Tetranectin as a potential novel prognostic biomarker in anthracycline-related cardiac dysfunction.

To assess the association of serum tetranectin levels with cardiac remodeling parameters and to evaluate its prognostic role in women with anthracycline-related cardiac dysfunction (ARCD) and without previous cardiovascular diseases (CVD) during 24-month follow-up period. A total of 362 women with primary diagnosed breast cancer who were planned to be treated with anthracyclines were examined. At 12 months after chemotherapy completion, all women were examined and ARCD was diagnosed in 114 patients. After 24 months of follow-up, all patients with ARCD were divided into 2 groups: group 1 comprised women with the adverse course of ARCD (n = 54), group 2 comprised those without it (n = 60). In group 1, the levels of tetranectin were lower than group 2 by 27.6% (p < 0.001) and the patients without ARCD by 33.7% (p < 0.001). In group 1, the levels of tetranectin decreased (p < 0.001) from 11.8 (7.1; 14.3) to 9.02 (5.3; 14.6) pg/mL at 24 months. Moreover, in group 2 (p = 0.871) and in patients without ARCD (p = 0.716), they did not change. The tetranectin values were the independent predictor (odds ratio 7.08; p < 0.001) and its levels ≤ 15/9 ng/mL (AUC = 0.764; p < 0.001) were identified as the predictors for the adverse course of ARCD. NT-proBNP levels did not show the prognostic role, but the addition of NT-proBNP improved prognostic value of analysis (AUC = 0.954; p = 0.002). The cut-off values of tetranectin were established as predictor for adverse course of ARCD, when NT-proBNP was not. The combined use of tetranectin and NT-proBNP demonstrated higher diagnostic value for prediction of adverse outcomes.

Exogenous Tetranectin Alleviates Pre-formed-fibrils-induced Synucleinopathies in SH-SY5Y Cells by Activating the Plasminogen Activation System.

Parkinson's disease (PD) is a common neurodegenerative disease. Previously we identified tetranectin (TN) as a differentially expressed protein in the cerebrospinal fluid of PD patients, and we were surprised to find that TN knockout mice developed PD features. However, the specific role of TN in PD has not been clarified. In this study, we aimed to determine the effect of exogenous TN on cellular PD models and elucidate the underlying mechanisms. We found that exogenous TN could alleviate pre-formed-fibrils (PFFs)-induced synucleinopathies in SH-SY5Y cells and reduce the cell-to-cell transmission of α-synuclein (SYN). We also found that TN can promote the degradation of SYN by plasmin, which may account for its effect on cellular PD models. Moreover, administration of SYN/PFFs decreased the expression of TN and increased the expression of plasminogen activator inhibitor-1 (PAI-1) in SH-SY5Y cells, thereby reducing plasmin activity. Our findings depict a possible SYN-TN-plasmin interaction in which elevated levels of extracellular SYN monomers and aggregates in PD diminish the production of TN and PAI-1. Such changes lead to a reduced plasmin activity, which in turn reduces the degradation of extracellular SYN, thus forming a vicious cycle.

Adipogenic function of mouse tetranectin and identification of its functional domain.

Tetranectin (TN), a plasminogen (Plg) binding protein, enhances the Plg activator (PA)-catalyzed activation of Plg to plasmin (Pln). Previously, TN was identified as an adipogenic serum protein, which promotes adipocyte differentiation. In the present study, we investigated the adipogenic function of mouse TN using recombinant proteins (rmTNs) in full-length and domain-truncated forms. Adipocyte differentiation in TN-depleted-FBS-media was significantly enhanced by rmTNs supplementation. The adipogenic effect of rmTNs was found to be dependent on the presence of a Plg binding domain, indicating the domain is essential for the adipogenic function of mTN. In addition, these results suggested the involvement of Plg activation, however Plg, PA and Pln appeared to have no direct effect on adipocyte differentiation. This study demonstrates the adipogenic function of mTN, which is dependent on the Plg binding domain as its functional domain.

Inverse changes in plasma tetranectin and titin levels in patients with type 2 diabetes mellitus: a potential predictor of acute myocardial infarction?

