído/metabolismo , Transducción de Señal , Familia de Aldehído Deshidrogenasa 1 , Animales , RatonesRESUMEN
OBJECTIVES: To explore the relationship between aldehye dehydrogenase-1 (ALDH-1) and biological characteristics of the stable ALDH-1 knock-down Hela cell lines. MATERIALS AND METHODS: Transfected Hela cells with lentiviral vector were utilized and puromycin was used to screen and achieve the stable cell lines. The interference efficiency was calculated by performing qRT-PCR to detect the expression of ALDH-1 of three groups including the interfering group, the negative control (NC) group, and the normal Hela group. CCK-8 assays were used to detect the OD value of each group. The coloning formation rate of each group was detected by colony formation assay. cell cycle distribution and apoptosis of each group were also detected by employing cell cycle and apoptosis experiments. Results: The stable ALDH-1 knock-down Hela cell lines were successfully obtained after two weeks' screening; compared with the NC and Hela group, the ALDH-1 expression level of interfering group was 1.05 ± 0.10 (both p < 0.05), whose silencing efficiency was 80.59%. CCK-8 assays verified that the mean OD value of interfering group was lower than that of NC and Hela group. Additionally, colony formation assays showed that the coloning efficiency of interfering group was lower than that of NC and Hela group. Cell cycle experiments proved that the proportion of G0/G1 phase of interfering group was higher than that of the other two groups, while the proportion of S phase was lower. Cell apoptosis assays indicated that the apoptosis rate of interfering group was the highest. CONCLUSIONS: Constructing stable interfering Hela cell lines with lentiviral vectors was successful and worthy of promotion. ALDH-1 plays an important role in promoting cell growth and proliferation, maintaining cell cycle and inhibiting Hela cell apoptosis.
Asunto(s)
Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Apoptosis , Ciclo Celular , Femenino , Células HeLa , Humanos , Isoenzimas/genética , Retinal-Deshidrogenasa/genética , TransfecciónRESUMEN
Pancreatic cancer (PC) remains a highly lethal malignancy due to its unusual chemoresistance and high aggressiveness. A subpopulation of pancreatic tumor cells, known as cancer stem cells (CSCs), is considered responsible not only for tumor-maintenance, but also for its widespread metastasis and therapeutic failure. Here we investigated the role of p-21 activated kinase 4 (PAK4) in driving PC stemness properties. Our data demonstrate that triple-positive (CD24(+)/CD44(+)/EpCAM(+)) subpopulation of pancreatic CSCs exhibits greater level of PAK4 as compared to triple-negative (CD24(-)/CD44(-)/EpCAM(-)) cells. Moreover, PAK4 silencing in PC cells leads to diminished fraction of CD24, CD44, and EpCAM positive cells. Furthermore, we show that PAK4-silenced PC cells exhibit decreased sphere-forming ability and increased chemosensitivity to gemcitabine toxicity. PAK4 expression is also associated with enhanced levels of stemness-associated transcription factors (Oct4/Nanog/Sox2 and KLF4). Furthermore, our data show decreased nuclear accumulation and transcriptional activity of STAT3 in PAK4-silenced PC cells and restitution of its activity leads to restoration of stem cell phenotypes. Together, our findings deliver first experimental evidence for the involvement of PAK4 in PC stemness and support its clinical utility as a novel therapeutic target in PC.
