Ang-2 Levels Research Articles

Serum levels of angiopoietin 2 mRNA in the mortality outcome prediction of septic shock.

It has been reported that angiopoietin 2 (Ang-2) plays an integral role in the pathophysiology of sepsis and many other inflammatory diseases. However, the specific role of Ang-2 in septic shock has not been defined. The aim of the present study was to assess the predictive value of serum Ang-2 in patients with septic shock. Clinical data of 85 patients with septic shock and 10 healthy controls admitted to the intensive care unit with a diagnosis of septic shock were collected between January 2020 and October 2020 at Tongji Hospital (Wuhan, China). The serum levels of Ang-2 mRNA were quantified using a quantitative real-time PCR assay. Ang-2, SOFA and APACHE II scores were retrospectively analyzed in relation to 28-day mortality. The area under the receiver operating characteristic (ROC) curve (AUC) was used to discriminate the accuracy of the prediction. Mean Ang-2 mRNA levels in the patients with septic shock were significantly higher than those in the healthy controls (P<0.05), and the Ang-2 levels showed a downwards trend over time following treatment. The three indicators (AUCs, SEMs, P-values) were Ang-2 (0.82, 0.03, P<0.01), SOFA score (0.76, 0.04, P<0.01), and APACHE II score (0.73, 0.04, P<0.01). The present study confirmed that Ang-2 mRNA levels were significantly elevated in septic shock. The Ang-2 mRNA level at ICU admission in a patient with septic shock could be a predictive biomarker for mortality.

Inhibitory Potential of Shen-Shuai-Ling Formulation on Renal Interstitial Fibrosis via Upregulation of PLZF.

Background Renal interstitial fibrosis (RIF) is an important cause of kidney disease, which seriously affects people's health. As a traditional Chinese medicine, Shen-Shuai-Ling Formulation (SSLF) has obvious kidney function. However, the therapeutic effect of SSLF on RIF and its molecular mechanism are still unclear. Methods First, the potential targets and pathways of SSLF for RIF were predicted by network pharmacology, and then, the binding of luteolin and target protein to SSLF was verified by molecular docking and Co-IP experiments. Finally, the effects of SSLF and luteolin on PLZF and (Pro) renin receptor (PRR) were verified by western blot and qPCR experiments. Angiotensin (Ang)-1, Ang-2, and transforming growth factor-β (TGF-β) were the indexes of renal interstitial fibrosis. Results Through the drug-active component-target network diagram, we found that luteolin has the most connections, and promyelocytic leukemia zinc finger (PLZF) is the target protein. GO analysis and KEGG pathway analysis of targets were performed using Cytoscape ClueGO. Molecular docking experiments and Co-IP are used to prove that luteolin and PLZF can be combined. Western blot and qPCR results showed that both SSLF and luteolin significantly upregulated the expression of PLZF and decreased the levels of PRR, Ang-1, Ang-2, and TGF-β. The overexpression of PLZF decreased the expression of PRR, the knockdown of PLZF increased the expression of PRR, and the overexpression of PRR decreased the expression of Ang-1, Ang-2, and TGF-β. Conclusions SSLF inhibits PRR and renal interstitial fibers by the upregulation of PLZF levels.

Early modulation of Angiopoietin-2 plasma levels predicts benefit from regorafenib in patients with metastatic colorectal cancer

BackgroundNo biomarkers are currently available to predict the efficacy of trifluridine/tipiracil (FTD/TPI) in chemorefractory metastatic colorectal cancer. The multicohort REGOLAND study aims at exploring and validating circulating markers potentially able to predict benefit from regorafenib in this setting. Material and methodsIn the retrospective ‘regorafenib exploratory cohort’, including 105 patients treated with regorafenib, baseline (d1) plasma levels of angiogenesis-related biomarkers and their early modulation after 15 days (d15) of treatment were investigated for correlation with clinical outcome. Based on a pre-specified statistical hypothesis, main retrospective findings were prospectively challenged in the ‘regorafenib validation cohort’, including 100 patients treated with regorafenib. Prospectively validated putative biomarkers were then assessed in the control ‘FTD/TPI cohort’, including 93 patients treated with FTD/TPI. ResultsIn the ‘regorafenib exploratory cohort’, the early (d15) increase of Angiopoietin-2 (Ang-2) was associated with longer progression-free survival (HR:0.57 [95%CI:0.38–0.88], P = 0.004) and a trend towards longer OS (HR:0.74 [95%CI:0.48–1.14], P = 0.165), than the early decrease. Similar results were prospectively confirmed in the ‘regorafenib validation cohort’ (HR for progression-free survival:0.72 [95%CI:0.48–1.08], P = 0.095; HR for OS:0.77 [95%CI:0.51–1.16], P = 0.204). No predictive impact was shown for the early modulation of Ang-2 in the ‘FTD/TPI cohort’. High baseline Ang-2 levels predict poor prognosis in all the investigated cohorts, independently of other clinical prognostic variables. ConclusionsThe early modulation of circulating Ang-2 predicts the efficacy of regorafenib. Baseline Ang-2 plasma levels are an independent prognostic biomarker in chemorefractory metastatic colorectal cancer.