An early intervention using biomarkers to predict acute myocardial infarction (AMI) will effectively reduce global heart attack incidence, particularly among high-risk patients with type 2 diabetes mellitus (T2DM). This study attempted to identify potential biomarkers by detecting changes in the levels of plasma proteins in T2DM patients following onset of AMI in comparison with those without AMI. Volunteer T2DM patients without AMI (control; n=10) and T2DM patients with AMI (n=10) were recruited. Plasma samples from these patients were evaluated via two-dimensional gel electrophoresis (2DE) to screen for proteins with level changes between the two groups. The abundance of spots on gel images was analyzed using Progenesis SameSpots and subjected to false discovery rate (FDR) analysis. Protein spots with statistically significant changes of at least 1.5 fold were selected for mass spectrometry (MS) analysis. Due to strong cardiac connections, tetranectin and titin were evaluated by enzymelinked immunosorbent assay (ELISA). The adjusted P-values and fold changes between the two groups resulted in identification of 34 protein spots with significantly altered abundance. Upon MS analysis, 17 plasma proteins were identified: tetranectin, titin, clusterin, haptoglobin, myosin-13, zinc fnger protein 445, DNA repair protein RAD50, serum albumin, apolipoprotein A-IV, caspase-6, aminoacyl tRNA synthase complex-interacting multifunctional protein 1, serotransferrin, retinol-binding protein 4, transthyretin, alpha-1-antitrypsin, apolipoprotein A-I and serum amyloid A. Comparable patterns of changes in tetranectin and titin between the control and AMI groups were confirmed using ELISA. In summary, tetranectin and titin in plasma appeared to be closely associated with the onset of AMI among T2DM patients and can be used as potential biomarkers for prediction of a cardiac event, though this requires validation in a prospective cohort study.

Identification of tetranectin as adipogenic serum protein.

Fetal bovine serum (FBS) is an essential culture supplement for adipocyte differentiation of various adipogenic precursor cells. Adipocyte differentiation greatly varies depending on the type of serum in the differentiation medium. In this study, we found that FBS supported adipocyte differentiation of 3T3-L1 cells to a significantly higher extent than other types of bovine serum such as adult bovine serum (ABS). This differential adipogenic effect of bovine serum was shown to be due to the protein contents of bovine sera, indicating the presence of an adipogenic protein(s) in FBS. Serum proteome analysis identified tetranectin as an adipogenic protein. The adipogenic effect of tetranectin was confirmed by supplementation of FBS-containing differentiation medium with anti-tetranectin antibody, which suppressed adipocyte differentiation of 3T3-L1 cells. These results demonstrate that tetranectin is an adipogenic serum protein mediating the adipogenic effect of FBS.

Insight into the function of tetranectin in human diseases: A review and prospects for tetranectin-targeted disease treatment.

Tetranectin (TN), a serum protein, is closely associated with different types of cancers. TN binds plasminogen and promotes the proteolytic activation of plasminogen into plasmin, which suggests that TN is involved in remodeling the extracellular matrix and cancer tissues during cancer development. TN is also associated with other diseases, such as developmental disorders, cardiovascular diseases, neurological diseases, inflammation, and diabetes. Although the functional mechanism of TN in diseases is not fully elucidated, TN binds different proteins, such as structural protein, a growth factor, and a transcription regulator. Moreover, TN changes and regulates protein functions, indicating that TN-binding proteins mediate the association between TN and diseases. This review summarizes the current knowledge of TN-associated diseases and TN functions with TN-binding proteins in different diseases. In addition, potential TN-targeted disease treatment by inhibiting the interaction between TN and its binding proteins is discussed.

Identification of novel inhibitors of tetranectin-plasminogen interaction to suppress breast cancer invasion: an integrated computational and cell-based investigation.

