Plasma TIMP-1 Research Articles

Abstract 4077: Crucial factors of the inflammatory microenvironment (IL-1β/TNF-α/TIMP-1) promote the selection of highly malignant hemopoietic clone of myelofibrosis

Abstract Myelofibrosis (MF) is a clonal disorder of the hemopoietic stem/progenitor cells (HSPCs). Together with molecular abnormalities (mutations in JAK2, Calreticulin (CALR) and MPL genes), chronic inflammation is the major hallmark of MF pathogenesis. It is argued that the up-regulated production of pro-inflammatory cytokines selects for the malignant clone. However, the key players linking inflammation and cancer in MF are still poorly known. Here, we investigated the in vitro effects of crucial factors of the inflammatory microenvironment (Interleukin (IL)-1β, Tumor Necrosis Factor (TNF)-α, Tissue Inhibitor of Metalloproteinases (TIMP)-1 and ATP) on the functional behaviour of MF-derived HSPCs. Plasma TIMP-1, TNF-α and IL-1β were measured by ELISA assay. Circulating CD34+ cells from MF patients (JAK2V617F (10 cases) and CALR (10 cases) mutations) or cord blood (CB; 8 samples) were functionally characterized after incubation with or without ATP (1000 μM), IL-1β (10 ng/mL), TNF-α (10 ng/mL) or TIMP-1 (100 ng/mL) (alone or in combination). Cells were then analyzed for survival/apoptosis (Annexin-V/Propidium Iodide staining), cell cycle and clonogenic capacity. Migration was assessed towards a C-X-C motif chemokine 12 (CXCL12) gradient in the presence or absence of the pro-inflammatory factors. MF-derived CD34+ cells were co-cultured with normal mesenchymal stromal cells (MSCs) in the presence of the pro-inflammatory factors and then evaluated for survival. Regardless mutation status, the plasma levels of IL-1β, TNF-α and TIMP-1 are increased in MF patients and the presence of pro-inflammatory cytokines confers a survival advantage of MF-derived CD34+/CD34+CD38- cells. In addition, MF-derived CD34+ cells promoted cell-cycle progression to the S-phase. Whereas in the JAK2V617F mutated group, the addition of IL-1β or TNF-α ± TIMP-1 impaired the erythroid clonogenic output of the CALR mutated patients. Of note, IL-1β + TNF-α ± TIMP-1 highly promoted the in vitro migration of MF-derived CD34+ cells in the presence of CXCL12. Intriguingly, MF-derived CD34+ cells revealed increased clonogenic ability after migration toward CXCL12 and IL-1β + TNF-α ± TIMP-1, suggesting a pivotal role of inflammation in the selection of the malignant clone. Finally, co-cultures of normal MSCs with MF-derived CD34+ cells sustained the survival effects of IL-1β+TNF-α±TIMP-1. Overall, our data indicate that the behavior of the MF-derived HSPCs can be influenced by regulatory signals provided by the microenvironment through the cooperation between various pro-inflammatory factors. The interplay of these pro-inflammatory factors promotes and selects the circulating MF-derived HSPCs with higher proliferative activity, clonogenic potential and migration ability. Targeting these micro-environmental interactions may be a clinically relevant approach. Citation Format: Daria Sollazzo, Dorian Forte, Nicola Polverelli, Marco Romano, Margherita Perricone, Sofia Fatica, Francesco Lapi, Emanuela Ottaviani, Martinelli Giovanni, Nicola Vianelli, Michele Cavo, Maria Pantaleo, Francesca Palandri, Lucia Catani. Crucial factors of the inflammatory microenvironment (IL-1β/TNF-α/TIMP-1) promote the selection of highly malignant hemopoietic clone of myelofibrosis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4077.

Expression of MMP and TIMP mRNA in peripheral blood leukocytes of patients with invasive ductal carcinoma of the breast.

An imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) appears critical for tumor progression and metastasis. This study aimed to determine whether gene expression of MMP1, MMP2, MMP9, TIMP1 and TIMP3 and the MMP/TIMP expression ratio in peripheral blood leukocytes (PBLs) and the MMP1 and TIMP1 contents or MMP1/TIMP1 ratio in plasma were associated with clinicopathological characteristics in invasive ductal carcinoma (IDC) of the breast. Blood samples were collected from women newly diagnosed with IDC who had not received prior treatment (n = 102). Gene expression in PBLs was analyzed by quantitative real-time polymerase chain reaction. Concentrations of MMP1 and TIMP1 in plasma were measured using ELISA. In univariate analysis the expression levels of MMP2 and TIMP1 mRNA were significantly higher in premenopausal compared to postmenopausal patients (p<0.001 and p = 0.014, respectively). MMP2 mRNA expression negatively correlated with age (p<0.001, r = -0.43). We found that the MMP2/TIMP3 expression ratio was significantly higher in women after menopause (p = 0.007). The MMP2/TIMP1 expression ratio was higher in human epidermal growth factor receptor 2 (HER2)-positive patients (p = 0.022). Low-grade tumors had significantly lower MMP1/TIMP1 and MMP2/TIMP1 expression ratios (p = 0.047 and p = 0.048, respectively). TIMP1 plasma concentration was significantly higher in small tumors compared with T2-T3 tumors (p = 0.013). These findings reveal an important association between tumor characteristics and expression ratios of MMP1/TIMP1 and MMP2/TIMP1 in PBLs and TIMP1 concentration in plasma. Menopausal status may influence the mRNA expression levels of MMP2 and TIMP1 as well as the MMP2/TIMP3 expression ratio in IDC of the breast.

41 Circulating level of TIMP-4 is elevated in preeclampsia

Introduction The MMPs (matrix metalloproteinases) and theirs endogenous inhibitors (proteins that bind MMP active sites preventing proteolytic cleavage of substrates: TIMP-1, -2, -3 and -4) play important roles in the cardiovascular system regulation and some studies have been demonstrated imbalance among MMPs and TIMPs in preeclampsia compared to healthy pregnancy. Even though the TIMPs present many similarities, they show several structural and biochemical differences. While the circulating levels of TIMP-1 and TIMP-2 (TIMP-3 is not soluble) have already been investigated in plasma/serum of preeclampsia pregnant compared to healthy pregnant, this comparison about TIMP-4 level is unknown. Importantly, TIMP-4 is expressed mainly in cardiovascular structures, and some studies have demonstrated high levels of TIMP-4 in some cardiovascular disorders, such as dilated cardiomyopathy and atherosclerosis; nevertheless of these evidences its function in cardiovascular biology and pathology is poorly understood. Aims To compare plasma levels of TIMP-4 between preeclampsia (PE) and healthy pregnant (HP) and to compare TIMP-4 values through of gestation periods (1st, 2nd and 3rd trimester) and post-partum in HP ( n = 18). Methods and results TIMP-4 was measured by ELISA. Pregnant were recruited in two different locations: Study 1 from Minas Gerais State (PE = 43 and HP = 36) and Study 2 from Sao Paulo State (PE = 20 and HP = 20). We found elevated plasma TIMP-4 levels in PE compared to HP in both in Study 1 and Study 2 ( P = 0.0046 and P = 0.0007, respectively). In addition, in the Study 2 we performed correlation among TIMP-4 levels and clinical parameters. We found positive correlation with serum liver enzymes (ALT, r = 0.84, P = 0.004; and AST, r = 0.51, P = 0.02) and proteinuria ( r = 0.66, P = 0.01); and negative correlations with newborn weight ( r = −0.45, P = 0.04). Finally, we observed lack of significant differences among gestational age and postpartum periods in HP group. Conclusions PE present elevated levels of circulating TIMP-4 compared to HP and these levels are correlated with clinical parameters of disease. Finacial Support This study was funded by the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), and the Fundacao o de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP-Brazil).

Perioperative time course of matrix metalloproteinase-9 (MMP-9), its tissue inhibitor TIMP-1 & S100B protein in carotid surgery.

Background & objectives:Ischaemic stroke is a life burdening disease for which carotid endarterectomy (CEA) is considered a gold standard intervention. Pro-inflammatory markers like matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) and S-100 Beta (S100B) may have a role in the early inflammation and cognitive decline following CEA. This study was aimed to describe the perioperative time courses and correlations between of MMP-9, TIMP-1 and S100B following CEA.Methods:Fifty four patients scheduled for CEA were enrolled. Blood samples were collected at four time points, T1: preoperative, T2: 60 min after cross-clamp release, T3: first postoperative morning, T4: third postoperative morning. Twenty atherosclerotic patients were included as controls. Plasma MMP-9, TIMP-1 and S100B levels were estimated by ELISA.Results:TIMP-1 was decreased significantly in the CEA group (P<0.01). Plasma MMP-9 was elevated and remained elevated from T1-4 in the CEA group (P<0.05) with a marked elevation in T3 compared to T1 (P<0.05). MMP-9/TIMP-1 was elevated in the CEA group and increased further by T2 and T3 (P<0.05). S100B was elevated on T2 and decreased on T3-4 compared to T1.Interpretation & conclusions:Our study provides information on the dynamic changes of MMP-9-TIMP-1 system and S100B in the perioperative period. Preoperative reduction of TIMP-1 might be predictive for shunt requirement but future studies are required for verification.

Matrix Metalloproteinases in Alzheimer's Disease and Concurrent Cerebral Microbleeds.