The Role of Angiopoietin-2 on Plasma Leakage in the Critical Phase and Convalescence of Dengue Virus Infection in Children

Abstract&#x0D; Background: The worst complication of dengue haemorrhagic fever (DHF) that lead to death is hypovolemic shock due to plasma leakage from intravascular to extravascular space caused by endothelial dysfunction. This study is aimed to analyze the difference of Ang-2 level in dengue haemorrhagic fever patients related to plasma leakage between critical phase and convalescence.&#x0D; &#x0D; Aim: One hundred and ten children with DHF confirmed serologically with positive IgM anti-dengue or IgM and IgG anti-dengue were recruited in this study. We examine the clinical manifestations, laboratory parameters (platelet count, AST and ALT levels), and molecular test (Ang-2). The molecular indicator was tested by Enzyme-Linked Immunosorbent Assay (ELISA). Data were collected between July – November 2019. This is an observational study with cross-sectional comparative design.&#x0D; &#x0D; Results: In our study, we found correlation of clinical symptoms (abdominal pain, persistent vomit, and hepatomegaly) with the occurrence of plasma leakage in DHF patients. We also found that Ang-2 level is significantly higher in children with DHF during critical phase compared to convalescence (p&lt; 0.01).&#x0D; &#x0D; Conclusion: There is a significant difference of Ang-2 level in DHF patients during critical phase and convalescence.&#x0D; &#x0D; Keywords: Dengue hemorrhagic fever, children, Ang-2, critical phase, convalescence

Association of Pre-Transplant Angiopoietin-2 Index with the Risk of Acute Graft-Versus-Host Disease after Hematopoietic Stem Cell Transplantation

Objective:Angiogenic factors (AFs) released under endothelial stress are reflective of tissue healing, while some may also contribute to tissue damage/inflammation. We investigated whether alterations in the pre-transplant levels of AFs were associated with the risk of acute graft-versus-host disease (aGvHD).Materials and Methods:The pre-conditioning plasma levels of angiopoietin-2 (Ang2), endoglin, and follistatin were measured for 37 patients together with inflammatory markers. The index values defined were evaluated to better identify the alterations.Results:The patients had higher pre-conditioning levels of Ang2, endoglin, and follistatin compared to controls. The patients with aGvHD had higher Ang2 index and lower albumin index scores in comparison to those without aGvHD. Multivariate analysis revealed that the pre-transplant Ang2 index was an independent risk factor for aGvHD development.Conclusion:Pre-transplant evaluation of plasma Ang2 levels along with inflammatory status even before conditioning is associated with endothelial vulnerability. The pre-transplant Ang2 index could be a promising candidate to estimate the risk of aGvHD.

Clinical significance of differential serum-signatures for early prediction of severe dengue among Eastern Indian patients.

Dengue infection can result in simple dengue fever or life-threatening severe dengue. Early identification of severe patients is needed for proper disease management. Dengue infection was screened among 168 symptomatic patients by qRT-PCR, anti-dengue IgM, and IgG ELISA. Dengue patients were categorized according to WHO classification. Viral load and dengue serotypes were determined by qRT-PCR. Levels of acute-phase-proteins (SAP, SAA2; CRP and ApoA1), endothelial (Ang2, VEGF), coagulation (fibrinogen) markers were determined by sandwich ELISA/immunoturbidimetry/western-blotting. Hepatic (ALT, AST, ALP) and other blood biochemical parameters were studied by autoanalyzer and haematology cell counter. Statistical analysis and protein-protein-interaction network were performed by GraphPad-Prism and STRINGS database, respectively. Among 87 dengue patients, significantly higher levels of Ang2, VEGF, CRP, SAA2, ApoA1, AST, ALT, and AST/ALT ratio and low level of fibrinogen were detected in severe-dengue cases compared to dengue without warning-signs, with seven of them severely altered during febrile-phase. Higher fold-change of Ang2 and VEGF as well as decreased fibrinogen were observed among patients with haemorrhagic-manifestation, clinical-fluid accumulation and thrombocytopenia. Functional network analysis predicted Ang2, VEGF, and CRP to be functionally and physically connected and SAA2 and ApoA1 to be functioning together. Correlation analyses also validated this connectivity by a strong positive correlation between Ang2, VEGF, and CRP. PCA analysis followed by hierarchical clustering heatmap analysis segregated severe-dengue patients from the rest, with VEGF, Ang2, ApoA1, AST, and ALT clearly distinguishing the severe-dengue group. Thus, serum levels of VEGF, Ang2, ApoA1, AST, and ALT might act as potential biomarkers for predicting dengue severity during the early stage.

