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標題: | 針對心肺相關疾病開發新穎治療策略 Developing New Therapeutic Approaches for the Treatment of Cardiopulmonary Disease |
其他標題: | Developing New Therapeutic Approaches for the Treatment of Cardiopulmonary Disease |
作者: | 洪佐宜 Zuo-Yi Hong |
指導教授: | 魏子堂 Tzu-Tang Wei |
關鍵字: | 特發性肺纖維化,β型干擾素,網孔型噴霧器,人類肺纖維母細胞,主動脈瘤及剝離,誘導性多功能幹細胞,人類誘導性多功能幹細胞之條件培養液,細胞外囊泡,凍乾培養液, Idiopathic pulmonary fibrosis (IPF),Interferon-beta (IFN-β),Mesh nebulizer,Human lung fibroblast (HLF),Aortic aneurysm and dissection (AAD),Induced pluripotent stem cells (iPSCs),Human induced pluripotent stem cell-derived conditioned medium (hiPSC-CM),Extracellular vesicles (EVs),Lyophilized medium, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 第一部分:
特發性肺纖維化 (idiopathic pulmonary fibrosis, IPF) 為十分嚴重的肺部疾病,其特徵是肺部組織會產生瘢痕並導致呼吸困難。至今,IPF患者的治療選擇有限且病患存活率低。β型干擾素 (IFN-β) 目前已獲得美國食品藥物管理局 (FDA) 批准,使用於復發型多發性硬化症之治療,而許多研究也發現IFN-β具有IPF治療潛力。然而,最近研究指出:臨床上所使用的IFN-β無法改善患者總體存活率,其中一個可能原因為IFN-β通過靜脈或皮下注射的方式投予,順利到達肺臟的IFN-β藥量有限。在目前的藥物輸送系統中,藥物透過霧化方式已證明對於呼吸系統疾病具有較好的療效。然而,噴射型噴霧器 (jet nebulizer) 和超聲波噴霧器 (ultrasonic nebulizer) 在霧化過程中可能會引起蛋白質結構產生變化進而影響蛋白質藥物(如IFN-β)的治療效果。相比之下,網孔型噴霧器 (mesh nebulizer) 可能較不影響到蛋白質藥物完整性及原本的生物學功能。 本研究顯示,於人類肺纖維母細胞 (human lung fibroblast, HLF) 增殖測定實驗中,不論是透過博來黴素 (bleomycin, BLM) 劑量反應或是轉化生長因子β (transforming growth factor-beta, TGF-β) 誘導的HLF細胞增殖反應,皆能藉由給予原型IFN-β和氣霧化IFN-β所逆轉。此外,使用網孔型噴霧器製出的氣霧化IFN-β也可顯著降低TGF-β處理所誘導的HLF細胞遷移及分化成肌纖維母細胞之能力。有趣的是,透過網孔型噴霧器製出的氣霧化IFN-β在TGF-β誘導的HLF細胞增殖反應中的抑制效果甚至更優於原型IFN-β。我們的研究揭示了IFN-β在使用網孔型噴霧器後治療IPF的潛力,並提升其治療功效。 第二部分: 主動脈瘤及剝離 (aortic aneurysm and dissection (AAD)) 是廣泛存在的心血管疾病,其嚴重性會導致病患隨時有危及生命的疑慮。然而,AAD在出現併發症前難以檢測和診斷這種危險、無聲的疾病,而且全球的AAD發生率仍然不斷攀升中。許多研究都在探討AAD和以下幾個因素的關聯性,並試圖找到防止這種疾病進展的策略,包括:彈性蛋白及膠原蛋白的結構降解、基質金屬蛋白酶 (matrix metalloproteinase, MMP) 的表現量、平滑肌細胞 (smooth muscle cell, SMC) 損失、細胞外基質斷裂 (extracellular matrix, ECM)、轉化生長因子β信號級聯放大 (TGF-β signaling cascade)。然而,現今臨床上大多只能透過特定藥物或手術治療,例如:給予β受體阻滯劑 (beta blocker)、血管張力素II型受體拮抗劑 (angiotensin II receptor blocker)、他汀類藥物 (statin) 或進行胸主動脈瘤開胸修補術 (open thoracic aneurysm repair)、血管腔內胸主動脈瘤修復手術 (thoracic endovascular aortic repair)。 誘導性多功能幹細胞 (induced pluripotent stem cells, iPSCs) 是一種能從體細胞表達特定胚胎轉錄因子的組合進而重新編程的多能幹細胞,而其最大的優勢為:以自體為來源,促使iPSCs作為細胞治療上擁有取之不盡、用之不竭的來源,並且也不會出現免疫排斥或是倫理問題等諸多疑慮。此外,研究也發現人類誘導性多功能幹細胞之條件培養液 (human induced pluripotent stem cell-derived conditioned medium (hiPSC-CM) 具有治療疾病及病症之潛力,而原因可能來自於iPSCs分泌的細胞外囊泡 (extracellular vesicles, EVs) 擁有某些特定的影響。本研究發現:hiPSC-CM經冷凍乾燥機凍乾後能保留大部分營養成分。我們進一步透過動物實驗指出:老鼠的主動脈瘤總體發生率以及彈性纖維 (elastic fiber) 損失在給予hiPSC-CM後皆有下降之趨勢,揭示了hiPSC-CM應用於AAD的治療潛力,為此棘手的疾病提供一項新穎的預防策略。 Part One: Idiopathic pulmonary fibrosis (IPF) is a severe lung disease that results in breathing difficulty characterized by lung scarring. Nowadays, patients with IPF have limited therapeutic options and unfortunately exhibit poor survival. Interferon-beta (IFN-β) has been approved by the US Food and Drug Administration (FDA) for the treatment of relapsing multiple sclerosis, while it has also been found to exhibit therapeutic potential in IPF. However, recent studies showed that the clinical use of IFN-β did not show an improvement in overall survival. The delivery of IFN-β through intravenous or subcutaneous injection may explain the possible reason that IFN-β is not successfully delivered to the lungs. Among