研發新頴核酸載體平台於調控基因表現與抑制之應用

Development of Nucleic-Acid Carrier Platform for Gene Expression and Silencing

曾士傑(S-Ja Tseng)

指導教授 ： 湯學成

國立清華大學/工學院/化學工程學系/博士(2010年)

https://doi.org/10.6843/NTHU.2010.00174

摘要

天然細胞外間質(extracellular matrices, ECM)已廣泛應用於組織工程與再生醫學。在市售的ECM之中，以豬體中取出的小腸黏膜組織下層(small intestinal submucosa, SIS)對於燒燙傷修補和組織重建具有明顯療效。由於SIS中含有負電之醣胺素分子，故本研究藉由靜電作用力讓帶正電之核酸傳遞載體(polymer/DNA奈米複合體)吸附於SIS上，將SIS與核酸傳遞結合以作為調控基因表現之平台。當奈米複合體與細胞接觸時，可將所包覆之DNA攜帶入細胞中，進行特定基因表現。以生醫材料平台調控基因傳遞至標的細胞，具有專一性且可以大幅提升基因傳遞效率，不管是組織工程或疾病治療均可使用這個平台。平台建立後，需要與低毒性之高分子核酸傳遞載體結合。本研究藉由現今已廣泛應用於組織工程之聚(酯-胺基甲酸酯) (poly(ester urethane), PEU)與聚乙二醇(poly(ethylene glycol), PEG)合成出生物可分解性、毒性低且溶於水之聚(胺基-酯-乙二醇-胺基甲酸酯) (poly(amino-ester-glycol-urethane), PaEGU)，其可與核酸物質自組裝成高度專一性帶且正電之PaEGU/DNA或PaEGU/siRNA奈米複合體，使核酸傳遞入細胞中並抵擋生物體內酵素將其分解，亦可將大幅提升核酸傳遞效率。以建立新穎核酸傳遞之平台，應用於組織工程或疾病治療。

關鍵字

生物可分解高分子 ； 細胞毒性 ； 核酸傳遞 ； 基因表現 ； 基因抑制

並列摘要

Naturally occurring extracellular matrices (ECMs) have shown great potential in clinical applications as tissue substrates to facilitate tissue repair and regeneration. Among the commercialized ECMs, porcine small intestinal submucosa (SIS) has been used in patients for wound treatment and soft tissue reconstruction. However, there have been no reports exploring the electrostatic properties of SIS as a substrate to control localized nucleic acid delivery. We have demonstrated that the negatively charged glycosaminoglycan (GAG) content in SIS was able to associate with the cationic polymer/DNA polyplexes through electrostatic adsorption and led to transfection upon cellular adhesion. However, a major challenge to the development of localized nucleic acid delivery is the design of suitable vectors with low cytotoxicity. Poly(ester urethane) (PEU) is a class of biodegradable polymer that has been applied as tissue-engineering scaffolds with minimal cytotoxicity in vitro and in vivo. We have developed a method of incorporating tertiary amines and poly(ethylene glycol) (PEG) into PEU to synthesize soluble poly(amino ester glycol urethane) (PaEGU) as a novel platform for nucleic acid delivery. PaEGU can condense DNA or siRNA into nano-scale polyplexes to enter cells through endocytosis, which can be a useful tool to work with ECMs for localized gene expression and silencing applications.

並列關鍵字

biodegradable polymer ； cytotoxicity ； nucleic acid delivery ； gene expression ； gene silencing

參考文獻

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