Identification of the germline-derived GNAS gain-of-function mutations and clarification of the novel mechanism leading to GNAS loss-of-function

2019 Fiscal Year Annual Research Report

• Project/Area Number: 17H04204
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: 緒方 勤 浜松医科大学, 医学部, 教授 (40169173)
• Co-Investigator(Kenkyū-buntansha): 深見 真紀 国立研究開発法人国立成育医療研究センター, 分子内分泌研究部, 部長 (40265872)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: GNAS / 抗利尿不適合性腎症候群 / 機能亢進 / 偽性副甲状腺機能低下症 / 機能低下

Outline of Annual Research Achievements

GNASは、Gタンパク質共役型受容体の細胞内シグナル伝達を介在し、腎集合管を含む多くの組織では両親性発現を示すが、腎近位尿細管などではA/B-DMRの制御により母性発現を示す。
本研究の目的は、(1)われわれが常染色体優性の抗利尿不適合性腎症候群を呈する2家系で同定した新規GNAS変異が、世界初の機能亢進型生殖細胞変異であることを示すこと（研究項目１）、(2) われわれが常染色体優性の偽性副甲状腺機能低下症を示す2家系で同定した過去に報告の無いGNAS-STX16領域のゲノム構造異常やレトロトランスポゾン挿入が、A/B-DMR単独のエピ変異を発症する機序を解明すること（研究項目2）である。
（研究項目１）抗利尿不適合性腎症候群で同定されたp.F68_G70delとp.M255V変異のタンパク構造解析を行い、p.F68_G70delの評価はその部位から困難であったが、p.M255V変異は間接的にGTPase作用を阻害してGNAS機能亢進を招くことが判明した。ついでAVPR2を用いたルシフェラーゼ解析でp.F68_G70delとp.M255V変異軽度機能亢進作用を有することを見出した。その程度は、p.M255V変異において明瞭であった。また、p.F68_G70delを導入したモデルマウスが、患者と同様、腎からの自由水排泄障害を呈することを見出し、p.M255Vを導入したモデルマウスが在胎時では生存するものの生後間もなく死亡することを明らかとした。これらのデータにより米国腎臓病学会雑誌に論文が掲載され、Editorial commentsが寄せられた。
（研究項目2）新規GNAS機能低下発症機序の解明では、レトロトランスポゾン挿入家系でdCAS9-VP64システムを用いた検討を行ったが、exonizationを示す明確なデータは得られず、未知の原因に起因することが想定された。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] IGF2 Mutations: Report of Five Cases, Review of the Literature, and Comparison with H19/IGF2:IG-DMR Epimutations. (2020)
  • Author(s): Masunaga Y, Inoue T, Yamoto K, Fujisawa Y, Sato Y, Kawashima-Sonoyama Y, Morisada N, Iijima K, Ohata Y, Namba N, Suzumura H, Kuribayashi R, Yamaguchi Y, Yoshihashi H, Fukami M, Saitsu H, Kagami M, Ogata T*
  • Journal Title: J Clin Endocrinol Metab Volume: 105 Pages: pii: dgz034
  • DOI: 10.1210/clinem/dgz034.
  • Peer Reviewed
• [Journal Article] De novo ZBTB7A variant in a patient with macrocephaly, intellectual disability, and sleep apnea: implications for the phenotypic development in 19p13.3 microdeletions. (2020)
  • Author(s): Ohishi A, Masunaga Y, Iijima S, Yamoto K, Kato F, Fukami M, Saitsu H, Ogata T*
  • Journal Title: J Hum Genet Volume: 65 Pages: 181-186
  • DOI: 10.1038/s10038-019-0690-5
  • Peer Reviewed
• [Journal Article] Erythrokeratodermia variabilis et progressiva with a rare GJB3 mutation. (2020)
  • Author(s): Imura K, Ikeya S, Ogata T, Tokura Y
  • Journal Title: J Dermatol Volume: 47 Pages: e111-e113
  • DOI: 10.1111/1346-8138.15206
  • Peer Reviewed
• [Journal Article] POLR3A variants in striatal involvement without diffuse hypomyelination (2020)
  • Author(s): Hiraide T, Kobota K, Kono Y, Watanabe S, Matsubayashi T, Nakashima M, Kaname T, Fukao T, Shimozawa N, Ogata T, Saitsu H*
  • Journal Title: Brain Dev Volume: 42 Pages: 363-368
  • DOI: 10.1016/j.braindev.2019.12.012.
