title: Tubers from patients with tuberous sclerosis complex are characterized by changes in microtubule biology through ROCK2 signalling.
abstract: Most patients with tuberous sclerosis complex (TSC) develop cortical tubers that cause severe neurological disabilities. It has been suggested that defects in neuronal differentiation and/or migration underlie the appearance of tubers. However, the precise molecular alterations remain largely unknown. Here, by combining cytological and immunohistochemical analyses of tubers from nine TSC patients (four of them diagnosed with TSC2 germline mutations), we show that alteration of microtubule biology through ROCK2 signalling contributes to TSC neuropathology. All tubers showed a larger number of binucleated neurons than expected relative to control cortex. An excess of normal and altered cytokinetic figures was also commonly observed. Analysis of centrosomal markers suggested increased microtubule nucleation capacity, which was supported by the analysis of an expression dataset from cortical tubers and control cortex, and subsequently linked to under-expression of Rho-associated coiled-coil containing kinase 2 (ROCK2). Thus, augmented microtubule nucleation capacity was observed in mouse embryonic fibroblasts and human fibroblasts deficient in the Tsc2/TSC2 gene product, tuberin. Consistent with ROCK2 under-expression, microtubule acetylation was found to be increased with tuberin deficiency; this alteration was abrogated by rapamycin treatment and mimicked by HDAC6 inhibition. Together, the results of this study support the hypothesis that loss of TSC2 expression can alter microtubule organization and dynamics, which, in turn, deregulate cell division and potentially impair neuronal differentiation.
authors.affiliation: Institute of Neuropathology, University Hospital Bellvitge, University of Barcelona, Bellvitge Institute for Biomedical Research (IDIBELL), CIBERNED, L'Hospitalet del Llobregat, Barcelona, Catalonia, Spain.
authors.forename: Isidre
authors.initials: I
authors.lastname: Ferrer
authors.forename: Pooja
authors.initials: P
authors.lastname: Mohan
authors.forename: Helen
authors.initials: H
authors.lastname: Chen
authors.forename: Joan
authors.initials: J
authors.lastname: Castellsague
authors.forename: Laia
authors.initials: L
authors.lastname: Gómez-Baldó
authors.forename: Marga
authors.initials: M
authors.lastname: Carmona
authors.forename: Nadia
authors.initials: N
authors.lastname: García
authors.forename: Helena
authors.initials: H
authors.lastname: Aguilar
authors.forename: Jihong
authors.initials: J
authors.lastname: Jiang
authors.forename: Margaretha
authors.initials: M
authors.lastname: Skowron
authors.forename: Mark
authors.initials: M
authors.lastname: Nellist
authors.forename: Israel
authors.initials: I
authors.lastname: Ampuero
authors.forename: Antonio
authors.initials: A
authors.lastname: Russi
authors.forename: Conxi
authors.initials: C
authors.lastname: Lázaro
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Maxwell
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Pujana
mesh_terms: D000818:Animals; D016022:Case-Control Studies; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D048749:Cytokinesis; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology; D020022:Genetic Predisposition to Disease; D018095:Germ-Line Mutation; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D000076222:Mechanistic Target of Rapamycin Complex 1; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D046912:Multiprotein Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D055495:Neurogenesis; D009474:Neurons / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D014162:Transfection; D014402:Tuberous Sclerosis / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D000077005:Tuberous Sclerosis Complex 2 Protein; D014404:Tubulin / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000172:deficiency / Q000235:genetics; D054460:rho-Associated Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D056572:Histone Deacetylase Inhibitors; D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; D047428:Protein Kinase Inhibitors; C118136:TPPP protein, human; C000624653:TSC2 protein, human; C582625:TUBG1 protein, human; C000624654:Tsc2 protein, mouse; D000077005:Tuberous Sclerosis Complex 2 Protein; D014404:Tubulin; D025521:Tumor Suppressor Proteins; D000076222:Mechanistic Target of Rapamycin Complex 1; C515852:ROCK2 protein, human; C515855:Rock2 protein, mouse; D058570:TOR Serine-Threonine Kinases; D054460:rho-Associated Kinases; C433811:HDAC6 protein, human; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
keywords: ROCK2; TSC2; cytokinesis; microtubule; tuberous sclerosis complex