2018-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/645882摘要：CD248，也稱為endosialin或tumor endothelial marker 1，屬於第一型跨膜醣蛋白。正常狀況 下，CD248在出生之後主要表現在腎臟與子宮。過去我們利用lacZ敲入(knock-in, KI〕技術已發現在 Cd248K1/+小鼠出生後，Cd248在大多數的器官内表現都降低下來，唯獨在腎臟内表現會持續存在， 尤其是表現在腎小球系膜細胞與血管周邊細胞。在人類的疾病中，目前已知CD248會增加表現於慢 性腎病間質的肌肉纖維母細胞，且其表現量與腎臟的存活負相關。除此之外，許多研究也證實CD248 在腫瘤與關節炎組織中也增加表現量。最近實驗室的研究證實利用DNA疫苗來抑制CD248可以抑制 腫瘤生長。我們的研究一直關注腎臟的損傷與後續功能惡化，成果之一證實血管周邊細胞是腎損 傷之後造成腎臟結疤纖維化的主要細胞，而腎臟血管周邊細胞正是表現CD248的細胞。因此在利用 同型合子剔除Cd248的小鼠〔即同型合子敲入lacZ基因的小鼠，Cd248K//K/〕進行的初步實驗中，我們 發現單侧輸尿管阻塞(UUO〕引起的腎臟纖維化與巨嗜細胞數目會因Cd248剔除而減少。在右侧腎臟 切除合併左侧缺血-再灌流損傷(UNx+IRI)的急性腎損傷模式中，我們也發現剔除Cd248呈現出減少 損傷嚴重度與保護腎功能的趨勢。在探討CD248對於腎臟損傷的角色機轉時，我們的初步發現包括 了 Cd248剔除會減少促進纖維化表型的巨嗜細胞。雖然其他團隊少數的研究指出CD248可能與血小 板衍生的生長因子受體〔PDGFR)受刺激後產生的細胞生長訊息有交互影響，但我們的初步實驗結 果並不支持此一主張，反而指向CD248可能是血管周邊細胞用來與單核球/巨嗜細胞交互對話的醣 蛋白，可能會影響血液循環中的單核球被吸引至受損傷的腎臟並影響單核球/巨嗜細胞的功能分 化。因此我們要提出這一個三年計畫，首先要進一步確認CD248對於至少三種腎臟纖維化模式的影 響，包括UUO，單侧IRI，與腎毒性腎炎(nephrotoxic nephritis〕模式以探討血管周邊細胞與系膜細 胞表現CD248的作用等。然後我們也要進一步再確認CD248對於至少兩種急性腎損傷模式的影響， 包括UNx+IRI，與cisplatin腎病變模式等。我們將利用動物與細胞實驗釐清CD248如何影響吸引單 核球進入損傷的腎臟之中與CD248如何影響單核球/巨嗜細胞的功能分化。同時，為了能進一步臨 床運用的可能，我們將製造出能中和CD248訊息的抗體，並測試對於各種腎病模式的治療效果。我 們相信此研究的設計與可行性將可充分訓練參與的年輕科學家，促進他們未來在生醫研究與治療 的職業生涯發展。而本研究所關注的CD248中和抗體也極有潛力在腎損傷，癌症，與關節炎等疾病 發展成為一個全新有效的治療。<br> Abstract: CD248, also known as endosialin or tumor endothelial marker 1, is a type I transmembrane glycoprotein that is expressed in stromal cells of normal kidneys and uterus after birth. Using lacZ knock-in (KI/+) mice we have shown that Cd248 expression decreases in most organs but increases and persists in postnatal kidneys, specifically in glomerular mesangial cells and pericytes/perivascular fibroblasts. In human chronic kidney disease (CKD), upregulated expression of CD248 is shown in myofibroblasts, which is negatively associated with renal survival. In addition, upregulated expression of CD248 has also been shown in cancers and inflammatory joints. Targeting CD248 by DNA vaccination has shown the beneficial effect on regressing cancer growth in laboratory. Many studies including ours have identified CD248+ pericytes as the major source of precursors of scar-producing myofibroblasts after acute kidney injury (AKI) and during progressive kidney fibrosis. Using Cd248 knockout mice (Cd248KI/KI) our preliminary data have shown less fibrosis and macrophages in unilateral ureteral obstruction (UUO) kidneys of Cd248KI/KI mice. In addition, our ongoing experiments in AKI model induced by right uni-nephrectomy and ischemia-reperfusion injury of left kidney (UNx+IRI) are also showing the trend of reno-protective effect of Cd248 disruption. In mechanism study, less pro-fibrotic macrophages were noted in UUO kidneys of Cd248KI/KI mice. Although limited experimental data suggest that CD248 is involved in the cross-talk with the post-receptor signalling activated by platelet-derived growth factors, our preliminary data rather support a role in monocyte recruitment and functional polarization of monocytes/macrophages during kidney injury. Therefore in this 3-year study, we will first confirm the role of CD248 in mouse models of progressive kidney fibrosis, including UUO, unilateral IRI and nephrotoxic nephritis using Cd248KI/KI mice. We will then study and confirm the effect of CD248 in mouse models of AKI including UNx+IRI and cisplatin nephropathy. We will then clarify the functional roles of CD248 in recruiting monocytes and polarization of monocytes/macrophages in injured kidneys and cultured cells. To advance the possible CD248 targeting in clinical diseases we will generate CD248 neutralizing antibody for versatile application in our kidney disease models. We believe this study will not only help young scientists to develop their careers, but also foster the development of neutralizing antibodies to target kidney disease and many clinical diseases including cancers and inflammatory diseases in modern medicine.CD248巨嗜細胞肌肉纖維母細胞單核球血管周邊細胞腎臟CD248macrophagemyofibroblastmonocytepericytekidney