Asunto(s)
Células Madre Neoplásicas/química , Neoplasias Pancreáticas/patología , Factor de Transcripción STAT3/fisiología , Transducción de Señal/fisiología , Quinasas p21 Activadas/fisiología , Familia de Aldehído Deshidrogenasa 1 , Antígenos de Neoplasias/análisis , Antígeno CD24/análisis , Moléculas de Adhesión Celular/análisis , Línea Celular Tumoral , Molécula de Adhesión Celular Epitelial , Humanos , Receptores de Hialuranos/análisis , Isoenzimas/fisiología , Factor 4 Similar a Kruppel , Fenotipo , Retinal-Deshidrogenasa/fisiologíaRESUMEN
Despite the introduction of 13-cis-retinoic acid (13-cis-RA) into the current chemotherapy, more than half of high-risk neuroblastoma patients have experienced tumor relapses driven by chemoresistant cancer stem cells (CSCs) that can be isolated by their ability to grow as spheres. Although aldehyde dehydrogenase (ALDH) has been used to characterize CSCs in certain cancers, ALDH remains elusive in neuroblastoma. In the present study, we determined ALDH activity and expression of its 19 isoforms in spheres and parental cells of neuroblastoma. ALDH activity and several ALDH isoforms were consistently induced in spheres of different neuroblastoma cells. While ALDH1A2, ALDH1L1 and ALDH3B2 expression was consistently induced in spheres and associated with the sphere and colony formation, only ALDH1A2 expression was significantly correlated with the poor prognosis of neuroblastoma patients. ALDH1A2 expression was further associated with the growth and undifferentiation of neuroblastoma xenografts and the resistance of neuroblastoma cells to 13-cis-RA. These results suggest that ALDH1A2 is involved in the regulation of CSC properties in neuroblastoma.
Asunto(s)
Células Madre Neoplásicas/fisiología , Neuroblastoma/patología , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Animales , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Células Cultivadas , Resistencia a Antineoplásicos/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Isotretinoína/farmacología , Isotretinoína/uso terapéutico , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Células Madre Neoplásicas/patología , Neuroblastoma/genética , ARN Interferente Pequeño/farmacología , Retinal-Deshidrogenasa/antagonistas & inhibidores , Esferoides Celulares/efectos de los fármacos , Esferoides Celulares/metabolismo , Esferoides Celulares/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The authors present a review of the literature to illustrate the progress of ALDH-1 in gynecological malignancies.
Asunto(s)
Neoplasias de los Genitales Femeninos/enzimología , Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Neoplasias Endometriales/enzimología , Femenino , Humanos , Isoenzimas/análisis , Neoplasias Ováricas/enzimología , Pronóstico , Retinal-Deshidrogenasa/análisis , Neoplasias del Cuello Uterino/enzimologíaRESUMEN
UNLABELLED: Aldehyde dehydrogenase 1 (ALDH1) has been regarded as a breast cancer stem cell marker. Several studies have reported that ALDH1 expression is associated with poor prognosis in breast cancer. We aimed, therefore, to determine the prognostic value of ALDH1 expression and its association with several biomarkers in breast cancer tissue using immunohistochemistry. Furthermore, we investigated the characteristics of and differences between cellular and stromal expression of ALDH1. We performed tissue microarray (TMA) analysis of 425 breast cancer tissue samples collected during surgery. Immunohistochemical staining was then performed to measure the expression of ALDH1 and other breast cancer biomarkers. Statistical analysis of the relationship between ALDH1 expression and clinicopathologic characteristics was performed for 390 TMA samples. We found that ALDH1 was expressed in 71 cases (18.2%) in the tumor cells and/or stroma. Of these cases, 38 (9.7%) showed ALDH1 expression in tumor cells and 38 (9.7%) showed ALDH1 expression in the stroma. ALDH1 expression was significantly associated with markers of a poor prognosis, such as young age, estrogen receptor negativity, progesterone receptor negativity, a high histological grade, and a high Ki-67 index. However, ALDH1 expression was not associated with p53, transforming growth factor-beta, Gli-1, YKL-40, or sonic hedgehog expression status. With regard to the expression site, the clinical characteristics did not differ between cases of cellular expression and those of stromal expression. However, ALDH1 expression in tumor cells was correlated with hormone receptor status, histological grade, molecular subtype, epidermal growth factor receptor expression status, and cytokeratin 5/6 expression status while stromal expression of ALDH1 was only correlated with hormone receptor status. Overall, these findings suggest that ALDH1 expression in tumor tissue is associated with a biologically aggressive phenotype. KEYWORDS: ALDH1, biologically aggressive, breast cancer.
