title: RNA Sequencing Analysis of Intracranial Aneurysm Walls Reveals Involvement of Lysosomes and Immunoglobulins in Rupture.
abstract: Analyzing genes involved in development and rupture of intracranial aneurysms can enhance knowledge about the pathogenesis of aneurysms, and identify new treatment strategies. We compared gene expression between ruptured and unruptured aneurysms and control intracranial arteries.
  
  
  
  We determined expression levels with RNA sequencing. Applying a multivariate negative binomial model, we identified genes that were differentially expressed between 44 aneurysms and 16 control arteries, and between 22 ruptured and 21 unruptured aneurysms. The differential expression of 8 relevant and highly significant genes was validated using digital polymerase chain reaction. Pathway analysis was used to identify enriched pathways. We also analyzed genes with an extreme pattern of differential expression: only expressed in 1 condition without any expression in the other.
  
  
  
  We found 229 differentially expressed genes in aneurysms versus controls and 1489 in ruptured versus unruptured aneurysms. The differential expression of all 8 genes selected for digital polymerase chain reaction validation was confirmed. Extracellular matrix pathways were enriched in aneurysms versus controls, whereas pathways involved in immune response and the lysosome pathway were enriched in ruptured versus unruptured aneurysms. Immunoglobulin genes were expressed in aneurysms, but showed no expression in controls.
  
  
  
  For rupture of intracranial aneurysms, we identified the lysosome pathway as a new pathway and found further evidence for the role of the immune response. Our results also point toward a role for immunoglobulins in the pathogenesis of aneurysms. Immune-modifying drugs are, therefore, interesting candidate treatment strategies in the prevention of aneurysm development and rupture.
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.). rachelkleinloog@gmail.com.
authors.forename: Rachel
authors.initials: R
authors.lastname: Kleinloog
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Bon H
authors.initials: BH
authors.lastname: Verweij
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Pieter
authors.initials: P
authors.lastname: van der Vlies
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Patrick
authors.initials: P
authors.lastname: Deelen
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Morris A
authors.initials: MA
authors.lastname: Swertz
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Louis
authors.initials: L
authors.lastname: de Muynck
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Philip
authors.initials: P
authors.lastname: Van Damme
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Giuliani
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Luca
authors.initials: L
authors.lastname: Regli
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Albert
authors.initials: A
authors.lastname: van der Zwan
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Jan W
authors.initials: JW
authors.lastname: Berkelbach van der Sprenkel
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: K Sen
authors.initials: KS
authors.lastname: Han
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Peter
authors.initials: P
authors.lastname: Gosselaar
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Peter C
authors.initials: PC
authors.lastname: van Rijen
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Emine
authors.initials: E
authors.lastname: Korkmaz
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Jan A
authors.initials: JA
authors.lastname: Post
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Gabriel J E
authors.initials: GJ
authors.lastname: Rinkel
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Jan H
authors.initials: JH
authors.lastname: Veldink
authors.affiliation: From the Department of Neurology and Neurosurgery, Brain Center Rudolf Magnus, University Medical Center Utrecht, Utrecht, The Netherlands (R.K., B.H.V., F.G., A.v.d.Z., J.W.B.v.d.S., K.S.H., P.G., P.C.v.R., G.J.E.R., J.H.V., Y.M.R.); Department of Genetics (P.v.d.V., P.D., M.A.S.) and Genomics Coordination Center (P.D., M.A.S.), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands; Laboratory of Neurobiology, Vesalius Research Center, VIB, Leuven, Belgium (L.d.M., P.V.D.); Department of Neurosurgery, University Hospital Zurich, Zurich, Switzerland (L.R.); and Cell Biology, Department of Biology, Science Faculty, Utrecht University, Utrecht, The Netherlands (E.K., J.A.P.).
authors.forename: Ynte M
authors.initials: YM
authors.lastname: Ruigrok
mesh_terms: D017542:Aneurysm, Ruptured / Q000235:genetics*; D005109:Extracellular Matrix / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics*; D002532:Intracranial Aneurysm / Q000235:genetics*; D008247:Lysosomes / Q000235:genetics*; D008297:Male; D053858:Metabolic Networks and Pathways; D008875:Middle Aged; D017423:Sequence Analysis, RNA / Q000379:methods*
chemical_list: D007136:Immunoglobulins
keywords: aneurysm; extracellular matrix; gene expression; lysosomes; subarachnoid hemorrhage