  • Peer Reviewed
• [Journal Article] A de novo TOP2B variant associated with global developmental delay and autism spectrum disorder. (2020)
  • Author(s): Hiraide T, Watanabe S, Matsybayashi T, Yanagi K, Nakashima M, Ogata T, Saitsu H*
  • Journal Title: Mol Genet Genomic Med Volume: 8 Pages: e1145
  • DOI: org/10.1002/mgg3.11
  • Peer Reviewed / Open Access
• [Journal Article] (Epi)genetic defects of MKRN3 are rare in Asian patients with central precocious puberty. (2019)
  • Author(s): Suzuki E, Shima H, Kagami M, Soneda S, Tanaka T, Yatsuga S, Nishioka J, Oto Y, Kamiya T, Naiki Y, Ogata T, Fujisawa Y, Nakamura A, Kawashima S, Morikawa S, Horikawa R, Sano S, Fukami M*
  • Journal Title: Hum Genome Var Volume: 21 Pages: 7
  • DOI: 10.1038/s41439-019-0039-9
  • Peer Reviewed / Open Access
• [Journal Article] Identification of de novo CSNK2A1 and CSNK2B variants in cases of global developmental delay with seizures. (2019)
  • Author(s): Nakashima M, Tohyama J, Nakagawa E, Watanabe Y, Siew CG, Kwong CS, Yamoto K, Hiraide T, Fukuda T, Kaname T, Nakabayashi K, Hata K, Ogata T, Saitsu H, Matsumoto N*
  • Journal Title: J Hum Genet Volume: 64 Pages: 313-322
  • DOI: 10.1038/s10038-018-0559-z.
  • Peer Reviewed
• [Journal Article] Coexistence of a CAV3 mutation and a DMD deletion in a family with complex muscular diseases. (2019)
  • Author(s): Hiraide T, Ogata T, Watanabe S, Nakashima M, Fukuda T, Saitsu H*
  • Journal Title: Brain Dev Volume: 41 Pages: 474-479
  • DOI: 10.1016/j.braindev.2019.01.005.
  • Peer Reviewed
• [Journal Article] Association of four imprinting disorders and ART. (2019)
  • Author(s): Hattori H, Hiura H, Kitamura A, Miyauchi N, Kobayashi N, Takahashi S, Okae H, Kyono K, Kagami M, Ogata T, Arima T*
  • Journal Title: Clin Epigenetics Volume: 11 Pages: 21
  • DOI: 10.1186/s13148-019-0623-3.
  • Peer Reviewed
• [Journal Article] Germline-derived gain-of-function variants of Gsa-coding GNAS gene identified in nephrogenic syndrome of inappropriate antidiuresis. (2019)
  • Author(s): Miyado M, Fukami M, Takada S, Terao M, Nakabayashi K, Hata K, Matsubara Y, Tanaka Y, Sasaki G, Nagasaki K, Shiina M, Ogata K, Masunaga Y, Saitsu H, Ogata T*
  • Journal Title: J Am Soc Nephrol Volume: 30 Pages: 877-889
  • DOI: 10.1681/ASN.2018121268.
  • Peer Reviewed
• [Journal Article] Exploring the unique function of imprinting control centers in the PWS/AS-responsible region: finding from array-based methylation analysis in cases with variously sized microdeletions (2019)
  • Author(s): Matsubara K, Itoh M, Shimizu K, Saito S, Enomoto K, Nakabayashi K, Hata K, Kurosawa K, Ogata T, Fukami M, Kagami M.
  • Journal Title: Clin Epigenetics Volume: 11 Pages: 36
  • DOI: 10.1186/s13148-019-0633-1.
  • Peer Reviewed / Open Access
• [Journal Article] Temple syndrome in a patient with variably methylated CpGs at the primary MEG3/DLK1:IG-DMR and severely hypomethylated CpGs at the secondary MEG3:TSS-DMR. (2019)
  • Author(s): Kagami M, Yanagisawa A, Ota M, Matsuoka K, Nakamura A, Matsubara K, Nakabayashi K, Takada S, Fukami M, Ogata T*
  • Journal Title: Clin Epigenetics Volume: 11 Pages: 42
  • DOI: 10.1186/s13148-019-0640-2.