Asunto(s)
Neoplasias de la Mama/patología , Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Adulto , Anciano , Anciano de 80 o más Años , Familia de Aldehído Deshidrogenasa 1 , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/mortalidad , Femenino , Humanos , Isoenzimas/análisis , Persona de Mediana Edad , Receptores de Estrógenos/análisis , Retinal-Deshidrogenasa/análisis , Estudios Retrospectivos , Análisis de Matrices TisularesRESUMEN
The enteric nervous system (ENS) forms from the neural crest-derived precursors that colonize the bowel before differentiating into a network of neurons and glia that control intestinal function. Retinoids are essential for normal ENS development, but the role of retinoic acid (RA) metabolism in development remains incompletely understood. Because RA is produced locally in the tissues where it acts by stimulating RAR and RXR receptors, RA signaling during development is absolutely dependent on the rate of RA synthesis and degradation. RA is produced by three different enzymes called retinaldehyde dehydrogenases (RALDH1, RALDH2 and RALDH3) that are all expressed in the developing bowel. To determine the relative importance of these enzymes for ENS development, we analyzed whole mount preparations of adult (8-12-week old) myenteric and submucosal plexus stained with NADPH diaphorase (neurons and neurites), anti-TuJ1 (neurons and neurites), anti-HuC/HuD (neurons), and anti-S100ß (glia) in an allelic series of mice with mutations in Raldh1, Raldh2, and Raldh3. We found that Raldh1-/-, Raldh2+/-, Raldh3+/- (R1(KO)R2(Het)R3(Het)) mutant mice had a reduced colon myenteric neuron density, reduced colon myenteric neuron to glia ratio, reduced colon submucosal neuron density, and increased colon myenteric fibers per neuron when compared to the wild type (WT; Raldh1WT, Raldh2WT, Raldh3WT) mice. These defects are unlikely to be due to defective ENS precursor migration since R1(KO)R2(Het)R3(KO) mice had increased enteric neuron progenitor migration into the distal colon compared to WT during development. RALDH mutant mice also have reduced contractility in the colon compared to WT mice. These data suggest that RALDH1, RALDH2 and RALDH3 each contribute to ENS development and function.
Asunto(s)
Aldehído Oxidorreductasas/fisiología , Colon/inervación , Sistema Nervioso Entérico/metabolismo , Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Animales , Movimiento Celular , Colon/enzimología , Suplementos Dietéticos , Regulación del Desarrollo de la Expresión Génica , Ratones , Ratones Endogámicos C57BL , Mutación , Neuroglía/citología , Neuronas/metabolismo , FenotipoRESUMEN
The elucidation of drug resistance mechanisms is important in the development of clinical therapies for the treatment of leukemia. To study the drug resistance mechanisms, protein expression profiles of 1-ß-D-arabinofuranosylcytosine (AraC)-sensitive K562 (K562S) cells and AraC-resistant K562 (K562AC) cells were compared using two-dimensional fluorescence difference gel electrophoresis. In a comparison of protein expression profiles, 2073 protein spots were found to be altered, and 15 proteins of them were remarkably altered. These proteins were identified by mass spectrometry. The most differently expressed proteins were aldehyde dehydrogenase 1 family member A2 (ALDH1A2) and vimentin. Both proteins were verified using reverse transcriptase polymerase chain reaction and Western blot analysis. ALDH1A2 protein was found to be effective in AraC resistance. ALDH1A2 knock-down induced sensitivity to AraC treatment in K562AC cells, and ALDH1A2 overexpressed K562S cells acquired the AraC resistance. Furthermore, the findings also suggest that ALDH1A2 expression is increased after the appearance of AraC resistance in clinical cases. These results will be helpful in understanding the mechanism of AraC resistance.
