Role Of Monocyte Chemoattractant Protein-1 Research Articles

The role of gene expression hypoxia-inducible factor-1a and serum level monocyte chemoattractant protein-1 in the incidence of chronic myeloid leukemia in Iraqi patients

Abstract: BACKGROUND: Chronic myeloid leukemia (CML) is a hematopoietic stem cell malignancy described by a translocation between chromosomes 9 and 22. There are many factors genetic or nongenetic effect on disease progression such as growth factors and transcription factors act as oncogenes or tumor suppressor genes. OBJECTIVES: The purpose of this research was to investigate the role of hypoxia-inducible factor (HIF1A) gene expression with CML, as well as the role of monocyte chemoattractant protein-1 (MCP-1) as a predictive biomarker on disease progression. MATERIALS AND METHODS: The current study consists of three groups: first group includes 50 newly diagnosed CML patients (females 22 and males 28), second group consists of 50 CML patients treated with tyrosine kinase inhibitor (TKI) with a complete molecular response (p210 BCR-ABL transcript levels ≤0.1% IS) (female 25 and male 25), and third group included another 50 apparently healthy volunteers (female 20 and male 30). The patients were admitted from the National Center of Hematology/Mustansiriyah University. All patients are diagnosed according to a complete blood count (CBC), a bone marrow examination, and a BCR-ABL gene test. RESULTS: Reverse transcription-quantitative polymerase chain reaction was applied to assess the expression levels of the HIF-1A gene and serum level of MCP1 by enzyme-linked immunosorbent assay. The results displayed downregulated of the HIF1A gene messenger RNA in CML patients in comparison to the controls group, as well as no statistically significant link was discovered when the fold of expression was correlated with the age and gender of CML patients. CONCLUSION: HIF1-alpha gene has an important role in pathological pathways such as angiogenesis. According to this study, HIF1-alpha gene is not an appropriate prognostic biomarker for detecting the risk of CML as well as MCP1 is thought to be a predictor of CML progression.

Role of MCP-1 as an inflammatory biomarker in nephropathy.

The Monocyte chemoattractant protein-1 (MCP-1), also referred to as chemokine ligand 2 (CCL2), belongs to the extensive chemokine family and serves as a crucial mediator of innate immunity and tissue inflammation. It has a notable impact on inflammatory conditions affecting the kidneys. Upon binding to its receptor, MCP-1 can induce lymphocytes and NK cells' homing, migration, activation, differentiation, and development while promoting monocytes' and macrophages' infiltration, thereby facilitating kidney disease-related inflammation. As a biomarker for kidney disease, MCP-1 has made notable advancements in primary kidney diseases such as crescentic glomerulonephritis, chronic glomerulonephritis, primary glomerulopathy, idiopathic proteinuria glomerulopathy, acute kidney injury; secondary kidney diseases like diabetic nephropathy and lupus nephritis; hereditary kidney diseases including autosomal dominant polycystic kidney disease and sickle cell kidney disease. MCP-1 not only predicts the occurrence, progression, prognosis of the disease but is also closely associated with the severity and stage of nephropathy. When renal tissue is stimulated or experiences significant damage, the expression of MCP-1 increases, demonstrating a direct correlation with the severity of renal injury.

The Role of Monocyte Chemoattractant Protein-1 (MCP-1) in Diabetic Kidney Disease

Monocyte chemoattractant protein-1 plays an important role in kidney disease. In several studies, the role of MCP-1 as a urine biomarker has been extensively studied. In many kidney diseases, there are elevated levels of MCP-1 in the kidney tissue and this is an important cause of monocyte infiltration in the pathogenesis of kidney damage. Elevated urinary MCP-1 levels have been able to predict outcome in proliferative kidney diseases such as lupus nephritis. However, in several studies it was also found that there was an increase in MCP-1 in diabetic kidney disease. Macrophages have even appeared early in diabetic kidney disease and are associated with the progression of kidney disease. Urinary MCP-1 levels are related to the degree of leukocyte infiltration in the tubulointerstitial. This supports inflammatory factors as part of the pathogenesis of diabetic kidney disease.

Monocyte chemoattractant protein 1 plasma concentration in blood from varicose veins decreases under venoactive drug treatment.

