RESUMO
PURPOSE: Fibroblast growth factor receptor 2 (FGFR2) and human epidermal growth factor receptor 2 (HER2) proteins are both molecular targets for cancer therapy. The objective of this study was to evaluate the expression status of FGFR2 and HER2 in patients with gastric cancer (GC) or colorectal cancer (CRC). METHODS: Archived tumor tissue samples from patients with histologically-confirmed GC or CRC suitable for chemotherapy were analyzed for FGFR2 and HER2 expression using immunohistochemistry and fluorescence in situ hybridization (HER2 in CRC only). RESULTS: A total of 176 GC patients and 389 CRC patients were enrolled. Among patients with GC, 25.6% were FGFR2-positive and 26.1% were HER2-positive. Among patients with CRC, 2.9% were FGFR2-positive and 16.2% were HER2-positive. No clear relationship was found between FGFR2 and HER2 status in either GC or CRC. In GC, FGFR2 and HER2 statuses did not differ between different primary cancer locations, whereas there were some differences between histological types. Based on FGFR2- and/or HER2-positive status, 117 patients were identified as potentially suitable for inclusion in clinical trials of therapeutic agents targeting the relevant protein (GC = 45, CRC = 72; FGFR = 56, HER2 = 62), of whom 7 were eventually enrolled into such clinical trials. CONCLUSIONS: This study indicated the prevalence of FGFR2 and HER2 in GC and CRC in the Japanese population. The screening performed in this study could be useful for identifying eligible patients for future clinical trials of agents targeting these proteins. TRIAL REGISTRATION: Clinical trial registration Japic CTI No.: JapicCTI-163380. https://www. CLINICALTRIALS: jp/cti-user/trial/ShowDirect.jsp?directLink=RNlzx1PPCuT.PrVNPxPRwA .
Assuntos
Neoplasias Colorretais , Neoplasias Gástricas , Neoplasias Colorretais/genética , Humanos , Hibridização in Situ Fluorescente , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/genética , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/metabolismo , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/uso terapêutico , Neoplasias Gástricas/genéticaRESUMO
Cellular cholesterol homeostasis involves sterol sensing at the endoplasmic reticulum (ER) and sterol export from the plasma membrane (PM). Sterol sensing at the ER requires efficient sterol delivery from the PM; however, the macromolecules that facilitate retrograde sterol transport at the PM have not been identified. ATP-binding cassette transporter A1 (ABCA1) mediates cholesterol and phospholipid export to apolipoprotein A-I for the assembly of high density lipoprotein (HDL). Mutations in ABCA1 cause Tangier disease, a familial HDL deficiency. Several lines of clinical and experimental evidence suggest a second function of ABCA1 in cellular cholesterol homeostasis in addition to mediating cholesterol efflux. Here, we report the unexpected finding that ABCA1 also plays a key role in facilitating retrograde sterol transport from the PM to the ER for sterol sensing. Deficiency in ABCA1 delays sterol esterification at the ER and activates the SREBP-2 cleavage pathway. The intrinsic ATPase activity in ABCA1 is required to facilitate retrograde sterol transport. ABCA1 deficiency causes alternation of PM composition and hampers a clathrin-independent endocytic activity that is required for ER sterol sensing. Our finding identifies ABCA1 as a key macromolecule facilitating bidirectional sterol movement at the PM and shows that ABCA1 controls retrograde sterol transport by modulating a certain clathrin-independent endocytic process.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/metabolismo , Retículo Endoplasmático/metabolismo , Esteróis/metabolismo , Animais , Transporte Biológico , Células Cultivadas , Metabolismo dos Lipídeos , Camundongos , Proteína de Ligação a Elemento Regulador de Esterol 2/metabolismoRESUMO
ABCB4, which is specifically expressed on the canalicular membrane of hepatocytes, exports phosphatidylcholine (PC) into bile. Because SM depletion increases cellular PC content and stimulates PC and cholesterol efflux by ABCA1, a key transporter involved in generation of HDL, we predicted that SM depletion also stimulates PC efflux through ABCB4. To test this prediction, we compared the lipid efflux activity of ABCB4 and ABCA1 under SM depletion induced by two different types of inhibitors for SM synthesis, myriocin and (1R,3S)-N-(3-hydroxy-1-hydroxymethyl-3-phenylpropyl)dodecanamide, in human embryonic kidney 293 and baby hamster kidney cells. Unexpectedly, SM depletion exerted opposite effects on ABCB4 and ABCA1, suppressing PC efflux through ABCB4 while stimulating efflux through ABCA1. Both ABCB4 and ABCA1 were recovered from Triton-X-100-soluble membranes, but ABCB4 was mainly recovered from CHAPS-insoluble SM-rich membranes, whereas ABCA1 was recovered from CHAPS-soluble membranes. These results suggest that a SM-rich membrane environment is required for ABCB4 to function. ABCB4 must have evolved to exert its maximum activity in the SM-rich membrane environment of the canalicular membrane, where it transports PC as the physiological substrate.