Abstract 163: Potential Therapeutic Effects Of Angiopoietin-2 Inhibition On Rescuing Hereditary Hemorrhagic Telangiectasia Associated Cerebrovascular Defects

Introduction: Hereditary Hemorrhagic Telangiectasia (HHT) is a vascular disease characterized by arteriovenous malformations (AVMs), which are abnormal, enlarged connections between arteries and veins that are prone to hemorrhage. AVMs often form in the brain and can lead to stroke. Mutations in endoglin (Eng), activin receptor-like kinase 1 (Alk1) or smad-related protein 4 (Smad4) cause HHT. Endothelial cell (EC) specific knock out of Alk1, Eng, or Smad4 in mice mimics HHT vascular defects, including AVM formation. We showed that Smad4 directly repressed transcription of the critical pro-angiogenic factor Angiopoietin-2 (Ang2) in ECs and that in Smad4-HHT mice, Ang2 /ANG2 levels are robustly and ectopically increased in ECs. By inhibiting Ang2, we prevented and rescued AVMs and normalized retinal vasculature. Since Smad4 is a central transcription factor in the TGF-β signaling pathway and a downstream effector of the Eng and Alk1 co-receptors, we hypothesized that Ang2 inhibition may universally normalize HHT vascular phenotypes. Methods: Blue latex dye perfusion methods were used to visualize and characterize cerebrovascular defects in HHT mouse models (n=5~7). RNA-seq was performed on isolated brain ECs (iBECs) to further define molecular phenotypic characteristics (n=5). In situ hybridization on brain slices (n=3) and ELISA on whole brain lysate (n=3~9) were performed to confirm elevated Ang2 RNA and protein levels. Ang2 inhibitor was delivered via intraperitoneal injection, followed by cerebrovascular assessments (n=5~9). Results: We found that brain AVMs are present in Smad4-, Alk1-, and Eng-HHT mutants, with all models displaying elevated Ang2 levels. RNA-seq of iBECs in Smad4-, Alk1-, and Eng-HHT models reveals 5502, 5307, and 1659 differentially expressed genes in respective mutants compared to controls. Data set comparisons revealed common, increased pro-angiogenic genes, which helped to define the HHT-specific angiogenic profile. Moreover, Ang2 inhibition decreased Smad4-HHT’s brain vessel diameter and Alk1-HHT’s brain vessel density, and prevented brain AVM formation in Eng-HHT mice. In conclusion, Ang2 inhibition partially normalized cerebrovascular defects in Smad4- and Alk1-HHT mice, and completely in Eng-HHT mice.

Angiogenic factors as prognostic markers in neuroendocrine neoplasms

Angiogenic markers in neuroendocrine neoplasms (NENs) have recently received increasing attention, but their clinical role remains unclear. The aim of this study was to evaluate the role of angiogenic markers in NEN aggressiveness and prognosis. We performed a prospective observational study including 46 consecutive patients with proven NENs of pulmonary (45.65%) and gastro-entero-pancreatic (GEP) (54.35%) origin and 29 healthy controls. Circulating pro-angiogenic factors were measured by ELISA assay. ANG2 tissue expression was evaluated in a subgroup of ten patients by immunohistochemistry. The study demonstrated a significantly higher level of ANG2, ANG1, sTIE2, and PROK2 in patients affected by NENs compared to controls. In the NENs' group we measured that: (i) ANG2 levels were higher in poorly vs well-differentiated NENs: 4.85 (2.75-7.42) vs 3.16 (1.66-6.36) ng/ml, p = 0.046 and in tumor stage 3-4 compared to stage 1-2: 4.24 (2.66-8.72) vs 2.73 (1.53-5.70), p = 0.044; (ii) ANG2 and PROK2 were significantly higher in patents with progressive disease compared to stable disease: ANG2 = 6.26 (3.98-10.99) vs 2.73 (1.65-4.36) pg/ml, p = 0.001; PROK2 = 29.19 (28.42-32.25) vs 28.37 (28.14-28.91) pg/ml, p = 0.035. Immunohistochemistry confirmed ANG2 expression in tumor specimens. We documented higher levels of angiogenic markers in NENs, with an association between ANG2 serum levels and NENs morphology and staging. In both GEP and lung NENs, ANG2 and PROK2 are higher in case of tumor progression, suggesting a potential role as prognostic markers in NENs patients.

ENAMPT neutralization reduces preclinical ARDS severity via rectified NFkB and Akt/mTORC2 signaling

