胰腺導管腺癌（PDAC）一直被視為最嚴重的疾病之一，由於其診斷性較差並高侵略性的遷移能力。除了腫瘤本身的惡性，其微環境（microenvironment）與腫瘤之交互作用能促使腫瘤生長並抑制免疫系統對腫瘤的攻擊，進而保護腫瘤免於各種診斷及治療手段。為了改善腫瘤微環境，在本研究中，我們將針對胰腺導管腺癌進行標靶治療。此外，本研究開發一奈米顆粒包覆一氧化氮供體（NO donor）藥物而後與腫瘤標靶綴合發展成為一新型胰腺腫瘤治療劑以調控腫瘤微環境進而誘導其與周遭腫瘤細胞產生凋亡（apoptosis）。結果表明，在體外和體內實驗中，我們的標靶載體擁有對腫瘤細胞的高度專一性 ，並且於腫瘤處具良好的累積量。同時，在細胞存活率和生化途徑調控的探討都顯示，此藥物的毒性測試和對蛋白質表現之效果。我們也透過生物分佈試驗來驗證奈米載體的標靶能力 。此外，我們藉由靶向奈米載體來傳遞藥物，證實能有效的增加腫瘤吸收，並能有效地抑制腫瘤生長。未來，此靶向奈米載體可參與臨床試驗，實際應用於胰臟癌病人。

Pancreatic ductal adenocarcinoma (PDAC) has been regarded as one of the most devastating diseases, attributed to its poor diagnosability and aggressively migrating ability. Aside from its malignancy, the interaction between the tumor and its microenvironment can promote tumor progression and diminish the attack of immune system, protect the tumor from chemotherapy. In order to ameliorate the tumor microenvironment, we developed a tumor targeting nanoparticle loaded with a nitric oxide (NO) donor, serving as a prospective therapeutic agent to regulate and further induce apoptosis in the tumor microenvironment. Our result indicated that the targeting nanoparticles bound to tumor cells in vitro as well as accumulation in tumor site specifically. Cytotoxicity and biological regulation were evaluated via viability assay and western blotting. Biodistribution of the tumor-specific nanoparticles were also studied to authenticate the targeting effect. Furthermore, tumor uptake was enhanced when exposed to the NO donor delivered by the targeting nanoparticles, elevating therapeutic efficacy. In the future, this targeted therapy can potentiate cancer treatment against pancreatic cancer.

Table of Contents
摘要 I
Abstract II
Acknowledgement III
Abbreviation IV
Table of Figures 2
Table of Tables 4
Introduction 5
1.1 Pancreatic cancer 5
1.2 Stromal microenvironment in pancreatic tumor 6
1.3 Anti-cancer treatments targeting stroma 8
1.3.1 Chemotherapy in PDAC 8
1.3.2 Stromal structure focused therapies 8
1.4 Nitric oxide and its biological functions 9
1.5 Nitric oxide donors 10
1.6 Drug delivery for NO donors 12
Specific Aims 14
Materials and methods 15
3.1 Materials and equipment 15
3.1.1 Materials 15
3.1.2 Equipment 16
3.2 Cell culture 17
3.3 Mice and orthotopic tumor model establishment 17
3.4 Cell viability 17
3.5 SEt decomposition 18
3.6 qRT-PCR 18
3.7 Western blot analysis 19
3.8 Phage display 20
3.9 ELISA assay 21
3.10 Homing assay 21
3.11 Preparation of nanoparticles 22
3.12 Drug encapsulation efficiency 23
3.13 Characterization of nanoparticles 23
3.14 Cellular uptake 23
3.15 Competition assay 23
3.16 Biodistribution 24
3.17 Immunofluorescence 24
3.18 Tumor perfusion and ECM inhibition 25
3.19 Statistics 25
Results 26
4.1 In vitro cytotoxicity of SEt 26
4.2 The effect of NO on the expression of fibrotic markers in PSCs 28
4.3 Isolation of PSC-specific phage in vivo and in vitro 32
4.4 Verification and characterization of selected phage peptide sequences 34
4.5 Uptake of PLGA nanoparticles modified with the PSC targeted peptides in PSCs in vitro and in the orthotopic PDAC model in vivo 39
4.6 Modulation of tumor microenvironment using SEt delivered by stroma-targeted NPs 44
Discussion and future work 46
Reference 48

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