Assuntos
Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Membrana Celular/metabolismo , Fosfatidilcolinas/metabolismo , Esfingomielinas/metabolismo , Transportador 1 de Cassete de Ligação de ATP/antagonistas & inibidores , Transportador 1 de Cassete de Ligação de ATP/genética , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/antagonistas & inibidores , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Animais , Transporte Biológico Ativo/fisiologia , Membrana Celular/genética , Cricetinae , Células HEK293 , Humanos , Fosfatidilcolinas/genética , Esfingomielinas/genéticaRESUMO
Although human MDR1 and MDR3 share 86% similarity in their amino acid sequences and are predicted to share conserved domains for drug recognition, their physiological transport substrates are quite different: MDR1 transports xenobiotics and confers multidrug resistance, while MDR3 exports phosphatidylcholine into bile. Although MDR1 shows high ATPase activity, attempts to demonstrate the ATPase activity of human MDR3 have not succeeded. Therefore, it is possible that the difference in the functions of these proteins is caused by their different ATPase activities. To test this hypothesis, a chimera protein containing the transmembrane domains (TMDs) of MDR1 and the nucleotide binding domains (NBDs) of MDR3 was constructed and analyzed. The chimera protein was expressed on the plasma membrane and conferred resistance against vinblastine and paclitaxel, indicating that MDR3 NBDs can support drug transport. Vanadate-induced ADP trapping of MDR3 NBDs in the chimera protein was stimulated by verapamil as was MDR1 NBDs. The purified chimera protein showed drug-stimulated ATPase activity like MDR1, while its Vmax was more than 10-times lower than MDR1. These results demonstrate that the low ATPase activity of human MDR3 cannot account for the difference in the functions of these proteins, and furthermore, that TMDs determine the features of NBDs. To our knowledge, this is the first study analyzing the features of human MDR3 NBDs.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/química , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Adenosina Trifosfatases/química , Adenosina Trifosfatases/metabolismo , Nucleotídeos/metabolismo , Proteínas Recombinantes/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Adenosina Trifosfatases/genética , Sequência de Aminoácidos , Linhagem Celular , Membrana Celular/metabolismo , Resistência a Múltiplos Medicamentos , Humanos , Dados de Sequência Molecular , Paclitaxel/farmacologia , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Verapamil/farmacologia , Vimblastina/farmacologiaRESUMO
BACKGROUND: Tyrosine kinase inhibitors (TKIs) are effective for the treatment of non-small cell lung cancer (NSCLC) patients with activating mutations of the epidermal growth factor receptor (EGFR), but responses are not durable as tumors develop resistance. DS-1205c is a novel, specific, orally bioavailable, small-molecule AXL receptor TKI. In preclinical studies, DS-1205c restored TKI antitumor activity in a TKI acquired-resistance EGFR-mutant NSCLC tumor xenograft model. METHODS: This first-in-human, multicenter, open-label Phase 1 study (registered at ClinicalTrials.gov: NCT03599518) primarily evaluated the safety and tolerability of combination therapy with DS-1205c and gefitinib in Japanese patients with metastatic or unresectable EGFR-mutant NSCLC and tumor progression during treatment with EGFR-TKIs. Patients (n = 20) received DS-1205c monotherapy (200-1200 mg twice daily [BID]) in a 7-day safety monitoring period before combination DS-1205c/gefitinib (250 mg once daily) in 21-day cycles. RESULTS: The observed common treatment-emergent adverse events (TEAEs) were increased aspartate aminotransferase (35%), increased alanine aminotransferase (30%), rash maculo-papular (30%), and diarrhea (25%). No serious TEAEs were reported. Plasma concentrations and pharmacokinetic parameters of DS-1205a (free form of DS-1205c) were unaffected by concomitant administration of gefitinib. No patient achieved a complete or partial response and 5 patients (25%) had stable disease. CONCLUSION: DS-1205c was generally safe and well tolerated at all dose levels, but the safety profile of ≤800 mg BID was more favorable than 1200 mg BID. The recommended dose for dose-expansion cohorts of DS-1205c in combination therapy with gefitinib was 800 mg BID.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Gefitinibe/uso terapêutico , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Inibidores de Proteínas Quinases/efeitos adversos , Mutação , Receptores ErbB/genéticaRESUMO