Vein-specific inflammation leads to vascular smooth muscle cells proliferation and extracellular matrix degradation of vein wall. This process is known as remodeling and is promoted by "trapped" leukocytes. Monocyte chemoattractant protein 1 (MCP-1) is a chemokine responsible for trafficking of leukocytes from blood to vein wall. The aim of this study was to measure the MCP-1 concentration in varicose veins blood before and after venoactive drug therapy and to compare it with a concentration of blood from varicose veins of subjects who did not receive drug treatment. Non-randomized comparative study was conducted on 30 patients with primary varicose veins. 20 patients of the study group received diosmin 900 mg/hesperidin 100 mg once daily. 10 controls received no treatment. MCP-1 level was measured (pg/mL) in the blood from varicose veins twice, at the day of inclusion and after 60 days. Legs discomfort related to chronic venous disease (CVD) symptoms was measured with 10-cm Visual Analogue Scale (VAS) at inclusion and at completion of the study. Median (interquartile range, IQR) MCP-1 concentrations in treatment and control groups at inclusion were 171.9 (124.4-216.0) and 157.0 (120.1-163.1), resp., P=0.285. After 60 days of treatment MCP-1 level decreased, but non-significantly to 152.3 (124.1-178.3). In patients who did not receive treatment chemokine level slightly increased to 163.0 (134.0-172.9). Median changes over time were -6.6 (-30.9-7.4) and 10.6 (-3.7-19.2) in the study and control groups, resp. (P=0.048). After 60 days in 12 of 19 and 2 of 9 patients of treatments and control groups MCP-1 decreased (P=0.103). Odds ratio for MCP-1 decreasing was 9.5 (95% CI 1.1-81.5, P=0.043) for those who received venoactive drug. Mean (± standard deviation [SD]) legs discomfort significantly dropped in the study group from 5.7 (±2.5) to 1.9 (±2.2) (P=0.0003), while in controls no changes were registered: 3.4 (±1.3) and 3.5 (± 1.4), resp., P=0.28). Mean difference of VAS at baseline and at follow-up was -3.5 (±2.6) and 0.9 (±2.1), resp. (P<0.0001). Plasma concentration of MCP-1 in varicose veins blood demonstrates a tendency to decrease under two months treatment with a venoactive drug. Future studies are needed to reveal a possible role of MCP-1 as a target considering its role in varicose veins pathogenesis.

Role of MCP-1 in the developments of diabetic nephropathy in patients with type 2 diabetes mellitus patients

Several complications may be developed during infection with type 2 diabetes mellitus. One of the most is diabetic nephropathy, leading to end-stage renal disease in more than 30% of diabetes mellitus. Monocyte, which is derived from blood circulation to the tissue the maintains an inflammatory state. Several factors and chemokines regulate the activity of monocytes. One of them is Monocyte chemoattractant protein-1 (MCP-1) which is released by several adipocytes that help attract monocytes during inflammation. Patients and methods: the samples were collected from patients who attended diabetes and endocrinology center in Al-Sader Teaching Hospital in Najaf between January 2019 to April 2019. This study involves 300 subjects. One hundred fifty of them were diagnosed with type 2 diabetes mellitus. According to their albumin/creatinine ratio (ACR), these patients were classified into three groups, including 50 patients with severe nephropathy, 50 patients with moderate nephropathy, and 50 patients with mild nephropathy. The control group involved 150 subjects who appeared healthy and the same age as the patients' group. Result: a significant increase of MCP-1 level was found in patients with mild, moderate and severe diabetic patients compared with healthy control.

Mesenchymal Stem Cells Enhance Chemotaxis of Activated T Cells through the CCL2-CCR2 Axis In Vitro.

Activation and migration of donor T cells to the host target organs are critical mechanisms in the pathogenesis of graft-versus-host disease (GVHD). The role of monocyte chemoattractant protein-1 (MCP-1/CCL2) and its receptor CCR2 in the recruitment of T cells during immune or inflammatory response is also well known. For elucidation of the mechanism of the therapeutic effect of human bone marrow derived-mesenchymal stem cells (MSC) in GVHD, we studied the effect of these cells on migration of activated donor T cells through the CCL2-CCR2 axis in vitro. MSC were expanded from donors' bone marrow mononuclear cells. After co-culturing of IL-2-activated T cells with allogeneic MSC at different ratios, the levels of CCL2 in supernatants were measured by ELISA, and CCR2 expression in CD4+/CD8+ T cells subsets were detected by flow cytometry. The effect of MSC on the migration of activated T cells in the Transwell system was studied in the absence or presence of CCL2. Our results show that CCL2 levels in supernatants of co-cultures were significantly higher than in MSC monoculture and this increase depended on the number of MSC. MSC inhibited proliferation of T cells, but did not change the percentages of CD4+ and CD8+ T cells subsets. MSC can up-regulate the CCR2 expression in CD8+ subsets rather than in CD4+ subsets; MSC enhanced migration of IL-2-activated T cells to CCL2 by increasing the expression of CCR2. The data demonstrate that MSC can enhance chemotaxis of cytokine-activated T cells through the CCL2-CCR2 axis in vitro.