Tetranectin-plasminogen interaction plays a defining role in extracellular matrix degradation, enabling tumor cell invasion and metastasis. This interaction occurs via the carbohydrate recognition domain (CRD) and Kringle 4 domain of tetranectin and plasminogen, respectively, leading to activation of the plasminogen-cascade that triggers the proteolytic processes. Thus targeting this interaction represents an important strategy to suppress tumor cell migration and invasion. In this direction, we attempted to target the CRD of tetranectin to inhibit its interaction with the Kringle-4 domain of plasminogen using natural bioactive compounds. A cheminformatics pipeline for drug designing and screening was utilized to obtain lead compound(s) that exhibit conformationally and energetically viable CRD binding. Out of 206 compounds screened, diosgenin and scytonemin displayed the most favorable interactions with CRD. Short-term molecular dynamics simulations of 20 ns were employed to further study the conformational stability of both compounds with tetranectin CRD which reflected at the increased stability of diosgenin in the CRD binding pocket compared to scytonemin. Finally, an extended molecular dynamic simulation of 100 ns affirmed the robust and stable interaction of diosgenin with CRD. Furthermore, diosgenin was observed to exert a pronounced anti-proliferative effect on high tetranectin-expressing MDA-MB-231 breast cancer cells. The inhibitory effect of diosgenin on the tetranectin-plasminogen interaction was corroborated by the reduced migration and invasiveness of MDA-MB-231 cells under diosgenin treatment. Overall the study presents an alternate and safer approach to impede breast cancer metastasis and delineates the novel anti-metastatic activity of diosgenin.Communicated by Ramaswamy H. Sarma.

Quantitative proteomics analysis of COVID-19 patients: Fetuin-A and tetranectin as potential modulators of innate immune responses.

Treatment of severe cases of coronavirus disease 2019 (COVID-19) is extremely important to minimize death and end-organ damage. Here we performed a proteomic analysis of plasma samples from mild, moderate and severe COVID-19 patients. Analysis revealed differentially expressed proteins and different therapeutic potential targets related to innate immune responses such as fetuin-A, tetranectin (TN) and paraoxonase-1 (PON1). Furthermore, protein changes in plasma showed dysregulation of complement and coagulation cascades in COVID-19 patients compared to healthy controls. In conclusion, our proteomics data suggested fetuin-A and TN as potential targets that might be used for diagnosis as well as signatures for a better understanding of the pathogenesis of COVID-19 disease.

Discovery and validation of a protein biomarker for the diagnosis and classification of disease severity of major depressive disorder.

BACKGROUND AND AIMS: Diagnosis and classification of disease severity of major depressive disorder (MDD) are determined through a doctor's consultation and questionnaire-based rating scale. This study aimed to identify and validate a serum protein biomarker for diagnosing and classifying the disease severity of MDD. MATERIALS AND METHODS: Based on the Hamilton Depression Rating Scale (HAMD) score, participants were divided into control, mild, moderate, and severe groups. Samples prepared from collected sera were analyzed using non-targeted qualitative and targeted quantitative tools to identify potential biomarkers. RESULTS: Four proteins were selected as biomarker candidates, which showed statistically significant consistent tendencies depending on MDD severity. Among them, tetranectin was the only validated protein in the quantitative analysis that showed the same decreasing tendency as that in the qualitative analysis. Furthermore, tetranectin showed fair discrimination performance between the control and MDD group. CONCLUSIONS: Tetranectin may be a novel potential biomarker for diagnosing and classifying the severity of MDD, though further verification and validation studies of its efficacy are needed.

Tetranectin targeting by epigallocatechin gallate suppresses colon cancer cell proliferation.

Tetranectin is a serum protein that binds to plasminogen and enhances its proteolytic activation, which underlies the involvement of tetranectin in the development of several carcinomas including colon cancer. In the present study, structure-based in silico screening of natural products showed that epigallocatechin gallate with anticancer effects binds to tetranectin. Binding to epigallocatechin gallate to tetranectin was confirmed by intrinsic fluorescence quenching assays and isothermal titration calorimetry. Furthermore, epigallocatechin gallate efficiently inhibited the activity of tetranectin to enhance the activation of plasminogen. We also found that tetranectin enhanced the proliferation of CT-26 colon cancer cells. Epigallocatechin gallate showed its cytotoxic effect on CT-26 cells due to its binding to tetranectin and the consequent suppression of the cell proliferation. These results demonstrate that the anticancer effect of epigallocatechin gallate is mediated, at least in part, by inhibiting tetranectin as a binding target.

Maternal psychiatric status and infant wheezing: The role of maternal hormones and cord blood cytokines.