BACKGROUND: Postprandial hyperlipidemia partially refers to the postprandial accumulation of chylomicrons and chylomicron remnants (CM-R). Many in vitro studies have shown that CM-R has highly atherogenic properties, but consensus is lacking on whether CM-R accumulation correlates with the development of atherosclerotic cardiovascular diseases. We investigated the correlation between CM-R accumulation and the prevalence of coronary artery disease (CAD). DESIGN: Subjects who received a coronary angiography and did not take any lipid-lowering drugs (n = 189) were enrolled. Subjects with coronary artery stenosis (≥ 75%) were diagnosed as CAD. Biochemical markers for glucose and lipid metabolism including fasting apolipoprotein (apo) B-48 concentration were compared between CAD patients (n = 96) and age-, sex-, and body mass index (BMI)-matched non-CAD subjects without overt coronary stenosis (< 75%) (n = 67). We tried to determine which metabolic parameters were correlated with the prevalence of CAD by multiple logistic regression analysis, and whether or not the combination of high apo B-48 and other coronary risk factors (high triglyceride, low HDL-C, high HbA1c or low adiponectin levels) increased the prevalence of CAD. RESULTS: Fasting serum apo B-48 levels were significantly higher in CAD patients than in non-CAD subjects (3·9 ± 2·4 vs. 6·9 ± 2·6 µg/mL, P < 0·0001) and had the most significant correlation with the existence of CAD. The clustering of high fasting apo B-48 levels (> 4·34 µg/mL, the cut-off value) and other coronary risk factors were found to be associated with a stronger risk of CAD compared with single high fasting apo B-48 levels. CONCLUSION: Fasting serum apo B-48 levels significantly correlated with the prevalence of CAD.
Assuntos
Apolipoproteína B-48/sangue , Doença da Artéria Coronariana/sangue , Hiperglicemia/sangue , Lipídeos/sangue , Idoso , Doença da Artéria Coronariana/fisiopatologia , Jejum , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Prevalência , Fatores de Risco , Estatística como AssuntoRESUMO
CD147 is an immunoglobulin-like receptor that is highly expressed in various cancers and involved in the growth, metastasis, and activation of inflammatory pathways via interactions with various functional molecules, such as integrins, CD44, and monocarboxylate transporters. Through screening of CD147-targeting antibodies with antitumor efficacy, we discovered a novel rat monoclonal antibody #147D. This humanized IgG4-formatted antibody, h4#147D, showed potent antitumor efficacy in xenograft mouse models harboring the human PDAC cell line MIA PaCa-2, HCC cell line Hep G2, and CML cell line KU812, which featured low sensitivity to the corresponding standard-of-care drugs (gemcitabine, sorafenib, and imatinib, respectively). An analysis of tumor cells derived from MIA PaCa-2 xenograft mice treated with h4#147D revealed that cell surface expression of CD147 and its binding partners, including CD44 and integrin α3ß1/α6ß1, was significantly reduced by h4#147D. Inhibition of focal adhesion kinase (FAK), activation of multiple stress responsible signal proteins such as c-JunN-terminal kinase (JNK) and mitogen-activated protein kinase p38 (p38MAPK), and expression of SMAD4, as well as activation of caspase-3 were obviously observed in the tumor cells, suggesting that h4#147D induced tumor shrinkage by inducing multiple stress responsible signals. These results suggest that the anti-CD147 antibody h4#147D offers promise as a new antibody drug candidate.
RESUMO
Lipoproteins in the plasma transport lipids to various tissues through the bloodstream. In an analysis based on specific gravity using ultracentrifugation, these can be divided into 4 main fractions: chylomicrons, VLDL, LDL, and HDL. Metabolism of the triglyceride-rich (TG-rich) lipoproteins, chylomicrons and VLDL, begins with the hydrolysis of TGs by lipoprotein lipase (LPL) and proceeds through intermediate metabolites (remnants). Those resulting from the former are referred to as chylomicron remnants, and those from the latter as VLDL remnants. Both types of remnant are enriched in cholesteryl esters and apolipoprotein E (apoE), and, moreover, they are referred to as atherogenic lipoproteins that readily accumulate in the arterial walls. At the research level, the analytical methods for remnants such as electrophoresis, ultracentrifugation, gel filtration, etc., are not necessarily simple. One of the methods used as a clinical laboratory test for the quantification of remnant lipoproteins is the remnant-like particle-cholesterol (RLP-C) assay. The significance of the assay as an evaluation for arteriosclerosis is well recognized internationally. Recently, a new method for measuring remnant lipoprotein cholesterol (RemL-C) has been developed that uses a reagent consisting of enzymes and a surfactant. With this method, measurements simply involve an automated analyzer in the same manner as for the homogeneous assay for HDL-C or LDL-C and have been shown to correlate well with values using conventional RLP-C reagents. Measurements can be made simply without the need for special equipment in a short period of time, about 10 minutes. Furthermore, this means that measurements can be made in this way with higher reproducibility and precision. Nevertheless, where the principles behind the two assays are different, discrepancies in the measured values can be identified in some cases. It is also important to understand the characteristics of both methods when using them. After detailing lipoprotein remnant measurement methods, we point out the significance of lipoprotein remnant measurements under conditions that have been the focus of recent particular attention, such as postprandial hyperlipidemia and metabolic syndrome.
