Endothelial Cell Processing and Alternatively Spliced Transcripts of Factor VIII: Potential Implications for Coagulation Cascades and Pulmonary Hypertension
| العنوان: | Endothelial Cell Processing and Alternatively Spliced Transcripts of Factor VIII: Potential Implications for Coagulation Cascades and Pulmonary Hypertension |
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| المؤلفون: | Michael Laffan, Claire L. Shovlin, Richard A. Manning, Graeme M. Birdsey, Fatima S Govani, Gillian Angus, IG Mollet, Francesco Mauri, Grace N Okoli, Kay Elderfield |
| المصدر: | PLoS ONE e9154 PLoS ONE, Vol 5, Iss 2, p e9154 (2010) |
| بيانات النشر: | Public Library of Science, 2010. |
| سنة النشر: | 2010 |
| مصطلحات موضوعية: | Gene isoform, CD31, congenital, hereditary, and neonatal diseases and abnormalities, Transcription, Genetic, Hypertension, Pulmonary, Hematology/Coagulation Disorders, lcsh:Medicine, Enzyme-Linked Immunosorbent Assay, Biology, Pulmonary Artery, Flow cytometry, Von Willebrand factor, hemic and lymphatic diseases, von Willebrand Factor, medicine, Weibel–Palade body, Humans, Protein Isoforms, lcsh:Science, Blood Coagulation, Lung, Cells, Cultured, Cardiovascular Disorders/Pulmonary Vascular Diseases, Multidisciplinary, Factor VIII, Microscopy, Confocal, medicine.diagnostic_test, Base Sequence, Reverse Transcriptase Polymerase Chain Reaction, lcsh:R, Endothelial Cells, Exons, Molecular Biology/Transcription Initiation and Activation, medicine.disease, Flow Cytometry, Molecular biology, Pulmonary hypertension, Immunohistochemistry, Endothelial stem cell, Alternative Splicing, medicine.anatomical_structure, biology.protein, lcsh:Q, Transcription Initiation Site, Research Article, Signal Transduction |
| الوصف: | Background Coagulation factor VIII (FVIII) deficiency leads to haemophilia A. Conversely, elevated plasma levels are a strong predictor of recurrent venous thromboemboli and pulmonary hypertension phenotypes in which in situ thromboses are implicated. Extrahepatic sources of plasma FVIII are implicated, but have remained elusive. Methodology/Principal Findings Immunohistochemistry of normal human lung tissue, and confocal microscopy, flow cytometry, and ELISA quantification of conditioned media from normal primary endothelial cells were used to examine endothelial expression of FVIII and coexpression with von Willebrand Factor (vWF), which protects secreted FVIII heavy chain from rapid proteloysis. FVIII transcripts predicted from database mining were identified by rt-PCR and sequencing. FVIII mAb-reactive material was demonstrated in CD31+ endothelial cells in normal human lung tissue, and in primary pulmonary artery, pulmonary microvascular, and dermal microvascular endothelial cells. In pulmonary endothelial cells, this protein occasionally colocalized with vWF, centered on Weibel Palade bodies. Pulmonary artery and pulmonary microvascular endothelial cells secreted low levels of FVIII and vWF to conditioned media, and demonstrated cell surface expression of FVIII and vWF Ab–reacting proteins compared to an isotype control. Four endothelial splice isoforms were identified. Two utilize transcription start sites in alternate 5′ exons within the int22h-1 repeat responsible for intron 22 inversions in 40% of severe haemophiliacs. A reciprocal relationship between the presence of short isoforms and full-length FVIII transcript suggested potential splice-switching mechanisms. Conclusions/Significance The pulmonary endothelium is confirmed as a site of FVIII secretion, with evidence of synthesis, cell surface expression, and coexpression with vWF. There is complex alternate transcription initiation from the FVIII gene. These findings provide a framework for future research on the regulation and perturbation of FVIII synthesis, and of potential relevance to haemophilia, thromboses, and pulmonary hypertensive states. |
| اللغة: | English |
| تدمد: | 1932-6203 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e42152af5ae1076a29340ddbd7747da2 http://europepmc.org/articles/PMC2823490 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....e42152af5ae1076a29340ddbd7747da2 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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