PMID- 32457579 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20211222 IS - 1662-5102 (Print) IS - 1662-5102 (Electronic) IS - 1662-5102 (Linking) VI - 14 DP - 2020 TI - Multimodal Analysis of STRADA Function in Brain Development. PG - 122 LID - 10.3389/fncel.2020.00122 [doi] LID - 122 AB - mTORopathies are a heterogeneous group of neurological disorders characterized by malformations of cortical development (MCD), enhanced cellular mechanistic target of rapamycin (mTOR) signaling, and epilepsy that results from mutations in mTOR pathway regulatory genes. Homozygous mutations (del exon 9-13) in the pseudokinase STE20-related kinase adaptor alpha (STRAD-alpha; STRADA), an mTOR modulator, are associated with Pretzel Syndrome (PS), a neurodevelopmental disorder within the Old Order Mennonite Community characterized by megalencephaly, intellectual disability, and intractable epilepsy. To study the cellular mechanisms of STRADA loss, we generated CRISPR-edited Strada mouse N2a cells, a germline mouse Strada knockout (KO-/-) strain, and induced pluripotent stem cell (iPSC)-derived neurons from PS individuals harboring the STRADA founder mutation. Strada KO in vitro leads to enhanced mTOR signaling and iPSC-derived neurons from PS individuals exhibit enhanced cell size and mTOR signaling activation, as well as subtle alterations in electrical firing properties e.g., increased input resistance, a more depolarized resting membrane potential, and decreased threshold for action potential (AP) generation. Strada-/- mice exhibit high rates of perinatal mortality and out of more than 100 litters yielding both WT and heterozygous pups, only eight Strada-/- animals survived past P5. Strada-/- mice are hypotonic and tremulous. Histopathological examination (n = 5 mice) revealed normal gross brain organization and lamination but all had ventriculomegaly. Ectopic neurons were seen in all five Strada-/- brains within the subcortical white matter mirroring what is observed in human PS brain tissue. These distinct experimental platforms demonstrate that STRADA modulates mTOR signaling and is a key regulator of cell size, neuronal excitability, and cortical lamination. CI - Copyright (c) 2020 Dang, Glanowska, Iffland, Barnes, Baybis, Liu, Patino, Vaid, Streicher, Parker, Kim, Moon, Henry, Murphy, Sutton, Parent and Crino. FAU - Dang, Louis T AU - Dang LT AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. AD - Department of Pediatrics, Michigan Medicine, Ann Arbor, MI, United States. FAU - Glanowska, Katarzyna M AU - Glanowska KM AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. AD - Michigan Neuroscience Institute, Michigan Medicine, Ann Arbor, MI, United States. FAU - Iffland Ii, Philip H AU - Iffland Ii PH AD - Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, United States. FAU - Barnes, Allan E AU - Barnes AE AD - Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, United States. FAU - Baybis, Marianna AU - Baybis M AD - Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, United States. FAU - Liu, Yu AU - Liu Y AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Patino, Gustavo AU - Patino G AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Vaid, Shivanshi AU - Vaid S AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. AD - Department of Pediatrics, Michigan Medicine, Ann Arbor, MI, United States. FAU - Streicher, Alexandra M AU - Streicher AM AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Parker, Whitney E AU - Parker WE AD - Department of Neurosurgery, Weill-Cornell Medical Center, New York, NY, United States. FAU - Kim, Seonhee AU - Kim S AD - Louis Katz School of Medicine, Temple University, Philadelphia, PA, United States. FAU - Moon, Uk Yeol AU - Moon UY AD - Louis Katz School of Medicine, Temple University, Philadelphia, PA, United States. FAU - Henry, Frederick E AU - Henry FE AD - Michigan Neuroscience Institute, Michigan Medicine, Ann Arbor, MI, United States. AD - Department of Molecular, and Integrative Physiology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Murphy, Geoffrey G AU - Murphy GG AD - Michigan Neuroscience Institute, Michigan Medicine, Ann Arbor, MI, United States. AD - Department of Molecular, and Integrative Physiology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Sutton, Michael AU - Sutton M AD - Michigan Neuroscience Institute, Michigan Medicine, Ann Arbor, MI, United States. AD - Department of Molecular, and Integrative Physiology, Michigan Medicine, Ann Arbor, MI, United States. FAU - Parent, Jack M AU - Parent JM AD - Department of Neurology, Michigan Medicine, Ann Arbor, MI, United States. AD - Neurology Service, VA Ann Arbor Healthcare System, Ann Arbor, MI, United States. FAU - Crino, Peter B AU - Crino PB AD - Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, United States. LA - eng GR - K08 NS109289/NS/NINDS NIH HHS/United States GR - R01 NS097498/NS/NINDS NIH HHS/United States PT - Journal Article DEP - 20200508 PL - Switzerland TA - Front Cell Neurosci JT - Frontiers in cellular neuroscience JID - 101477935 PMC - PMC7227375 OTO - NOTNLM OT - epilepsy OT - iPSC OT - mTOR OT - megalencephaly OT - mouse OT - seizure EDAT- 2020/05/28 06:00 MHDA- 2020/05/28 06:01 PMCR- 2020/01/01 CRDT- 2020/05/28 06:00 PHST- 2019/12/06 00:00 [received] PHST- 2020/04/14 00:00 [accepted] PHST- 2020/05/28 06:00 [entrez] PHST- 2020/05/28 06:00 [pubmed] PHST- 2020/05/28 06:01 [medline] PHST- 2020/01/01 00:00 [pmc-release] AID - 10.3389/fncel.2020.00122 [doi] PST - epublish SO - Front Cell Neurosci. 2020 May 8;14:122. doi: 10.3389/fncel.2020.00122. eCollection 2020.