Assuntos
Aterosclerose/diagnóstico , Colesterol/análise , Hiperlipoproteinemia Tipo II/diagnóstico , Lipoproteínas/metabolismo , Triglicerídeos/análise , Aterosclerose/etiologia , Biomarcadores/análise , Cromatografia em Gel , Técnicas de Laboratório Clínico/métodos , Humanos , Hiperlipoproteinemia Tipo II/etiologia , Lipoproteínas/análise , Síndrome Metabólica/diagnóstico , Síndrome Metabólica/etiologia , Kit de Reagentes para Diagnóstico , Tensoativos , UltracentrifugaçãoRESUMO
A spontaneous, morphological variation 'sango' was observed in the progeny of a Pleurotus pulmonarius (Fr.) Quél. wild-type basidiocarp (also known as fruiting body) collected from the field. This variant developed wart- and coral-like structures instead of normal basidiocarps. Microscopic analysis showed that the sango phenotype had defects in the differentiation of the pileus and hymenium. Basidiocarp phenotypic data analysis in the progenies revealed that the sango trait is a heritable mutation character controlled by a single recessive gene. This mutation locus was mapped on linkage group III of a previously constructed genetic linkage map by amplified fragment length polymorphism (AFLP) technique in P. pulmonarius. Four AFLP markers identified by bulked segregant analysis showed linkage to the sango mutation locus, with the genetic distance ranging from 0 to 2.1 cM. Of these markers, one marker was co-segregated with the sango mutation locus. This knowledge will be a useful foundation for practical breeding as well as for elucidating molecular mechanisms in basidiocarp development of main edible mushrooms.
Assuntos
Carpóforos/genética , Genes Fúngicos , Genes Recessivos , Mutação , Pleurotus/genética , Característica Quantitativa Herdável , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Carpóforos/metabolismo , Carpóforos/ultraestrutura , Ligação Genética , Loci Gênicos , Marcadores Genéticos , Fenótipo , Pleurotus/metabolismo , Pleurotus/ultraestruturaRESUMO
Plasma high density lipoprotein (HDL)-cholesterol levels are inversely correlated to the risk of atherosclerotic cardiovascular diseases. Reverse cholesterol transport (RCT) is one of the major protective systems against atherosclerosis, in which HDL particles play a crucial role to carry cholesterol derived from peripheral tissues to the liver. Recently, ATP-binding cassette transporters (ABCA1, ABCG1) and scavenger receptor (SR-BI) have been identified as important membrane receptors to generate HDL by removing cholesterol from foam cells. Adiponectin (APN) secreted from adipocytes is one of the important molecules to inhibit the development of atherosclerosis. Epidemiological studies have revealed a positive correlation between plasma HDL-cholesterol and APN concentrations in humans, although its mechanism has not been clarified. Therefore, in the present study, we investigated the role of APN on RCT, in particular, cellular cholesterol efflux from human monocyte-derived and APN-knockout (APN-KO) mice macrophages. APN up-regulated the expression of ABCA1 in human macrophages, respectively. ApoA-1-mediated cholesterol efflux from macrophages was also increased by APN treatment. Furthermore, the mRNA expression of LXRalpha and PPARgamma was increased by APN. In APN-KO mice, the expression of ABCA1, LXRalpha, PPARgamma, and apoA-I-mediated cholesterol efflux was decreased compared with wild-type mice. In summary, APN might protect against atherosclerosis by increasing apoA-I-mediated cholesterol efflux from macrophages through ABCA1-dependent pathway by the activation of LXRalpha and PPARgamma.
