2015-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647084摘要：71 型腸病毒為一新興致命性病毒，特別好發於亞太地區。71 型腸病毒感染會造成典型的手足口症、腦脊髓炎、肺水腫、類小兒麻痺症癱瘓或其他神經系統壞損徵狀。截至目前，並無有效之藥物或疫苗可對抗71 型腸病毒。因此，徹底瞭解71 型腸病毒感染後宿主防禦機制以及病毒致病機轉，對於臨床上發展抗病毒治療方式是有其必要性與迫切性。本研究團隊在過去發現71 型腸病毒可透過調控宿主的轉錄物質，如mRNA 與miRNA，進而關閉宿主蛋白質合成機制，逃拖宿主的免疫攻擊，並促進病毒自身的複製。除了mRNA 及miRNA 的變化，宿主在病毒感染時會產生另外兩種轉錄物質，包括替代異構體mRNA 以及長片段非編碼核糖核酸LincRNAs。RNA 選擇性裁剪為一種重要生物程序，根據不同RNA 裁切位點，細胞可以從單一基因產生多種蛋白質因而增加哺乳類基因組的複雜度。從過去的研究指出，單一基因可以產生出多種替代異構體mRNA，彼此間的調控呈現互相拮抗的情形。長片段非編碼核糖核酸LincRNA 是一種長度大於200 個核苷酸且非編碼之RNA。最近的研究指出，長片段非編碼核糖核酸能夠參與調控表觀基因體及基因表現。宿主長片段非編碼核糖核酸的表現趨勢亦會受病毒感染所改變。另一方面，免疫反應是人體中相當重要的防禦系統，特別於病毒感染。T 細胞受器在辨識細胞內病原體和清除被感染的細胞扮演關鍵性角色。病毒感染能夠塑型局部與全身性T細胞庫。適當T 細胞受器庫可用於攻擊71 型腸病毒達到防禦作用，然而，過度T 細胞受器庫反應可能會造成宿主組織壞損並導致無法復原之後遺症。因此，調節T 細胞受器庫是發展免疫治療一項具潛能且可行策略。於本計劃中，我們希望能藉由新一代定序儀分析異構體轉換以及長片段非編碼核糖核酸於71 型腸病毒致病機轉中的角色。此外，我們利用馴化後之71 型腸病毒作為樣本，探討T 細胞受器庫再病毒感染後之變化並找尋過度放大之T 細胞受器族群，可作為免疫治療標的。<br> Abstract: Enterovirus 71 (EV71) is a newly emerging life-threatening pathogen particularly in the Asia-Pacific regionrecently and EV71 infection causes typical hand-foot-and-mouth disease, encephalomyelitis, pulmonaryedema, poliomyelitis-like paralysis or even neurologic and psychiatric effects. At present, there are noantivirals or vaccines available against EV71. Understandings of viral pathogenesis and host defensemechanisms in EV71 infection thoroughly are urgent for antiviral therapies development clinically.Previously we found EV71 can govern host protein synthesis, escape host immune attacks and furtherfacilitate viral propagation by regulating host transcripts such as mRNAs and miRNAs. Besides both RNAs ,the host cells also response to stimuli by the other two types of transcriptional molecules, at least, alternativemRNA variants and lincRNAs. Alternative RNA splicing is an important biological process wherein cellsgenerate multiple protein products from a single gene, thereby contributing to the complexity of mammaliangenomes. It was found that antagonistic regulation might occur between different isoforms derived from thesame gene. LincRNA is one kind of non-protein coding RNAs with longer than 200 nucleotides. Recently itis discovered that lincRNAs are involved in the regulation of epigenetic modification and gene expressionand virus infection alters the expression of host lincRNAs. On the other hand, the immune response is themost important defense system particularly on the virus infection. T-cell receptor (TCR) plays a critical rolein recognizing intracellular pathogens and initiating the destruction cascade of infected cells. Virus infectionwas found to shape the local and systemic T cell repertoire. The proper response of TCR repertoire isnecessary for immune attack against EV71 while overwhelmed immune response might damage hosts andcause unrecoverable sequela. Hence, manipulation of TCR repertoire may be a potential strategy fordevelopment of immunotherapy.In this proposal, we would like to thoroughly understand the transcriptional regulation of EV71 infectionthrough characterizing the novel isoform switches and lincRNAs critical for EV71 pathogenesis by nextgeneration sequencing (NGS) RNA sequencing strategy. We also explore the TCR repertoire of variousseverities in mouse-adapted EV71 model and find the potential expansive clones for immunotherapy targets.71 型腸病毒異構體轉換長片段非編碼核糖核酸T 細胞免疫庫EV71isoform switchlincRNATCRimmune repertoire