{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T01:56:05Z","timestamp":1760234165104,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,4,16]],"date-time":"2021-04-16T00:00:00Z","timestamp":1618531200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["NS103981"],"award-info":[{"award-number":["NS103981"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Tuberous sclerosis complex (TSC) is a genetic disorder caused by inactivating mutations in TSC1 (hamartin) or TSC2 (tuberin), crucial negative regulators of the mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway. TSC affects multiple organs including the brain. The neurologic manifestation is characterized by cortical tubers, subependymal nodules (SEN), and subependymal giant cell astrocytoma (SEGA) in brain. SEGAs may result in hydrocephalus in TSC patients and mTORC1 inhibitors are the current recommended therapy for SEGA. Nevertheless, a major limitation in the research for SEGA is the lack of cell lines or animal models for mechanistic investigations and development of novel therapy. In this study, we generated TSC1-deficient neural cells from spontaneously immortalized mouse astrocytes in an attempt to mimic human SEGA. The TSC1-deficient cells exhibit mTORC1 hyperactivation and characteristics of transition from astrocytes to neural stem\/progenitor cell phenotypes. Rapamycin efficiently decreased mTORC1 activity of these TSC1-deficient cells in vitro. In vivo, TSC1-deficient cells could form SEGA-like tumors and Rapamycin treatment decreased tumor growth. Collectively, our study generates a novel SEGA-like cell line that is invaluable for studying mTORC1-driven molecular and pathological alterations in neurologic tissue. These SEGA-like cells also provide opportunities for the development of novel therapeutic strategy for TSC patients with SEGA.<\/jats:p>","DOI":"10.3390\/ijms22084116","type":"journal-article","created":{"date-parts":[[2021,4,16]],"date-time":"2021-04-16T10:20:11Z","timestamp":1618568411000},"page":"4116","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The Characterization of a Subependymal Giant Astrocytoma-Like Cell Line from Murine Astrocyte with mTORC1 Hyperactivation"],"prefix":"10.3390","volume":"22","author":[{"given":"Xin","family":"Tang","sequence":"first","affiliation":[{"name":"Department of Cancer Biology, University of Cincinnati College Medicine, Cincinnati, OH 45267, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabrielle","family":"Angst","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, University of Cincinnati College Medicine, Cincinnati, OH 45267, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Haas","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, University of Cincinnati College Medicine, Cincinnati, OH 45267, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fuchun","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, University of Cincinnati College Medicine, Cincinnati, OH 45267, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chenran","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Cancer Biology, University of Cincinnati College Medicine, Cincinnati, OH 45267, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.pedhc.2006.05.004","article-title":"Tuberous Sclerosis Complex: A Review","volume":"21","author":"Leung","year":"2007","journal-title":"J. Pediatr. Health Care"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1093\/jnen\/63.12.1236","article-title":"Pathogenesis of Tuberous Sclerosis Subependymal Giant Cell Astrocytomas: Biallelic Inactivation ofTSC1orTSC2Leads to mTOR Activation","volume":"63","author":"Chan","year":"2004","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1007\/s11940-011-0123-z","article-title":"Subependymal Giant Cell Astrocytoma (SEGA) Treatment Update","volume":"13","author":"Campen","year":"2011","journal-title":"Curr. Treat. Options Neurol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1101\/gad.16750211","article-title":"Tsc1 mutant neural stem\/progenitor cells exhibit migration deficits and give rise to subependymal lesions in the lateral ventricle","volume":"25","author":"Zhou","year":"2011","journal-title":"Genes Dev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1038\/s42255-019-0137-5","article-title":"Autophagic lipid metabolism sustains mTORC1 activity in TSC-deficient neural stem cells","volume":"1","author":"Wang","year":"2019","journal-title":"Nat. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1688","DOI":"10.1172\/JCI96342","article-title":"Tuberous sclerosis complex\u2013associated CNS abnormalities depend on hyperactivation of mTORC1 and Akt","volume":"128","author":"Zordan","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1038\/s41591-018-0139-y","article-title":"Genetically engineered human cortical spheroid models of tuberous sclerosis","volume":"24","author":"Blair","year":"2018","journal-title":"Nat. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13229-021-00417-x","article-title":"Mind the translational gap: Using iPS cell models to bridge from genetic discoveries to perturbed pathways and therapeutic targets","volume":"12","author":"Pintacuda","year":"2021","journal-title":"Mol. Autism"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/j.pediatrneurol.2013.12.004","article-title":"Surgical Treatment of Subependymal Giant Cell Astrocytoma in Tuberous Sclerosis Complex Patients","volume":"50","author":"Kotulska","year":"2014","journal-title":"Pediatr. Neurol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1002\/ana.20784","article-title":"Rapamycin causes regression of astrocytomas in tuberous sclerosis complex","volume":"59","author":"Franz","year":"2006","journal-title":"Ann. Neurol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1801","DOI":"10.1056\/NEJMoa1001671","article-title":"Everolimus for Subependymal Giant-Cell Astrocytomas in Tuberous Sclerosis","volume":"363","author":"Krueger","year":"2010","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12883-021-02160-5","article-title":"Experience using mTOR inhibitors for subependymal giant cell astrocytoma in tuberous sclerosis complex at a single facility","volume":"21","author":"Tomoto","year":"2021","journal-title":"BMC Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/S0140-6736(12)61134-9","article-title":"Efficacy and safety of everolimus for subependymal giant cell astrocytomas associated with tuberous sclerosis complex (EXIST-1): A multicentre, randomised, placebo-controlled phase 3 trial","volume":"381","author":"Franz","year":"2013","journal-title":"Lancet"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s13041-016-0222-6","article-title":"Everolimus improves neuropsychiatric symptoms in a patient with tuberous sclerosis carrying a novel TSC2 mutation","volume":"9","author":"Hwang","year":"2016","journal-title":"Mol. Brain"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"951","DOI":"10.2147\/TCRM.S145630","article-title":"Everolimus for the treatment of refractory seizures associated with tuberous sclerosis complex (TSC): Current perspectives","volume":"15","author":"Overwater","year":"2019","journal-title":"Ther. Clin. Risk Manag."