Assuntos
Adiponectina/fisiologia , Aterosclerose/imunologia , HDL-Colesterol/metabolismo , Macrófagos Peritoneais/metabolismo , Transportador 1 de Cassete de Ligação de ATP , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/biossíntese , Adiponectina/genética , Animais , Apolipoproteína A-I/metabolismo , Transporte Biológico , Células Cultivadas , HDL-Colesterol/sangue , Proteínas de Ligação a DNA/biossíntese , Humanos , Receptores X do Fígado , Camundongos , Camundongos Knockout , Receptores Nucleares Órfãos , PPAR gama/biossíntese , Receptores Citoplasmáticos e Nucleares/biossíntese , Receptores Depuradores Classe B/biossínteseRESUMO
AIM: Homogeneous assay reagents for the determination of low-density lipoprotein cholesterol (LDL-C) have been available from several manufacturers. However, there has been considerable controversy due to uncertainty regarding their reactivity with intermediate-density lipoprotein (IDL), which is detected at an especially high frequency in patients with type III hyperlipemia. In this study, we examined the reactivity of a homogeneous assay, Cholestest LDL (R) (CT-LDL), with hyperlipemic sera that were classified according to the WHO system. METHODS: Sera from 6 normolipidemic and 22 hyperlipidemic patients classified according to the WHO system were used for this study. All serum specimens were fractionated by the ultracentrifugation method of Hatch and Lees, and subjected to lipid and protein measurements. RESULTS: The percent bias of values measured by CT-LDL relative to those determined by the ultracentrifugation method was calculated and compared to the lipid/protein ratios of each lipoprotein fraction. Consequently, the coefficient of correlation between the bias and the Triglyceride/Total cholesterol (TG/TC) ratio in the IDL fraction was 0.742. There were also correlations with the TG/TC ratio and the apo-lipoprotein B/Total Cholesterol ratio in the LDL fraction and in the LDL+IDL fraction, respectively. CONCLUSION: Further testing will be required in order to know more about the clinical condition of hyperlipidemic patients, since CT-LDL may react differently with some beta lipoproteins having a diverse lipid/protein composition compared to those in normolipidemic specimens.
Assuntos
Bioensaio/normas , LDL-Colesterol/sangue , Hiperlipidemias/sangue , Hiperlipidemias/diagnóstico , Colesterol/sangue , Humanos , Lipoproteínas/sangue , Reprodutibilidade dos Testes , Triglicerídeos/sangue , UltracentrifugaçãoRESUMO
The quality and quantity of high-density lipoprotein (HDL) in blood plasma are important for preventing coronary artery disease. ATP-binding cassette protein A1 (ABCA1) and apolipoprotein A-I (apoA-I) play essential roles in nascent HDL formation, but controversy persists regarding the mechanism by which nascent HDL is generated. In the "direct loading model", apoA-I acquires lipids directly from ABCA1 while it is bound to the transporter. By contrast, in the "indirect model", apoA-I acquires lipids from the specific membrane domains created by ABCA1. In this study, we found that trypsin treatment causes rapid release of phosphatidylcholine (PC) and cholesterol from BHK/ABCA1 cells, and that the time course of lipid release coincides with those of trypsin digestion of extracellular domains (ECDs) of surface ABCA1 and of release of ECD fragments into the medium. This trypsin-dependent lipid release was dependent on ABCA1 ATPase activity, and did not occur in cells that express ABCG1, which exports lipids like ABCA1 but does not have large ECDs. These results suggest that the trypsin-sensitive sites on the cell surface are the large ECDs of ABCA1, and that lipids transported by ABCA1 are temporarily sequestered within the ECDs during nascent HDL formation.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/metabolismo , Colesterol/metabolismo , Lipoproteínas HDL/biossíntese , Fosfatidilcolinas/metabolismo , Domínios e Motivos de Interação entre Proteínas , Transportador 1 de Cassete de Ligação de ATP/química , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Transporte Biológico , Linhagem Celular Transformada , Membrana Celular/metabolismo , Humanos , Metabolismo dos Lipídeos , Modelos Biológicos , Ligação ProteicaRESUMO
