2021-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/654166代謝重整已經在免疫細胞的發育和免疫反應中扮演重要的角色。而細胞代謝包含合成和分解代謝對於樹突狀細胞（DCs）的分化和功能調節都有被報導過。糖解作用、氧化磷酸化（OXPHOS）和脂肪酸氧化（FAO）的代謝重整可以調節DC的功能，以增強或降低其針對感染和抗腫瘤免疫的免疫反應。然而，代謝酶在調節DC發育中的角色仍不清楚。為了解決這個問題，我們建立了一個幹細胞及先驅細胞株（iHSPC），並使用它來篩選代謝酶和DC發育有關的基因，並使用慢病毒去敲低cDC或pDC中大量表現的基因。我們已經找到Cox6a2和Atp5d這兩個參與線粒體電子傳遞鏈（ETC）的基因會影響pDC發育。和非靶向對照組shLacZ相比，敲低其中一個基因都可以顯著降低pDC發育的頻率。任一基因的敲低也降低細胞ATP的產生，這很可能是由於Cox6a2和Atp5d所產生的蛋白分別是ETC複合物IV（CIV）和CV的成分。經由免疫基因體計劃中（immgen.org）RNA-seq數據庫的分析顯示， Cox6a2不僅在pDC中高度表現，而且和其他免疫細胞（包括共同淋巴先驅細胞，B細胞和T細胞）也是相對高度表達，顯示Cox6a2可能在pDC具有重要的作用。因此，此子計畫目的研究目標在探討Cox6a2如何調節pDC的發育和功能。 Metabolic reprogramming has become central events in governing homeostasis of immune cells and immune response. Cellular metabolism, including anabolic and catabolic metabolism, has been reported to regulate the differentiation and function of dendritic cells (DCs). Metabolic rewiring of glycolysis, oxidative phosphorylation (OXPHOS), and fatty acid oxidation (FAO) are also shown to modulate the function of DCs either to gear up or lower down their immune response against infections and anti-tumor immunity. However, the effect of metabolic enzymes in regulating DC development remain uncharacterized. To address this question, we have established an immortalized stem and progenitor cell line (iHSPC) and used it to screen genes that encode metabolic enzymes and are preferentially expressed either in cDC or pDC using lentivirus-mediated gene knockdown. To this end, we have identified Cox6a2 and Atp5d, two genes encode proteins involved in mitochondrial respiratory electron transport chain (ETC), as critical molecules for pDC development. Knockdown of each gene resulted in significantly reduced pDC frequency as compared to shLacZ, a non-targeting control. Knockdown of either gene also negatively impacted the production cellular ATP, which was most likely due to the fact that Cox6a2 and Atp5d encodes a component of the complex IV (CIV) and V, respectively, of the ETC. Analysis of RNA-seq database in The Immunological Genome Project (immgen.org) revealed that Cox6a2 was not only preferentially expressed in pDC compared to cDC, it was also highly expressed among other immune cells, including common lymphoid progenitor (CLP), B cells and T cells, suggesting that Cox6a2 may have important role in pDC. Therefore, the specific objectives of this subproject are aimed to study how Cox6a2 regulate the development and functions of pDC.漿狀樹突細胞粒線體Cox6a2代謝mitochondriametabolismPlasmacytoid dendritic cell