2015-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647722摘要：全球約有1.7億人口為慢性C型肝炎病毒感染者。根據世界衛生組織估計，每年有超過640萬以上的人遭受到急性C型肝炎病毒感染。急性C型肝炎病毒感染後有50-80%的患者會轉變為慢性C型肝炎，慢性C型肝炎患者中有五分之一的人最終變成肝硬化，之後每年有1-5%的患者產生肝細胞癌。因此C型肝炎病毒感染是一個影響全球醫療、公共衛生與社會經濟的重要健康課題。 長效型干擾素(pegylated interferon)合併口服雷巴威林(ribavirin)為目前大部分亞洲慢性C肝患者的標準治療，然而其療效仍有侷限，並且伴隨許多不良反應。因此許多學者進行研究，嘗試尋找可以調控C肝治療的反應因子，一方面除了能藉此對慢性C型肝炎的病理機制及病程進展更加了解外，也希望能夠藉由調控這些因子來改善目前以干擾素為基礎的C肝標準治療，減少患者治療時產生的不良反應以提高療效。 干擾素（interferon, IFNs）除了可以直接作用於宿主細胞，誘發抗病毒基因表現(IFN-stimulated response genes，ISGs)，使細胞進入抗病毒狀態（antiviral state）並抑制病毒的複製外，還可以抑制細胞生長、改變細胞分化時期、影響細胞週期、逆轉惡性瘤形態以及干擾致癌基因的表現。最近的研究中還發現第二型干擾素( type II)：干擾素-γ(IFN-γ)可以誘發腫瘤抑制因子--死亡相關蛋白激酶（DAPK:Death associated protein kinase）的表現。DAPK為一維持細胞正常生理的重要蛋白，除了與細胞凋亡（apoptosis）、促死因子、腫瘤轉移、及細胞轉型（transformation）有關外，目前的研究還發現DAPK在T淋巴細胞活化功能的調控上扮演重要角色。值得注意的是，吾人先前的研究中發現，第一型干擾素( type I)：干擾素-α (IFN-α)也會誘發DAPK的表現。因此DAPK蛋白是否在慢性C型肝炎病毒感染中扮演重要角色，影響干擾素訊息傳導、先天免疫作用以及C肝病毒清除，甚至影響抗C肝病毒藥物的效果就值得探討。然而目前關於DAPK蛋白在干擾素與C肝病毒的交互作用中所扮演的角色仍不清楚。C肝病毒蛋白、C肝病毒核醣核酸是否可以與DAPK產生交互作用也仍未知。此外，干擾素所誘發表現的DAPK蛋白是否可以有效的降低C型肝炎病毒複制及基因表現，也仍待釐清。因此吾人將探討DAPK蛋白對於抗C肝病毒藥物治療效果的影響，以及DAPK基因多形性在臨床上的可能應用，並進一步探討DAPK蛋白、干擾素以及C肝病毒蛋白在分子層面上的可能交互作用與機轉。 吾人研究假說如下：干擾素所誘發的DAPK蛋白可以抑制C肝病毒複製，而C肝病人DAPK的基因表現與C型肝炎病毒量有關，並且可以影響C肝患者治療的效果。 在此，吾人提出三年之研究計畫。第一年為病例對照的研究設計(case-control study)，將比較慢性 C 型肝炎基因型第一型患者接受長效型干擾素與雷巴威林合併治療後，100 位治療後有持續病毒反應者(SVR)與 50 位無反應者(Non-SVR)，其周邊血液單核細胞中 DAPK 蛋白及 RNA 表現與抗病毒治療反應的關係。第二年將比較這 150 例慢性 C 型肝炎基因型第一型患者在接受長效型干擾素與雷巴威林合併治療後，DAPK 相關單一核苷酸多形性與抗病毒藥物治療反應，以及相關臨床因子的關聯性。第三年為基礎研究，將探討干擾素誘發表現的 DAPK 蛋白對 C 肝病毒的影響，其分子機轉、訊息傳遞以及調控活化的方式，並進一步探討 C 肝病毒蛋白與 DAPK 蛋白的交互作用。希望能以此研究所獲得的知識應用於臨床醫藥衛生科學上，使 C 肝病毒感染的預防與治療更臻完善。<br> Abstract: Chronic hepatitis C virus (HCV) infection affects more than 180 million people worldwide, and is an important etiology of chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). The clinical manifestations of chronic HCV infection vary widely. In addition to hepatic injury, ample experimental, clinical and epidemiological data have demonstrated a close linkage between chronic HCV infection and metabolic derangements. Chronic HCV infection has been shown to be an independent predictor of metabolic abnormality such as insulin resistance, steatosis and diabetes mellitus, and metabolic profiles can also affect treatment outcomes in chronic hepatitis C patients. Although HCV can be efficiently eradicated in chronic hepatitis C (CHC) patients by peginterferon (PEG-IFN) plus ribavirin (RBV) combination therapy, the current standard of care for HCV in most Asian countries. However, the regimen has many undesirable side effects, is expensive and not effective in a substantial proportion of patients. Therefore, identifying modifiable factors that may improve therapeutic efficacy and reduce adverse events is clinically important. HCV is known to extensively exploit host factors to escape the immediate host defenses detecting and clearing pathogens from infected cells and tissues. Therefore, studying the nature and mechanisms of these HCV-host interactions has the potential to identify new targets for therapeutic intervention, as well as develop rational vaccine and adjuvant development aiming at protecting against HCV infection. Interferons are pleiotropic cytokines that exhibit important biologic activities, including antiviral, antitumor, immunomodulatory effects, and important to HCV clearnce. IFN-α/β and IFN-γ may induce the transcription of interferon-stimulated genes (ISGs) and affect some other genes which depend on the IFN type, the cellular target, and the nature of the infection/host challenge. Recent study has identified Death-associated protein kinase-1 (DAPK) as a factor that regulates apoptosis in response to interferon-γ. DAPK is an important kinase that can regulate diverse biological signals, including membrane blebbing, autophagy, growth factor-induced survival, tumour necrosis factor-mediated cell death, cancer development and ischaemic-induced neuronal cell death. Current study has also demonstrated that DAPK is a novel T cell regulator on TCR-activated NF-κB and T cell activation. Of note, our unpublished data find that interferon-α may induce the expression of DAPK. Because the key role of interferon-α on chronic HCV infection, it is important to examine the interactions between DAPK and HCV infection. Therefore, we would like to study the interactions of DAPK, interferon and HCV by examining the role of DAPK on HCV during interferon treatment and the usefulness of DAPK polymorphisms in a clinical setting. We hypothesize that DAPK may inhibit HCV replication, and the expression of DAPK may affect Hepatitis C viral titers, and therapeutic efficacy of CHC patients. Herein, we propose a 3-year study to examine to the interaction between HCV and DAPK. In the first year study, we will examine the protein and RNA expressions of DAPK in peripheral blood mononuclear cells between 100 responders and 50 non-responders of genotype 1 CHC patients receiving pegylated interferon-based treatment. In the second year study, we will examine the asociations of different DAPK polymorphisms and the antiviral response in these 150 genotype 1 CHC patients. In the third year study, we will explore the interactions involved in DAPK and HCV by studying the molecular mechanisms, signal transduction, and epigenetic regulation of DAPK and HCV proteins with and without interferon treatment.C型肝炎病毒細胞膜受體胰島素抗性葡萄糖代謝脂肪代謝hepatitis C virusentry factorsinsulin resistanceglucose metabolismlipid metabolism