Asunto(s)
Antimetabolitos Antineoplásicos/farmacología , Citarabina/farmacología , Resistencia a Antineoplásicos/genética , Regulación Leucémica de la Expresión Génica , Leucemia/enzimología , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Citarabina/administración & dosificación , Doxorrubicina/farmacología , Electroforesis en Gel Bidimensional/métodos , Inducción Enzimática , Humanos , Idarrubicina/administración & dosificación , Células K562/efectos de los fármacos , Células K562/metabolismo , Leucemia/sangre , Leucemia/tratamiento farmacológico , Leucemia/genética , Leucemia Eritroblástica Aguda/enzimología , Leucemia Eritroblástica Aguda/patología , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Profármacos/farmacología , Interferencia de ARN , ARN Interferente Pequeño/farmacología , Retinal-Deshidrogenasa/biosíntesis , Retinal-Deshidrogenasa/genética , Transfección , Regulación hacia ArribaRESUMEN
Mechanisms for sex- and depot-specific fat formation are unclear. We investigated the role of retinoic acid (RA) production by aldehyde dehydrogenase 1 (Aldh1a1, -a2, and -a3), the major RA-producing enzymes, on sex-specific fat depot formation. Female Aldh1a1(-/-) mice, but not males, were resistant to high-fat (HF) diet-induced visceral adipose formation, whereas subcutaneous fat was reduced similarly in both groups. Sexual dimorphism in visceral fat (VF) was attributable to elevated adipose triglyceride lipase (Atgl) protein expression localized in clusters of multilocular uncoupling protein 1 (Ucp1)-positive cells in female Aldh1a1(-/-) mice compared with males. Estrogen decreased Aldh1a3 expression, limiting conversion of retinaldehyde (Rald) to RA. Rald effectively induced Atgl levels via nongenomic mechanisms, demonstrating indirect regulation by estrogen. Experiments in transgenic mice expressing an RA receptor response element (RARE-lacZ) revealed HF diet-induced RARE activation in VF of females but not males. In humans, stromal cells isolated from VF of obese subjects also expressed higher levels of Aldh1 enzymes compared with lean subjects. Our data suggest that an HF diet mediates VF formation through a sex-specific autocrine Aldh1 switch, in which Rald-mediated lipolysis in Ucp1-positive visceral adipocytes is replaced by RA-mediated lipid accumulation. Our data suggest that Aldh1 is a potential target for sex-specific antiobesity therapy.
Asunto(s)
Adiposidad , Grasa Intraabdominal/metabolismo , Isoenzimas/fisiología , Retinal-Deshidrogenasa/fisiología , Caracteres Sexuales , Células 3T3-L1 , Familia de Aldehído Deshidrogenasa 1 , Animales , Dieta Alta en Grasa , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
Aldehyde dehydrogenase 1 (ALDH1) activity has now been employed successfully as a cancer stem cells (CSCs) marker in various tumors. We wanted to investigate whether ALDH1 can be used as a putative CSCs marker and a powerful prognostic factor in nasopharyngeal carcinoma (NPC). Here, we isolated ALDH1-positive cells from human NPC cell lines (5-8F and CNE2) and found that ALDH1-positive cancer cells grew faster and had higher clone formation efficiency (0.435±0.15; 0.431±0.025 vs. 0.131±0.007; 0.121±0.126), differentiation capability and had higher migration (233.00±5.29; 228.60±9.34 vs. 123.20±7.70 vs. 97.20±6.61) than those of ALDH1-negative cancer cells in vitro. In addition, ALDH1- positive cancer cells formed significantly more tumor spheres. Our in vivo experiments showed that only 5×10(3) ALDH1-positive NPC cells were required to induce tumors. Notably, ALDH1-positive cells are enriched in the side-population (SP) cells, and stem cells-like genes OCT4, BMI1, KLF4 and SOX2 are preferentially expressed in ALDH1-positive cells. Immunohistochemical results showed that the expression of ALDH1 correlated significantly with T classification (P=0.011), N classification (P=0.005), M classification (P=0.002) and clinical stage (P=0.001). Interestingly, ALDH1 expression in the tumor correlated significantly with epithelial-mesenchymal transition (EMT) markers including vimentin expression and loss of E-cadherin (P=0.003 and 0.008, respectively). Furthermore, multivariate analyses showed that ALDH1 expression was an independent prognostic indicator (P=0.032). Taken together, for the first time, we demonstrate that ALDH1 could be a novel stem cells marker and a valuable predictor of poor survival NPC.