Effects of Acute Fructose Loading on Markers of Inflammation-A Pilot Study.

Inflammation plays a role in development of diabetic complications. The postprandial state has been linked to chronic low grade inflammation. We therefore aimed to investigate the acute effects of fructose loading, with and without a pizza, on metabolic and inflammatory markers in patients with type 2 diabetes (T2D) (n = 7) and in healthy subjects (HS) (n = 6), age 47–76 years. Drinks consumed were blueberry drink (18 g fructose), Coca-Cola (17.5 g fructose), and fructose drink (35 g fructose). The levels of glucose, insulin, insulin-like growth factor binding protein-1 (IGFBP-1) and inflammatory markers: Interleukin-6 (IL-6), Monocyte chemoattractant protein-1 (MCP-1), Interleukin-18 (IL-18), Intercellular Adhesion Molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and bacterial lipopolysaccharides (LPS) were analyzed in blood. The postprandial responses were assessed using Wilcoxon’s matched-pairs test, Friedman’s ANOVA and Mann–Whitney U test. There was no difference in baseline levels of inflammatory markers between the groups. In T2D, MCP-1 decreased following blueberry drink and Coca-Cola (p = 0.02), Coca-Cola + pizza and fructose + pizza (p = 0.03). In HS, IL-6 increased following blueberry + pizza and fructose + pizza (p = 0.03), there was a decrease in MCP-1 following blueberry drink and Coca-Cola (p = 0.03), and in ICAM-1 following blueberry + pizza (p = 0.03). These results may indicate a role for MCP-1 as a link between postprandial state and diabetes complications, however further mechanistic studies on larger population of patients with T2D are needed for confirmation of these results.

Exploring the role of monocyte chemoattractant protein-1 in fibroblast-like synovial cells in rheumatoid arthritis

BackgroundRheumatoid arthritis (RA) is a chronic systemic inflammatory disease with persistent synovitis. In the present study, the impact of monocyte chemoattractant protein-1 (MCP-1) was explored to determine methods for the diagnosis and treatment of RA.MethodsFirst, fibroblast-like synoviocytes (FLSs) were obtained from a collagen-induced rat RA model. Next, MCP-1-overexpression plasmid and small interfering RNA were transfected into human and rat FLSs. Cell Counting Kit-8 (CCK-8), Transwell migration and flow cytometry assays were used to analyze cell proliferation, migration and apoptosis of FLSs following MCP-1 transfections, respectively. Furthermore, western blotting was used to analyze the expression levels of p-P38, p-PI3K, PI3K, CD31, VEGF, TNF-α and IL-β in FLSs following MCP-1 transfection. In addition, reverse transcription-quantitative PCR and ELISAs were used to analyze the expression levels of C-reactive protein (CRP), estrogen receptor, MCP-1 and pentraxin-3 in patients with clinical RA, followed by correlation analysis of clinical data. Finally, expression validation, diagnostic and protein-protein interaction (PPI) network analysis of MCP-1 were performed.ResultsMCP-1 promoted FLS proliferation and migration, and affected the apoptosis of FLSs. In addition, the expression levels of p-P38, p-PI3K, PI3K, CD31, VEGF, TNF-α and IL-β were also affected by MCP-1. In patients with clinical RA, the expression level of MCP-1 was increased. Moreover, CRP expression level was significantly up-regulated in RA. Clinically, MCP-1 was strongly correlated with tender joint count, swollen joint count, visual analog scale for general health and disease activity score 28 (DAS28)-MCP-1, and was moderately correlated with DAS28 and DAS28-CRP. PPI analysis showed that MCP-1 mainly interacted with other inflammatory cytokines.ConclusionIn conclusion, MCP-1 may play a significant regulatory role in RA, and could be used as a measurement index of clinical RA activity.