Despite encouraging preclinical data, therapies to reduce ARDS mortality remains a globally unmet need, including during the COVID-19 pandemic. We previously identified extracellular nicotinamide phosphoribosyltransferase (eNAMPT) as a novel damage-associated molecular pattern protein (DAMP) via TLR4 ligation which regulates inflammatory cascade activation. eNAMPT is tightly linked to human ARDS by biomarker and genotyping studies in ARDS subjects. We now hypothesize that an eNAMPT-neutralizing mAb will significantly reduce the severity of ARDS lung inflammatory lung injury in diverse preclinical rat and porcine models. Sprague Dawley rats received eNAMPT mAb intravenously following exposure to intratracheal lipopolysaccharide (LPS) or to a traumatic blast (125 kPa) but prior to initiation of ventilator-induced lung injury (VILI) (4 h). Yucatan minipigs received intravenous eNAMPT mAb 2 h after initiation of septic shock and VILI (12 h). Each rat/porcine ARDS/VILI model was strongly associated with evidence of severe inflammatory lung injury with NFkB pathway activation and marked dysregulation of the Akt/mTORC2 signaling pathway. eNAMPT neutralization dramatically reduced inflammatory indices and the severity of lung injury in each rat/porcine ARDS/VILI model (~ 50% reduction) including reduction in serum lactate, and plasma levels of eNAMPT, IL-6, TNFα and Ang-2. The eNAMPT mAb further rectified NFkB pathway activation and preserved the Akt/mTORC2 signaling pathway. These results strongly support targeting the eNAMPT/TLR4 inflammatory pathway as a potential ARDS strategy to reduce inflammatory lung injury and ARDS mortality.

Plasma Angiopoietin-2 as a Promising Biomarker for the Prognosis of Acute Pulmonary Embolism.

Endothelial damage is one of the pathogenic conditions of acute pulmonary embolism (APE). The essential role of angiogenin, ribonuclease A family, member 2 (Ang-2) in APE remains unclear. This study aimed to investigate the predictive value of Ang-2 in the clinical outcomes of patients with APE. Plasma Ang-2 levels were measured by an enzyme-linked immunosorbent assay kit using a DuoSet methodology in 118 APE patients and 53 healthy controls. Baseline data relevant to mortality over time were obtained from hospital databases or by patient's follow-up (median follow-up time: 25.0 ± 13.2 months). The main outcome was all-cause mortality. Plasma Ang-2 level was significantly higher in APE patients than in healthy controls (p < 0.001). Patients dying during the first 30 days presented higher baseline levels of Ang-2 than the survivors (p < 0.001). Patients dying during the follow-up also showed higher baseline levels of Ang-2 than the survivors (p < 0.001). The multi-variable logistic regression analysis showed that the N-terminal propeptide of B-type natriuretic peptide (NT-proBNP) [odds ratio (OR): 19.8; 95% CI: 1.5 - 255.8; p = 0.022] and Ang-2 (OR: 9.9; 95% CI: 1.4 - 70.5; p = 0.022) emerged as independent predictors of the 30-day mortality. Furthermore, the multivariable Cox's regression identified plasma Ang-2 [hazards ratio (HR): 1.35; 95% CI: 1.10 - 1.66; p = 0.004] as an independent predictor of long-term mortality in patients with APE. A high circulating level of Ang-2 can be considered as an independent predictor for the poor outcome of APE and may serve as a biomarker for the risk stratification in patients with APE.

DYNAMICS OF FIBROTIC CHANGES IN THE LIVER AFTER THE SUCCESSFUL ERADICATION OF HEPATITIS C VIRUS IN PATIENTS WITH NAFLD.

The aim: To assess the dynamics of serum levels of angiopoietin-2 and transforming growth factor-β1 in patients with chronic hepatitis C (CHC) with concomitant nonalcoholic fatty liver disease (NAFLD) after successful DAAs. Materials and methods: 82 patients with CHC were examined, of which 56 were diagnosed with NAFLD and increased body weight. Ang-2, TGF-β1, leptin, adiponectin, and the degree of liver fibrosis were determined for all participants. The patients were divided into groups: 1 gr. (n=23) - CHС + increased body weight + hepatic steatosis, 2 gr. (n=33) - СНС + increased body weight + nonalcoholic steatohepatitis, 3rd gr. (n=26) - CHC. All patients received DAAs for 12 weeks. Results: From 82 patients F3-4 had 31 people, F1-2 - 25, F0-1 - 11, F0 -15 patients. F3-4 and steatosis S2-3 (p<0.05) was more common in patients of 2 gr.. Serum Ang-2 levels were higher (p<0.05) in patients of 2 gr. with F3-4, than in patients with F0-2. Fibrosis regression occurred more often in patients with 1 and 3 gr. with F1-2 than in patients 2 gr. and F3-4 and was accompanied by a decrease in Ang-2 and TGF-β1 levels. Conclusions: High levels of Ang-2 and TGF-β1 are registered in patients with CHC+NAFLD, which correlate with the degree of liver fibrosis and significantly decrease after successful DAAs in patients with low initial stages of liver fibrosis and normal body weight.

Impact of Hypertension and Physical Fitness on SARS-COV-2 and Related Consequences. (Possible Mechanisms with Focusing on ACE2).

Hypertension disease as an absolute risk factor of Covid-19 disease has been well-proven in recent evidence. The factors such as the use of antihypertensive drugs, protein expression, and compensatory axes resulted in hypertension disease playing very important roles in the occurrence of this problem. In this review study, we first attempted to investigate the higher chance reason for Covid-19 disease in people with high blood pressure; then we examined the related mechanisms, and finally, we reported the differences and similarities between people with high blood pressure and athletes. All in all, we concluded that people who exercise regularly, the same as hypertensive patients (Compensatory mechanism) are more susceptible to COVID-19 infection due to the high concentration of ACE2 (Physiological mechanism) caused by exercise adaptation, but for the low level of ANG2 (Systematic and gene expression) these individuals (Active subjects) indicate fewer complications and severity symptoms of COVID-19 such as dyspnea, hospitalization and, heart disease compared with hypertensive patients.

