Tuberous sclerosis complex (TSC) is a rare autosomal dominant disorder causing benign tumors in the brain and other vital organs. The genes implicated in disease development are TSC1 and TSC2. Here, we have performed mutational analysis followed by a genotype-phenotype correlation study based on the clinical characteristics of the affected individuals. Twenty unrelated probands or families from Greece have been analyzed, of whom 13 had definite TSC, whereas another 7 had a possible TSC diagnosis. Using direct sequencing, we have identified pathogenic mutations in 13 patients/families (6 in TSC1 and 7 in TSC2), 5 of which were novel. The mutation identification rate for patients with definite TSC was 85%, but only 29% for the ones with a possible TSC diagnosis. Multiplex ligation-dependent probe amplification (MLPA) did not reveal any genomic rearrangements in TSC1 and TSC2 in the samples with no mutations identified. In general, TSC2 disease was more severe than TSC1, with more subependymal giant cell astrocytomas and angiomyolipomas, higher incidence of pharmacoresistant epileptic seizures, and more severe neuropsychiatric disorders. To our knowledge, this is the first comprehensive TSC1 and TSC2 mutational analysis carried out in TSC patients in Greece.

• MeSH
• delece genu MeSH
• dítě MeSH
• dospělí MeSH
• exony MeSH
• genetické asociační studie MeSH
• hamartin genetika   MeSH
• lidé MeSH
• missense mutace MeSH
• mutační analýza DNA MeSH
• rodokmen MeSH
• terciární struktura proteinů MeSH
• tuberin genetika   MeSH
• tuberózní skleróza genetika   patologie   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Řecko MeSH
• Názvy látek
• hamartin MeSH
• tuberin MeSH

OBJECTIVE: To study the association between timing and characteristics of the first electroencephalography (EEG) with epileptiform discharges (ED-EEG) and epilepsy and neurodevelopment at 24 months in infants with tuberous sclerosis complex (TSC). METHODS: Patients enrolled in the prospective Epileptogenesis in a genetic model of epilepsy - Tuberous sclerosis complex (EPISTOP) trial, had serial EEG monitoring until the age of 24 months. The timing and characteristics of the first ED-EEG were studied in relation to clinical outcome. Epilepsy-related outcomes were analyzed separately in a conventionally followed group (initiation of vigabatrin after seizure onset) and a preventive group (initiation of vigabatrin before seizures, but after appearance of interictal epileptiform discharges [IEDs]). RESULTS: Eighty-three infants with TSC were enrolled at a median age of 28 days (interquartile range [IQR] 14-54). Seventy-nine of 83 patients (95%) developed epileptiform discharges at a median age of 77 days (IQR 23-111). Patients with a pathogenic TSC2 variant were significantly younger (P-value .009) at first ED-EEG and more frequently had multifocal IED (P-value .042) than patients with a pathogenic TSC1 variant. A younger age at first ED-EEG was significantly associated with lower cognitive (P-value .010), language (P-value .001), and motor (P-value .013) developmental quotients at 24 months. In the conventional group, 48 of 60 developed seizures. In this group, the presence of focal slowing on the first ED-EEG was predictive of earlier seizure onset (P-value .030). Earlier recording of epileptiform discharges (P-value .019), especially when multifocal (P-value .026) was associated with higher risk of drug-resistant epilepsy. In the preventive group, timing, distribution of IED, or focal slowing, was not associated with the epilepsy outcomes. However, when multifocal IEDs were present on the first ED-EEG, preventive treatment delayed the onset of seizures significantly (P-value <.001). SIGNIFICANCE: Early EEG findings help to identify TSC infants at risk of severe epilepsy and neurodevelopmental delay and those who may benefit from preventive treatment with vigabatrin.