Abstract P771: Monocyte-Chemoattractant Protein-1 Levels in Human Carotid Atherosclerosis Associate With Hallmarks of Plaque Vulnerability

Background: Monocyte chemoattractant protein-1 (MCP-1) is a chemokine recruiting monocytes to the atherosclerotic plaque. Experimental, genetic, and epidemiological data support a key role of MCP-1 in atherosclerosis. Yet, the translational potential of targeting MCP-1 signaling for lowering vascular risk is limited by the lack of data on plaque MCP-1 activity in human atherosclerosis. Methods: We measured MCP-1 levels in human plaque samples from 1,199 patients undergoing carotid endarterectomy from the Athero-Express Biobank. We explored associations of plaque MCP-1 levels with histopathological features of plaque vulnerability, clinical plaque instability (symptomatic vs. asymptomatic plaque), molecular markers of plaque inflammation and remodeling, and with incident vascular events up to three years after plaque removal. Results: MCP-1 plaque levels were associated with individual histopathological hallmarks of plaque vulnerability (large lipid core, low collagen, high macrophage burden, low smooth muscle cell burden, intraplaque hemorrhage), as well as with a cumulative vulnerability index (range 0-5, beta: 0.42, 95%CI: 0.30-0.53, p=5.4x10 -13 ) independently of age, sex, and conventional vascular risk factors. Furthermore, MCP-1 levels were higher among patients with symptomatic, as compared to asymptomatic plaques (p=0.0001) and were associated with the levels of pro-inflammatory cytokines involved in leukocyte adhesion, as well as with matrix metalloproteinase activity in the plaque. In the follow-up analyses, MCP-1 levels were associated with a higher risk of peri-procedural events (up to 30 days after surgery). Conclusions: Our findings highlight a role of MCP-1 in human plaque vulnerability, the leading mechanism underlying vascular events like stroke and myocardial infarction. As such, they suggest that interfering with MCP-1 signaling in patients with established atherosclerosis could lower vascular risk.

Мезенхимные стволовые клетки усиливают хемотаксис активированных Т-клеток in vitro через функциональную ось CCL2-CCR2

Activation and migration of donor T cells to the host target organs are critical mechanisms in the pathogenesis of graft-versus-host disease (GVHD). The role of monocyte chemoattractant protein-1 (MCP-1/CCL2) and its receptor CCR2 in the recruitment of T cells during immune or inflammatory response is also well known. For elucidation of the mechanism of the therapeutic effect of human bone marrow derived-mesenchymal stem cells (MSC) in GVHD, we studied the effect of these cells on migration of activated donor T cells through the CCL2-CCR2 axis in vitro. MSC were expanded from donors' bone marrow mononuclear cells. After co-culturing of IL-2-activated T cells with allogeneic MSC at different ratios, the levels of CCL2 in supernatants were measured by ELISA, and CCR2 expression in CD4+/CD8+ T cells subsets were detected by flow cytometry. The effect of MSC on the migration of activated T cells in the Transwell system was studied in the absence or presence of CCL2. Our results show that CCL2 levels in supernatants of co-cultures were significantly higher than in MSC monoculture and this increase depended on the number of MSC. MSC inhibited proliferation of T cells, but did not change the percentages of CD4+ and CD8+ T cells subsets. MSC can up-regulate the CCR2 expression in CD8+ subsets rather than in CD4+ subsets; MSC enhanced migration of IL-2-activated T cells to CCL2 by increasing the expression of CCR2. The data demonstrate that MSC can enhance chemotaxis of cytokine-activated T cells through the CCL2-CCR2 axis in vitro.

The Prevalence Role of Monocyte Chemoattractant Protein-1 in Hashimoto's Thyroiditis via Various Stimuli Mechanisms.

Hashimoto's thyroiditis (HT) is an autoimmune disease occurred at any age especially in the 30 and 50 years of patient, with time thyroid gland is lost its function, as the lymphocytes produce inflammatory cytokines, have a direct effect on the thyroid gland function. The monocyte chemoattractant protein (MCP) produced by the podocyte and monocyte in response to these stimuli and has a role in the feedback based on thyroid hormones therefore MCP is increased secretion depending on thyroid hormones; also it's increased with enlargement of adipocyte tissue, i.e., in patients with high body mass index (BMI), there is a relationship between Hashimoto's disease and obesity, that has an effect on the thyroid gland function. Obesity have important role on thyrostimulated hormones (TSHs), as it has a great effect on metabolism and expansion of the thyroid gland; researchers have previously found that obese individuals have higher serum TSH levels. The results of the study have reported that MCP 1, TSH, and BMI levels were highly significant increased in sera of G2 compared with G1, while T3 and T4 levels were highly significant decreased in sera of G2 compared with G1 in referring to G1: Healthy control group while G2: HT patients.