The Dual Role of ANG-2 in Pancreatic Cancer

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is well-known for its inflammatory nature. Inflammation leads to secretion of Angiopoietin-2 (Ang-2), which works in vascular remodeling. Increased Ang-2 can be detected in endothelial cells and in the circulation in various cancers. High circulating Ang-2 is a marker for shorter survival in PDAC. Different from other tumours, in addition to the endothelial expression, PDAC cells themselves express Ang-2. The correlation between tumour cell Ang-2 expression and survival is still unknown, and the aim of this study was to look into this relationship. Method: Ang-2 immunohistochemistry (IHC) staining was done on 158 PDAC patient samples and staining intensity scored by two examiners (range: negative (0) to strongly positive (3)). We measured circulating Ang-2 levels in sera of 196 patients with ELISA (DuoSet methodology). The results were assessed with respect to clinical data and patient outcomes. Results: Neither circulating Ang-2 nor tumour expression was affected by patient characteristics. Disease specific survival (DSS) was shorter in patients with a negative to weak tumour Ang-2 expression (0.95 years, IQR 1.99), when compared to those with an intermediate to high expression (1.84 years, IQR 2.73, p<0.017). In contrast, higher than median (≥2.72ng/ml) circulating Ang-2 associated with shorter DSS (1.49 years, IQR 2.02) when compared to patients with low circulating Ang-2 (2.17years, IQR 2.49, p=0.002). Conclusion: Ang-2 has a dual role in PDAC with respect to survival. Further studies are needed to find the mechanisms that cause tumour cell Ang-2 expression and its beneficial association with respect to survival.

Up-regulated miR-204-5p promoted the migration, invasion, and angiogenesis of endothelial progenitor cells to enhance the thrombolysis of rats with deep venous thrombosis by targeting SPRED1

Deep venous thrombosis (DVT) endangers human health. Endothelial progenitor cells (EPCs) were proven to promote thrombolysis and miR-204-5p was discovered to be low-expressed in DVT patients. This study concentrated on exploring whether miR-204-5p had a regulatory effect on EPCs and DVT. Concretely, the expression of miR-204-5p in DVT patients’ blood was detected by qRT-PCR. The target of miR-204-5p was predicted by bioinformatics and verified by dual-luciferase reporter assay. After rat EPCs were isolated, identified, and transfected with miR-204-5p agomiR, antagomiR, or SPRED1 plasmids, the viability, migration, invasion, and tube formation of EPCs were detected by MTT, wound healing, Transwell, and tube formation assays, respectively. MiR-204-5p, SPRED1, p-PI3K, PI3K, p-AKT, AKT, VEGFA, and Ang1 expressions in EPCs were measured by qRT-PCR or Western blot. EPCs transfected with miR-204-5p overexpression lentivirus plasmid were injected into the DVT rat model. The histopathology of the thrombus and the homing of EPCs to thrombus in the DVT rats were observed by hematoxylin-eosin staining and confocal microscopy, respectively. We found that miR-204-5p was low-expressed in DVT patients and SPRED1 was a target gene of miR-204-5p. MiR-204-5p agomiR promoted the viability, migration, invasion, and tube formation of EPCs, the levels of VEGFA and Ang1 and the activation of PI3K/AKT pathway in EPCs, while miR-204-5p antagomiR and SPRED1 worked oppositely. SPRED1 reversed the effect of miR-204-5p agomiR on EPCs. Up-regulated miR-204-5p inhibited thrombosis and promoted EPCs homing to thrombus in DVT rats. Collectively, up-regulated miR-204-5p enhanced the angiogenesis of EPCs and thrombolysis in DVT rats by targeting SPRED1.

A Comparative Study of Serum Angiogenic Biomarkers in Cirrhosis and Hepatocellular Carcinoma