  • Peer Reviewed / Open Access
• [Journal Article] MYRF haploinsufficiency causes 46,XY and 46,XX disorders of sex development. (2019)
  • Author(s): Hamanaka K, Takata A, Uchiyama Y, Miyatake S, Miyake N, Mitsuhashi S, Iwama K, Fujita A, Imagawa E, Alkanaq AN, Koshimizu E, Azuma Y, Nakashima M, Mizuguchi T, Saitsu H, Yuka Wada, Minami S, Katoh-Fukui Y, Masunaga Y, Fukami M, Hasegawa T, Ogata T, Matsumoto N*
  • Journal Title: Hum Mol Genet Volume: 28 Pages: 2319-2329
  • DOI: 10.1093/hmg/ddz066
• [Journal Article] Unbalanced Y;7 translocation between two low-similarity sequences leading to SRY-positive 45,X-testicular disorders of sex development. (2019)
  • Author(s): Uehara E, Hattori A, Shima H, Ishiguro A, Abe Y, Ogata T, Ogawa E, Fukami M*
  • Journal Title: Cytogenet Genome Res Volume: 158 Pages: 115-120
  • DOI: 10.1159/000501378
  • Peer Reviewed
• [Journal Article] Japanese Study Group of Insulin Therapy for Childhood and Adolescent Diabetes.: KLF11 (MODY7) variant in a family clinically diagnosed with early childhood-onset type 1B diabetes. (2019)
  • Author(s): Ushijima K, Narumi S, Ogata T, Yokota I, Sugihara S, Kaname T, Horikawa Y, Matsubara Y, Fukami M*, Kawamura T
  • Journal Title: Pediatr Diabetes Volume: 20 Pages: 712-719
  • DOI: 10.1111/pedi.12868.
  • Peer Reviewed
• [Journal Article] Aneuploid rescue precedes X chromosome inactivation and increases the incidence of its skewness by reducing the size of the embryonic progenitor cell pool. (2019)
  • Author(s): Yoshida T, Miyado M, Mikami M, Suzuki E, Kinjo K, Matsubara K, Ogata T, Akutsu H, Kagami M, Fukami M*
  • Journal Title: Hum Reprod Volume: 34 Pages: 1762-1769
  • DOI: 10.1093/humrep/dez117
  • Peer Reviewed
• [Journal Article] Comprehensive clinical and molecular studies in split hand/foot malformation: identification of two plausible candidate genes (LRP6 and UBA2). (2019)
  • Author(s): Yamoto K, Saitsu H, Nishimura G, Kosaki R, Takayama S, Haga N, Tonoki H, Okumura A, Horii E, Okamoto N, Suzumura H, Ikegawa S, Kato F, Fujisawa Y, Nagata E, Takada S, Fukami M, Ogata T*
  • Journal Title: Eur J Hum Gene Volume: 27 Pages: 1845-1857
  • DOI: 10.1038/s41431-019-0473-7.
  • Peer Reviewed
• [Journal Article] De novo AFF3 variant in a patient with mesomelic dysplasia with foot malformation. (2019)
  • Author(s): Shimizu D, Sakamoto R, Yamoto K, Saitsu H, Fukami M, Nishimura G, Ogata T*
  • Journal Title: J Hum Genet Volume: 64 Pages: 1041-1044
  • DOI: 10.1038/s10038-019-0650-0.
  • Peer Reviewed
• [Presentation] 世界初の生殖細胞由来GNAS機能亢進型バリアントの同定 (2020)
  • Author(s): 宮戸真美，深見真紀，緒方勤
  • Organizer: 第30回バゾプレシン研究会
• [Presentation] Germline-derived Gain-of-Function Variants of Gsα-coding GNAS Gene Identified in Nephrogenic Syndrome of Inappropriate Antidiuresis: The First report (2019)
  • Author(s): Fukami M, Miyado M, Takada S, Sasaki G, K Nagasaki, Masunaga M, Saitsu H, Ogata T
  • Organizer: The 58th Annual Meeting of European Pediatric Endocrinology
  • Int'l Joint Research
• [Presentation] Germline-derived gain-of-function variants of Gsα-coding GNAS gene identified in nephrogenic syndrome of inappropriate antidiuresis: the first report (2019)
  • Author(s): Miyado M, Fukami M, Matsubara Y, Tanaka Y, Sasaki G, Nagasaki K, Masunaga Y, Saitsu H and Ogata T.
  • Organizer: 第53回日本小児内分泌学会学術集会