A Shared Pathogenic Mechanism for Valproic Acid and SHROOM3 Knockout in a Brain Organoid Model of Neural Tube Defects

被引：10

作者：

Takla, Taylor N. ^{[1
]}

Luo, Jinghui ^{[1
]}

Sudyk, Roksolana ^{[1
]}

Huang, Joy ^{[1
]}

Walker, John Clayton ^{[1
]}

Vora, Neeta L. ^{[2
]}

Sexton, Jonathan Z. ^{[3
,4
,5
]}

Parent, Jack M. ^{[1
,6
,7
]}

Tidball, Andrew M. ^{[1
]}

机构：

[1] Univ Michigan, Med Sch, Dept Neurol, Ann Arbor, MI 48109 USA

[2] Univ N Carolina, Sch Med, Dept Obstet & Gynecol, Div Maternal Fetal Med, Chapel Hill, NC 27599 USA

[3] Univ Michigan, Med Sch, Dept Internal Med, Ann Arbor, MI 48109 USA

[4] Univ Michigan, Coll Pharm, Dept Med Chem, Ann Arbor, MI 48109 USA

[5] Univ Michigan, Ctr Drug Repurposing, Ann Arbor, MI 48109 USA

[6] Univ Michigan, Michigan Neurosci Inst, Med Sch, Ann Arbor, MI 48109 USA

[7] VA Ann Arbor Healthcare Syst, Ann Arbor, MI 48105 USA

来源：

CELLS | 2023年 / 12卷 / 13期

关键词：

neuroteratogen; anencephaly; induced pluripotent stem cells; APICAL CONSTRICTION; MUTATIONS; MALFORMATIONS; TOXICITY; POLARITY; CELSR1; MOUSE;

D O I：

10.3390/cells12131697

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Neural tube defects (NTDs), including anencephaly and spina bifida, are common major malformations of fetal development resulting from incomplete closure of the neural tube. These conditions lead to either universal death (anencephaly) or severe lifelong complications (spina bifida). Despite hundreds of genetic mouse models of neural tube defect phenotypes, the genetics of human NTDs are poorly understood. Furthermore, pharmaceuticals, such as antiseizure medications, have been found clinically to increase the risk of NTDs when administered during pregnancy. Therefore, a model that recapitulates human neurodevelopment would be of immense benefit to understand the genetics underlying NTDs and identify teratogenic mechanisms. Using our self-organizing single rosette cortical organoid (SOSR-COs) system, we have developed a high-throughput image analysis pipeline for evaluating the SOSR-CO structure for NTD-like phenotypes. Similar to small molecule inhibition of apical constriction, the antiseizure medication valproic acid (VPA), a known cause of NTDs, increases the apical lumen size and apical cell surface area in a dose-responsive manner. GSK3 & beta; and HDAC inhibitors caused similar lumen expansion; however, RNA sequencing suggests VPA does not inhibit GSK3 & beta; at these concentrations. The knockout of SHROOM3, a well-known NTD-related gene, also caused expansion of the lumen, as well as reduced f-actin polarization. The increased lumen sizes were caused by reduced cell apical constriction, suggesting that impingement of this process is a shared mechanism for VPA treatment and SHROOM3-KO, two well-known causes of NTDs. Our system allows the rapid identification of NTD-like phenotypes for both compounds and genetic variants and should prove useful for understanding specific NTD mechanisms and predicting drug teratogenicity.

引用

页数：21