Simple SummaryThe progression of liver disease is accompanied by pathological angiogenesis, a prerequisite for the development of HCC. In this paper, we analyzed the clinical significance of serum angiogenic markers VEGF, Ang-1, Ang-2, angiopoietin receptor Tie1/2, HGF, and PECAM-1 in 62 patients with liver disease, out of which 33 were diagnosed with HCC and 29 with liver cirrhosis without signs of neoplasia. Biomarkers levels were investigated as a function of “Model for End-Stage Liver Disease” (MELD) score and Fibrosis Index (FI). HCC patients showed higher HGF levels than ones with cirrhosis, while high Ang-1 levels appeared to have a protective role in HCC as well as prognostic significance; we also found a strong correlation between HGF levels, Ang-2, and VEGF levels, further supporting their role in tumor angiogenesis. Due to the complexity of angiogenesis and the small size of the study group, further investigations are widely desired especially in the era of immunotherapy and HCC-targeted anti-angiogenic drugs.Background: Hepatocellular carcinoma (HCC) is a global health problem associated with chronic liver disease. Its pathogenesis varies according to the underlying etiological factors, although in most cases it develops from liver cirrhosis. The disease progression is accompanied by pathological angiogenesis, which is a prerequisite that favors the development of HCC. Aims: This study aims at contributing to our understanding of the role of angiogenic factors in the progression of liver disease. For this purpose, we evaluate the clinical significance of serum angiogenic markers (VEGF, Ang-1, Ang-2, the angiopoietin receptor Tie1/2, HGF, and PECAM-1) first in cirrhotic and HCC patients separately, and then comparing cirrhotic patients with and without HCC. Materials and Methods: We enrolled 62 patients, out of whom 33 were diagnosed with HCC and 29 with liver cirrhosis without signs of neoplasia. Patients underwent venous blood sampling before and after receiving treatments for the diagnosed disease. Serum markers were evaluated using ELISA assays for Tie1 and the Bio-Plex Multiplex system for the remaining ones. Biomarker levels were investigated as a function of clinical scores for disease staging (MELD and Fibrosis Index, FI). Results: In cirrhotic patients, Ang-1 and Ang-2 correlate with MELD (ρAng-1 = −0.73, p = 2E−5) and FI (ρAng-1 = −0.52, p = 7E−3, ρAng-2 = 0.53, p = 3E−3). A reduction of Ang-2 levels (p = 0.047) and of the Ang-2/Ang-1 ratio (p = 0.031) is observed in cirrhotic patients diagnosed with viral hepatitis after antiviral treatments. In HCC patients, Ang-1 negatively correlates with FI (ρ = −0.63, p = 1E−4), and PECAM-1 positively correlates with MELD (ρ = 0.44, p = 0.01). A significant Ang-1 reduction was observed in deceased patients during the study compared to ones who survived (p = 0.01). In HCC patients, VEGF levels were increased after tumor treatment (p = 0.037). Notably, HGF levels in cirrhotic patients with HCC are significantly raised (p = 0.017) compared to that in those without HCC. Conclusions: Our results suggest that serum angiogenic markers, with emphasis on Ang-1/2, can contribute to the development of quantitative tools for liver disease staging and therapy monitoring. The comparison between cirrhotic patients with and without HCC suggests that HGF levels are potentially useful for monitoring the insurgence of HCC after a cirrhosis diagnosis. High Ang-1 levels in HCC patients appear to have a protective role as well as prognostic significance.

313. Host Protein Biomarkers Predicting Severity of Lung Damage due to COVID-19

BackgroundBiomarkers to predict the severity of lung damage due to COVID-19 are urgently needed to inform management and treatment decisions. Our objective was to investigate the predictive value of host proteins for worsening respiratory failure in one of the by COVID-19 most affected and diverse patient populations in the US.MethodsWe performed a prospective single-center cross-sectional study of 34 adult patients admitted to Montefiore Medical Center in the Bronx, New York, for respiratory symptoms due to PCR-confirmed COVID-19. Exclusion criteria were age < 21, history of prior SARS-CoV-2 infection, and/or underlying severe chronic lung diseases requiring home O2 and/or high dose steroids. We stratified and compared patients by whether they developed worsening respiratory failure, necessitating transfer to the intensive care unit (ICU) during their hospital stay. Using a custom Luminex Assay, we measured hospital admission serum concentrations of 8 host proteins, representing respiratory-associated epithelial (RAGE, SP-D, CC16), endothelial (Ang-2, vWF), and immune pathways (S100A12, ICAM-1, VCAM-1).ResultsExcept for race and WHO COVID-19 scores, demographics, co-morbidities, symptoms, and symptom duration were not statistically significantly different between patients requiring transfer to the ICU (n=15) and non-ICU patients (n=19). Higher log-transformed levels for 5/8 proteins (S100A12, ICAM-1, Ang-2, RAGE, SP-D) showed significant or marginally significant increased cause-specific hazard for ICU transfer (n=15). Estimated cumulative incidence functions further showed a significantly or near significantly increased risk for ICU transfer for patients with above the median values of S100A12 or ICAM-1 (p=0.013), Ang-2 (p=0.056) or RAGE (p=0.077), respectively (Figure 1). Host proteins predicting need for ICU transfer did not correlate strongly with other clinical laboratory markers for COVID-19 severity (CRP, LDH, D-Dimer, Fibrinogen, Ferritin).Figure 1. Patients with above median levels of host protein markers S100A12, ICAM-1, Ang-2, and RAGE have a significantly or near significantly increased risk for severe respiratory failure requiring transfer to the ICU. Comparison of estimated cumulative incidence at 7 days post admission for host protein markers above and below median levels for (A) S10012 (median 96,675 pg/ml); (B) ICAM-1 (median (1,192,277 pg/ml); (C) Ang-2 (median 3463 pg/ml); (D) RAGE (median 6356 pg/ml); and (E) SP-D (median 11,832 pg/ml).ConclusionThese results suggest that host proteins have additional predictive value for the severity of COVID-19-associated lung damage at time of presentation to the hospital.Disclosures Inessa Gendlina, Nothing to disclose

