Prostasin and hepatocyte growth factor B in factor VIIa generation: Serine protease knockdowns in zebrafish

Khandekar, Gauri; Iyer, Neha; Jagadeeswaran, Pudur

doi:10.1002/rth2.12428

Cited by 8 publications

(8 citation statements)

References 20 publications

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“…Prostasin (PRSS8), a serine protease that not only regulates the epithelial sodium channel but is also expressed in endothelial and smooth muscle cells (Zhu et al., 2008), was identified as one of the most upregulated proteins in patients from the Severe group when compared to the UI Control group in both sets of EVs (Figures 3b & 4; Supplementary Figures 4C & 5). Interestingly, prostasin has recently been reported to directly or indirectly convert coagulation Factor VII to its active form (Khandekar et al., 2020).…”

Section: Resultsmentioning

confidence: 99%

Extracellular vesicle‐mediated endothelial apoptosis and EV‐associated proteins correlate with COVID‐19 disease severity

Krishnamachary

Cook

Kumar

et al. 2021

J of Extracellular Vesicle

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Coronavirus disease-2019 (COVID-19), caused by the novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has lead to a global pandemic with a rising toll in infections and deaths. Better understanding of its pathogenesis will greatly improve the outcomes and treatment of affected patients. Here we compared the inflammatory and cardiovascular disease-related protein cargo of circulating large and small extracellular vesicles (EVs) from 84 hospitalized patients infected with SARS-CoV-2 with different stages of disease severity. Our findings reveal significant enrichment of proinflammatory, procoagulation, immunoregulatory and tissueremodelling protein signatures in EVs, which remarkably distinguished symptomatic COVID-19 patients from uninfected controls with matched comorbidities and delineated those with moderate disease from those who were critically ill. Specifically, EN-RAGE, followed by TF and IL-18R1, showed the strongest correlation with disease severity and length of hospitalization. Importantly, EVs from COVID-19 patients induced apoptosis of pulmonary microvascular endothelial cells in the order of disease severity. In conclusion, our findings support a role for EVs in the pathogenesis of COVID-19 disease and underpin the development of EV-based approaches to predicting disease severity, determining need for patient hospitalization and identifying new therapeutic targets. K E Y WO R D S endothelial injury, inflammation, SARS-CoV-2, thrombosis  INTRODUCTIONThe coronavirus disease-2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) (Dashboard WCDC-2020; Wu et al., 2020) is marked by endothelial dysfunction and dysregulated immune responses (Huertas et al., 2020). Like the SARS-CoV pathogen that led to epidemics in 2002 and 2003, SARS-CoV-2 enters cells via binding of its spike protein to angiotensin converting enzyme 2 (ACE2) receptors. ACE2 receptors are abundantly present in pulmonary alveolar type II and endothelial cells, thereby making the lungs and pulmonary vasculature susceptible to SARS-CoV-2-induced inflammation and injury (Zhao et al., 2020). Further, alveolar capillary micro-thrombi and endothelial damage with evidence of intracellular virus have been noted on post-mortem analysis of infected lungs (Ackermann et al., 2020). While there is growing evidence that endothelial injury, vascular remodelling and coagulopathy are key consequences of COVID-19 infection, it remains unclear how the virus induces these changes. Extracellular vesicles (EVs) carry proteins, coding and non-coding RNA, DNA fragments and lipids, which facilitate cross-talk between cells. The transfer of EV cargo plays aThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Section: Resultsmentioning

confidence: 99%

Extracellular vesicle‐mediated endothelial apoptosis and EV‐associated proteins correlate with COVID‐19 disease severity

Krishnamachary

Cook

Kumar

et al. 2021

J of Extracellular Vesicle

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“…In addition, this relationship was maintained when metastatic recurrence was excluded as a possible underlying cause of thrombosis. The interaction between HPN and coagulation in this type of cancer could be explained by the ability of HPN to activate proteins such as factor VII, XII and IX of the coagulation cascade (6,7,49), although none of these studies have been done in humans. Another key fact to explain the association between HPN and thrombosis is that, although this protein is described as a transmembrane protease, there are studies that have identified it in the serum of patients with CRC (16).…”

Section: Discussionmentioning

confidence: 99%

Implication of Hepsin from primary tumor in the prognosis of colorectal cancer patients

