2021-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/673145我們闡明腺苷酸激酶4 (Ak4) 主要表現在M1巨噬細胞。此外， Ak4在M1巨噬細胞中對於維持ATP的恆定相當重要。重要的是，在M1巨噬細胞中敲弱Ak4顯著的減少發炎基因的表現，包含Il1b、Il6、Tnfa、Nos2與Cybb。因此，我們的數據顯示在巨噬細胞中有一個潛在的機制連結能量代謝與發炎。Ak4位在粒線體的基質中，且維持ATP/ADP/AMP的恆定。然而，關鍵問題是為什麼Ak4的上調對巨噬細胞發炎是至關重要，許多的可能性皆參與其中。第一，Ak4通過消耗ATP和AMP產生ADP的可逆反應中，可能會影響巨噬細胞發炎所必需的粒線體訊號。第二，Ak4介導的AMP消耗可能抑制AMPK的活性，從而影響特定M1基因的轉錄控制與發炎細胞激素的產生。第三，Ak4可能影響粒線體的呼吸作用與ROS的產生，從而在M1的轉錄程序中改變Hif-1信號。因此，我們研究的主要目標是(1)比較Ak4缺失與野生型(WT)M1巨噬細胞ATP/ADP/AMP的恆定(2)在M1巨噬細胞中測定AMPK的活化、Hif1-a的信號與轉錄程序受Ak4調控(3)比較Ak4缺失與WT的M1巨噬細胞中，粒線體恆定與生物再造(4)分析在M1巨噬細胞中Ak4介導的能量代謝(5)研究Ak4缺失的M1巨噬細胞所介導的發炎與抗癌免疫力的影響。我們將會 (1)與子計畫一共同合作，研究在M1巨噬細胞中測定Ak4所參與的ATP/ADP/AMP的恆定與發炎；(2)與子計畫三共同合作，測定是否EGFR TKI能反轉巨噬細胞缺失Ak4的功能；與(3)與子計畫四共同合作，探討Ak4在樹突細胞發育與功能所扮演的角色。 We have demonstrated that adenylate kinase 4 (Ak4) is preferentially expressed in M1 macrophages. Furthermore, Ak4 is critical for maintaining ATP homeostasis in M1 macrophages. Importantly, knockdown of Ak4 markedly reduces the expression of inflammation genes, including Il1b, Il6, Tnfa, Nos2, and Cybb, in M1 macrophages. Therefore, our data depict a potential mechanism linking energy metabolism and inflammation in macrophages. Ak4, located in mitochondria matrix, maintains ATP/ADP/AMP homeostasis. However, the key question is why Ak4 up-regulation is so critical for macrophage inflammation. Several possibilities might be involved. First, the catalytic function of Ak4 in the reversible reaction of ADP production through the consumption of ATP and AMP indeed affects mitochondrial cues necessary for macrophage inflammation. Second, Ak4-mediated consumption of AMP might suppress AMPK activity, thereby affecting transcriptional control of specific M1 gene signature and inflammatory cytokine production. Third, Ak4 might regulate mitochondrial respiratory function and ROS production, thereby altering Hif-1 signal in transcriptional programming specified for M1 function. Therefore, our specific aims of this study are (1) to investigate the ATP/ADP/AMP homeostasis in Ak4-deficient M1 macrophage compare to WT M1 macrophage; (2) to examine the AMPK activation, Hif-1 signal and transcriptional programming regulated by Ak4 in M1 macrophage; (3) to characterize the mitochondria homeostasis and biogenesis in Ak4-deficient M1 macrophage compare to WT M1 macrophage; (4) to analyse the energy metabolism mediated by Ak4 in M1 macrophage and (5) to study the impact of in Ak4-deficiency in M1 macrophage mediated inflammation and anti-cancer immunity. We will collaborate with (1) sub-project 1 to examine the involvement of Ak4 in ATP/ADP/AMP homeostasis and inflammasome of M1 macrophage; (2) subproject 3 to test whether EGFR TKI can rescue the function of AK4 knockout (KO) macrophages, and (3) subproject 4 to investigate the role of Ak4 in DC development and function.腺&#33527酸激&#372384巨噬細胞免疫adenylate kinase 4macrophage