RATIONALE: Maternal psychosocial stress might be associated with development of allergic diseases in the offspring. OBJECTIVES: To evaluate the association of maternal depression and anxiety with ever wheezing and recurrent wheezing among infants and to assess the role of maternal hypothalamo-pituatary-adrenal axis changes and fetal immune response in this association. METHODS: This study encompasses two designs; cohort design was developed to evaluate the association of prenatal depression with development of wheezing in infants while nested case-control design was used to assess the role of maternal cortisol and tetranectin and cord blood interleukin 13 and interferon γ. RESULTS: We enrolled 697 pregnant women. Elementary school graduate mother (odds ratio [OR] = 1.5, p = .06), maternal smoking during pregnancy (OR = 3.4, p = .001), familial history of asthma (OR = 2.7, p < .001) increased the risk of ever wheezing. Elementary school graduate mother (OR = 2.6, p = .002), maternal smoking during pregnancy (OR = 4.8, p < .001) and familial history of asthma (OR = 1.7, p = .01) increased the risk of recurrent wheezing. Maternal previous psychiatric disease, or Edinburgh Postnatal Depression Scale or Spielberger State-Trait Anxiety Inventory scores were not associated with wheezing. Maternal tetranectin levels were significantly higher among never wheezers compared to the ever wheezers (264.3 ± 274.8 vs. 201.6 ± 299.7, p = .04). CONCLUSIONS: In conclusion, the major risk factors for ever wheezing and recurrent wheezing were maternal smoking, level of education and family history of asthma. However, maternal depression and anxiety were not determined as risk factors for wheezing. Maternal tetranectin carries potential as a biomarker for wheezing in the infant.

Docosahexaenoic Acid Suppresses Expression of Adipogenic Tetranectin through Sterol Regulatory Element-Binding Protein and Forkhead Box O Protein in Pigs.

Tetranectin (TN), a plasminogen-binding protein originally involved in fibrinolysis and bone formation, was later identified as a secreted adipokine from human and rat adipocytes and positively correlated with adipogenesis and lipid metabolism in adipocytes. To elucidate the nutritional regulation of adipogenic TN from diets containing different sources of fatty acids (saturated, n-6, n-3) in adipocytes, we cloned the coding region of porcine TN from a cDNA library and analyzed tissue expressions in weaned piglets fed with 2% soybean oil (SB, enriched in n-6 fatty acids), docosahexaenoic acid oil (DHA, an n-3 fatty acid) or beef tallow (BT, enriched in saturated and n-9 fatty acids) for 30 d. Compared with tissues in the BT- or SB-fed group, expression of TN was reduced in the adipose, liver and lung tissues from the DHA-fed group, accompanied with lowered plasma levels of triglycerides and cholesterols. This in vivo reduction was also confirmed in porcine primary differentiated adipocytes supplemented with DHA in vitro. Then, promoter analysis was performed. A 1956-bp putative porcine TN promoter was cloned and transcription binding sites for sterol regulatory-element binding protein (SREBP)-1c or forkhead box O proteins (FoxO) were predicted on the TN promoter. Mutating binding sites on porcine TN promoters showed that transcriptional suppression of TN by DHA on promoter activity was dependent on specific response elements for SREBP-1c or FoxO. The inhibited luciferase promoter activity by DHA on the TN promoter coincides with reduced gene expression of TN, SREBP-1c, and FoxO1 in human embryonic kidney HEK293T cells supplemented with DHA. To conclude, our current study demonstrated that the adipogenic TN was negatively regulated by nutritional modulation of DHA both in pigs in vivo and in humans/pigs in vitro. The transcriptional suppression by DHA on TN expression was partly through SREBP-1c or FoxO. Therefore, down-regulation of adipogenic tetranectin associated with fibrinolysis and adipogenesis may contribute to the beneficial effects of DHA on ameliorating obesity-induced metabolic syndromes such as atherosclerosis and adipose dysfunctions.

Identification of tetranectin as a potential biomarker for metastatic oral cancer.

