2018-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/705500摘要：分枝桿菌引起之感染於台灣及全球多個國家都有不斷增加之趨勢。其中皮膚及軟組織為僅次於肺部之最常見感染部位，其臨床及組織病理學表現隨宿主免疫力及菌種的不同而呈現個別差異，除缺乏典型的臨床症狀可作為診斷依據外，傳統實驗室方法診斷率偏低，再加上治療須使用多種藥物組合，不僅療程冗長且藥物副作用大，復因不同菌種 (甚至亞種) 對藥物之感受性不盡相同，因此這類感染已成為當今感染症診治之棘手課題。然而，目前分枝桿菌感染之相關研究均集中在肺部感染，鮮少有肺外感染之探討；此外，前瞻式及結合基礎臨床資源之轉譯研究更是付之闕如。緣此，為解決這些臨床診斷及治療上面臨之困境，我們擬橫向結合臨床、分子微生物學、病理學及免疫學等各種面向，並以病人檢體、細胞株培養暨動物實驗等研究模式做縱向串接，期能找出提昇診斷及治療成功率之新方向。本研究計畫之目的為：(一) 延續申請人過去所成功建立之分枝桿菌皮膚及軟組織感染之臨床及分子生物學研究，本計劃首度透過前瞻式收集病人皮膚或軟組織切片檢體，以聚合酶鏈鎖反應 (PCR) 偵測皮膚組織檢體內是否具有分枝桿菌，並輔以定序分析來鑑別菌種，特別是最常引起人類皮膚及軟組織感染之M. marinum, M. avium complex及M. abscessus complex (包括傳統實驗方法無法區分之兩種重要臨床亞種—M. abscessus 與 M. massiliense)，再深入比較不同菌株所引起之皮膚及軟組織感染之臨床特性、嚴重程度、與抗生素效果之差異。(二) 研究同時分析病人血中是否會產生對抗分枝桿菌醣類胺基酸脂質鍵結物 (glycopeptolipid) 之特定免疫球蛋白，並詳實比較此血清學診斷方法結合分子生物學診斷是否較傳統培養及病理診斷具有更快捷、更敏感及更準確之診斷優勢。(三) 研究同步藉由角質細胞株培養實驗，全面分析不同菌種感染皮膚之速度、細胞內存活能力、活性氧化物質誘發程度、細胞激素、抗微生物胜肽等之改變情形。(四) 計畫最後將根據臨床及細胞實驗分析之結果，創建動物模型來相互驗證並提供各種有潛力改善臨床診治之轉譯研究平台。　　藉由整合基礎與臨床研究所得之豐碩成果，我們期能針對分枝桿菌皮膚及軟組織感染提供更快速及精確之診斷方法，並闡明不同分枝桿菌與宿主間之交互作用，協助臨床醫師擬訂更精準化之治療策略，對改善病人預後有莫大的參考價值與貢獻。<br> Abstract: Non-tuberculous mycobacterial (NTM) infections are increasingly reported in Taiwan and worldwide. Among them, skin and soft tissue infections (SSTIs) are the most common extrapulmonary diseases. Diagnosis of NTM SSTIs can be challenging due to diverse clinical presentation, low yield from cultured specimens, and nonspecific histopathology on tissue biopsy. Despite aggressive medical treatment that often necessitates prolonged combination therapy with frequent drug toxicities, responses may be suboptimal with poor outcomes. The precise differentiation of the etiologic organisms is crucial for the clinicians since the treatment response and antimicrobial susceptibility differ among species and even subspecies. However, previous studies on NTM infections primarily focused on pulmonary disease, while there is a dearth of knowledge about the clinical, histopathological and immunological characteristics of NTM SSTIs. Large-scale, prospective and translational studies are exceedingly scarce. Given the unmet medical needs of rapid diagnosis, precise speciation and better treatment strategies, we will conduct a multifaceted study based on our established patient cohort and study methods, encompassing clinical investigations together with in vitro and in vivo models to better decipher the distinctive clinical features, histopathological characteristics, and the immunopathogenesis triggered by different strains of NTM in the cutaneous microenvironment. This study is comprised of 4 specific aims. First, we will prospectively collect biopsy specimens from patients with protracted SSTIs for PCR and targeted sequencing to enhance diagnosis. We will then compare the demographics, clinical characteristics, histopathological features and antimicrobial susceptibility patterns of various strains of NTM, with special focus on the most predominant NTM causing SSTIs, namely M. marinum, M. avium complex, M. abscessus complex (particularly subspecies M. abscessus and M. massiliense which are indistinguishable from conventional laboratory methods). Second, serum will be obtained to measure the level of anti-glycopeptolipid immunoglobulins. The performance of such molecular assays will be compared with that of conventional culture and histopathological examination. Third, we will investigate the infectivity, differences in the productions of reactive oxygen species (ROS) and proinflammatory cytokines in human keratinocytes infected with various species of NTM. Finally, we seek to establish a mouse model to decipher host-microbe interactions in vivo, and to serve as a platform to verify hypothesis generated from clinical and in vitro studies. The knowledge derived from this integrated, in-depth study can facilitate clinical diagnosis and formulation of individualized patient care. Moreover, the present study can accelerate progress toward clinical implementation of precision medicine in management of NTM infection and improve disease outcome.分枝桿菌皮膚及軟組織感染免疫組織病理學nontuberculous mycobacterial infectionNTMskin and soft tissue infectionsimmune responsehistopathological