• Klíčová slova
• EEG, epilepsy, epileptogenesis, neurodevelopment, tuberous sclerosis complex,
• MeSH
• antikonvulziva terapeutické užití   MeSH
• časná diagnóza * MeSH
• elektroencefalografie MeSH
• epilepsie diagnóza   farmakoterapie   etiologie   MeSH
• hamartin genetika   MeSH
• kojenec MeSH
• lidé MeSH
• novorozenec MeSH
• tuberin genetika   MeSH
• tuberózní skleróza komplikace   diagnóza   genetika   MeSH
• vigabatrin terapeutické užití   MeSH
• vývojové poruchy u dětí epidemiologie   etiologie   MeSH
• Check Tag
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antikonvulziva MeSH
• hamartin MeSH
• TSC1 protein, human MeSH Prohlížeč
• TSC2 protein, human MeSH Prohlížeč
• tuberin MeSH
• vigabatrin MeSH

PURPOSE: To perform comprehensive genotyping of TSC1 and TSC2 in a cohort of 94 infants with tuberous sclerosis complex (TSC) and correlate with clinical manifestations. METHODS: Infants were enrolled at age <4 months, and subject to intensive clinical monitoring including electroencephalography (EEG), brain magnetic resonance imaging (MRI), and neuropsychological assessment. Targeted massively parallel sequencing (MPS), genome sequencing, and multiplex ligation-dependent probe amplification (MLPA) were used for variant detection in TSC1/TSC2. RESULTS: Pathogenic variants in TSC1 or TSC2 were identified in 93 of 94 (99%) subjects, with 23 in TSC1 and 70 in TSC2. Nine (10%) subjects had mosaicism. Eight of 24 clinical features assessed at age 2 years were significantly less frequent in those with TSC1 versus TSC2 variants including cortical tubers, hypomelanotic macules, facial angiofibroma, renal cysts, drug-resistant epilepsy, developmental delay, subependymal giant cell astrocytoma, and median seizure-free survival. Additionally, quantitative brain MRI analysis showed a marked difference in tuber and subependymal nodule/giant cell astrocytoma volume for TSC1 versus TSC2. CONCLUSION: TSC2 pathogenic variants are associated with a more severe clinical phenotype than mosaic TSC2 or TSC1 variants in TSC infants. Early assessment of gene variant status and mosaicism might have benefit for clinical management in infants and young children with TSC.

• Klíčová slova
• TSC1, TSC2, clinical manifestations, mosaicism, tuberous sclerosis complex (TSC),
• MeSH
• fenotyp MeSH
• hamartin genetika   MeSH
• kojenec MeSH
• lidé MeSH
• mozaicismus MeSH
• mutace MeSH
• předškolní dítě MeSH
• tuberin genetika   MeSH
• tuberózní skleróza * diagnostické zobrazování   genetika   MeSH
• Check Tag
• kojenec MeSH
• lidé MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hamartin MeSH
• tuberin MeSH

UNLABELLED: Tuberous sclerosis is a disease with an autosomal dominant pattern of inheritance which is characterized by the development of benign tumours in many tissues and organs. Clinical signs are extremely variable, causing mutations in the gene TSC1 or TSC2. Complex formed by the products of the TSC genes regulates cell growth and proliferation by inhibition of mTORC1 signalling. Early diagnosis of TSC is very important to plan appropriate perinatal care. Using ultrasound and eventually MRI it is possible in the prenatal period to capture the following major features of tuberous sclerosis: cardiac rhabdomyo-ma, subependymal nodules, cortical tubers and renal angiomyolipomas. In connection with the syndrome of contiguous genes TSC2 / PKD1 can also be detect foetal renal cysts. Often these TSC-associated lesions represent an incidental finding during a routine ultrasound. In the period from the 20th week of pregnancy it is most often found cardiac rhabdomyoma/s as the first marker suggestive of tuberous sclerosis. In the case, where one of the parents is a carrier of already identified mutation in the TSC gene, it is possible to carry out targeted genetic testing of a sample of DNA isolated from cells of chorionic villi, amniocytes or tissue from aborted foe-tuses. Significantly more time consuming is to perform molecular analysis of the TSC genes in foetuses with suspected tuberous sclerosis without the occurrence of illness in the family. After finding a causal mutation and its confirmation, it is possible to offer genetic testing for other persons at risk, prenatal (eventually preimplantation) diagnosis for future pregnancies. It is also necessary to consider the possibility of gonadal mosaicism. DESIGN: Review of the literature.

