小児の脳障害に関連する遺伝子群を解明するための新しいシステムの開発

• Project/Area Number: 11170236
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tottori University
• Principal Investigator: 難波 栄二 鳥取大学, 遺伝子実験施設, 助教授 (40237631)
• Co-Investigator(Kenkyū-buntansha): 小枝 達也 鳥取大学, 教育地域科学部, 教授 (70225390) ; 岡 明 鳥取大学, 医学部, 助教授 (00251273)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥5,000,000 (Direct Cost: ¥5,000,000); Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000); Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
• Keywords: 小児 / 脳障害 / 遺伝的背景 / 単一遺伝 / 多因子遺伝 / 自閉症 / セロトニン / インプリンティング / 発達障害 / 脳 / 遺伝子 / 遺伝子診断 / 精神遅滞 / ゲノムインプリンティング

Research Abstract

【目的】小児の脳障害には様々な遺伝子が関係するがその多くは末だ解明されていない。そこで本研究は、単一遺伝子病やさらに頻度の高い疾患も含め、それらに関連する遺伝群を解明することを目的とした。
【単一遺伝子病】結節性硬化症をはじめとする単一遺伝子病の解析システムは確立でき、様々な脳障害を引き起こす患者の遺伝子異常が解明できた(研究発表参照)。【自閉症】頻度の高い自閉症に焦点をあてた。本疾患は多因子遺伝と考えられ新たなアプローチが必要であった。自閉症の患者や正常対照のDNAサンプルは鳥取大学および東京大学倫理委員会の承認のもと、インフォームドコンセントを得て収集した。(1)関連遺伝子解析:11のセロトニン代謝関連遺伝子についてプロモーター領域多型やSingle Nucleotide Polymorphisms(SNPs)を検討した。17SNPsを解析したところ4つのSNPs(HTR1A,HTR1B,HTR1D,HTR6)で多型が認められた。従来示唆されていたセロトニントランスポータープロモーター領域多型の関連はなく、HTR1B,HTR1D,HTR6との関連も示されなかった。しかし、HTR1A遺伝子が関連する可能性が示された。(2)インプリンテイング機構からのアプローチ:インプリンティング遺伝子を単離できるシステムとしてマウスA9細胞にヒト染色体を1本のみ保持するライブラリーを構築し研究を進めた。本システムを用いて19番染色体上のPEG3のインプリンティング機構が解析できた。さらにセロトニン関連遺伝子がインプリンティング機構と関連するかどうかを検討した。
HTR1D,HTR1A,HTR1B,HTR1E,HTR5A,HTR3A,HTR2Aの遺伝子発現を検討したが、いずれもインプリンティング機構との関連は示されなかった。