Potential Role of Monocyte Chemoattractant Protein-1 in Monitoring Disease Progression and Response to Treatment in Overactive Bladder Patients.

PurposeTo compare urinary levels of monocyte chemoattractant protein-1 (MCP-1), an inflammatory cytokine, in healthy controls and overactive bladder (OAB) patients, to correlate changes in urinary MCP-1 with OAB treatment response and symptom severity, and to study the diagnostic potential of MCP-1 for OAB, as well as the efficacy of MCP-1 as a potential biomarker for different phenotypes of OAB.MethodsWe used enzyme-linked immunosorbent assay to measure normalized urinary MCP-1 levels in 56 individuals (43 OAB patients and 13 controls). We assessed the OAB patients at 3 visits with 2 validated symptom severity questionnaires (OAB-V8 and Patient Perception of Bladder Condition).ResultsThe mean pretreatment urinary MCP-1 level at visit 1 (229.2-pg/mg creatinine) was significantly greater than the MCP-1 levels at visit 3 in both the treatment (107.0-pg/mg creatinine) (P<0.001) and control (52.35-pg/mg creatinine) groups (P<0.001). Average OAB symptom severity decreased significantly from visit 1 (baseline) to visits 2 (4 weeks) and 3 (12–14 weeks) and was significantly correlated with urinary MCP-1 levels. Urinary MCP-1 levels dropped significantly (P=0.002) posttreatment in patients whose symptom severity improved by >30%, whereas nonresponders displayed no significant MCP-1 decrease (P=0.164). The receiver operating characteristic analysis of the OAB visit 1 and control groups produced an area under the curve of 0.891. We found no significant differences in sex, race, or age between the OAB and control groups.ConclusionsMCP-1 levels differed significantly between the control and OAB groups and were closely correlated with symptom severity and treatment response. The good diagnostic accuracy of MCP-1 for OAB suggests the potential usage of MCP-1 for OAB diagnosis. The varying response of urinary MCP-1 levels to treatment may indicate at least 2 potential phenotypes of OAB. MCP-1, in combination with other biomarkers and symptom severity questionnaires, could potentially aid in developing a patient-centered OAB treatment approach.

Monocyte Chemoattractant Protein-1 promotes cancer cell migration via c-Raf/MAPK/AP-1 pathway and MMP-9 production in osteosarcoma

BackgroundOsteosarcoma is generally reported among younger individuals and has a very poor prognosis, particularly for the development of metastasis. However, more effective metastatic biomarkers and therapeutic methods are absent. Monocyte chemoattractant protein-1 (MCP-1) is involved in cancer progression and inflammatory recruitment. Although previous studies have reported higher serum MCP-1 levels in patients with osteosarcoma, the role of MCP-1 in osteosarcoma progression remains to be addressed.MethodsThe osteosarcoma cell migratory ability was assessed by transwell migration assay. The MCP-1 and MMP-9 expression levels were analyzed by Western blot and qPCR. The signal activation was conducted by Western blot. The in vivo mouse experiment and tumor tissue array were performed to confirm our findings in vitro.ResultsThe present study demonstrates that MCP-1 regulates cell mobility through matrix metalloproteinase (MMP)-9 expression in osteosarcoma cells. Moreover, MCP-1 promotes MMP-9 expression, cell migration, and cell invasion by mediating CCR2, c-Raf, MAPK, and AP-1 signal transduction. Using MCP-1 knockdown stable cell lines, we found that MCP-1 knockdown reduces MMP-9 expression and cell mobility. Finally, we found high MCP-1 expression levels in osteosarcoma specimens.ConclusionsOur results provide prognostic value of MCP-1 in osteosarcoma by promoting MMP-9 expression.

Factors related to peripheral arterial disease in patients undergoing hemodialysis: the potential role of monocyte chemoattractant protein-1.