• Klíčová slova
• TSC1, TSC2, foetal rhabdomyoma, mosaicism., sequencing, tuberous sclerosis complex,
• MeSH
• chromozomální aberace MeSH
• detekce genetických nosičů MeSH
• dominantní geny genetika   MeSH
• genetické testování MeSH
• hamartin MeSH
• lidé MeSH
• mozaicismus MeSH
• mutační analýza DNA MeSH
• nádorové supresorové proteiny genetika   MeSH
• náhodný nález MeSH
• odběr choriových klků MeSH
• prenatální péče * MeSH
• těhotenství MeSH
• tuberin MeSH
• tuberózní skleróza diagnóza   genetika   MeSH
• ultrasonografie prenatální MeSH
• ultrasonografie MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hamartin MeSH
• nádorové supresorové proteiny MeSH
• TSC1 protein, human MeSH Prohlížeč
• TSC2 protein, human MeSH Prohlížeč
• tuberin MeSH

Tuberous sclerosis complex (TSC) is a dominant autosomal genetic disorder caused by loss-of-function mutations in TSC1 and TSC2, which lead to constitutive activation of the mammalian target of rapamycin C1 (mTORC1) with its decoupling from regulatory inputs. Because mTORC1 integrates an array of molecular signals controlling protein synthesis and energy metabolism, its unrestrained activation inflates cell growth and division, resulting in the development of benign tumors in the brain and other organs. In humans, brain malformations typically manifest through a range of neuropsychiatric symptoms, among which mental retardation, intellectual disabilities with signs of autism, and refractory seizures, which are the most prominent. TSC in the rat brain presents the first-rate approximation of cellular and molecular pathology of the human brain, showing many instructive characteristics. Nevertheless, the developmental profile and distribution of lesions in the rat brain, with neurophysiological and behavioral manifestation, deviate considerably from humans, raising numerous research and translational questions. In this study, we revisit brain TSC in human and Eker rats to relate their histopathological, electrophysiological, and neurobehavioral characteristics. We discuss shared and distinct aspects of the pathology and consider factors contributing to phenotypic discrepancies. Given the shared genetic cause and molecular pathology, phenotypic deviations suggest an incomplete understanding of the disease. Narrowing the knowledge gap in the future should not only improve the characterization of the TSC rat model but also explain considerable variability in the clinical manifestation of the disease in humans.

• Klíčová slova
• TSC1, TSC2, autism spectrum disorders, hamartoma, mTOR signaling, neoplasia, refractory epilepsy,
• MeSH
• druhová specificita MeSH
• duševní poruchy genetika   patologie   psychologie   MeSH
• fenotyp * MeSH
• hamartin genetika   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• mozek patologie   MeSH
• TOR serin-threoninkinasy genetika   MeSH
• tuberin genetika   MeSH
• tuberózní skleróza genetika   patologie   psychologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• hamartin MeSH
• MTOR protein, human MeSH Prohlížeč
• TOR serin-threoninkinasy MeSH
• tuberin MeSH

Tuberous Sclerosis Complex (TSC) is a rare genetic disorder that results from a mutation in the TSC1 or TSC2 genes leading to constitutive activation of the mechanistic target of rapamycin complex 1 (mTORC1). TSC is associated with autism, intellectual disability and severe epilepsy. Cortical tubers are believed to represent the neuropathological substrates of these disabling manifestations in TSC. In the presented study we used high-throughput RNA sequencing in combination with systems-based computational approaches to investigate the complexity of the TSC molecular network. Overall we detected 438 differentially expressed genes and 991 differentially expressed small non-coding RNAs in cortical tubers compared to autopsy control brain tissue. We observed increased expression of genes associated with inflammatory, innate and adaptive immune responses. In contrast, we observed a down-regulation of genes associated with neurogenesis and glutamate receptor signaling. MicroRNAs represented the largest class of over-expressed small non-coding RNA species in tubers. In particular, our analysis revealed that the miR-34 family (including miR-34a, miR-34b and miR-34c) was significantly over-expressed. Functional studies demonstrated the ability of miR-34b to modulate neurite outgrowth in mouse primary hippocampal neuronal cultures. This study provides new insights into the TSC transcriptomic network along with the identification of potential new treatment targets.