Matrix metalloproteinases (MMPs) are a family of enzymes able to degrade components of the extracellular matrix, which is important for normal blood-brain barrier function. Their function is regulated by tissue inhibitors of matrix metalloproteinases (TIMPs). We investigated whether MMPs and TIMPs in cerebrospinal fluid (CSF) and plasma were altered in Alzheimer's disease (AD) and vascular dementia (VaD), and whether this effect was modified by presence of cerebral micro-bleeds in AD patients. In addition, we assessed associations of MMPs and TIMPs with CSF amyloid-β(1-42) (Aβ42), tau, and tau phosphorylated at threonine-181 (p-tau). We measured MMP2, MMP9, and MMP10, and TIMP1 and TIMP2 in CSF and plasma of 52 AD patients, 26 matched controls, and 24 VaD patients. AD patients showed higher plasma MMP2 levels compared to VaD patients (p < 0.05), and higher CSF MMP10 levels compared to controls (p < 0.05). Microbleeds in AD were associated with lower CSF TIMP1, TIMP2 and MMP9 in a dose-response relation. In addition, CSF MMP2 was associated with p-tau (St.B 0.23, p < 0.05), and CSF MMP10 with tau (St.B 0.38, p < 0.001) and p-tau (St.B 0.40, p < 0.001). Our findings suggest involvement of MMP2 and MMP10 in AD pathology. Lower levels of TIMPs in AD patients with microbleeds suggest less MMP inhibition in patients with concurrent cerebral microbleeds, which may hypothetically lead to a more vulnerable blood-brain barrier in these patients.

Increased expression of metalloproteinase-2 and -9 (MMP-2, MMP-9), tissue inhibitor of metalloproteinase-1 and -2 (TIMP-1, TIMP-2), and EMMPRIN (CD147) in multiple myeloma

Introduction: Activity of metalloproteinases (MMP) is controlled both by specific tissue inhibitors (TIMP) and activators (extracellular matrix metalloproteinase inducer, EMMPRIN). There are few data available concerning concentration the bone marrow of MMP-2, MMP-9, TIMP-1, and TIMP-2, or EMMPRIM expression by bone marrow mesenchymal stromal cells (BMSCs) in patients with multiple myeloma (MM).Patients and methods: We studied 40 newly diagnosed, untreated patients: 18 males and 22 females with de novo MM and 11 healthy controls. Bone marrow was collected prior to therapy. BMSCs were derived by culturing bone marrow cells on MesenCult. Protein concentrations were determined in bone marrow plasma and culture supernatants by ELISA. EMMPRIN expression by BMSCs was assessed by flow cytometry.Results: The median concentrations of MMP-9, TIMP-1, and TIMP-2 in both marrow plasma and culture supernatants were significantly higher in MM patients than controls.Conclusion: EMMPRIN expression and ratios MMP-9/TIMP-1 and MMP-2/TIMP-2 were higher in MM patients, our results demonstrate that in MM patients MMP-2 and MMP-9 are secreted in higher amounts and are not balanced by inhibitors.

Matrix metalloproteinases and their tissue inhibitor after reperfused ST-elevation myocardial infarction treated with doxycycline. Insights from the TIPTOP trial

BackgroundThe TIPTOP (Early Short-term Doxycycline Therapy In Patients with Acute Myocardial Infarction and Left Ventricular Dysfunction to Prevent The Ominous Progression to Adverse Remodelling) trial demonstrated that a timely, short-term therapy with doxycycline is able to reduce LV dilation, and both infarct size and severity in patients treated with primary percutaneous intervention (pPCI) for a first ST-elevation myocardial infarction (STEMI) and left ventricular (LV) dysfunction. In this secondary, pre-defined analysis of the TIPTOP trial we evaluated the relationship between doxycycline and plasma levels of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). MethodsIn 106 of the 110 (96%) patients enrolled in the TIPTOP trial, plasma MMPs and TIMPs were measured at baseline, and at post-STEMI days 1, 7, 30 and 180. To evaluate the remodeling process, 2D-Echo studies were performed at baseline and at 6months. A 99mTc-SPECT was performed to evaluate the 6-month infarct size and severity. ResultsDoxycycline therapy was independently related to higher plasma TIMP-2 levels at day 7 (p<0.05). Plasma TIMP-2 levels above the median value at day 7 were correlated with the 6-month smaller infarct size (3% [0%–16%] vs. 12% [0%–30%], p=0.002) and severity (0.55 [0.44–0.64] vs. 0.45 [0.29–0.60], p=0.002), and LV dilation (−1ml/m2 [from −7ml/m2 to 9ml/m2] vs. 3ml/m2 [from −2ml/m2 to 19ml/m2], p=0.04), compared to their counterpart. ConclusionsIn this clinical setting, doxycycline therapy results in higher plasma levels of TIMP-2 which, in turn, inversely correlate with 6month infarct size and severity as well as LV dilation.