Plasma high density lipoprotein (HDL)-cholesterol levels are inversely correlated with the incidence of cardiovascular diseases. HDL is mainly assembled in the liver through the ATP-binding cassette transporter (ABCA1) pathway. In humans, plasma HDL-cholesterol levels are positively correlated with plasma adiponectin (APN) concentrations. Recently, we reported that APN enhanced apolipoprotein A-I (apoA-I) secretion and ABCA1 expression in HepG2 cells. In the present study, we investigated HDL assembly in APN-knockout (KO) mice. The apoA-I protein levels in plasma and liver were reduced in APN-KO mice compared with wild-type-mice. The ABCA1 expression in liver was also decreased in APN-KO mice. APN deficiency might cause the impaired HDL assembly by decreasing ABCA1 expression and apoA-I synthesis in the liver.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Adiponectina/fisiologia , Apolipoproteína A-I/metabolismo , Lipoproteínas HDL/metabolismo , Fígado/metabolismo , Transportador 1 de Cassete de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Adiponectina/genética , Animais , Apolipoproteína A-I/sangue , Lipoproteínas HDL/sangue , Lipoproteínas VLDL/sangue , Camundongos , Camundongos KnockoutRESUMO
OBJECTIVE: The contribution of visceral fat accumulation to the development of coronary heart disease was previously reported, but the relation between visceral fat accumulation and serum lipoprotein subclasses was unknown. METHODS AND RESULTS: We examined the relation of lipoprotein subclasses with visceral fat accumulation in 62 male subjects (aged 22 to 67 years) with visceral fat syndrome or obesity. Cholesterol levels in very low-density, low-density, and high-density lipoprotein subclasses (VLDL, LDL, and HDL) were determined by computer-assisted high-performance liquid chromatography. Subcutaneous fat area and visceral fat area were measured by computed tomographic scanning. There was no significant correlation between the subcutaneous fat area and the cholesterol levels in all lipoprotein subclasses. In contrast, the visceral fat area was correlated positively (P<0.002) with VLDL and LDL subclasses, except for large LDL, but negatively (P<0.001) with those in large and medium HDL subclasses. The observed positive correlations of small and very small LDL subclasses remained significant (P<0.005) after adjustment for serum cholesterol, triglycerides, HDL cholesterol, and LDL cholesterol, respectively, but a significant negative correlation (P<0.005) of large LDL was obtained after adjustment for LDL cholesterol. CONCLUSIONS: These findings indicate that this simple high-performance liquid chromatography method may be applied for easy detection and evaluation of abnormal distribution of lipoprotein subclasses.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Lipoproteínas/análise , Lipoproteínas/sangue , Obesidade/sangue , Obesidade/patologia , Tecido Adiposo/patologia , Adulto , Idoso , Biomarcadores/análise , Biomarcadores/sangue , Doença das Coronárias/sangue , Doença das Coronárias/epidemiologia , Doença das Coronárias/patologia , Humanos , Lipoproteínas HDL/análise , Lipoproteínas HDL/sangue , Lipoproteínas LDL/análise , Lipoproteínas LDL/sangue , Lipoproteínas VLDL/análise , Lipoproteínas VLDL/sangue , Masculino , Pessoa de Meia-Idade , Obesidade/epidemiologia , Fatores de Risco , VíscerasRESUMO
BACKGROUND: Takayasu arteritis (TA) is a chronic vasculitis that primarily affects large elastic arteries. Monitoring of disease activity is crucial because the disease tends to progress despite treatment with glucocorticoid and/or immunosuppressive agents. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) have generally been used as disease activity markers, but these are nonspecific inflammatory markers and lack the sensitivity and specificity to accurately monitor the disease status. Given the histological findings characterized by destruction of elastic fibers, we hypothesized that matrix metalloproteinases (MMPs) could be useful as markers of disease activity in TA. METHODS AND RESULTS: A consecutive series of 25 patients with TA were enrolled in this study. According to the National Institutes of Health criteria of disease activity, 11 were in an active phase and the remaining 14 were in remission. Circulating levels of MMP-2, MMP-3, and MMP-9 were determined by ELISA in all patients with TA and controls. MMP-2 levels were higher in patients with TA than in controls, but no correlation was found between serum MMP-2 and disease activity score. In contrast, MMP-3 and MMP-9 levels in patients with active disease were higher than in patients in remission and controls, and a positive correlation was demonstrated between circulating levels of MMP-3 or MMP-9 and disease activity score. The high levels of MMP-3 and MMP-9 improved when patients underwent remission. CONCLUSIONS: The present results indicate that MMP-2 can be helpful in diagnosing TA and that MMP-3 and MMP-9 can be used as activity markers for TA.