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5129","DOI":"10.1038\/onc.2012.640","article-title":"Senescence and aging: The critical roles of p53","volume":"32","author":"Rufini","year":"2013","journal-title":"Oncogene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"re2","DOI":"10.1126\/scisignal.2005326","article-title":"Transcriptional regulation of the stress response by mTOR","volume":"7","author":"Aramburu","year":"2014","journal-title":"Sci. Signal."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1177\/08830738040190091401","article-title":"Mouse models of tuberous sclerosis complex","volume":"19","author":"Scheidenhelm","year":"2004","journal-title":"J. Child Neurol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.celrep.2016.02.090","article-title":"mTORC1 Inhibition Corrects Neurodevelopmental and Synaptic Alterations in a Human Stem Cell Model of Tuberous Sclerosis","volume":"15","author":"Costa","year":"2016","journal-title":"Cell Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/j.stemcr.2017.02.020","article-title":"Abnormal Neural Progenitor Cells Differentiated from Induced Pluripotent Stem Cells Partially Mimicked Development of TSC2 Neurological Abnormalities","volume":"8","author":"Li","year":"2017","journal-title":"Stem Cell Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1056\/NEJMcibr1406613","article-title":"Molecular Basis of Giant Cells in Tuberous Sclerosis Complex","volume":"371","author":"Kwiatkowski","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1007\/s004010050438","article-title":"Immunohistochemical characterization of subependymal giant cell astrocytomas","volume":"91","author":"Lopes","year":"1996","journal-title":"Acta Neuropathol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.stem.2011.09.008","article-title":"Sustained Activation of mTOR Pathway in Embryonic Neural Stem Cells Leads to Development of Tuberous Sclerosis Complex-Associated Lesions","volume":"9","author":"Magri","year":"2011","journal-title":"Cell Stem Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1007\/s11626-999-0072-y","article-title":"A novel human astrocyte cell line (A735) with astrocyte-specific neurotransmitter function","volume":"35","author":"Price","year":"1999","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Rusnakova, V., Honsa, P., Dzamba, D., St\u00e5hlberg, A., Kubista, M., and Anderova, M. (2013). Heterogeneity of Astrocytes: From Development to Injury\u2014Single Cell Gene Expression. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069734"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1007\/BF00691876","article-title":"Subependymal giant-cell tumor: Astrocytic or neuronal?","volume":"60","author":"Nakamura","year":"1983","journal-title":"Acta Neuropathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/S0165-4608(98)00140-X","article-title":"Cytogenetic changes in two cases of subependymal giant-cell astrocytoma","volume":"109","author":"Zakrzewski","year":"1999","journal-title":"Cancer Genet. Cytogenet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1002\/ana.10579","article-title":"Markers of cellular proliferation are expressed in cortical tubers","volume":"53","author":"Lee","year":"2003","journal-title":"Ann. Neurol."},{"key":"ref_29","first-page":"39","article-title":"Subependymal giant cell astrocytomas with atypical histological features mimicking malignant gliomas","volume":"49","author":"Grajkowska","year":"2011","journal-title":"Folia Neuropathol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1038\/s41576-019-0205-4","article-title":"Engineering adeno-associated virus vectors for gene therapy","volume":"21","author":"Li","year":"2020","journal-title":"Nat. Rev. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Cheah, P.-S., Prabhakar, S., Yellen, D., Beauchamp, R.L., Zhang, X., Kasamatsu, S., Bronson, R.T., Thiele, E.A., Kwiatkowski, D.J., and Stemmer-Rachamimov, A. (2021). Gene therapy for tuberous sclerosis complex type 2 in a mouse model by delivery of AAV9 encoding a condensed form of tuberin. Sci. Adv., 7.","DOI":"10.1126\/sciadv.abb1703"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2063","DOI":"10.1002\/glia.22577","article-title":"Traumatic scratch injury in astrocytes triggers calcium influx to activate the JNK\/c-Jun\/AP-1 pathway and switch on GFAP expression","volume":"61","author":"Gao","year":"2013","journal-title":"Glia"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2581","DOI":"10.1083\/jcb.201609093","article-title":"Autophagy gene FIP200 in neural progenitors non\u2013cell autonomously controls differentiation by regulating microglia","volume":"216","author":"Wang","year":"2017","journal-title":"J. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1038\/nn.3365","article-title":"FIP200 is required for maintenance and differentiation of postnatal neural stem cells","volume":"16","author":"Wang","year":"2013","journal-title":"Nat. Neurosci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1083\/jcb.201507023","article-title":"Elevated p62\/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide","volume":"212","author":"Wang","year":"2016","journal-title":"J. Cell Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"29398","DOI":"10.1074\/jbc.M110.141440","article-title":"Regulation of Integrin beta 1 recycling to lipid rafts by Rab1a to promote cell migration","volume":"285","author":"Wang","year":"2010","journal-title":"J. Biol. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/8\/4116\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:48:41Z","timestamp":1760161721000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/8\/4116"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,16]]},"references-count":36,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["ijms22084116"],"URL":"https:\/\/doi.org\/10.3390\/ijms22084116","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,4,16]]}}}