文獻(xiàn)：cRGD target liposome delivery system promoted immunogenic cell death through enhanced anticancer potency of a thymidine conjugate under UVA activation as a cancer vaccine

文獻(xiàn)鏈接：https://www.sciencedirect.com/science/article/pii/S022352341930145X

作者：

Rong Yang 1, Zhiwei Wang 1, Ying Yuan, Ting Qian, Qibing Zhou

摘要：

Conventional chemotherapeutic and photodynamic therapy have recently been shown to also elicit immune response against cancer through the immunogenic cell death mechanism, which can be potentially translated into effective cancer vaccines. However, there are few studies on the potential of nanodelivery system to promote the immunogenic cell death as a cancer vaccine. We reported here that cRGD target liposome delivery system was capable to promote the immunogenic cell death through enhanced potency of a thymidine conjugate post UVA activation as a cancer vaccine. Liposomes and cRGD target liposomes were found to significantly increase the cellular accumulation of the thymidine conjugate and subsequently translated into enhanced cytotoxic potency after UVA activation. More importantly, cRGD target liposomes of the thymidine conjugate markedly promoted the early detection of immunogenic cell death markers including ATP, HMGB1 and calreticulin. Subsequent in vivo vaccination?challenge study confirmed effective tumor growth inhibition by the cRGD liposomal thymidine conjugate and UVA treated cancer cells as the cancer vaccine. In addition, liposomes and cRGD target liposomes alone did not shown any induction of the immunogenic cell death markers, revealing the adjuvant nature of the nanodelivery system.

DSPE-PEG-cRGD的合成

向溶于2:3甲醇和水（500μL）的DSPE-PEG馬來酰亞胺（3.64mg）中加入磷酸緩沖鹽水（PBS，200?μL).所得混合物在室溫下反應(yīng)過夜，然后用10?mL甲醇。用制備型HPLC C18柱（內(nèi)徑20mm）分離所需產(chǎn)物?×?150?mm，GS-120-10-C18-AP，和0.1%三氟乙酸在MeOH中的洗脫液（3μL/min）。收集的DSPE-PEG-cRGD為脫色油狀物（1.96mg，44%收率），并通過1H NMR和質(zhì)譜分析進(jìn)行確認(rèn)。

對于cRGD靶脂質(zhì)體，通過硬脂酰馬來酰亞胺的共軛硫醇加成合成了硬脂酰-PEG-cRGD共軛物（DSPE-PEG-cRGD），該共軛物易于通過HPLC分離分離，并通過1H NMR和MS分析得到證實(shí)）。cRGD脂質(zhì)體的制備方法與DSPC脂質(zhì)體相似。為了優(yōu)化cRGD靶向的有效性，在兩種具有代表性的癌癥細(xì)胞系MCF-7和Bel-7402上研究了DSPE-PEG-cRGD比DSPE-PEG比例增加的脂質(zhì)體?細(xì)胞。

MCF-7?免疫熒光研究首次證實(shí)，細(xì)胞表面cRGD靶αv和β3整合素水平高于Bel-7402?細(xì)胞。然后基于2Cl化合物的熒光特性，通過熒光顯微鏡分析評估2Cl化合物在這兩個(gè)細(xì)胞系中的積累。顯然，在MCF-7中發(fā)現(xiàn)了最高的熒光?與具有較低量的非靶向脂質(zhì)體和2Cl對照的細(xì)胞相比，具有高量cRGD脂質(zhì)的細(xì)胞。

在貝爾-7402?高比率cRGD脂質(zhì)體似乎比其他脂質(zhì)體產(chǎn)生更高的熒光強(qiáng)度。DSPC脂質(zhì)體系統(tǒng)中DSPE-PEG-cRGD量的進(jìn)一步增加導(dǎo)致納米顆粒尺寸增加超過100?nm和2Cl化合物負(fù)載量差。因此，cRGD脂質(zhì)體被優(yōu)化為含0.15?μmol/mL DSPE-PEG-cRGD用于以下表征和生物學(xué)研究。空和2Cl負(fù)載的cRGD脂質(zhì)體的平均粒徑為90 nm，ζ電位分別顯著增加至+14.3和+15.9 mV。

所得帶正電的cRGD脂質(zhì)體與非靶向脂質(zhì)體和其他報(bào)道的通常帶負(fù)電的脂質(zhì)體形成對比，這可能是由于與PEG部分連接的長疏水烷基硫醇鏈的存在。2Cl的裝載效率為8.3?nmol/mL為81.2%。此外，空的和負(fù)載的cRGD靶脂質(zhì)體在超過72小時(shí)的血清存在下是穩(wěn)定的?h。最后，TEM分析證實(shí)了2Cl納米顆粒、DSPC脂質(zhì)體和cRGD-DSPC脂質(zhì)體在沒有和有2Cl負(fù)載的情況下的生產(chǎn)，其流體動(dòng)力學(xué)尺寸是一致的。

相關(guān)推薦產(chǎn)品：

DSPE-PEG-Glucose

DSPE-PEG-Glutamic acid

DSPE-PEG-Glycine triarginine

DSPE-PEG-His

DSPE-PEG-Mannose,DSPE-PEG-Man

DSPE-PEG-Methyltetrazine

DSPE-PEG-MTX

DSPE-PEG-nano gold

DSPE-PEG-NB

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