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Genome Medicine

, 8:94

First Online: 21 September 2016Received: 20 April 2016Accepted: 18 August 2016DOI: 10.1186-s13073-016-0347-3

Cite this article as: Grabole, N., Zhang, J.D., Aigner, S. et al. Genome Med 2016 8: 94. doi:10.1186-s13073-016-0347-3


BackgroundTuberous sclerosis complex TSC is a genetic disease characterized by benign tumor growths in multiple organs and neurological symptoms induced by mTOR hyperfunction. Because the molecular pathology is highly complex and the etiology poorly understood, we employed a defined human neuronal model with a single mTOR activating mutation to dissect the disease-relevant molecular responses driving the neuropathology and suggest new targets for treatment.

MethodsWe investigate the disease phenotype of TSC by neural differentiation of a human stem cell model that had been deleted for TSC2 by genome editing. Comprehensive genomic analysis was performed by RNA sequencing and ribosome profiling to obtain a detailed genome-wide description of alterations on both the transcriptional and translational level. The molecular effect of mTOR inhibitors used in the clinic was monitored and comparison to published data from patient biopsies and mouse models highlights key pathogenic processes.

ResultsTSC2-deficient neural stem cells showed severely reduced neuronal maturation and characteristics of astrogliosis instead. Transcriptome analysis indicated an active inflammatory response and increased metabolic activity, whereas at the level of translation ribosomal transcripts showed a 5’UTR motif-mediated increase in ribosome occupancy. Further, we observed enhanced protein synthesis rates of angiogenic growth factors. Treatment with mTOR inhibitors corrected translational alterations but transcriptional dysfunction persisted.

ConclusionsOur results extend the understanding of the molecular pathophysiology of TSC brain lesions, and suggest phenotype-tailored pharmacological treatment strategies.

KeywordsAstrocytes mTOR Stem cell disease modeling TSC2 Tuberous sclerosis AbbreviationsATPAdenosine triphosphate

BCABicinchoninic assay

cDNAComplementary DNA


DMSODimethyl sulfoxide

DNADesoxyribonuceic acid

ECLEnhanced chemiluminescence

ESCEmbryonic stem cell

FDRFalse discovery rate

GSEAGene set enrichment analysis

HRPHorseradish peroxidase

LOHLoss of heterozygosity

mTORMechanistic target of rapamycin

NSCNeural stem cell


PBSPhosphate buffered saline

PCRPolymerase chain reaction


qPCRQuantitative PCR

RNARibonucleic acid


rRNARibosomal RNA

SDS-PAGESodium dodecyl sulphate-polyacrylamide gel electrophoresis

SEGASubependymal giant cell astrocytoma

TOPTerminal oligo-pyrimidine

tRNATransfer RNA

TSCTuberous sclerosis complex

UTRUntranslated region

Electronic supplementary materialThe online version of this article doi:10.1186-s13073-016-0347-3 contains supplementary material, which is available to authorized users.

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Autor: Nils Grabole - Jitao David Zhang - Stefan Aigner - Nadine Ruderisch - Veronica Costa - Felix C. Weber - Michel Theron - N


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