BACKGROUND: Tuberous sclerosis complex (TSC), a multi-system genetic disorder often associated with autism spectrum disorder (ASD), is caused by mutations of TSC1 or TSC2, which lead to constitutive overactivation of mammalian target of rapamycin (mTOR). In several Tsc1+/- and Tsc2+/- animal models, cognitive and social behavior deficits were reversed by mTOR inhibitors. However, phase II studies have not shown amelioration of ASD and cognitive deficits in individuals with TSC during mTOR inhibitor therapy. We asked here if developmental epilepsy, common in the majority of individuals with TSC but absent in most animal models, could explain the discrepancy. METHODS: At postnatal day P12, developmental status epilepticus (DSE) was induced in male Tsc2+/- (Eker) and wild-type rats, establishing four experimental groups including controls. In adult animals (n = 36), the behavior was assessed in the paradigms of social interaction test, elevated plus-maze, light-dark test, Y-maze, and novel object recognition. The testing was carried out before medication (T1), during a 2-week treatment with the mTOR inhibitor everolimus (T2) and after an 8-week washing-out (T3). Electroencephalographic (EEG) activity was recorded in a separate set of animals (n = 18). RESULTS: Both Tsc2+/- mutation and DSE caused social behavior deficits and epileptiform EEG abnormalities (T1). Everolimus led to a persistent improvement of the social deficit induced by Tsc2+/-, while deficits related to DSE did not respond to everolimus (T2, T3). CONCLUSIONS: These findings may contribute to an explanation why ASD symptoms in individuals with TSC, where comorbid early-onset epilepsy is common, were not reliably ameliorated by mTOR inhibitors in clinical studies.

1st Faculty of Medicine Charles University Prague Czech Republic

2nd Faculty of Medicine Charles University Prague Czech Republic

Department of Child and Adolescent Psychiatry University Hospital and Medical Faculty Carl Gustav Carus Technical University of Dresden Dresden Germany

Department of Child and Adolescent Psychiatry University Medical Center Göttingen Von Siebold Str 5 37075 Göttingen Germany

Department of Developmental Epileptology Institute of Physiology CAS Prague Czech Republic

Department of Psychiatry University Hospital and Medical Faculty Carl Gustav Carus Technical University of Dresden Dresden Germany

Division of Child and Adolescent Psychiatry University of Cape Town Cape Town South Africa

Faculty of Science Charles University Prague Czech Republic

National Institute of Mental Health Topolova 748 250 67 Klecany Czech Republic

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Behavioral phenotyping identifies autism-like repetitive stereotypies in a Tsc2 haploinsufficient rat model