Assuntos
Metaloproteinases da Matriz/sangue , Arterite de Takayasu/diagnóstico , Arterite de Takayasu/enzimologia , Adulto , Idoso , Biomarcadores/sangue , Progressão da Doença , Feminino , Humanos , Masculino , Metaloproteinase 2 da Matriz/sangue , Metaloproteinase 3 da Matriz/sangue , Metaloproteinase 9 da Matriz/sangue , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Valores de Referência , Indução de Remissão , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Arterite de Takayasu/sangue , Inibidor Tecidual de Metaloproteinase-1/sangueRESUMO
OBJECTIVE: Many cell types in atherosclerotic lesions are thought to have various biological abnormalities, such as impaired lipid homeostasis and slow cell proliferation, which may be related to senescence at cellular and individual levels. One of the common characteristics of senescent cells in vitro is the alteration of actin cytoskeletons, which have been reported to be involved in the intracellular transport of lipids. Recently, we raised the hypothesis that Cdc42, which is a member of the Rho-GTPase family and is known to play an important role in actin dynamics, might be important in cellular lipid transport. METHODS AND RESULTS: In the present study, we found that the protein expression levels and GTP-binding activities of Cdc42 were decreased in aged human skin fibroblasts. Moreover, we found the intracellular kinetics of Golgi-associated lipids to be retarded in these cells, which was demonstrated by the fluorescence recovery after photobleaching (FRAP) technique and the use of N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-6-aminohexanoyl-D-erythro-sphingosine as a tracer. To correlate the decreased expression of Cdc42 with the retarded FRAP, we complemented the amount of wild-type c-myc-tagged Cdc42Hs (myc-Cdc42Hs-WT) by adenovirus-mediated gene transfer. We further tested the effect of the dominant-active form (myc-Cdc42Hs-DA, V12Cdc42Hs) or dominant-negative form (myc-Cdc42Hs-DN, N17Cdc42Hs) of Cdc42Hs on FRAP. Introduction of myc-Cdc42Hs-WT or myc-Cdc42Hs-DA recovered the retarded FRAP in the aged fibroblasts. Conversely, control fibroblasts infected with myc-Cdc42Hs-DN exhibited significantly retarded FRAP. CONCLUSIONS: These data clearly indicate that the expression of Cdc42, a small G protein, is decreased in the aged cells in close association with the retarded intracellular lipid transport. The present study demonstrates a possible function of Cdc42 in the mediation of intracellular lipid transport.
Assuntos
Fibroblastos/enzimologia , Fibroblastos/metabolismo , Membranas Intracelulares/metabolismo , Metabolismo dos Lipídeos , Pele/citologia , Proteína cdc42 de Ligação ao GTP/biossíntese , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Transporte Biológico Ativo/fisiologia , Células Cultivadas , Feminino , Fibroblastos/citologia , Recuperação de Fluorescência Após Fotodegradação/métodos , Humanos , Pessoa de Meia-Idade , Vesículas Transportadoras/fisiologia , Proteínas de Transporte Vesicular/metabolismoRESUMO
Remnant lipoproteins are known to promote atherosclerosis especially in patients with type III hyperlipoproteinemia (HLP). In the current study, the effects of atorvastatin were investigated with special reference to the exogenous and endogenous apolipoprotein (apo) B-containing lipoprotein metabolism in type III HLP. Four Japanese male patients with type III HLP associated with homozygous apoE2 were studied. One-month administration of atorvastatin (20 mg once daily), after a 4-week dietary run-in, strikingly reduced serum total cholesterol and triglyceride (TG) levels by 52 (P<0.01) and 56% (P<0.05), respectively. Atorvastatin further decreased remnant-like particle (RLP)-cholesterol by 73% and RLP-TG by 65% (P<0.05), respectively. Distribution analysis by polyacrylamide gel disc electrophoresis clearly showed that atorvastatin diminished very low-, intermediate- and low-density lipoprotein particles. The relative particle diameter of intermediate-density lipoprotein became smaller after atorvastatin treatment (P<0.01). Furthermore, ultracentrifugal analysis demonstrated that atorvastatin significantly decreased cholesterol, TG and phospholipid concentrations in all apoB-containing lipoprotein fractions and very low-density lipoprotein (VLDL)-cholesterol/serum TG ratio (P<0.05), implying atorvastatin-induced reduction of beta-VLDL. Finally, newly developed assays of apoB-48 and apoB-100 revealed that atorvastatin markedly reduced these apolipoproteins by 43 and 52%, respectively (P<0.01), suggesting that atorvastatin decreased the number of both exogenous and endogenous apoB-containing lipoproteins. Taken together, atorvastatin improves remnant lipoprotein metabolism in type III HLP both in quality and in quantity. Atorvastatin can be one of the optimal options for the treatment of patients with type III HLP.