Peripheral arterial disease (PAD) is substantially prevalent among patients in the end stage of renal disease (ESRD). We considered factors related to peripheral arterial disease in patients undergoing hemodialysis including the important role of monocyte chemoattractant protein-1 (MCP-1) serum concentrations. We studied 150 patients in on-line-predilution hemodiafiltration. Dialysis sufficiency was defined by Kt/V for urea. PAD was defined using clinical criteria, ankle-brachial index and Doppler ultrasound in the lower limbs. MCP-1 serum concentrations were measured using enzyme-linked immunoabsorbed assay (ΕLISA). We performed chi-square tests and logistic regression analysis to investigate risk factors for the prevalence of PAD in these patients including MCP-1 serum concentrations. The patients with manifested PAD had elevated MCP-1, higher BP, higher arterial stiffness markers, higher markers of malnutrition, uncontrolled metabolic acidosis, bone disease and lower obtained dialysis adequacy than the patients without PAD. The association between PAD manifestation and high MCP-1 was found significant (x2 = 9.6, p = 0.001). The built logistic regression analysis showed that the high MCP-1 increased the risk for PAD 3.2 (95% C.I 1.3-8.2) folds after adjustment for confounders. PAD was also significantly associated with non-administration of vitamin D agents during dialysis (x2 = 3.5, p = 0.04).Malnutrition, low-grade inflammation mainly defined by high MCP-1 serum concentrations, metabolic acidosis and bone disease were included in significant predictors for peripheral arterial disease in patients undergoing hemodiafiltration. The obtained dialysis sufficiency and the therapy during dialysis sessions seem to play an additional role in the demonstration of peripheral vascular disease in these patients.

Role of Monocyte Chemoattractant Protein-1 for Diagnosing Acute Myocardial Infarction

Background: Acute myocardial infarction (AMI) controlled and promoted by inflammation within the coronary plaque. Monocyte chemoattractant protein-1 (MCP-1) is a pro-inflammatory mediator, that’s playing a major role in plaque rupture. This study aimed to assess the diagnostic performance of MCP-1 for early diagnosis of AMI among chest pain (CP) patients. Methods: MCP-1 and cardiac Troponin I (cTnI) were performed for all studied patients. The receiver operating characteristic (ROC) curve was used to assess the diagnostic accuracy of biomarkers. Results: The baseline level of MCP-1 has a good capacity for discriminating patients with AMI from non-coronary chest pain (NCCP), stable angina (SA), and unstable angina (UA) patients with efficiency 91%, 91%, and 83%; respectively. Area under the curves (AUCs) of MCP-1 for diagnosis AMI patients at 0-6 hours and > 6-12 hours after onset time of CP were 0.69 (P 12-24 hours, cTnI has AUC 0.93 (P < 0.0001) compared with MCP-1 0.74 (P < 0.0001). In conclusion: Independent early baseline MCP-1 has given sufficient diagnostic information for patients with AMI.

Role of MCP-1 and CCR2 in ethanol-induced neuroinflammation and neurodegeneration in the developing brain

BackgroundNeuroinflammation and microglial activation have been implicated in both alcohol use disorders (AUD) and fetal alcohol spectrum disorders (FASD). Chemokine monocyte chemoattractant protein 1 (MCP-1) and its receptor C-C chemokine receptor type 2 (CCR2) are critical mediators of neuroinflammation and microglial activation. FASD is the leading cause of mental retardation, and one of the most devastating outcomes of FASD is the loss of neurons in the central nervous system (CNS). The underlying molecular mechanisms, however, remain unclear. We hypothesize that MCP-1/CCR2 signaling mediates ethanol-induced neuroinflammation and microglial activation, which exacerbates neurodegeneration in the developing brain.MethodsC57BL/6 mice and mice deficient of MCP-1 (MCP-1−/−) and CCR2 (CCR2−/−) were exposed to ethanol on postnatal day 4 (PD4). Neuroinflammation, and microglial activation, and neurodegeneration in the brain were evaluated by immunohistochemistry and immunoblotting. A neuronal and microglial co-culture system was used to evaluate the role of microglia and MCP-1/CCR2 signaling in ethanol-induced neurodegeneration. Specific inhibitors were employed to delineate the involved signaling pathways.ResultsEthanol-induced microglial activation, neuroinflammation, and a drastic increase in the mRNA and protein levels of MCP-1. Treatment of Bindarit (MCP-1 synthesis inhibitor) and RS504393 (CCR2 antagonist) significantly reduced ethanol-induced microglia activation/neuroinflammation, and neuroapoptosis in the developing brain. MCP-1−/− and CCR2−/− mice were more resistant to ethanol-induced neuroapoptosis. Moreover, ethanol plus MCP-1 caused more neuronal death in a neuron/microglia co-culture system than neuronal culture alone, and Bindarit and RS504393 attenuated ethanol-induced neuronal death in the co-culture system. Ethanol activated TLR4 and GSK3β, two key mediators of microglial activation in the brain and cultured microglial cells (SIM-A9). Blocking MCP-1/CCR2 signaling attenuated ethanol-induced activation of TLR4 and GSK3β.ConclusionMCP-1/CCR2 signaling played an important role in ethanol-induced microglial activation/neuroinflammation and neurodegeneration in the developing brain. The effects may be mediated by the interaction among MCP-1/CCR2 signaling, TLR4, and GSK3β.