Asunto(s)
Isoenzimas/fisiología , Neoplasias Nasofaríngeas/enzimología , Células Madre Neoplásicas/enzimología , Retinal-Deshidrogenasa/fisiología , Adolescente , Adulto , Anciano , Familia de Aldehído Deshidrogenasa 1 , Animales , Biomarcadores , Carcinoma , Línea Celular Tumoral , Transición Epitelial-Mesenquimal , Femenino , Humanos , Isoenzimas/análisis , Factor 4 Similar a Kruppel , Masculino , Ratones , Persona de Mediana Edad , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/mortalidad , Neoplasias Nasofaríngeas/patología , Estadificación de Neoplasias , Retinal-Deshidrogenasa/análisisRESUMEN
HIV-1 preferentially infects activated CD4(+) T cells expressing α4ß7 integrin and conventional vaccination approaches non-selectively induce immune responses including α4ß7(high) CD4(+) T cells, suggesting that current candidate AIDS vaccines may produce more target cells for HIV-1 and paradoxically enhance HIV-1 infection. Thus it remains a challenge to selectively induce robust anti-HIV immunity without the unwanted HIV-1 susceptible α4ß77(high) CD4(+)+ T cells. Here we describe a vaccination strategy that targets ALDH1a2, a retinoic acid producing enzyme in dendritic cells (DCs). Silencing ALDH1a2 in DCs enhanced the maturation and production of proinflammatory cytokines of DCs and promoted Th1/Th2 differentiation while suppressing Treg. ALDH1a2-silenced DCs effectively downregulated the expression of guthoming receptors α4ß77 and CCR9 on activated T and B lymphocytes. Consequently, intranasal immunization of a lentiviral vaccine encoding ALDH1a2 shRNA and HIV-1 gp140 redirected gp140-specific mucosal T cell and antibody responses from the gut to the vaginal tract, while dramatically enhancing systemic gp140-specific immune responses. We further demonstrated that silencing ALDH1a2 in human DCs resulted in downregulation of ß7 expression on activated autologous CD4(+) T cells. Hence this study provides a unique and effective strategy to induce α4ß7(low) anti-HIV immune responses.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Retinal-Deshidrogenasa/fisiología , Vacunación/métodos , Vagina/inmunología , Familia de Aldehído Deshidrogenasa 1 , Animales , Linfocitos B/inmunología , Linfocitos B/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Citocinas/metabolismo , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Femenino , Silenciador del Gen , Infecciones por VIH/metabolismo , Infecciones por VIH/prevención & control , Integrinas/metabolismo , Ratones , Ratones Endogámicos BALB CRESUMEN
Retinaldehyde dehydrogenases (RALDH) catalyze the synthesis of the regulatory factor retinoic acid (RA). Cultured astrocytes express several of the RALDH enzyme family, and it has been assumed that this can be extrapolated to astrocytes in vivo. However, this study finds that few astrocytes in the rodent brain express detectable RALDH enzymes, and only when these cells are grown in culture are these enzymes upregulated. Factors controlling the expression of the RALDHs in cultured astrocytes were explored to determine possible reasons for differences between in vitro versus in vivo expression. Retinoids were found to feedback to suppress several of the RALDHs, and physiological levels of retinoids may be one route by which astrocytic RALDHs are maintained at low levels. In the case of RALDH2, in vivo reduction of vitamin A levels in rats resulted in an increase in astrocyte RALDH2 expression in the hippocampus. Other factors though are likely to control RALDH expression. A shift in astrocytic RALDH subcellular localization is a potential mechanism for regulating RA signaling. Under conditions of vitamin A deficiency, RALDH2 protein moved from the cytoplasm to the nucleus where it may synthesize RA at the site of the nuclear RA receptors. Similarly, in conditions of oxidative stress RALDH1 and RALDH2 moved from the cytoplasm to a predominantly nuclear position. Thus, the RALDHs have been revealed to be dynamic in their expression in astrocytes where they may maintain retinoid homeostasis in the brain.