• MeSH
• dítě MeSH
• dospělí MeSH
• epilepsie genetika   MeSH
• genetická transkripce genetika   MeSH
• hamartin genetika   MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mechanistické cílové místo rapamycinového komplexu 1 genetika   MeSH
• mikro RNA genetika   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mozková kůra fyziologie   MeSH
• mutace genetika   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neurony fyziologie   MeSH
• předškolní dítě MeSH
• signální transdukce genetika   MeSH
• tuberin genetika   MeSH
• tuberózní skleróza genetika   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• myši MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hamartin MeSH
• mechanistické cílové místo rapamycinového komplexu 1 MeSH
• mikro RNA MeSH
• MIRN34a microRNA, mouse MeSH Prohlížeč
• tuberin MeSH

BACKGROUND: Tuberous sclerosis is an autosomal-dominant disease characterised by development of benign growth - hamartomas in different organs. Disorder is caused by mutations affecting either of the tumor-suppressor genes, TSC1 or TSC2. Quest for causing mutations is very difficult due to their random distribution over the genes. METHODS AND RESULTS: Article refers on accomplishment of the first tuberous sclerosis prenatal diagnostics in Czech Republic based on knowledge of causing mutation. Foetal DNA sample, obtained in 13th week from Q435X family pregnant woman, was analyzed by DGGE method. CONCLUSIONS: Examination excluded presence of tested TSC1 gene defect in an offspring.

• MeSH
• hamartin MeSH
• lidé MeSH
• mutace * MeSH
• nádorové supresorové proteiny genetika   MeSH
• prenatální diagnóza * MeSH
• rodokmen MeSH
• těhotenství MeSH
• tuberin MeSH
• tuberózní skleróza diagnóza   MeSH
• tumor supresorové geny * MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hamartin MeSH
• nádorové supresorové proteiny MeSH
• TSC1 protein, human MeSH Prohlížeč
• TSC2 protein, human MeSH Prohlížeč
• tuberin MeSH

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.

• Klíčová slova
• Autism spectrum disorders, Developmental status epilepticus, Everolimus, TSC, Tuberous sclerosis complex, mTOR,
• MeSH
• autistická porucha * MeSH
• haploinsuficience MeSH
• krysa rodu Rattus MeSH
• status epilepticus * MeSH
• TOR serin-threoninkinasy genetika   MeSH
• tuberin genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• mTOR protein, rat MeSH Prohlížeč
• TOR serin-threoninkinasy MeSH
• Tsc2 protein, rat MeSH Prohlížeč
• tuberin MeSH

Pathogenic mutations in the genes associated with tuberous sclerosis complex (TSC)/mTOR pathway are linked to histologically diverse renal cell neoplasms, including eosinophilic solid and cystic renal cell carcinoma (ESC RCC), low grade oncocytic tumor (LOT), eosinophilic vacuolated tumor (EVT), and xanthomatous giant cell renal cell carcinoma (XGC RCC). Here, we validate a TSC2 immunohistochemistry (IHC) assay by comparison to genomic data in these neoplasms. Automated TSC2 IHC was performed on formalin-fixed paraffin embedded (FFPE) tissues from 38 genetically-confirmed TSC/mTOR-associated renal tumors (6 ESCs, 16 EVTs, 13 LOTs, 2 XGC and 1 clear cell RCC) and visually scored in a semi-dichotomous fashion compared to internal control tissue. The positive predictive value (PPV) of TSC2 protein loss for underlying pathogenic mutation in TSC2 was 92% (11/12), while the negative predictive value (NPV) of intact TSC2 by IHC for lack of underlying pathogenic mutation in TSC2 was 81% (21/26). Intact TSC2 by IHC was 95% (21/22) specific for absence of underlying pathogenic TSC2 mutation. All the cases lacking TSC2 mutation with intact TSC2 protein had an underlying mutation in TSC1, MTOR or PIK3CA. Loss of TSC2 was 77% (10/13) sensitive for underlying TSC2 truncation mutations and 33% (1/3) sensitive for underlying TSC2 missense mutations. Overall, 73% (8/11) tumors with TSC2 IHC loss and underlying pathogenic alterations in TSC2 showed heterogeneous protein loss, with rare interspersed positively staining tumor cells. These data support TSC2 IHC as a potentially useful assay for the diagnostic workup of renal tumors suspected to belong to the TSC/mTOR-associated subgroups.