Assuntos
Anticolesterolemiantes/uso terapêutico , Apolipoproteínas B/sangue , Ácidos Heptanoicos/uso terapêutico , Hiperlipoproteinemia Tipo III/sangue , Hiperlipoproteinemia Tipo III/tratamento farmacológico , Lipoproteínas/sangue , Pirróis/uso terapêutico , Idoso , Apolipoproteína B-100 , Apolipoproteína B-48 , Apolipoproteínas/sangue , Atorvastatina , Colesterol/sangue , Humanos , Lipídeos/sangue , Lipoproteínas/química , Masculino , Pessoa de Meia-Idade , Concentração Osmolar , Triglicerídeos/sangueRESUMO
A patient with cholesteryl ester transfer protein (CETP) deficiency presents with marked hyperalphalipoproteinemia (HALP). To investigate the contribution of CETP deficiency to the cause of HALP (HDL-C> or =1.94 mmol/l, 75 mg/dl), we investigated the CETP activities and the prevalence of genetic CETP mutations among 624 Japanese HALP subjects. The subjects were screened for four known genetic CETP mutations (intron 14 splicing defect (In14), exon 15 missense mutation (Ex15), intron 10 splicing defect (In10) and exon 6 nonsense mutation (Ex6)). We found the frequency of the patients with reduced CETP activity (<75% of normal controls) to be 55.5 and 64.1% in a high HDL group (1.94< or =HDL-C<2.59 mmol/l) and a marked HALP group (HDL-C> or =2.59 mmol/l, 100 mg/dl), respectively. At least one of the four mutations was identified in 65.7% of subjects with reduced CETP activities and 57.5% of subjects with marked HALP. The In14 and Ex15 mutations were very common in HALP subjects and the frequency of In10 mutation and Ex6 mutation was quite low. To investigate the impact of genetic CETP mutation on the phenotypes, we compared the plasma lipid levels and CETP activities between the subjects with two common mutations. All In14 homozygotes showed marked HALP, while marked HALP is less frequent (64.3%) in Ex15 homozygotes. HDL-C levels in Ex15 heterozygotes were significantly higher than those of In14 heterozygotes, suggesting the mutation has dominant negative effects on CETP activity in vivo. Some cases with In14 (5.7%) or Ex15 (7.2%) mutation showed low HDL-C levels. We conclude that CETP deficiency is a major cause of HALP; nevertheless CETP deficiency is not necessarily HALP.
Assuntos
Apolipoproteínas/sangue , Proteínas de Transporte/genética , Colesterol/sangue , Glicoproteínas , Hiperlipoproteinemias/genética , Adulto , Idoso , Proteínas de Transporte/metabolismo , Proteínas de Transferência de Ésteres de Colesterol , Feminino , Humanos , Hiperlipoproteinemias/epidemiologia , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Fenótipo , PrevalênciaRESUMO
'Visceral fat syndrome' is a clinical entity compatible with Syndrome X, Deadly quartet, and insulin resistance syndrome. All of these entities express the pathophysiology of multiple risk factor syndrome in which multiple risks of arteriosclerosis, i.e., hypertension, hyperlipidemia, and glucose intolerance, cluster in an individual. The accumulation of visceral fat plays a crucial role in the pathogenesis of visceral fat syndrome. On the other hand, familial combined hyperlipidemia (FCHL) is a common hyperlipidemia associated with premature arteriosclerosis. The pathogenesis of FCHL is not fully elucidated but the visceral fat accumulation may play an important role in the pathophysiology of FCHL. In this review, we will discuss the relationship between visceral fat syndrome and FCHL.
Assuntos
Hiperlipidemia Familiar Combinada/etiologia , Hipertrigliceridemia/complicações , Resistência à Insulina , Obesidade/complicações , Feminino , Humanos , Hiperlipidemia Familiar Combinada/genética , Masculino , Fatores de RiscoRESUMO
AIM: Lipoprotein lipase (LPL) deficiency is a rare autosomal recessive disorder characterized by severe hypertriglyceridemia. Similar clinical phenotypes have been reported with respect to defects in several LPL-associated proteins. However, it remains controversial whether severe hypertriglyceridemia itself is atherogenic. We herein present a case of LPL deficiency due to novel combined mutations of glycosylphosphatidylinositol (GPI)-anchored high-density lipoprotein (HDL)-binding protein 1 (GPIHBP1) in a patient with coronary artery disease (CAD). PATIENT: We evaluated a 54-year-old woman with severe hypertriglyceridemia and double vessel CAD. Although the LPL mass and activity in the postheparin plasma were extremely low, no mutations were detected in the LPL gene itself. RESULTS: Genetic analyses revealed that the patient had double homozygous mutations at 41 bp (c.41 G > T) and 202 bp (c.202 T > C) in the GPIHBP1 gene, resulting in C14F and C68R, respectively. Although the C14F/C68R GPIHBP1 exhibited a normal LPL-binding activity, the levels of mutant proteins were extremely reduced compared to those of the wild-type proteins in vitro. CONCLUSION: We found novel combined mutations of GPIHBP1 in a patient with hypertriglyceridemia and severe CAD. The present case provides important insight into the pathogenesis of severe hypertriglyceridemia associated with atherosclerosis.