Zaragoza-Huesca

Nieto-Olivares

García‐Molina

et al. 2022

Preprint

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BackgroundHepsin is a type II transmembrane serine protease whose functions include degradation of the extracellular matrix and activation of factor VII. In many tumors, hepsin dysregulation leads to tumor invasion by proteolysis of pericellular components. Colorectal cancer is a tumor with high thrombotic and metastatic risk, but the role of hepsin is unknown.ObjectivesTo study the correlations between hepsin expression in the primary tumor and different clinical-histopathological variables in patients with localized and metastatic colorectal cancer.MethodsA cohort of 287 patients, 169 with localized colorectal cancer and 118 with metastases at diagnosis, was recruited. Tissue microarrays were made from samples at diagnosis of primary tumors and stained with an anti-hepsin antibody. Hepsin levels were determined and the association with clinical-histopathological variables was evaluated using the chi-square and Kruskal-Wallis tests. Time-to-event results and cumulative incidence functions were calculated using Kaplan-Meier and Aalen-Johansen estimators, in combination with Cox and Fine&Gray multivariate models.ResultsIn patients with localized colorectal cancer, high intensity hepsin staining was associated with reduced 5-year disease-free survival (p-value = 0.16). In this context, according to multivariable Cox regression, medium and high intensity of hepsin expression versus low intensity was associated with an increased risk of metastatic relapse (hazard ratio 2.83, p-value=0.035, and hazard ratio 3.30, p-value=0.012, respectively), being a more accurate prognostic factor than classic histological variables. Also, in patients with localized colorectal cancer, the cumulative incidence of thrombosis at 5 years increased significantly with higher hepsin levels (p-value = 0.038). According to the multivariate Fine&Gray regression, the medium and high intensity of hepsin with respect to the low intensity, was associated with an increase in thrombotic risk (hazard ratio 7.71, p-value=0.043 and hazard ratio 9.02, p-value=0.028, respectively). This relationship between hepsin and thrombosis was independent of previous tumor relapse (p-value = 0.036). In patients with metastatic colorectal cancer, low intensity of hepsin was associated with a low degree of tumor differentiation (p-value <0.001) and spread to more than one metastatic site (p-value = 0.023).ConclusionHepsin is a potential biomarker of thrombosis and metastasis in patients with localized colorectal cancer. Validation in a larger series and clarification of the underlying mechanism may be helpful in evaluating these patients. In metastatic patients, Hepsin behaves in a paradoxical way with respect to differentiation and invasion processes.

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“…[32][33][34] Various examples include the two tlr4 genes, 35 four igf genes, 36 two copies of cytokine genes 37 , and two hgf genes. 38,39 The genome duplication may have also given rise to more than one f9 gene in the zebrafish genome whose functions remain uncharacterized. We identified three potential f9 genes in the zebrafish genome, namely, f9a, f9b, and f9l.…”

Section: Discussionmentioning

confidence: 99%

“…This duplication generated paralogous groups of many genes, lending the genes an opportunity to acquire new functions, take over part of the original function, or become inactivated 32‐34 . Various examples include the two tlr4 genes, 35 four igf genes, 36 two copies of cytokine genes 37 , and two hgf genes 38,39 . The genome duplication may have also given rise to more than one f9 gene in the zebrafish genome whose functions remain uncharacterized.…”

Section: Discussionmentioning

confidence: 99%

Identification of zebrafish ortholog for human coagulation factor IX and its age‐dependent expression

Iyer

Qaryoute

Kacham

et al. 2021

Journal of Thrombosis and Haemostasis

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Background: Coagulation factor IX (FIX) is a serine protease zymogen involved in the intrinsic blood coagulation pathway, and its deficiency causes hemophilia B. Zebrafish has three f9 genes, and the ortholog to human F9 is unknown.Objective: To identify the zebrafish ortholog to F9 using sequence analysis and piggyback knockdown technology.Methods: Gene and protein sequence analysis for three f9 genes, f9a, f9b, and f9l, present in the zebrafish genome was performed. In vivo and in vitro assays after knockdown of each gene and immunodepletion using specific antibodies were carried out.Results: Sequence analysis revealed that f9a and f9b are similar to human F9, whereas f9l is similar to human F10. RNA analysis showed an age-dependent increase in expression of all three genes. Zebrafish f9a gene knockdown and Fixa immunodepletion prolonged kinetic partial thromboplastin time (kPTT), whereas f9l knockdown and Fixl immunodepletion prolonged kPTT, kinetic prothrombin time, and kinetic Russell viper venom activation time. Laser-assisted venous thrombosis increased time to occlusion after f9a and f9l knockdown and antibody inhibition of Fixa and Fixl. Further, analysis of plasma proteins by mass spectrometry and immunohistochemistry detected all three proteins. Conclusions: Our findings suggest that zebrafish f9a has functional activity similar to human F9. Fixl is functionally similar to Fx. The age-dependent increases of these factors are comparable to those observed in mice and humans. Thus, the zebrafish model could be used to study factors involved in increasing f9a expression during aging. It could also be used to test whether normal human Factor IX and Factor IX Leyden promoter work in zebrafish background.

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Prostasin and hepatocyte growth factor B in factor VIIa generation: Serine protease knockdowns in zebrafish

Abstract: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

Cited by 8 publications

References 20 publications

Extracellular vesicle‐mediated endothelial apoptosis and EV‐associated proteins correlate with COVID‐19 disease severity

Extracellular vesicle‐mediated endothelial apoptosis and EV‐associated proteins correlate with COVID‐19 disease severity

Implication of Hepsin from primary tumor in the prognosis of colorectal cancer patients

Identification of zebrafish ortholog for human coagulation factor IX and its age‐dependent expression

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