Asunto(s)
Astrocitos/fisiología , Encéfalo/metabolismo , Retinal-Deshidrogenasa/fisiología , Tretinoina/metabolismo , Familia de Aldehído Deshidrogenasa 1 , Animales , Animales Recién Nacidos , Astrocitos/metabolismo , Células Cultivadas , Humanos , Masculino , Ratones , Ratones Transgénicos , Persona de Mediana Edad , Ratas , Ratas Sprague-Dawley , Retinal-Deshidrogenasa/biosíntesis , Retinal-Deshidrogenasa/genética , Deficiencia de Vitamina A/genética , Deficiencia de Vitamina A/metabolismoRESUMEN
Recent data link vitamin A and its retinoid metabolites to the regulation of adipogenesis, insulin sensitivity, and glucose homeostasis. Retinoid metabolism is tightly controlled by an enzymatic network in which retinaldehyde dehydrogenases (Aldh1-3) are the rate-limiting enzymes that convert retinaldehyde to retinoic acid. Aldh1a1-deficient mice are protected from diet-induced obesity and hence diabetes. Here we investigated whether Aldh1a1 and the retinoid axis regulate hepatic glucose and lipid metabolism independent of adiposity. The impact of Aldh1a1 and the retinoid pathway on glucose homeostasis and lipid metabolism was analyzed in hepatocytes in vitro and in chow-fed, weight-matched Aldh1a1-deficient vs. wild-type (WT) mice in vivo. Aldh1a1-deficient mice displayed significantly decreased fasting glucose concentrations compared with WT controls as a result of attenuated hepatic glucose production. Expression of key gluconeogenic enzymes as well as the activity of Forkhead box O1 was decreased in Aldh1a1-deficient vs. WT livers. In vitro, retinoid or cAMP agonist stimulation markedly induced gluconeogenesis in WT but not Aldh1a1-deficient primary hepatocytes. Aldh1a1 deficiency increased AMP-activated protein kinase α activity, decreased expression of lipogenic targets of AMP-activated protein kinase α and significantly attenuated hepatic triacylglycerol synthesis. In metabolic cage studies, lean Aldh1a1-deficient mice manifested enhanced oxygen consumption and reduced respiratory quotient vs. WT controls, consistent with increased expression of fatty acid oxidation markers in skeletal muscle. Taken together, this work establishes a role for retinoid metabolism in glucose homeostasis in vivo and for Aldh1a1 as a novel determinant of gluconeogenesis and lipid metabolism independent of adiposity.
Asunto(s)
Gluconeogénesis/fisiología , Isoenzimas/genética , Isoenzimas/fisiología , Lípidos/química , Hígado/metabolismo , Retinal-Deshidrogenasa/genética , Retinal-Deshidrogenasa/fisiología , Familia de Aldehído Deshidrogenasa 1 , Animales , Calorimetría/métodos , Cruzamientos Genéticos , Femenino , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead/metabolismo , Glucosa/metabolismo , Hepatocitos/citología , Homeostasis , Metabolismo de los Lípidos , Espectroscopía de Resonancia Magnética , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Consumo de Oxígeno , Isoformas de Proteínas , Triglicéridos/metabolismoRESUMEN
Aldehyde dehydrogenase 1 (ALDH1) has been shown to serve as a marker for cancer-initiating cells (CICs), but little is known about the regulation of the CIC functions of ALDH1+ cancer cells. We isolated ALDH1+ cells from human diffuse-type gastric carcinoma cells and characterized these cells using an Aldefluor assay. ALDH1+ cells constituted 5-8% of the human diffuse-type gastric carcinoma cells, OCUM-2MLN and HSC-39; were more tumourigenic than ALDH1- cells; and were able to self-renew and generate heterogeneous cell populations. Using gene expression microarray analyses, we identified REG4 (regenerating islet-derived family, member 4) as one of the genes up-regulated in ALDH1+ cells, and thus as a novel marker for ALDH1+ tumour cells. Induced expression of REG4 enhanced the colony-forming ability of OCUM-2MLN cells, while knockdown of REG4 inhibited the tumourigenic potential of ALDH1+ cells. We further found that TGF-ß signalling reduces the expression of ALDH1 and REG4, and the size of the ALDH1+ cell population. In human diffuse-type gastric carcinoma tissues, the expression of ALDH1 and REG4 correlated with each other, as assessed by immunohistochemistry, and ALDH1 expression correlated inversely with Smad3 phosphorylation as a measure of TGF-ß signalling. These findings illustrate that, in diffuse-type gastric carcinoma, REG4 is up-regulated in ALDH1+ CICs, and that the increased tumourigenic ability of ALDH1+ cells depends on REG4. Moreover, TGF-ß down-regulates ALDH1 and REG4 expression, which correlates with a reduction in CIC population size and tumourigenicity. Targeting REG4 in ALDH1+ CICs may provide a novel strategy in the treatment of diffuse-type gastric carcinoma.
