Identification of a gene regulatory network associated with prion replication.
Prions consist of aggregates of abnormal conformers of the cellular prion protein (PrP(C)). They propagate by recruiting host-encoded PrP(C) although the critical interacting proteins and the reasons for the differences in susceptibility of distinct cell lines and populations are unknown. We derived a lineage of cell lines with markedly differing susceptibilities, unexplained by PrP(C) expression differences, to identify such factors. Transcriptome analysis of prion-resistant revertants, isolated from highly susceptible cells, revealed a gene expression signature associated with susceptibility and modulated by differentiation. Several of these genes encode proteins with a role in extracellular matrix (ECM) remodelling, a compartment in which disease-related PrP is deposited. Silencing nine of these genes significantly increased susceptibility. Silencing of Papss2 led to undersulphated heparan sulphate and increased PrP(C) deposition at the ECM, concomitantly with increased prion propagation. Moreover, inhibition of fibronectin 1 binding to integrin α8 by RGD peptide inhibited metalloproteinases (MMP)-2/9 whilst increasing prion propagation. In summary, we have identified a gene regulatory network associated with prion propagation at the ECM and governed by the cellular differentiation state.
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Masue M
MM
Marbiah
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Anna
A
Harvey
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Billy T
BT
West
Department of Clinical Neuroscience, Karolinska Institute, Stockholm, Sweden.
Anais
A
Louzolo
Biomedical Communications, Terrence Donnelly Health Sciences Complex University of Toronto, Toronto, ON, Canada.
Priya
P
Banerjee
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Jack
J
Alden
Breakthrough Breast Cancer Research Unit, Research Oncology, Guy's Hospital, London, UK.
Anita
A
Grigoriadis
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Holger
H
Hummerich
Department of Biology, University of Virginia, Charlottesville, VA, USA.
Ho-Man
HM
Kan
Department of Biology, University of Virginia, Charlottesville, VA, USA.
Ying
Y
Cai
Department of Biology, University of Virginia, Charlottesville, VA, USA.
George S
GS
Bloom
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
Parmjit
P
Jat
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK.
John
J
Collinge
MRC Prion Unit and Department of Neurodegenerative Disease, UCL Institute of Neurology Queen Square, London, UK p.kloehn@prion.ucl.ac.uk.
Peter-Christian
PC
Klöhn
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D003001:Cloning, Molecular; D005109:Extracellular Matrix / Q000378:metabolism*; D005434:Flow Cytometry; D053263:Gene Regulatory Networks / Q000235:genetics*; D006801:Humans; D051379:Mice; D046228:Microarray Analysis; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D018621:PrPC Proteins / Q000235:genetics / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013053:Spectrophotometry; D059467:Transcriptome / Q000235:genetics*; D020703:ras GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism
C101002:IQGAP2 protein, human; D018621:PrPC Proteins; D011328:Prions; D034741:RNA, Small Interfering; D020703:ras GTPase-Activating Proteins
extracellular matrix; integrin; neurodegeneration; prion diseases; scrapie