PO-289 Identification of microenvironmental regulation and therapeutic targeting of ongenic EF-2 kinase in pancreatic cancer

IntroductionPancreatic cancer (PaCa) is one of the most aggressive and deadliest cancer with 6 months average survival rate. Recent studies indicate that the tumour microenvironment (TME) plays an important role in all stages of tumorigenesis in PaCa. Therefore, both tumour cells and their TME should be targeted for an effective treatment. Eukaryotic elongation factor 2 kinase (EF2K) is an enzyme which is overexpressed in cancer cells and plays a key role in cancer cell survival under stress conditions. There is no study that have shown the relationship between EF2K and TME. Thus, we investigated that the effects of EF2K expression on PaCa cell line (PANC1) and the TME.Material and methodsPANC1 cells were cocultured with monocytic THP-1 cells and macrophages were polarised from THP-1 cells. The changes in EF2K expression, cell migration and invasion were analysed using western blot, migration and invasion assays, respectively. The effects of coculture on monocyte chemoattractant protein-1 (MCP-1) levels, which is one of the most important chemokines in TME were analysed using ELISA. The role of MCP-1 on both EF2K and migration of PANC1 cells were investigated. The role of MCP-1 on monocyte-macrophage differentiation was also investigated. To determine the relationship between EF2K and MCP-1, EF2K was stably overexpressed in PANC1 cells and MCP-1 levels were measured using western blot. Then, EF2K was silenced using siRNA and the changes in MCP-1 expression levels, the ability of colony formation, migration and invasion of PANC1 cells were investigated. Pancreatic orthotopic tumour model was used to determine the in vivo effects of EF2K inhibition. In tumour tissues, the changes in expression levels of MCP-1 was measured using western blot and the presence of tumor-infiltrated pro-tumorigenic macrophages were shown using immunohistochemical staining.Results and discussionsThe interaction between PANC1 cells and macrophages caused an increase in EF2K and MCP-1 proteins and this interaction accelarated cell migration and invasion. In addition, it was found a bidirectional interaction between MCP-1 and EF2K. MCP-1 also caused differentiation of monocytes to pro-tumorigenic macrophages. In vitro silencing of EF2K decreased MCP-1 expression, cell invasion and migration. In vivo inhibition of EF2K also decreased MCP-1 expression, tumour volume and the number of tumor-infiltrated pro-tumorigenic macrophages.ConclusionTargeting both tumour cells and macrohages through EF2K inhibition might be a promising strategy for PaCa treatment.

Role of monocyte chemoattractant protein-1 in liver fibrosis with transient myeloproliferative disorder in down syndrome.

Liver fibrosis is a common complication associated with transient myeloproliferative disorder (TMD) in Down syndrome (DS). The exact molecular pathogenesis that regulates disease progression is largely unknown. We recently found serum and/or urinary monocyte chemoattractant protein‐1 (MCP‐1) as a novel biomarker of liver fibrosis. This study was an in vitro analysis to investigate the fibrogenic activity of MCP‐1 using the collagen‐producing LX‐2 human hepatic stellate cell line. We also examined the fibrogenic activity of serum from a male neonate with DS in whom late‐onset liver fibrosis developed even after the resolution of TMD. MCP‐1 stimulated both cell growth and collagen synthesis of LX‐2 in a dose‐dependent manner. Patient serum obtained during the active disease phase significantly up‐regulated fibrogenic activity, which was suppressed in the presence of MCP‐1‐blocking antibody. Transient transforming growth factor beta 1 stimulation primed LX‐2 to induce prolonged hypersecretion of MCP‐1 in the culture supernatant and in collagen synthesis, which was suppressed with MCP‐1 blocking antibody as well. Conclusion: MCP‐1 accounts for the prolonged activation of collagen‐producing hepatic stellate cells in both a paracrine and autocrine manner, thereby promoting liver fibrosis. Anti‐cytokine therapy targeting the fibrogenic cytokines of MCP‐1, for example, herbal medicine, could provide a new therapeutic intervention for liver fibrosis associated with TMD in DS. (Hepatology Communications 2018;2:230‐236)