Asunto(s)
Transformación Celular Neoplásica/patología , Regulación hacia Abajo/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/fisiología , Isoenzimas/fisiología , Lectinas Tipo C/fisiología , Células Madre Neoplásicas/fisiología , Retinal-Deshidrogenasa/fisiología , Neoplasias Gástricas/fisiopatología , Factor de Crecimiento Transformador beta/farmacología , Adenocarcinoma/patología , Adenocarcinoma/fisiopatología , Familia de Aldehído Deshidrogenasa 1 , Animales , Biomarcadores de Tumor/fisiología , Carcinoma de Células en Anillo de Sello/patología , Carcinoma de Células en Anillo de Sello/fisiopatología , Línea Celular Tumoral , Células Cultivadas , Modelos Animales de Enfermedad , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Células Madre Neoplásicas/patología , Proteínas Asociadas a Pancreatitis , Transducción de Señal/fisiología , Neoplasias Gástricas/patología , Factor de Crecimiento Transformador beta/fisiología , Trasplante Heterólogo , Regulación hacia Arriba/fisiologíaRESUMEN
Epithelial-to-mesenchymal transition (EMT) is an important driver of tumor invasion and metastasis, which causes many cancer deaths. Cancer stem cells (CSC) that maintain and initiate tumors have also been implicated in invasion and metastasis, but whether EMT is an important contributor to CSC function is unclear. In this study, we investigated whether a population of CSCs that have undergone EMT (EMT CSCs) exists in squamous cell carcinoma (SCC). We also determined whether a separate population of CSCs that retain epithelial characteristics (non-EMT CSCs) is also present. Our studies revealed that self-renewing CSCs in SCC include two biologically-distinct phenotypes. One phenotype, termed CD44(high)ESA(high), was proliferative and retained epithelial characteristics (non-EMT CSCs), whereas the other phenotype, termed CD44(high)ESA(low), was migratory and had mesenchymal traits characteristic of EMT CSCs. We found that non-EMT and EMT CSCs could switch their epithelial or mesenchymal traits to reconstitute the cellular heterogeneity which was characteristic of CSCs. However, the ability of EMT CSCs to switch to non-EMT character was restricted to cells that were also ALDH1(+), implying that only ALDH1(+) EMT cells had the ability to seed a new epithelial tumor. Taken together, our findings highlight the identification of two distinct CSC phenotypes and suggest a need to define therapeutic targets that can eradicate both of these variants to achieve effective SCC treatment.
Asunto(s)
Carcinoma de Células Escamosas/patología , Transición Epitelial-Mesenquimal/fisiología , Células Madre Neoplásicas/patología , Familia de Aldehído Deshidrogenasa 1 , Animales , Antígenos de Neoplasias/análisis , Moléculas de Adhesión Celular/análisis , Movimiento Celular , Proliferación Celular , Células Clonales/patología , Células Clonales/trasplante , Molécula de Adhesión Celular Epitelial , Humanos , Receptores de Hialuranos/análisis , Isoenzimas/fisiología , Ratones , Ratones Endogámicos NOD , Ratones SCID , Neoplasias de la Boca/patología , Proteínas de Neoplasias/análisis , Proteínas de Neoplasias/fisiología , Células Madre Neoplásicas/trasplante , Fenotipo , Lesiones Precancerosas/patología , Retinal-Deshidrogenasa/fisiología , Enfermedades de la Piel/patología , Neoplasias Cutáneas/patología , Trasplante HeterólogoRESUMEN
The vitamin A metabolite retinoic acid (RA) plays a crucial role in mucosal immune responses. We demonstrate in this study that RA-producing retinaldehyde dehydrogenase (RALDH) enzymes are postnatally induced in mesenteric lymph node (MLN) dendritic cells (DCs) and MLN stromal cells. RALDH enzyme activity in lamina propria-derived CD103(+) MLN-DCs did not depend on TLR signaling. Remarkably, RA itself could directly induce RALDH2 in both DCs and stromal cells in vitro. Furthermore, upon provision of a vitamin A-deficient diet, it was found that RA-mediated signaling was strongly reduced within the small intestines, while RALDH2 mRNA and RALDH enzyme activity in lamina propria DCs and MLN-DCs, as well as RALDH2 mRNA expression in MLN stromal cells, were strongly diminished. Moreover, supply of vitamin A to vitamin A-deficient mice restored RA-mediated signaling in the intestine and RALDH activity in lamina propria-derived CD103(+) MLN-DCs. Our results show that RA-dependent signaling within the intestine is indispensable for RALDH activity in the draining MLN.
