Elucidation of the pathogenesis of the pediatric intractable epilepsy using human cerebral and midbrain organoids

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K23952
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0902:General internal medicine and related fields
• Research Institution: Nara Medical University
• Principal Investigator: Kaoru Kinugawa 奈良県立医科大学, 医学部附属病院, 研究員 (90846677)
• Project Period (FY): 2019-08-30 – 2021-03-31
• Keywords: 脳幹オルガノイド / 中脳オルガノイド / 1細胞遺伝子発現解析 / 小児難治性てんかん / ZEB2遺伝子

Outline of Final Research Achievements

Recently, three-dimensional in vitro human brain models derived from human pluripotent stem cells, brain organoids, have been developed. Brain organoids technologies are thought to be useful for research on pediatric neurological disorders, such as epilepsy. The aim of this study was to elucidate the pathogenesis of Mowat-Wilson Syndrome using human brain organoids of cerebrum and brainstem. MWS is one of the pediatric intractable epilepsy due to deficits of ZEB2 (zinc finger E-box-binding homeobox 2) gene. We established a method to induce brainstem organoids. Various methods, such as immunohistochemistry and single-cell RNA-sequencing, suggested that brainstem organoids mimic the structure of human brainstem. We developed the ZEB2 knock-out human brain organoids of cerebrum and brainstem, and performed morphological analysis and quantitative PCR. These results suggested that brain organoids might be useful model to elucidate the pathogenesis of the pediatric neurological disorders.

Free Research Field

脳神経内科学

Academic Significance and Societal Importance of the Research Achievements

小児難治性てんかんなどの小児神経疾患は、胎生期の中枢神経系の異常によって生じる。その多くは発生メカニズムが十分に解明されておらず、故に根本的な治療法がなく、病態解明が早急に必要とされていた。ヒト脳疾患の研究には、げっ歯類が用いられてきたが、ヒト脳との構造の違いから、疾患の病態解明を行うのは困難であった。本研究ではヒト多能性幹細胞、CRISPER/Cas9によるゲノム編集で得たZEB2欠損ヒト多能性幹細胞から、ヒト脳幹オルガノイド及びZEB2欠損ヒト脳幹オルガノイドを樹立した。これら脳オルガノイドは、小児の中枢神経疾患の発生過程の再現や病態解明に有用なツールとなりうると考えられた。