870 Increased Soluble Cancer-Associated Biomarker Levels at Colonoscopy and Subsequent Long-Term Risk of Primary Cancer

Background: The majority of patients with colorectal cancer (CRC) have increased levels of soluble cancer-associated biomarkers at diagnosis of primary disease. Subjects with clean colorectum at colonoscopy may however, also have increased levels of such biomarkers. The aim of the present study was to evaluate a possible association between increased levels of soluble TIMP-1, CEA, CA19-9 and YKL-40 in subjects offered colorectal endoscopy without findings of neoplastic bowel lesions and subsequent long-term risk of developing a primary malignant disease. Methods: During 2004/05 4,509 subjects were included in a multicenter study with collection of blood samples before sigmoidoand/or colonoscopy due to symptoms of CRC. The aim was to evaluate a possible relation between certain biomarkers and subsequent findings at endoscopy. A subgroup of 1,025 subjects had findings of diverticula and was chosen as research group, because the median age was similar to the subjects diagnosed with colorectal neoplasia. The levels of TIMP-1, CEA and CA19-9 were determined in EDTA plasma at an automatic analysis platform Architect® at Abbotts Center of Excellence, Amsterdam and YKL-40 was determined on a validated commercially available ELISA platform (Quidell, USA) at Herlev Hospital. With cut-point 12.31.2012 the subjects, who have developed a primarymalignant disease, except skin cancer, were identified and a relation between the biomarker levels at endoscopy and long-term risk of developing malignancy was calculated. On the basis of calculated normal levels, the relation with the biomarkers was separated into three groups: 0 = none of the four increased; 1 = one of the four increased; 2 = two or more of the four increased. In addition, the time from endoscopy to development of malignancy within the first five years was included; the model also includes an analysis of competing risk of dead before development of malignancy. Results: In total, 152 of the 1,025 subjects with diverticula developed a primary malignant disease within the observation period. Univariate analyses of the biomarker levels in the three predefined groups showed that TIMP-1 (p=0.0004), CEA (p=0.009) and CA19-9 (p=0.028) were significantly associated with development of malignancy. In a subsequent multivariate analysis including the three biomarkers showed that increased levels were associated with development of subsequent malignancy (p=0.0014). The cumulated risk of developing a primary malignant disease within the first five years was: group 0: 0.075 (0.055-0.099); group 1: 0.088 (0.059-0.126); group 2: 0.167 (0.120-0.221). Conclusion: Increased levels of the soluble biomarkers plasma TIMP-1, CEA and CA19-9 at endoscopy with findings of diverticula are associated with subsequent risk of developing a primary malignant disease.

871 Adipose Derived Mesenchymal Stem Cells in Fistula Treatment - Prospective Study on Experimental Model of Perianal Crohn's Disease.

Background: The majority of patients with colorectal cancer (CRC) have increased levels of soluble cancer-associated biomarkers at diagnosis of primary disease. Subjects with clean colorectum at colonoscopy may however, also have increased levels of such biomarkers. The aim of the present study was to evaluate a possible association between increased levels of soluble TIMP-1, CEA, CA19-9 and YKL-40 in subjects offered colorectal endoscopy without findings of neoplastic bowel lesions and subsequent long-term risk of developing a primary malignant disease. Methods: During 2004/05 4,509 subjects were included in a multicenter study with collection of blood samples before sigmoidoand/or colonoscopy due to symptoms of CRC. The aim was to evaluate a possible relation between certain biomarkers and subsequent findings at endoscopy. A subgroup of 1,025 subjects had findings of diverticula and was chosen as research group, because the median age was similar to the subjects diagnosed with colorectal neoplasia. The levels of TIMP-1, CEA and CA19-9 were determined in EDTA plasma at an automatic analysis platform Architect® at Abbotts Center of Excellence, Amsterdam and YKL-40 was determined on a validated commercially available ELISA platform (Quidell, USA) at Herlev Hospital. With cut-point 12.31.2012 the subjects, who have developed a primarymalignant disease, except skin cancer, were identified and a relation between the biomarker levels at endoscopy and long-term risk of developing malignancy was calculated. On the basis of calculated normal levels, the relation with the biomarkers was separated into three groups: 0 = none of the four increased; 1 = one of the four increased; 2 = two or more of the four increased. In addition, the time from endoscopy to development of malignancy within the first five years was included; the model also includes an analysis of competing risk of dead before development of malignancy. Results: In total, 152 of the 1,025 subjects with diverticula developed a primary malignant disease within the observation period. Univariate analyses of the biomarker levels in the three predefined groups showed that TIMP-1 (p=0.0004), CEA (p=0.009) and CA19-9 (p=0.028) were significantly associated with development of malignancy. In a subsequent multivariate analysis including the three biomarkers showed that increased levels were associated with development of subsequent malignancy (p=0.0014). The cumulated risk of developing a primary malignant disease within the first five years was: group 0: 0.075 (0.055-0.099); group 1: 0.088 (0.059-0.126); group 2: 0.167 (0.120-0.221). Conclusion: Increased levels of the soluble biomarkers plasma TIMP-1, CEA and CA19-9 at endoscopy with findings of diverticula are associated with subsequent risk of developing a primary malignant disease.