Outcomes Based on Plasma Biomarkers for the Phase 3 CELESTIAL Trial of Cabozantinib versus Placebo in Advanced Hepatocellular Carcinoma

Introduction: Cabozantinib, an inhibitor of MET, AXL, and VEGF receptors, significantly improved overall survival (OS) and progression-free survival (PFS) versus placebo in patients with previously treated advanced hepatocellular carcinoma (HCC). In this exploratory analysis, outcomes were evaluated according to plasma biomarker levels. Methods: Baseline plasma levels were evaluated for MET, AXL, VEGFR2, HGF, GAS6, VEGF-A, PlGF, IL-8, EPO, ANG2, IGF-1, VEGF-C, and c-KIT for 674/707 randomized patients; and Week 4 levels were evaluated for MET, AXL, VEGFR2, HGF, GAS6, VEGF-A, PlGF, IL-8, and EPO for 614 patients. OS and PFS were analyzed by baseline levels as dichotomized or continuous variables and by on-treatment changes at Week 4 as continuous variables; biomarkers were considered potentially prognostic if p < 0.05 and predictive if p < 0.05 for the interaction between treatment and the biomarker. Multivariable analyses adjusting for clinical covariates were also performed. Results: In the placebo group, high levels of MET, HGF, GAS6, IL-8, and ANG2 and low levels of IGF-1 were associated with shorter OS in univariate and multivariable analyses; these associations were also observed for MET, IL-8, and ANG2 in the cabozantinib group. Hazard ratios for OS and PFS favored cabozantinib over the placebo at low and high baseline levels for all biomarkers. No baseline biomarkers were predictive of a treatment benefit. Cabozantinib promoted pharmacodynamic changes in several biomarkers, including increases in VEGF-A, PlGF, AXL, and GAS6 levels and decreases in VEGFR2 and HGF levels; these changes were not associated with OS or PFS. Conclusion: Cabozantinib improved OS and PFS versus placebo at high and low baseline concentrations for all biomarkers analyzed. Low baseline levels of MET, HGF, GAS6, IL-8, and ANG2 and high levels of IGF-1 were identified as potential favorable prognostic biomarkers for survival in previously treated advanced HCC. Although cabozantinib promoted pharmacodynamic changes in several biomarkers, these changes were not associated with survival.

Angiopoietin-2 as a predictor of fibrosis regression in chronic hepatitis C virus patients after direct-acting antiviral drugs

BackgroundHCV infection is a major health concern. Disease progression to fibrosis, cirrhosis, and HCC is aided by the persistence of inflammatory reactions and cellular damage. Moreover, angiogenesis was found to have a substantial pathogenic role in disease progression. Serum Angiopoietin-2 appears to be correlated with liver stiffness in chronic HCV and its elevation is linked to disease progression from chronic hepatitis to cirrhosis. The aim of this study was to evaluate the role of serum Angiopoietin-2 in the prediction of regression of fibrosis in chronic HCV patients receiving direct-acting antiviral agents. Forty Egyptian chronic HCV patients for whom direct-acting antiviral agent (DAA) therapy was planned were included. All patients underwent assessment twice, at baseline and at SVR12, for standard laboratory tests, measurement of fibrosis using FibroScan, FIB-4 and APRI scores, and Angiopoietin-2 level.ResultsStatistically significantly higher levels of baseline Ang-2 were detected with the progression of fibrosis stages with a p-value of <0.001. The best cutoff value of baseline Ang-2 in discrimination of liver cirrhosis (F4) from F0-F3 was > 630 pg/ml with 85.71% sensitivity and 84.85% specificity. A statistically significant decline of Ang-2 (from 464.3±237.2 pg/ml to 401.3±277.1 pg/ml) was noted after the achievement of SVR12 with a p-value < 0.001. Regression of liver fibrosis in this study is defined as a decrease of more than or equal to one stage in liver fibrosis. Lower baseline fibrosis stages and other non-invasive scoring systems (FIB-4 and APRI scores) were associated with regression of fibrosis following successful DAAs treatment. However, higher baseline Ang-2 levels were significantly associated with non-regression of fibrosis, and at a cutoff of >680 pg/ml, it might predict non-regression of fibrosis after successful eradication of HCV with DAAs with 93.33% sensitivity and 70% specificity.ConclusionsAngiopoietin-2 can be a useful predictor of fibrosis regression in chronic HCV patients receiving direct-acting antiviral agents. Elevated baseline Angiopoietin-2 and advanced fibrosis stages may predict non-regression of liver fibrosis.

Lactobacillus fermentum ZS09 Mediates Epithelial-Mesenchymal Transition (EMT) by Regulating the Transcriptional Activity of the Wnt/β-Catenin Signalling Pathway to Inhibit Colon Cancer Activity.