present drug delivery systems, aerosolized medication has demonstrated the enhanced efficacy of drugs for the treatment of respiratory diseases. Nevertheless, the degradation of protein drugs caused by the jet and ultrasonic nebulizers may reduce the therapeutic effects of those drugs, such as IFN-β. In the contrast, neither the structural integrity nor the biological function of nebulized IFN-β was compromised by the nebulization process within the mesh nebulizer. In bleomycin (BLM) dose-response or transforming growth factor-beta (TGF-β)-induced lung fibroblast proliferation assays, we found both parenteral IFN-β and IFN-β nebulized with the mesh nebulizer could rescue these effects. On top of that, nebulized IFN-β obtained with the mesh nebulizer considerably reduced the capacity of the migration and myofibroblast differentiation ability of TGF-β-treated HLF cells, and intriguingly showed more potent rescue effects than parenteral IFN-β in TGF-β-induced lung fibroblast proliferation. Our investigations revealed the potential efficacy of IFN-β in the treatment of IPF with the mesh nebulizer, demonstrating the higher efficiency of IFN-β. Part Two: Aortic aneurysm and dissection (AAD) are severe cardiovascular diseases worldwide that can cause life-threatening bleeding. Unfortunately, this parlous and silent disease is notoriously arduous to diagnose before cardiac complications, and the number of cases has increased. Nowadays, a great deal of research has focused on the structural degradation of elastin and collagen, matrix metalloproteinase (MMP) expression, progressive smooth muscle cell (SMC) loss, extracellular matrix (ECM) fragmentation, and transforming growth factor-beta (TGF-β) signaling cascade to explore a novel strategy to prevent AAD progression. However, there are only two methods clinically: specific medications (e.g., beta blockers, angiotensin II receptor blockers, and statins) and surgeries (e.g., open thoracic aneurysm repair and thoracic endovascular aortic repair) against this disease. Induced pluripotent stem cells (iPSCs) are a type of pluripotent stem cell that can be reprogrammed by expressing a combination of embryonic transcription factors from a somatic cell. The greatest advantage of iPSCs is their autologous source, which leads to utilizing iPSCs as an inexhaustible source for cell therapies without immune rejection or ethical issues. Besides, some studies have shown that human induced pluripotent stem cell-derived conditioned medium (hiPSC-CM) possesses therapeutic potential against diseases and disorders, which could be attributed to the extracellular vesicles (EVs) derived from iPSCs. In this study, hiPSC-CM was lyophilized by a freeze-dryer to retain most nutrients. With a lower overall incidence of aortic aneurysms and less dilatation of ascending thoracic aortas accompanied by diminished loss of elastic fibers in vivo, our investigations revealed the potential efficacy of hiPSC-CM in the treatment of AAD, providing new insight into preventing the progression of this intractable disease. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/83297 |
DOI: | 10.6342/NTU202300589 |
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顯示於系所單位: | 藥理學科所 |
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