Increased risk of stroke among hypertensive patients with abnormally short sleep duration: analysis of the national health interview survey

acute myocardial infarction .This study aimed to investigate the association between MMP-9 and TIMP-1 levels and the occurrence of LVH in resistant hypertensive patients (RHTN) and mild to moderate hypertensives (HTN). Methods:We measured plasma TIMP-1 and MMP-9 (ELISA) in 126 HTN subjects and 86 RHTN patients. LVH was analyzed by left ventricle (LV) mass index (LVMI). Measurements of the dimensions of the LV were performed using the American Society of Echocardiography recommendations. The Mann-whitney test was used to compare the levels of biomarkers of extracellular matrix and IMVE between the groups while Spearman’s correlation was used to determine the relationship between the variables. Results: Resistant hypertensive patients had higher prevalence of HVE (66%) compared to mild to moderate hypertensive (46%), p1⁄40.005. Higher levels of TIMP-1 (mean SD) were found in RHTN (93.75 29.09 pg/mL) compared with HTN individuals (86.24 27.53 pg/mL) while lower MMP-9/TIMP-1 relationship was observed in RHTN group (0.37 0.5 vs. 0.77 1.75, p1⁄40.015). No differences were found between MMP-9 levels between the groups. TIMP1 levels were inversely correlated with LVMI in RHTN group (r1⁄4 -0.31 p1⁄40.005). Conclusion: RHTN patients had higher prevalence of LVH and higher TIMP-1 plasma levels. Moreover, RHTN group presented lower MMP-9/ TIMP-1 ratio compared to HTN. These data suggest that changes in the MMP-9/TIMP-1 balance may play an important role in the structural and clinical manifestations of resistant hypertension.

Plasma levels of matrix metalloproteinase-2, -3, -10, and tissue inhibitor of metalloproteinase-1 are associated with vascular complications in patients with type 1 diabetes: the EURODIAB Prospective Complications Study

BackgroundImpaired regulation of extracellular matrix remodeling by matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMP) may contribute to vascular complications in patients with type 1 diabetes. We investigated associations between plasma MMP-1, −2, −3, −9, −10 and TIMP-1, and cardiovascular disease (CVD) or microvascular complications in type 1 diabetic patients. We also evaluated to which extent these associations could be explained by low-grade inflammation (LGI) or endothelial dysfunction (ED).Methods493 type 1 diabetes patients (39.5 ± 9.9 years old, 51% men) from the EURODIAB Prospective Complications Study were included. Linear regression analysis was applied to investigate differences in plasma levels of MMP-1, −2, −3, −9, −10, and TIMP-1 between patients with and without CVD, albuminuria or retinopathy. All analyses were adjusted for age, sex, duration of diabetes, Hba1c and additionally for other cardiovascular risk factors including LGI and ED.ResultsPatients with CVD (n = 118) showed significantly higher levels of TIMP-1 [β = 0.32 SD (95%CI: 0.12; 0.52)], but not of MMPs, than patients without CVD (n = 375). Higher plasma levels of MMP-2, MMP-3, MMP-10 and TIMP-1 were associated with higher levels of albuminuria (p-trends were 0.028, 0.004, 0.005 and 0.001, respectively). Severity of retinopathy was significantly associated with higher levels of MMP-2 (p-trend = 0.017). These associations remained significant after further adjustment for markers of LGI and ED.ConclusionsThese data support the hypothesis that impaired regulation of matrix remodeling by actions of MMP-2, -3 and-10 and TIMP-1 contributes to the pathogenesis of vascular complications in type 1 diabetes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12933-015-0195-2) contains supplementary material, which is available to authorized users.

TIMP-1 and CEA as biomarkers in third-line treatment with irinotecan and cetuximab for metastatic colorectal cancer.