The Relationship of Residual Renal Function with Cardiovascular Morbidity in Hemodialysis Patients and the Potential Role of Monocyte Chemoattractant Protein-1

Background: Residual renal function (RRF) provides several benefits to patients on dialysis. Monocyte chemoattractant protein-1 (MCP-1) plays an important role in atherosclerotic lesions. We considered the relationship between RRF and cardiovascular morbidity and the significant role of MCP-1 serum concentrations in hemodiafiltration (HDF) patients. Methods: We enrolled 76 patients on on-line HDF. RRF was defined by interdialytic urine output, and we studied the patients in two groups according to the preservation or not of urine output. MCP-1 levels were measured using enzyme-linked immunosorbent assay. χ² tests were applied for the association between RRF and left ventricular hypertrophy (LVH), coronary artery disease (CAD), peripheral artery disease (PAD), and systolic and diastolic cardiac dysfunction. We built an adjusted model using logistic regression analysis for the factors which might impact on the loss of urine output. Results: χ² tests showed a significant association between the loss of urine output and LVH, diastolic dysfunction, and PAD (χ² = 7.4, p = 0.007; χ² = 14.3, p = 0.001; χ² = 4.2, p = 0.03, respectively), although the association with CAD and systolic dysfunction was found to be nonsignificant. The patients without RRF had significantly higher MCP-1, and the urine volume was inversely associated with MCP-1 (r = -465, p = 0.03). In the built adjusted model, the elevated MCP-1 was found to be a significant predictor for the loss of RRF. Conclusion: The loss of RRF was significantly associated with LVH, diastolic dysfunction, and PAD in HDF patients. The increased MCP-1, affected by the lack of urine, may act as an additional underlying factor on this relationship, reflecting a progressive inflammation/oxidative stress condition.

Associations Between Deceased-Donor Urine MCP-1 and Kidney Transplant Outcomes

IntroductionExisting methods to predict recipient allograft function during deceased-donor kidney procurement are imprecise. Understanding the potential renal reparative role for monocyte chemoattractant protein-1 (MCP-1), a cytokine involved in macrophage recruitment after injury, might help to predict allograft outcomes.MethodsWe conducted a substudy of the multicenter prospective Deceased Donor Study cohort that evaluated deceased kidney donors from 5 organ procurement organizations from May 2010 to December 2013. We measured urine MCP-1 (uMCP-1) concentrations from donor samples collected at nephrectomy to determine associations with donor acute kidney injury (AKI), recipient delayed graft function (DGF), 6-month estimated glomerular filtration rate (eGFR), and graft failure. We also assessed perfusate MCP-1 concentrations from pumped kidneys for associations with DGF and 6-month eGFR.ResultsAKI occurred in 111 donors (9%). The median (interquartile range) uMCP-1 concentration was higher in donors with AKI compared with donors without AKI (1.35 [0.41–3.93] ng/ml vs. 0.32 [0.11–0.80] ng/ml, P < 0.001). DGF occurred in 756 recipients (31%), but uMCP-1 was not independently associated with DGF. Higher donor uMCP-1 concentrations were independently associated with a higher 6-month eGFR in those without DGF (0.77 [0.10–1.45] ml/min per 1.73 m2 per doubling of uMCP1). However, there were no independent associations between uMCP-1 and graft failure over a median follow-up of ∼2 years. Lastly, perfusate MCP-1 concentrations significantly increased during pump perfusion but were not associated with DGF or 6-month eGFR.DiscussionDonor uMCP-1 concentrations were modestly associated with higher recipient 6-month eGFR in those without DGF. However, the results suggest that donor uMCP-1 has minimal clinical utility given no associations with graft failure.