• Klíčová slova
• Genetic validation, Immunohistochemistry, Renal cell carcinoma, TSC2,
• MeSH
• imunohistochemie * MeSH
• karcinom z renálních buněk * genetika   patologie   MeSH
• lidé MeSH
• mutace MeSH
• nádorové biomarkery * genetika   analýza   MeSH
• nádorové supresorové proteiny * analýza   genetika   MeSH
• nádory ledvin * genetika   patologie   MeSH
• prediktivní hodnota testů MeSH
• reprodukovatelnost výsledků MeSH
• signální transdukce MeSH
• TOR serin-threoninkinasy * metabolismus   MeSH
• tuberin * genetika   MeSH
• tuberózní skleróza genetika   patologie   diagnóza   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Názvy látek
• MTOR protein, human MeSH Prohlížeč
• nádorové biomarkery * MeSH
• nádorové supresorové proteiny * MeSH
• TOR serin-threoninkinasy * MeSH
• TSC2 protein, human MeSH Prohlížeč
• tuberin * MeSH

Low-grade oncocytic tumour (LOT) of the kidney has recently emerged as a potential novel tumour type. Despite similarity to oncocytoma or eosinophilic chromophobe renal cell carcinoma, it shows diffuse keratin 7 immunohistochemistry (IHC) and negative KIT (CD117), which differs from both. We aimed to identify the molecular characteristics of these tumours. Seventeen tumours (one male, 16 female, nine previously published) fitting the original description of this entity (solid eosinophilic cell morphology, often with areas of tumour cells loosely stretched in oedematous stroma, and the above IHC features) were analysed with a next-generation sequencing panel of 324 cancer-associated genes from formalin-fixed, paraffin-embedded tissue. All tumours harboured at least one alteration in either TSC1 (n = 7, 41%), TSC2 (n = 2, 12%), MTOR (n = 5, 29%) or PIK3CA (n = 4, 24%). Four tumours harboured a second alteration, including two NF2, one each in conjunction with MTOR and TSC2 alterations, one PTEN with TSC1 alteration and one tumour with both MTOR and TSC1 alterations. No other renal cancer-related or recurring gene alterations were identified. In addition to the previously described IHC findings, 16 of 16 were positive for GATA3. Eleven patients with follow-up had no metastases or recurrent tumours. Recurrent tuberous sclerosis/MTOR pathway gene alterations in LOT support its consideration as a distinct morphological, immunohistochemical and genetic entity. PIK3CA is another pathway member that may be altered in these tumours. Further study will be necessary to determine whether tumour behaviour or syndromic associations differ from those of oncocytoma and chromophobe carcinoma, warranting different clinical consideration.

• Klíčová slova
• PIK3CA, MTOR, TSC1, TSC2, low-grade oncocytic tumour, oncocytoma,
• MeSH
• fosfatidylinositol-3-kinasy třídy I genetika   MeSH
• karcinom z renálních buněk * genetika   MeSH
• ledviny MeSH
• lidé MeSH
• lokální recidiva nádoru MeSH
• mutace MeSH
• nádory ledvin * genetika   MeSH
• oxyfilní adenom * genetika   MeSH
• TOR serin-threoninkinasy genetika   MeSH
• transkripční faktor GATA3 genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfatidylinositol-3-kinasy třídy I MeSH
• GATA3 protein, human MeSH Prohlížeč
• MTOR protein, human MeSH Prohlížeč
• PIK3CA protein, human MeSH Prohlížeč
• TOR serin-threoninkinasy MeSH
• transkripční faktor GATA3 MeSH