KRAS wild-type (wt) status determines indication for treatment with combination therapy, including epidermal growth factor receptor (EGFR) inhibitors, but still, the overall response rate in KRAS wt patients is less than 40 %. Consequently, the majority of patients will suffer from substantial side effects and no apparent benefit. Tissue inhibitor of metalloproteinases-1 is a glycoprotein, which regulates metalloproteinases and may consequently imply a central role in tumour progression. Furthermore, it is closely related to the EGFR regulation and has shown promising potential as a biomarker in colorectal cancer (CRC). The aim of the present study was to investigate the clinical value of TIMP-1 in patients with metastatic colorectal cancer (mCRC) treated with cetuximab and irinotecan. Patients with chemotherapy-resistant mCRC referred to third-line treatment with cetuximab (initial 400 mg/m(2) followed by weekly 250 mg/m(2))/irinotecan (350 mg/m(2) q3w) were prospectively included in the biomarker study, as previously published. Pre-treatment blood samples were collected, and plasma TIMP-1 was measured by a validated in-house ELISA assay. In addition, carcinoembryonic antigen (CEA) measurement was performed with a standardised method. A total of 107 patients were included in the biomarker study. The median baseline plasma TIMP-1 level was 271.1 ng/ml (range 65.9-1432 ng/ml) with no significant associations with baseline clinical characteristics. Median baseline plasma TIMP-1 levels were significantly higher in patients with early progression compared to patients who achieved disease control, 349 ng/ml (233-398 95 % confidence interval (CI)) and 215 ng/ml (155-289 95 % CI), respectively, p = 0.03, suggesting some association with treatment efficacy. When dividing patients according to TIMP-1 tertiles, the median progression-free survival (PFS) in patients with a high level of TIMP-1 was 2.4 months (95 % CI 2.1-4.1) compared to 3.3 months (95 % CI 2.1-6.2) and 4.7 months (95 % 3.2-7.6) in patients with intermediate or low levels, respectively. Analysis of TIMP-1 as a continuous variable revealed a shorter PFS associated with increasing levels of TIMP-1 (hazard ratio (HR) 1.36). These results translated into a significantly lower overall survival (OS) in patients with a high baseline TIMP-1 level (4.5 months (95 % CI 3.4-5.4)), compared to those with intermediate or low TIMP-1 levels (7.8 months (95 % CI 4.4-13.7) and 12.0 months (95 % CI 10.1-14.3), respectively, p < 0.0001). An 83 % higher hazard for death was revealed (HR = 1.83) with each twofold increase in the TIMP-1 level. Pre-treatment levels of CEA were not associated with any of the baseline characteristics (except primary tumour localisation) or to differences in PFS or OS. The rank correlation between CEA and TIMP-1 was r = 0.50, and a test for interaction between TIMP-1 and CEA (dichotomised at 5 ng/ml) in survival analysis was not significant (p = 0.18). A multivariate analysis for PFS and OS resulted in a model with significant contributions from TIMP-1, KRAS, and the number of metastatic sites. We have confirmed the potential prognostic value of TIMP-1 measurement prior to palliative chemotherapy for mCRC. However, validation in randomised trials will be essential with the perspective of establishing a potential predictive role of plasma TIMP-1 in this setting.

0281: Mitral stenosis: implication of metalloproteinase remodeling and calcification

mitral stenosis is characterized by pathological remodelling of valvular tissue but the molecular effectors involved in these processes are not well known. The role of matrix metalloproteinase MMP-9, MMP-3, and tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2 are investigated here. Subjects and methods: 235 patients with mitral stenosis and 150 healthy control subjects were recruited. MMP-9, MMP-3, TIMP-1 and TIMP-2 levels in plasma were measured using an ELISA assay. The plasma concentrations of MMP-3, TIMP-1 were significantly lower in patients compared with control group. MMP-9 rate is significantly increased in patients with mitral stenosis. In men, a negative correlation was observed between calcification degree and rate of MMP-9 (r=–0.484, p<0.001) and positive with MMP-3 (r=0.588, p<0.001). In women, positive correlation was found between MMP-9 and mitral area (r=0.387, p=0.002) and also between MMP3 and calcification degree (r=0.603, p<0.001). This study demonstrates the involvement of the MMP/TIMP system in ECM remodelling of stenosis mitral. We have shown a difference in level of MMP- 9 between the mitral stenosis and the control. MMP9 was involved in calcification of the mitrale valve.

Blood-based Biomarkers at Large Bowel Endoscopy and Prediction of Future Malignancies.

