間質幹細胞載體裝載 治療性反轉錄病毒應用於腦瘤治療

Mesenchymal Stem Cells as Cellular Vehicles Encapsulate Therapeutic Retroviruses Against Gliomas

楊宇軒(Yu-Hsuan Yang)

指導教授 ： 戴建國

國立中正大學/理學院/生命科學系分子生物研究所/碩士(2013年)

摘要

傳統上治療惡性神經膠質瘤不外乎手術切除、放射線治療與化學治療，然而這些治療方式對於病人的預後效果仍然有限。主要因為神經膠質瘤具有特別的浸潤性能力，侵入周遭的正常組織之中，所以難以徹底清除腫瘤而造成復發。近年來，間質幹細胞被研究發現其具有腫瘤的趨向性並可以作為腦瘤治療的攜帶性載體。我們發展出一個新穎的治療策略，用來治療惡性腦瘤，使用大鼠脂肪間質幹細胞(adipose-derived mesenchymal stem cells, A-MSC)為載體表現CD合併給予前藥5-FC毒殺細胞。收集A-MSC-CD培養後的培養液，其中包含了5-FC被CD轉換成的5-FU，用來與大鼠腦瘤細胞共培養，發現腦瘤細胞的生長明顯受到抑制，也觀察了A-MSC的移動能力。另外我們也嘗試分離出大鼠骨髓間質幹細胞(bone marrow-derived mesenchymal stem cells , BM-MSC)，作為RRV (replicating retrovirus vector)的細胞載體。結果顯示，從BM-MSC釋放出的RRV可以進一步感染腦瘤細胞，實現基因治療的目標，在動物實驗中也看到同樣的結果。未來我們將具有治療效果的間質幹細胞植入原位腦瘤模式小鼠中，希望作為細胞載體的BM-MSC可以穩定釋放RRV，且感染散佈在正常組織之中的腦瘤細胞並表現出治療基因，進而毒殺腫瘤細胞。期望於未來提供一個新的腦瘤治療方法。

關鍵字

複製行逆轉錄病毒 ； 間質幹細胞 ； 腦瘤

並列摘要

Although conventional therapies for malignant glioma such as surgical resection, radiotherapy and chemotherapy are available, the prognosis for patients with this disease remains extremely poor. Glioma has an exceptional infiltrative potential and integrate into abutting tissue, therefore, it is hard to completely cure glioma. Recently, mesenchymal stem cells (MSC) have been found possessing inherent tumor tropic properties and offering much promise as delivery vehicles for brain tumor therapy. We evaluated a new therapeutic strategy for malignant glioma, which combines intratumoral inoculation of rat adipose-derived MSC (A-MSC) expressing cytosine deaminase (CD) gene and 5-fluorocytosine (5-FC) treatments. Co-culture of rat glioma cells (RG2) with A-MSC-CD conditioned media which contain 5-FU converted from 5-FC resulted in a remarkable reduction in cell viability. The migratory ability of A-MSC toward glioma cells was demonstrated by transwell assay. We have also infected rat bone marrow-derived MSC (BM-MSC) with replicating retrovirus vector (RRV) developed in our lab. The result showed that RRV released from BM-MSC could further infect glioma cells, achieving persistent gene delivery to gliomas in vitro and in vivo. In the future we will exam the therapeutic effects of such MSCs in an orthotopic mouse model of human glioma. We wish the use of BM-MSC as the vehicle for dispersing RRV may provide a viable strategy for delivering a therapeutic payload to disseminated glioma burdens.

並列關鍵字

Replicating retrovirus ； Mesenchymal stem cells ； glioma

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