文獻：Efficient Delivery of an Antisense Oligodeoxyribonucleotide Formulated in Folate Receptor-targeted Liposomes

作者：SHIH-JIUAN CHIU, GUIDO MARCUCCI and ROBERT J. LEE

文獻鏈接：https://ar.iiarjournals.org/content/26/2A/1049.short

摘要：

Background: Folate receptors (FRs) are cellular surface markers for numerous solid tumors and myeloid leukemias. The aim of this study was to develop an antisense oligodeoxyribonucleotide (ODN) carrier targeting FR-overexpressing cancer cells using folate (FA) as the targeting moiety. G3139, a phosphorothioate antisense ODN against human bcl2 mRNA, was evaluated in this study. Materials and Methods: G3139-containing liposomes were prepared using an ethanol dilution method. For the targeted formulation, 0.5 mol% of folate-PEG-DSPE was incorporated as a targeting ligand into cationic liposomes composed of DC-Chol/egg PC/PEG-DSPE at 25:65:10 mol/mol. Particle size and surface charge were measured and cellular uptake was assessed by fluorescence microscopy and flow cytometry. The ODN-containing formulations were evaluated in FR+ KB cells for Bcl2 down-regulation measured by Western blot. The cytotoxicity of the formulations was determined by MTT assay. Results: The G3139-containing liposomes had an average diameter of 80-90 nm with high ODN entrapment efficiency (70-80%). Incorporation of the folate ligand did not significantly alter the particle size and entrapment efficiency. The formulation exhibited colloidal stability in a serum-containing environment. In uptake studies, the folate-targeted formulation showed ligand concentration-dependent uptake that was up to 6-fold more efficient than that of the non-targeted formulation (p<0.05). The uptake could be blocked by an excess amount of free folate, thus indicating an FR-dependent mechanism. Conclusion: FR-targeted G3139-containing liposomes showed promising transfection activity in KB cells. FR-targeted formulations were capable of specific targeting to FR-overexpressing cell lines and optimizing the amount of folate ligand in the liposomal formulation can result in more efficient antisense delivery.

本研究的目的是開發一種反義寡脫氧核糖核苷酸（ODN）載體，以葉酸（FA）為靶向部分，靶向過表達FR的細胞。本研究評估了針對人bcl2 mRNA的硫代反義寡核苷酸G3139。

材料和方法：采用乙醇稀釋法制備含G3139的脂質體。對于靶向制劑，以25:65:10 mol/mol的比例將0.5 mol%的葉酸-PEG-DSPE作為靶向配體摻入由DC-Chol/egg PC/PEG-DSPE組成的陽離子脂質體中。測量粒徑和表面電荷，并通過熒光顯微鏡和流式細胞術評估細胞攝取。通過蛋白質印跡法在FR+KB細胞中評估含ODN的制劑的Bcl2下調情況。通過MTT法測定制劑的細胞毒性。

結果：含G3139的脂質體平均直徑為80-90nm，ODN包封率高（70-80%）。葉酸配體的摻入沒有顯著改變粒徑和包封效率。該制劑在含血清的環境中表現出膠體穩定性。

在攝取研究中，葉酸靶向制劑顯示出配體濃度依賴性攝取，其效率比非靶向制劑高出6倍（p<0.05）。過量的游離葉酸可以阻斷攝取，從而表明FR依賴機制。

結論：含FR靶向G3139的脂質體在KB細胞中顯示出良好的轉染活性。FR靶向制劑能夠特異性靶向FR過表達的細胞系，優化脂質體制劑中葉酸配體的量可以導致更有效的反義遞送。

相關推薦：

Cy7-DSPE

Cy7-PEG-DOPE

Cy7-PEG-DPPE

DSPE-PEG-Cy7

DSPE-PEG-cRGD

DPPE-PEG-cRGD

DMPE-PEG-cRGD

DOPE-PEG-cRGD

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