Soluble cancer-related protein biomarker levels may be increased in subjects without findings at large bowel endoscopy performed due to symptoms associated with colorectal cancer. The present study focused on a possible association between increased biomarker levels in such subjects and subsequent development of malignant diseases. In a major study of 4,990 subjects undergoing large bowel endoscopy, 691 were without pathology and comorbidity. Plasma levels of TIMP-1, CEA, CA19-9, and YKL-40 were determined in samples collected just before endoscopy and compared with subsequent development of a malignant disease within a period of 7–8 years. The upper 90% limits of the reference levels of every single protein were used to differentiate between normal and increased levels. The levels were separated into three groups: 0, none of the biomarkers increased; 1, one biomarker increased; 2, two or more biomarkers increased. A total of 43 subjects developed a primary malignant disease in the observation period. Univariatly, increase of all four biomarkers was significantly associated with subsequent development of a malignant disease. A multivariate analysis showed that increased biomarker levels were associated with subsequent development of a malignant disease (P = 0.002). The cumulative risk of developing malignant disease within the first 5 years after endoscopy was group 0, 3.3%; group 1, 5.8%; group 2, 7.8%. It is concluded that increased levels of plasma TIMP-1, CEA, CA19-9, and serum YKL-40 at large bowel endoscopy without findings may be associated with an increased risk of developing a subsequent malignant disease.

Evaluation of plasma MMP-8, MMP-9 and TIMP-1 identifies candidate cardiometabolic risk marker in metabolic syndrome: results from double-blinded nested case–control study

AimsMatrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are dysregulated in metabolic syndrome (MetS) and associated with atherosclerosis and cardiovascular disease (CVD). Previous studies on the association between MMPs/TIMPs and MetS are controversial. We aimed to evaluate circulating MMP-8, MMP-9 and TIMP-1 in a group of MetS individuals and healthy controls to find the potential marker associated with MetS and its components. Methods243 MetS individuals participated in a nested case–control design, of whom 63 were excluded (study subjects for analysis n=180; 87 MetS cases, 93 controls). We employed the International Diabetes Federation criteria using national waist circumference cutoffs for case definition. Anthropometric and biochemical measurements were done using standard methods. ResultsPlasma MMP-8, TIMP-1, tumor necrosis factor-alpha (TNF-α), highly sensitive C-reactive protein (hs-CRP) and MMP-8/TIMP-1 ratio were significantly higher in MetS cases (P for all < 0.05). Each component of MetS except raised fasting plasma glucose positively correlated with MMP-8 and numbers of MetS components increased with higher MMP-8. In all regression models, MMP-8 was a significant predictor of MetS and in the final model the relationship persisted even after adjusting for pro-inflammatory cytokines hs-CRP and TNF-α (odds ratio=6.008, 95% confidence interval: 1.612–22.389, P=0.008). ConclusionStrong associations of MMP-8 with components of MetS in univariate, bivariate and multivariate models suggest plasma MMP-8 as a potential cardiometabolic risk marker for MetS. Higher MMP-8 in MetS is possibly mediated through mechanisms both dependent and independent of chronic low grade inflammation.

Tissue inhibitor of metalloproteinases (TIMP)-1 creates a premetastatic niche in the liver through SDF-1/CXCR4-dependent neutrophil recruitment in mice.

Due to its ability to inhibit prometastatic matrix metalloproteinases, tissue inhibitor of metalloproteinases (TIMP)-1 has been thought to suppress tumor metastasis. However, elevated systemic levels of TIMP-1 correlate with poor prognosis in cancer patients, suggesting a metastasis-stimulating role of TIMP-1. In colorectal cancer patients, tumor as well as plasma TIMP-1 levels were correlated with synchronous liver metastasis or distant metastasis-associated disease relapse. In mice, high systemic TIMP-1 levels increased the liver susceptibility towards metastasis by triggering the formation of a premetastatic niche. This promoted hepatic metastasis independent of origin or intrinsic metastatic potential of tumor cells. High systemic TIMP-1 led to increased hepatic SDF-1 levels, which in turn promoted recruitment of neutrophils to the liver. Both inhibition of SDF-1-mediated neutrophil recruitment and systemic depletion of neutrophils reduced TIMP-1-induced increased liver susceptibility towards metastasis. This indicates a crucial functional role of neutrophils in the TIMP-1-induced premetastatic niche. Our results identify TIMP-1 as an essential promoter of hepatic premetastatic niche formation.