Asunto(s)
Aldehído Oxidorreductasas/biosíntesis , Regulación de la Expresión Génica/inmunología , Mucosa Intestinal/enzimología , Intestino Delgado/enzimología , Ganglios Linfáticos/enzimología , Retinal-Deshidrogenasa/biosíntesis , Tretinoina/fisiología , Vitamina A/fisiología , Aldehído Oxidorreductasas/genética , Aldehído Oxidorreductasas/fisiología , Alimentación Animal , Animales , Células Dendríticas/enzimología , Células Dendríticas/inmunología , Células Dendríticas/patología , Mucosa Intestinal/inmunología , Mucosa Intestinal/patología , Intestino Delgado/inmunología , Intestino Delgado/patología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Mesenterio/enzimología , Mesenterio/inmunología , Mesenterio/patología , Ratones , Retinal-Deshidrogenasa/genética , Retinal-Deshidrogenasa/fisiología , Células del Estroma/enzimología , Células del Estroma/inmunología , Células del Estroma/patología , Vitamina A/administración & dosificación , Deficiencia de Vitamina A/enzimología , Deficiencia de Vitamina A/inmunología , Deficiencia de Vitamina A/patologíaRESUMEN
The isomerization of 11-cis retinal to all-trans retinal in photoreceptors is the first step in vision. For photoreceptors to function in constant light, the all-trans retinal must be converted back to 11-cis retinal via the enzymatic steps of the visual cycle. Within this cycle, all-trans retinal is reduced to all-trans retinol in photoreceptors and transported to the retinal pigment epithelium (RPE). In the RPE, all-trans retinol is converted to 11-cis retinol, and in the final enzymatic step, 11-cis retinol is oxidized to 11-cis retinal. The first and last steps of the classical visual cycle are reduction and oxidation reactions, respectively, that utilize retinol dehydrogenase (RDH) enzymes. The visual cycle RDHs have been extensively studied, but because multiple RDHs are capable of catalyzing each step, the exact RDHs responsible for each reaction remain unknown. Within rods, RDH8 is largely responsible for the reduction of all-trans retinal with possible assistance from RDH12. retSDR1 is thought to reduce all-trans retinal in cones. In the RPE, the oxidation of 11-cis retinol is carried out by RDH5 with possible help from RDH11 and RDH10. Here, we review the characteristics of each RDH in vitro and the findings from knockout models that suggest the roles for each in the visual cycle.
Asunto(s)
Retina/metabolismo , Retinal-Deshidrogenasa/fisiología , Visión Ocular/fisiología , Animales , Humanos , Ratones , Ratones Noqueados , Oxidación-Reducción , Epitelio Pigmentado de la Retina/metabolismoRESUMEN
In humans and mice, mutations in the Ret gene result in Hirschsprung's disease and renal defects. In the embryonic kidney, binding of Ret to its ligand, Gdnf, induces a program of epithelial cell remodeling that controls primary branch formation and branching morphogenesis within the kidney. Our previous studies showed that transcription factors belonging to the retinoic acid (RA) receptor family are crucial for controlling Ret expression in the ureteric bud; however, the mechanism by which retinoid-signaling acts has remained unclear. In the current study, we show that expression of a dominant-negative RA receptor in mouse ureteric bud cells abolishes Ret expression and Ret-dependent functions including ureteric bud formation and branching morphogenesis, indicating that RA-receptor signaling in ureteric bud cells is crucial for renal development. Conversely, we find that RA-receptor signaling in ureteric bud cells depends mainly on RA generated in nearby stromal cells by retinaldehyde dehydrogenase 2, an enzyme required for most fetal RA synthesis. Together, these studies suggest that renal development depends on paracrine RA signaling between stromal mesenchyme and ureteric bud cells that regulates Ret expression both during ureteric bud formation and within the developing collecting duct system.