Isolation and characterization of the new genes isolated from three diseases presenting with severe psychomotor retardation.

从三种表现为严重精神运动迟缓的疾病中分离出的新基因的分离和表征。

• 批准号: 15390332
• 负责人: WAKAMATSU Nobuaki
• 金额: $ 9.34万
• 依托单位: Institute for Developmental Research, Aichi Human Service Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390332/
• 关键词: mental retardation; gene; PLEKHA5; autosomal recessive; X-linked recessive; brain atrophy; 重度知的障害; Xq28; 伴性劣性遺伝; 常染色体劣性遺伝; PEPP2

1. Isolation of the causal gene from the patient presenting with reciprocal 6q16/12p12 translocation, epilepsy and severe psychomotor retardation.We identified a new gene, PLEKHA5, at the breaking point of the 12p12 by FISH, Southern blot and nucleotide sequencing analyses. There was no gene located at the 6q16 breaking point. To examine the role of this gene, we transfected Plekha5-speciflc siRNA into mouse Neuro2a cells. Transfected cells express about 40% of Plekha5 mRNA and showed limited extension of the neurites compared to cells undergoing Mock transfection (vector only) when treated with 20 μM retinoic acid. This finding suggests that PLEKHA5 is likely a causal gene of this disease. Plekha5 localized at the cytoplasm. Affinity purified PLEKHA5 showed a size of 130 kDa.2. Isolation of the causal gene from male patients in a family presenting with severe psychomotor retardation.We performed linkage analysis in 11 members of the family. Three of these eleven were patients. Maximal … More lod score, 1.10, was obtained at DXS1205 of Xq27. Nucleotide sequencing analysis was performed for 15 candidate genes at Xq27-ter from the patients' DNA, but there were no apparent mutations.3. Isolation of the causal gene from patients in a family presenting with rapid brain atrophy and severe psychomotor retardation.We performed linkage analysis in 10 members of the family. Four of these ten were patients. Maximal lod score, 4.75 was obtained between D2S163 and D2S2344 at 2q35-36. Nucleotide sequence analysis was performed for 16 candidate genes and an E320Q missense mutation was found in SLC19A3. The patients were homozygous for the mutation and their parents were heterozygous. We constructed an expression vector pC1-SLC19A3 (E320Q), that expressed SLCJ9A3 (E320Q) driven by a CMV promoter and examined the transporter activity of thiamine and biotin after transfection of this vector into HEK293 cells. However, there was no remarkable reduction of transporter activity. We are now producing transgenic mouse expressing Slc193 (E320Q) to further investigate the effects of this mutation. Less

1. 从患有 6q16/12p12 相互易位、癫痫和严重精神运动迟缓的患者中分离出致病基因。我们通过 FISH、Southern blot 和核苷酸测序分析在 12p12 断裂点鉴定了一个新基因 PLEKHA5。 6q16断裂点处没有基因。为了检查该基因的作用，我们将 Plekha5 特异性 siRNA 转染至小鼠 Neuro2a 细胞中。当用 20 μM 视黄酸处理时，与模拟转染（仅载体）的细胞相比，转染的细胞表达约 40% 的 Plekha5 mRNA，并且显示出有限的神经突延伸。这一发现表明 PLEKHA5 可能是这种疾病的致病基因。 Plekha5 定位于细胞质。亲和纯化的 PLEKHA5 显示大小为 130 kDa.2。从患有严重精神运动性迟缓的家庭中的男性患者中分离出致病基因。我们对该家庭的 11 名成员进行了连锁分析。这十一人中有三人是病人。最大 lod 分数 1.10，是在 Xq27 的 DXS1205 上获得的。对患者DNA中Xq27-ter位点的15个候选基因进行了核苷酸测序分析，未发现明显突变。 3．从患有快速脑萎缩和严重精神运动迟缓的家庭患者中分离出致病基因。我们对该家庭的 10 名成员进行了连锁分析。这十人中有四人是病人。 D2S163 和 D2S2344 之间在 2q35-36 处获得的最大 lod 得分为 4.75。对16个候选基因进行核苷酸序列分析，在SLC19A3中发现E320Q错义突变。患者的突变是纯合的，而他们的父母是杂合的。我们构建了表达载体pC1-SLC19A3 (E320Q)，该载体表达由CMV启动子驱动的SLCJ9A3 (E320Q)，并检测该载体转染至HEK293细胞后硫胺素和生物素的转运蛋白活性。然而，转运蛋白活性并没有显着减少。我们现在正在生产表达 Slc193 (E320Q) 的转基因小鼠，以进一步研究这种突变的影响。较少的

项目成果

MECP2遺伝子異常を伴うRett症候群の臨床症状について.

与 MECP2 基因异常相关的 Rett 综合征的临床症状。

• 发表时间: 2005
• 期刊: 脳と発達 37・1
• 作者: 三浦清邦;他
• 通讯作者: 他

Expression level of vascular endothelial growth factor-C and -A in cultured human oral squamous cell carcinoma correlates respectively with lymphatic metastasis and angiogenesis when transtplanted into nude mouse oral cavity.

培养的人口腔鳞状细胞癌中血管内皮生长因子-C和-A的表达水平分别与移植到裸鼠口腔后的淋巴转移和血管生成相关。

• 发表时间: 2006
• 期刊: Oncology Reports 15-4
• 作者: Nakazato T;et al.
• 通讯作者: et al.

Two cases of partial trisomy 21 (pter-q22. 1) without the major features of Down syndrome.

两例部分21三体（pter-q22.1），不具有唐氏综合症的主要特征。

• 发表时间: 2006
• 期刊: Am J Med Genet 140A
• 作者: Kondo Y;Mizuno S;Ohara K;Nakamura T;Yamada K;Yamamori S;Hayakawa C;Ishii T;Yamada Y;Wakamatsu N
• 通讯作者: Wakamatsu N

Comparison of the analyses to diagnose the carrier of the mutation in a Lesch-Nyhan family.

Lesch-Nyhan 家族中突变携带者诊断分析的比较。

• 发表时间: 2004
• 期刊: Japanese Journal of Pediatrics 57-5
• 作者: Igarashi N;et al.
• 通讯作者: et al.

Disruption in the hypoxanthine phosphoribosyltransferase gene caused by translocation in a patient with Lesch-Nyhan syndrome

Lesch-Nyhan 综合征患者易位导致次黄嘌呤磷酸核糖转移酶基因破坏

• 发表时间: 2004
• 期刊: Nucleosides Nucleotides Nucleic Acids 23
• 作者: Mizunuma M;Yamada Y;Yamada K;Santa S;Wakamatsu N;Kaneko K;Ogasawara N;Fujimori S
• 通讯作者: Fujimori S