2017-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/647577摘要：免疫系統與疾病的發生、進程、緩解、治療與預後息息相關，過度活化或抑制都不利。以癌症為例，癌細胞藉由抑制免疫系統於體內生長擴張，免疫治療就是再活化使其對癌細胞進行清除，有效提高存活。免疫系統於個體中可以特異、敏感且即時感應生理狀態，免疫全面(immuno-Repertoire)多樣來自T 細胞受器CDR3 區域在遭遇外源入侵物或內生細胞變異時轉錄V-(D)-J 重組所致。少數細胞產生癌變，免疫系統可以立即應變，一由CD8+T 細胞進行毒殺，二由抗原呈現細胞將癌細胞抗原呈現予CD4+T 細胞進行後天免疫反應；此時，特定免疫株(clone)會明顯擴張(clonal expansion)，造成多樣性因此降低，這過程較目前早期癌篩方法更為敏感、專一。本三年計畫主要利用新一代定序儀基因體策略與誘導式肺癌動物模型探討癌發過程早期至晚期體內免疫全面多樣頻譜變化，找出肺癌高特異性CDR3；再藉由立體模擬鑑定此序列酸結合的抗原胜肽鍊，最後於臨床樣本驗證此抗原做為早期診斷或是用藥效果追蹤的預測能力，或做為來發展抗體藥物治療的標的。特異性目標: 穩定量產純品系四環黴素誘導突變EGFR (L858R 及L858R/T790M) 肺癌小鼠；利用新一代定序儀分析誘導後早晚期與標靶治療產生抗性後週邊血液、肺臟、脾臟等免疫全面多樣；鑑定新穎性抗原以為診斷標記；利用臨床週邊血液驗證標記。藉由計畫執行，提供非侵入診斷、追蹤與預後的新穎策略。<br> Abstract: Immune system is highly involved in disease onset, progression, remission, therapy efficacy, and outcome.Over activated or silenced of immune system may harm to normal physiological homeostasis. Take cancerfor example, tumor cell can progress and metastasize by inhibiting endogenous immune function. Recently, ithas been focused on immunotherapy that blockage of immunosuppression caused by tumor cells cansignificantly reactivate immune and enhance overall survival in many cancers. Immune system canspecifically, sensitively and immediately sense the physiological condition due to immuno-Repertoire. Thediversity of immuno-Repertoire is a result of V-(D)-J rearrangement in CDR3 sequences of T cell or B cellreceptor (TCR or BCR) when exogenous pathogens invaded or endogenous cell transformed. Immune systemcan immediately response to a few cell undergone carcinogenesis by two strategies including sensitizingtumor antigen by CD8+ cytotoxic T cell for tumor clearance or antigen presented by antigen presenting cellfor activating CD4+ T cell followed by acquired immune response. After activation, specific TCR clone willbe significantly expanded and result in diversity reduction. This process is more sensitive and specific thanany other early cancer screening method. This three-year proposal aims to analyze immune-repertoire of lungcancer form early onset to late progression by Next-generation sequencing technology combined withinducible mutant EGFR driven lung cancer transgenic mouse model and identify lung cancer specific CDR3of TCR. Potential peptide chain generated from CDR3 sequence will be simulated by 3D computer modelingto identify most possible corresponding binding antigen from cancer cell. This antigen will be furthervalidated for prediction power in early detection and therapeutic monitor. Furthermore, it may be a potentialtherapeutic target for drug development in the future. The specific aims of this proposal are: 1. Stablegeneration of inducible mutant EGFR (including the strain carried sensitive mutation L858R and the straincarried resistant mutation L858R/T790M) driven syngeneic C57BL/6 transgenic mice. 2. Collect peripheralblood, spleen, and lung at early, middle, and late stage for immune-repertoire analysis by Next-generationsequencer. 3. Identification novel tumor specific antigen by binding model simulation. 3. Prediction powerevaluation for early lung cancer onset, drug resistant, and disease progression biomarker in peripheral blood.By the achievements of this proposal, we can provide not a only non-invasive diagnostic method withsensitivity and specificity but also a novel monitor strategies for treatment monitor and outcome prediction.Furthermore, according to novel tumor specific antigen identification, it can promote new era ofimmunotherapy.