Lymph node involvement is the most important predictor of survival rates in patients with oral squamous cell carcinoma (OSCC). A biomarker that can indicate lymph node metastasis would be valuable to classify patients with OSCC for optimal treatment. In this study, we have performed a serum proteomic analysis of OSCC using 2-D gel electrophoresis and liquid chromatography/tandem mass spectrometry. One of the down-regulated proteins in OSCC was identified as tetranectin, which is a protein encoded by the CLEC3B gene (C-type lectin domain family 3, member B). We further tested the protein level in serum and saliva from patients with lymph-node metastatic and primary OSCC. Tetranectin was found significantly under-expressed in both serum and saliva of metastatic OSCC compared to primary OSCC. Our results suggest that serum or saliva tetranectin may serve as a potential biomarker for metastatic OSCC. Other candidate serum biomarkers for OSCC included superoxide dismutase, ficolin 2, CD-5 antigen-like protein, RalA binding protein 1, plasma retinol-binding protein and transthyretin. Their clinical utility for OSCC detection remains to be further tested in cancer patients.

A rapid, early detection of oral squamous cell carcinoma: Real time PCR based detection of tetranectin.

The current study is focused on determining the mRNA expression levels of tetranectin, to detect oral squamous cell carcinoma (OSCC) and thus aiding in its classification at an early stage. RNA was isolated and cDNA synthesis was performed from the saliva samples of the patients and healthy individuals. A semiquantitative PCR based analysis was performed prior to quantitative and expression based analysis using Real time PCR. The study showed that the mRNA levels are much lesser in patients suffering from dysplastic and metastatic tumors as compared to healthy individuals (P≤0.05). This study can be a breakthrough in medical and dentistry studies. One of the most common malignant carcinomas, OSCC is a type of cancer of the mouth. Though surgical methods have been quite effective in delaying the metastasis, the detection methods using histology parameters are not very efficient and the disease is diagnosed generally in the last stages of the cancer. Tetranectin is a protein biomarker which has been used for detection of several cancers including oral cancer where the protein quantity is calculated.

Exogenous Tetranectin Protects Against 1-Methyl-4-Phenylpyridine-Induced Neurotoxicity by Inhibiting Apoptosis and Autophagy Through Ribosomal Protein S6 Kinase Beta-1.

BACKGROUND: Tetranectin is a secreted homotrimeric protein belonging to the C-type lectin family. Our previous studies found that tetranectin was not only related to, but also played a protective role in, Parkinson disease. In this study, we aim to illustrate the molecular mechanism of the secreted tetranectin. METHODS: We used exogenous tetranectin to investigate the function and molecular mechanism of secreted tetranectin in a 1-methyl-4-phenylpyridine (MPP+)-induced SH-SY5Y cell model. Cell viability and reactive oxygen species were measured to assess the protective effects of tetranectin against MPP+. Apoptosis was measured in several aspects, including Bcl-2/Bax expression, caspase-3/7 activity, annexin V staining, and nuclear morphology. Autophagy was measured as LC3 expression and autophagy flux. Moreover, we used cell immunofluorescence to detect the transport of tetranectin. Western blotting was performed to measure the phosphorylation level of ribosomal protein S6 kinase beta-1 (p70S6K1), and co-immunoprecipitation was applied to confirm the interaction between tetranectin and p70S6K1. RESULTS: The data showed exogenous tetranectin alleviated MPP+-induced toxicity, high reactive oxygen species levels, apoptosis, and autophagy and changed the phosphorylation level of p70S6K1. Immunofluorescence images suggested exogenous tetranectin could be taken into SH-SY5Y cells, and the co-immunoprecipitation experiment indicated tetranectin interacted with p70S6K1. CONCLUSIONS: Exogenous tetranectin protects against MPP+-induced neurotoxicity by promoting p70S6K1 phosphorylation once taken into SH-SY5Y cells.

High Intratumoral Expression of Tetranectin Associates with Poor Prognosis of Patients with Gastric Cancer after Gastrectomy.

