Factor VIII exhibits chaperone-dependent and glucose-regulated reversible amyloid formation in the endoplasmic reticulum.

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dc.contributor.authorPoothong, J.
dc.contributor.authorPottekat, A.
dc.contributor.authorSiriin, M.
dc.contributor.authorCampos, A.R.
dc.contributor.authorPaton, A.W.
dc.contributor.authorPaton, J.C.
dc.contributor.authorLagunas-Acosta, J.
dc.contributor.authorChen, Z.
dc.contributor.authorSwift, M.
dc.contributor.authorVolkmann, N.
dc.contributor.authorHanein, D.
dc.contributor.authorYong, J.
dc.contributor.authorKaufman, R.J.
dc.date.issued2020
dc.description.abstractHemophilia A, an X-linked bleeding disorder caused by deficiency of factor VIII (FVIII), is treated by protein replacement. Unfortunately, this regimen is costly due to the expense of producing recombinant FVIII as a consequence of its low-level secretion from mammalian host cells. FVIII expression activates the endoplasmic reticulum (ER) stress response, causes oxidative stress, and induces apoptosis. Importantly, little is known about the factors that cause protein misfolding and aggregation in metazoans. Here, we identified intrinsic and extrinsic factors that cause FVIII to form aggregates. We show that FVIII forms amyloid-like fibrils within the ER lumen upon increased FVIII synthesis or inhibition of glucose metabolism. Significantly, FVIII amyloids can be dissolved upon restoration of glucose metabolism to produce functional secreted FVIII. Two ER chaperone families and their cochaperones, immunoglobulin binding protein (BiP) and calnexin/calreticulin, promote FVIII solubility in the ER, where the former is also required for disaggregation. Ashort aggregation motif in the FVIII A1 domain (termed Aggron) is necessary and sufficient to seed b-sheet polymerization, and BiP binding to this Aggron prevents amyloidogenesis. Our findings provide novel insight into mechanisms that limit FVIII secretion and ER protein aggregation in general and have implication for ongoing hemophilia A gene-therapy clinical trials.
dc.description.statementofresponsibilityJuthakorn Poothong, Anita Pottekat, Marina Siirin, Alexandre Rosa Campos, Adrienne W. Paton, James C. Paton, Jacqueline Lagunas-Acosta, Zhouji Chen, Mark Swift, Niels Volkmann, Dorit Hanein, Jing Yong, and Randal J. Kaufman
dc.identifier.citationBlood, 2020; 135(21):1899-1911
dc.identifier.doi10.1182/blood.2019002867
dc.identifier.issn0006-4971
dc.identifier.issn1528-0020
dc.identifier.orcidPaton, J.C. [0000-0001-9807-5278]
dc.identifier.urihttps://hdl.handle.net/2440/146530
dc.language.isoen
dc.publisherAmerican Society of Hematology (ASH Publications)
dc.rights© 2020 by The American Society of Hematology
dc.source.urihttps://doi.org/10.1182/blood.2019002867
dc.subjectamyloid; bodily secretions; emergency service; hospital; endoplasmic reticulum; factor viii; gene therapy; glucose; hemophilia a; mammals; molecular chaperones; gene therapy; thrombosis; hemostasis
dc.subject.meshEndoplasmic Reticulum
dc.subject.meshHumans
dc.subject.meshAmyloid
dc.subject.meshGlucose
dc.subject.meshFactor VIII
dc.subject.meshMolecular Chaperones
dc.subject.meshHemostatics
dc.subject.meshSweetening Agents
dc.subject.meshHep G2 Cells
dc.subject.meshEndoplasmic Reticulum Stress
dc.titleFactor VIII exhibits chaperone-dependent and glucose-regulated reversible amyloid formation in the endoplasmic reticulum.
dc.typeJournal article
pubs.publication-statusPublished

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