2012-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647496摘要：精神分裂症是具有遺傳成分及高遺傳率的精神疾病。過去針對台灣精神分裂症家族的基因體掃描發現10q22.3 有遺傳連鎖的證據。進一步的細部定位發現有四個位於此處的基因，ANXA7, PPP3CB, DNAJC9, ZMYND17 與持續注意力及執行功能缺損的精神分裂症亞群有顯著的遺傳關聯性，且表現量在病患組與正常對照組也有顯著差異，以基因功能及基因表現與基因型及神經認知缺損的關係推論，ANXA7 和PPP3CB 是最可能的候選基因。其基因病理機轉可能與基因表現量的改變有關，進一步探尋在這兩個基因的調節區域 (regulatory element) 的功能性變異，以及基因在神經細胞及神經系統扮演的主要功能，是未來研究的重點。本計劃的目的為(1)使用直接定序的方法，找尋ANXA7 和PPP3CB 的危險性基因多型性；(2)以定基因型的方法，在大樣本證實這些危險基因多型性的關聯性；(3)以luciferase reporter assay, RNA stability assay, Electrophoretic mobility shift assay (EMSA)等方法，了解這些危險性基因多型性的可能的生物功能。第一年，我們計畫使用直接定序的方法，以之前研究已知的42 位帶有同型危險性基因型的病人樣本及50 位正常對照組為樣本，定序ANXA7 和PPP3CB 基因的所有exon及promoter region (至轉譯起始點前2kb)。第二年將以大樣本確認這些危險性基因多型性的關聯性。第三年我們將以luciferase reporter assay, RNA stability assay, EMSA等方法確認這些危險性基因多型性的生物功能。本計劃是創新且可行的，因為(1)我們有一系列關於10q 的遺傳連鎖及關聯性研究及基因表現研究成果；(2)樣本皆已收集完畢且可進行定序及基因型的工作；(3)我們有研究其他基因的危險多型性的生物功能的經驗，其方法學已建立。<br> Abstract: Schizophrenia is a neuropsychiatric illness having a strong genetic component and highheritability. A previous genome scan of Taiwanese schizophrenia families suggested linkageto chromosome 10q22.3. Our further fine mapping study has identified annexin A7(ANXA7), protein phosphatase 3 (formerly 2B), catalytic subunit, beta isoform (PPP3CB),DnaJ (Hsp40) homolog, subfamily C, member 9 (DNAJC9), and zinc finger, MYND-typecontaining 17 (ZMYND17) at this chromosome region as potential candidate genes forschizophrenia, especially in patients with deficits in sustained attention and executivefunction. We also found that there was significantly lower expression of ANXA7, PPP3CB,and DNAJC9 and significantly higher expression of ZMYND17 in EBV transformedlymphocytes of schizophrenic patients. Therefore, we suggest that the pathophysiology ofschizophrenia may be related to the reduced expression of ANXA7, PPP3CB, DNAJC9 andincreased expression of ZMYND17, which suggest that the functional variants may belocated in the regulatory elements of these genes. Of the four genes, ANXA7 and PPP3CBare the candidate genes for the first priority because of their biological relevance and otherassociation evidence.The specific aims of this project are: (1) searching for the risk polymorphisms ofANXA7 and PPP3CB through direct sequencing method in the original associated sample,(2) verification of the association evidence of the risk polymorphisms using genotypingmethod in a large case-control sample, (3) clarification of the possible biological function ofthese identified risk polymorphisms through luciferase reporter assay, RNA stability assay,and Electrophoretic mobility shift assay (EMSA).In the first year, we prepare to perform the mutational analysis for ANXA7 and PPP3CBgene using direct sequencing method. We prepare to sequence all the exons and promoterregions (2kb before the transcription start site). The samples for direct sequencing are 42independent patients, who carried the homozygous risk haplotype reported in our previous10q fine-mapping study, and 50 normal controls. In second year, we prepare to genotype theidentified risk polymorphisms in a large case-control sample to verify the associationevidence of the risk polymorphisms. In third year, we prepare to perform the biologicalfunctional studies for the risk polymorphisms using luciferase reporter assay, RNA stabilityassay and EMSA.The project is novel and feasible because (1) we have had a series of linkage andassociation and expression studies for the 10q candidate genes in our samples. (2) Thesamples have been collected and ready for sequencing and genotyping studies. (3) We havehad experiences for the biological functional study for the risk polymorphisms of othercandidate genes of schizophrenia.