Tetranectin, encoded by the clec3b gene, is a plasminogen kringle-4 binding protein that can be detected in the plasma and the extracellular matrix. In malignancies, tetranectin is thought to enhance proteolytic processes enabling tumor cells to invade and metastasize. Nevertheless, the prognostic value of tetranectin in gastric cancer remains elusive. In this study, we found the expression of tetranectin was decreased in gastric cancer. High intratumoral tetranectin level was positively associated with tumor invasion (P = 0.013), lymph node metastasis (P = 0.005), advanced TNM stage (P = 0.003) and shorter overall survival (OS) (P < 0.001) for patients with gastric cancer. Tetranectin expression was identified as an independent prognostic factor for poor OS, and combining tetranectin expression with other independent prognostic factors generated a predictive nomogram, which showed better prognostic efficiency for OS in patients with gastric cancer. In summary, our study suggests that intratumoral tetranectin is a potential independent unfavorable prognostic biomarker for OS of patients with gastric cancer after gastrectomy.

Tetranectin positive expression in tumour tissue leads to longer survival in Danish women with ovarian cancer. Results from the 'Malova' ovarian cancer study.

The primary objective of this study was to analyse Tetranectin (TN) expression in tumour tissues and TN serum concentration in 758 women with epithelial ovarian tumours. The second was to evaluate, whether TN tissue expression levels correlate with clinico-pathological parameters and prognosis of the disease. Using tissue arrays we analysed the expression levels in tissues from 166 women with borderline ovarian tumours (BOTs) and 592 women with ovarian cancer (OC). A panel of three antibodies was used for immunohistochemistry: a polyclonal and two monoclonal antibodies. Serum TN was measured using the polyclonal antibody A-371. Univariate survival analyses stratified for chemotherapy showed that positive tissue TN as demonstrated by the polyclonal antibody indicated a significantly longer overall survival (OS) (p = 0.0001) as well as cancer specific survival (CSS) (p < 0.0001). High serum TN was likewise found to imply longer OS (p < 0.0001) and CSS (p < 0.0001), whereas tissue staining with the two monoclonal antibodies failed to demonstrate any significant correlation with either survival type. Univariate Kaplan-Meier survival analysis performed on all OC cases showed a significantly longer OS (p = 0.0009) and CSS (p = 0.0006) for women with TN positive tumour tissue and in women with high serum TN levels (p < 0.0001 for both). However, in the multivariate Cox regression analysis, only serum TN was found to be an independent prognostic factor for OS (p = 0.01) and not for CSS (p = 0.08). In conclusion, our results predict that a positive TN expression of both tumour tissue and serum points to a more favourable outcome for OC patients.

Comparative analysis of serum proteomes: Identification of proteins associated with sciatica due to lumbar intervertebral disc herniation.

Lumbar intervertebral disc herniation (LDH) is one of the most common orthopedic conditions that can cause lower back pain and sciatica. However, the pathogenesis of LDH is poorly understood. The aim of the present study was to use proteomic analysis of blood samples to establish whether there are serum proteins associated with LDH, which may be useful in elucidating LDH pathogenesis. The ultimate aim was to develop a simple technique for the diagnosis of LDH based on the blood samples of patients with sciatica. The study used comparative analysis of serum proteomes associated with sciatica due to LDH. A total of 30 LDH patients with sciatica, receiving treatment between August and December 2007, were selected as the experimental group (or LDH group). A total of 2 ml of blood was obtained from each of the 30 patients in the LDH group and from 30 healthy volunteers, who constituted the control group. Two-dimensional electrophoresis of the blood samples was conducted, distinct protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and proteins associated with LDH were detected. An enzyme-linked immunosorbent assay (ELISA) was developed to screen for the LDH proteins and was tested on the sera of a second test and control group that included 10 patients with LDH and 10 healthy subjects, respectively. Based on signal intensity, the expression levels of 6 proteins on the dielectrophoretogram were found to be significantly associated with LDH. The identities of the LDH proteins were upregulated apolipoprotein-L1 (APO-L1) and two types of serum albumin precursors, and downregulated apolipoprotein M (APO-M), tetranectin (TN) and immunoglobulin light chain (IGL). Further ELISA experiments confirmed that there were increased serum levels of 4 out of the 6 proteins in patients with sciatica due to LDH, which was statistically different compared to the healthy subjects. In conclusion, these results suggest that serum APO-L1, TN, APO-M and IGL may serve as LDH biomarkers.