Increased Plasma Matrix Metalloproteinase-2 (MMP-2), Tissue Inhibitor of Proteinase-1 (TIMP-1), TIMP-2, and Urine MMP-2 Concentrations Correlate With Proteinuria in Renal Transplant Recipients

BackgroundThe most frequent cause of kidney allograft loss is chronic allograft injury, often with proteinuria as the clinical feature. Occurrence of proteinuria late after kidney transplantation is associated with worse graft function and patient survival. AimThe aim of the study was to assess plasma and urine matrix metalloproteinases (MMP-2 and MMP-9) and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) in proteinuric renal transplant recipients (RTRs). The factors were determined by enzyme-linked immunosorbent assay in 150 RTRs (51 women and 99 men), aged 49.2 ± 11.5 years, at mean 73.4 ± 41.2 months after kidney transplantation (range: 12 to 240 months). ResultsProteinuric RTRs compared with non-proteinuric RTRs had higher median plasma MMP-2 (P = .012), TIMP-1 (P = .0003), and TIMP-2 (P = .0021) concentrations, as well as higher urine MMP-2 (P < .0001) excretion. The presence of proteinuria had no impact on plasma MMP-9 and urine MMP-9, TIMP-1, and TIMP-2. Proteinuria and estimated daily proteinuria (uPr:uCr) correlated positively with plasma MMP-2 (rs = 0.226, P = .0054 and rs = 0.241, P = .003), TIMP-1 (rs = 0.305, P = .00015 and rs = 0.323, P = .000055), TIMP-2 (rs = 0.273, P = .0007 and rs = 0.269, P = .001) and urine MMP-2 (rs = 0.464, P < .0001 and rs = 0.487, P < .0001), respectively. Proteinuric RTRs had impaired graft function with higher median serum creatinine concentrations (1.91 [1.60–2.43] mg/dL versus 1.41 [1.20–1.65] mg/dL, P < .00001) and lower estimated glomerular filtration rate (36 [28–45] mL/min/1.73 m2 versus 53 [43–61] mL/min/1.73 m2, P < .00001) than RTRs without proteinuria. ConclusionsOur research revealed that in RTRs, proteinuria was significantly associated with increased concentrations of enzymes involved in extracellular matrix (ECM) degradation: plasma MMP-2, TIMP-1, TIMP-2, and urine MMP-2. Findings strongly emphasize increased plasma TIMPs in proteinuric RTRs that inhibit degradation of ECM by MMPs and favor excessive deposition of ECM proteins.

Tissue inhibitor of matrix metalloproteinase-1 polymorphism, plasma TIMP-1 levels, and antihypertensive therapy responsiveness in hypertensive disorders of pregnancy.

Tissue inhibitor of metalloproteinase (TIMP)-1 is a major endogenous inhibitor of matrix metalloproteinase (MMP)-9, which may affect the responsiveness to therapy in hypertensive disorders of pregnancy. We examined whether TIMP-1 polymorphism (g.-9830T>G, rs2070584) modifies plasma MMP-9 and TIMP-1 levels and the response to antihypertensive therapy in 596 pregnant: 206 patients with preeclampsia (PE), 183 patients with gestational hypertension (GH) and 207 healthy pregnant controls. We also studied the TIMP-3 polymorphism (g.-1296T>C, rs9619311). Plasma MMP-9 and TIMP-1 levels were measured by ELISA. GH patients with the GG genotype for the TIMP-1 polymorphism had lower MMP-9 levels and MMP-9/TIMP-1 ratios than those with the TT genotype. PE patients with the TG genotype had higher TIMP-1 levels. The G allele and the GG genotype were associated with PE and responsiveness to antihypertensive therapy in PE, but not in GH. Our results suggest that the TIMP-1 g.-9830T>G polymorphism not only promotes PE but also decreases the responses to antihypertensive therapy.

Benefit of EGFR-inhibition therapy for metastatic colorectal cancer patients with KRAS-mutated tumors and high plasma TIMP-1 level: Results from the NORDIC VII study.

3590 Background: RAS status is the only biomarker to select patients with metastatic colorectal cancer (mCRC) for therapy with an epidermal growth factor receptor (EGFR) inhibitor. However, not all...

An observational study of matrix metalloproteinase (MMP)-9 in cystic fibrosis

BackgroundIn cystic fibrosis (CF), cross-sectional studies have reported sputum matrix metalloproteinase (MMP)-9 to be elevated and negatively correlated with FEV1. This longitudinal study examined the association between MMP-9 and tissue inhibitors of metalloproteinases (TIMPs) to prognostic parameters in CF. MethodA cross-sectional survey of CF and control subjects; CF patients were followed up for a median of 49months. MMP-9 and TIMP-1 and TIMP-2 were quantified in sputum and plasma. ResultsSeventy-three patients with CF, median age 22years, and 40 controls were recruited. Fifty-three of these CF patients were followed up. Prospectively, in CF subjects, plasma MMP-9 activity was adversely associated with FEV1 (β −1.15 (95% CI −2.10, −0.20), p=0.019) and rate of FEV1 decline, and plasma TIMP-1 was adversely associated with mortality: hazard ratio 3.66 (1.91–7.04), p<0.001. ConclusionsThese associations further justify investigation of MMP-9 and TIMP-1 as biomarkers for short- to medium-term FEV1 decline and mortality in patients with CF.