Research Products

• [Publications] Murakami F: "Anxiety traits associated with a polymorphism in the serotonin transporter gene regulatory region in the Japanese."J Hum Genet. 44. 15-17 (1999)
• [Publications] Zhang H: "Novel TSC2 mutation in a patient with pulmonary tuberous sclerosis : lack of loss of heterozygosity in a lung cyst."Am J Med Genet. 82. 368-370 (1999)
• [Publications] Ueta E: "A rapid screening method by the polymerase chain reaction-single strand conformation polymorphism using fluorescent labeled primer on automatic sequencer for porcine ryanodine receptor gene mutation."J Animal Genet. 27. 7-13 (1999)
• [Publications] Kono Y: "Analysis of the CAG repeat number in patient with Huntington's disease."Intern Med. 38. 407-411 (1999)
• [Publications] Yamamoto T: "A novel mutation (A246T) in exon 6 of the proteolipid protein gene associated with connatal Pelizaeus-Merzbacher disease."Hum Mut. 14. 182 (1999)
• [Publications] Yamamoto T: "NPC1 gene mutations in Japanese patients with Niemann-Pick discase type C."Hum Genet. 105. 10-16 (1999)
• [Publications] Zhang H: "Mutational analysis of TSC1 and TSC2 genes in Japanese patients with tuberous selerosis complex."J Hum Genet. 44. 391-396 (1999)
• [Publications] Oka A: "The up-regulation of metabotropic glutamate receptor 5 (MGluR5) in Down's syndrome brains."Acta Neuropathol. 97. 275-278 (1999)
• [Publications] Yamamoto T: "A patient with cerebral palsy whose mother had a traffic accident during pregnancy : a diffuse axonal injury?"Brain Dev. 21. 334-336 (1999)
• [Publications] Kato M: "Sonic hedgehog signal peptide mutation in a patient with holoprosencephaly."Ann Neurol. 47. 514-516 (2000)
• [Publications] Imai J: "Polymorphism of the cytochrome P450 (CYP) 2C9 gene in Japanese epileptic patients : genetic analysis of the CYP2C9 locus."Pharmacogenetics. 10. 85-89 (2000)
• [Publications] Ikebuchi M: "The Arg1075His substitution in the FBN1 gene is clinieally innocent for Marfan syndrome."Hum Mut. 15. 298 (2000)
• [Publications] Chikumi H: "Fibrillin gene (FBN1) mutations in Japanese patients with Marfan syndrome."J Hum Genet. 45. 115-118 (2000)
• [Publications] Nagata K: "A novel interstitial deletion of KAL1 in a Japanese family with Kallmann syndrome."J Hum Genet. 45. 237-240 (2000)
• [Publications] Pipo JR: "Two novel serine repeat length polymorphisms (1043insS and 1043insSS) at exon 23 of the TSC1 gene."Hum Mut. 16. 375 (2000)
• [Publications] Saito Y: "Widespread expression of α-synuelein and tau immunoreactivity in Hallcrvorden-Spatz syndrome with protracted clinical course."J Neurol Sci. 177. 48-59 (2000)
• [Publications] Tanaka Y: "Congenital myotonic dystrophy : report of paternal transmission."Brain Dev. 22. 132-134 (2000)
• [Publications] Kurachi Y: "Rapid immunologic diagnosis of classic late infantile neuronal ceroid lipofuseinosis."Neurology. 54. 1676-1680 (2000)
• [Publications] Oka A: "Expression of DNA-dependent protein kinase catalytic subunit and Ku80 in developing human brains : implication of DNA-repair in neurogenesis."Neuosci Lett. 292. 167-170 (2000)
• [Publications] Shitoa M: "Cognitive and neurophysiological evaluation of Japanese dyslexia."Brain Dev. 22. 421-426 (2000)
• [Publications] Yamamoto T: "Genotype-phenotype relationship of Niermann-Pick disease Type C : a possibility correlation between clinical onset and levels of NPC1 protein in isolated skin fibroblasts."J Med Genet. 37. 707-711 (2000)
• [Publications] Tsukamoto H: "SSCP analysis by RT-PCR for the prenatal diagnosis of Niemann-Pick disease type C."Prenat Diagn. 21. 55-57 (2001)
• [Publications] Yamamoto T: "DEFECT 11 syndrome associated with agenesis of the corpus callosum."J Med Genet.. 38. E5 (2001)
• [Publications] Kato,M,: "Sonic hedgehog signal peptide mutation in a patient with holoprosencephaly"Ann.Neurol.. (in press).
• [Publications] Ohno,K.: "Molecular epidemiology of tuberous sclerosis"Gann Monograhp on Cancer Research. 46. 53-59 (1999)
• [Publications] Zhang,H.: "Mutational Analysis of TSC1 and TSC2 Genes in Japanese Patients with Tuberous Sclerosis Complex"J.Hum.Genet.. 44. 391-396 (1999)
• [Publications] Yamamoto,T.: "NPC1 gene mutations in Japanese patients with Niemann-Pick disease typeC"Hum.Genet.. 105. 10-16 (1999)
• [Publications] Kono,T.: "Analysis of the GAG repeat number in patient with Huntigton's disease"Internal Medicine. 38. 407-411 (1999)
• [Publications] Zhang,H.: "A novel TSC2 Mutation in a Patient With Pulmonary Tuberous Selerosis:Lacl of LOH in a Lung Cyst"Am.J.Med.Genet.. 82. 368-370 (1999)
• [Publications] Murakami,F.: "Anxiety traits associated with a polymorphism in the serotonin transporter gene regulatory region in the Japanese"J.Hum.Genet.. 44. 15-17 (1999)
• [Publications] Koeda,T.: "Distribution in normal subjects of performance in token test: a basic study for the diagnosis of learning disabilities"No To Hattatsu. 32(1). 25-28 (2000)
• [Publications] Oka,A.: "The early induction of cyclooxygenase2 associated with neurofibrillary degeneration in brains of patients with Fukuyama-type congenital muscular dystrophy"Neuropediatrics. 30(1). 34-37 (1999)