ObjectiveThe epithelial–mesenchymal transition (EMT) pathway can mediate tumour migration, and the occurrence of EMT is closely related to the Wnt/β-catenin signalling pathway. The purpose of this paper was to study the effect of Lactobacillus fermentum ZS09 (L. fermentum ZS09) on the EMT pathway in mouse with azoxymethane/dextran sulfate sodium salt (AOM/DSS) induced colon cancer and the potential underlying mechanism.Materials and MethodsIn this study, a mouse colon cancer model was established through intraperitoneal injection of 10 mg/kg azoxymethane (AOM) and three cycles of 2.5% dextran sulfate sodium salt (DSS) in the drinking water. H&E staining, enzyme-linked immunosorbent assay (ELISA), real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting (WB) were used to study the antitumour mechanisms of L. fermentum ZS09 through the EMT pathway.ResultsThe results of this study showed that compared with the model group, the high-dose L. fermentum ZS09 intervention group exhibited decreased serum levels of MMP-9, TNF-α, IL-6R, Ang-2 and VEGFR-2 and increased contents of DKK1 (P<0.05). The expression of Wnt/β-catenin signalling pathway-related genes (Dv1, GSK-3β, β-catenin, c-myc, cyclinD1, Vim, and MMP-9) was significantly reduced, and the gene expression levels of APC, CDH1, and Axin were increased. The levels of related proteins (β-catenin, N-cadherin, and VEGF) were downregulated, and the levels of p-β-catenin and E-cadherin were upregulated.ConclusionThe results indicate that L. fermentum ZS09 could inhibit EMT and angiogenesis pathways by inhibiting the Wnt/β-catenin signalling pathway, which could inhibit tumour metastasis.

The effect of Shendan Sanjie capsule on angiogenesis in mice with colitis associated cancer and mechanism

Objective: To investigate the effects and the mechanism of Shendansanjie capsules on angiogenesis of colitis associated cancer(CAC) mice. Methods: Azoxymethane and dextran sulfact sodium were used to construct a mice model with CAC. Ten mice were divided into the normal group, model group, Shendan Sanjie capsule group, MK-2206 group, and Shendan Sanjie capsule + IGF-1 group, respectively. Immunohistochemistry was used to detect the microvessel density (MVD) in the colon tissue of each group of mice. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the mRNA levels of basic fibroblast growth factor (bFGF) and angiopoietin 2 (Ang2) in colon tissue. Western blot was used to detect the expressions of Akt, p-Akt, vascular endothelial growth factor A (VEGFA), hypoxia-inducible factor-1α (HIF-1α). Results: The number of MVD in the colon tissue of mice in the model group, Shendan Sanjie capsule group, MK-2206 group, Shendan Sanjie capsule + IGF-1 group were 63.3±3.3, 36.6±2.3, 36.6±2.2, 50.3±2.5, significantly higher than 2.0±0.1 in the normal group (P<0.05). The number of MVD in Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group are lower than that in model group (P<0.05), while Shendan Sanjie capsule+ IGF-1 group is higher than Shendan Sanjie Capsule group (P<0.05). The relative expressions of bFGF mRNA in the colon cancer tissue of mice in the model group, Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group were 4.55±0.31, 2.46±0.37, 2.49±0.33, 3.34±0.21, respectively, and the relative mRNA expressions of Ang2 were 5.78±0.19, 2.21±0.14, 2.26±0.17 and 3.67±0.32, respectively, which were significantly higher than 1.01±0.05 and 0.99±0.07 in the normal group (P<0.05). The mRNA levels of bFGF and Ang2 in Shendan Sanjie capsule group, MK-2206 group and Shendan Sanjie capsule+ IGF-1 group were lower than those in the model group (P<0.05), while Shendan Sanjie capsule+ IGF-1 group is higher than Shendan Sanjie capsule group (P<0.05). The relative expression levels of p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissues of the model group were 4.75±0.18, 4.64±0.22 and 4.84±0.12, respectively, which were significantly higher than 1.01±0.07, 0.95± 0.08 and 0.98±0.05 in the normal group (P<0.05). The relative expressions of p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissues in the Shendan Sanjie capsule group were 2.24±0.22, 3.15±0.26 and 2.07±0.18, respectively, which were significantly lower than those in the model group (P<0.05). However, compared with the MK-2206 group, the difference was not statistically significant (P>0.05). The relative expression levels p-Akt/Akt, VEGFA and HIF-1α in colon cancer tissue of the Shendan Sanjie capsule+ IGF-1 group were 3.37±0.15, 4.02±0.11, 3.52±0.24, respectively, which were significantly higher than those in the Shendan Sanjie capsule group (P<0.05). Conclusion: Shendan Sanjie capsules may inhibit Akt/HIF-1α/VEGFA signaling pathway, and then reduce the expression of microvascular growth factors bFGF and Ang2, thereby inhibit the tumor angiogenesis of CAC.