Fingolimod-Conjugated Charge-Altering Releasable Transporters Efficiently and Specifically Deliver mRNA to Lymphocytes In Vivo and In Vitro.
Fingolimod-Conjugated Charge-Altering Releasable Transporters Efficiently and Specifically Deliver mRNA to Lymphocytes In Vivo and In Vitro.
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Fingolimod偶联的电荷改变可释放转运蛋白在体内和体外有效特异性地将mRNA递送至淋巴细胞。
DOI：
10.1021/acs.biomac.2c00469
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发表时间：
2022-07-11
影响因子：
6.2
通讯作者：
Levy, Ronald
中科院分区：
文献类型：
--
作者：
Testa, Stefano;Haabeth, Ole A. W.;Blake, Timothy R.;Del Castillo, Trevor J.;Czerwinski, Debra K.;Rajapaksa, Ranjani;Wender, Paul A.;Waymouth, Robert M.;Levy, Ronald
电荷改变可释放转运体（CART）是一类寡核苷酸递送载体，其显示出在体外和体内均有效递送信使RNA（mRNA）。在这里，我们利用CART合成的化学多功能性来产生含有小分子药物芬戈莫德（FTY 720）的CART，作为通过特异性配体-受体相互作用增加淋巴细胞中mRNA递送和表达的策略。芬戈莫德是FDA批准的小分子药物，在体内磷酸化后，与鞘氨醇-1-磷酸受体1（S1 P1）结合，该受体在淋巴细胞上高度表达。与非芬戈莫德偶联类似物相比，芬戈莫德偶联CART在体外实现了对活化的人和鼠T和B淋巴细胞的上级转染。当细胞在转染前暴露于游离芬戈莫德时，芬戈莫德缀合的CART的较高转染丧失。在体内，与缺乏芬戈莫德的CART相比，芬戈莫德偶联的CART显示向脾脏边缘区B细胞和NK细胞的mRNA递送增加。此外，与游离芬戈莫德类似，芬戈莫德-CART介导的mRNA递送诱导外周血T细胞耗竭。因此，我们表明，CART与经验证的小分子的功能化可以增加感兴趣的细胞群体的转染，同时赋予CART的缀合的小分子的一些靶向性质。
Charge-altering releasable transporters (CARTs) are a class of oligonucleotide delivery vehicles shown to be effective for delivery of messenger RNA (mRNA) both in vitro and in vivo. Here, we exploited the chemical versatility of the CART synthesis to generate CARTs containing the small-molecule drug fingolimod (FTY720) as a strategy to increase mRNA delivery and expression in lymphocytes through a specific ligand−receptor interaction. Fingolimod is an FDA-approved small-molecule drug that, upon in vivo phosphorylation, binds to the sphingosine-1-phosphate receptor 1 (S1P1), which is highly expressed on lymphocytes. Compared to its non-fingolimod-conjugated analogue, the fingolimod-conjugated CART achieved superior transfection of activated human and murine T and B lymphocytes in vitro. The higher transfection of the fingolimod-conjugated CARTs was lost when cells were exposed to a free fingolimod before transfection. In vivo, the fingolimod-conjugated CART showed increased mRNA delivery to marginal zone B cells and NK cells in the spleen, relative to CARTs lacking fingolimod. Moreover, fingolimod-CART-mediated mRNA delivery induces peripheral blood T-cell depletion similar to free fingolimod. Thus, we show that functionalization of CARTs with a pharmacologically validated small molecule can increase transfection of a cellular population of interest while conferring some of the targeting properties of the conjugated small molecule to the CARTs.
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影响因子：
20.3
作者：
Kamphuis, Elisabeth;Junt, Tobias;Kalinke, Ulrich
通讯作者：
Kalinke, Ulrich
在体内用脂质纳米颗粒系统地传递了未修饰和假氨酸修饰的mRNA的功效和免疫原性。
影响因子：
14
作者：
Kauffman KJ;Mir FF;Jhunjhunwala S;Kaczmarek JC;Hurtado JE;Yang JH;Webber MJ;Kowalski PS;Heartlein MW;DeRosa F;Anderson DG
通讯作者：
Anderson DG
影响因子：
16.1
作者：
Batty CJ;Heise MT;Bachelder EM;Ainslie KM
通讯作者：
Ainslie KM
DOI：
10.1073/pnas.1810002115
发表时间：
2018-09-25
影响因子：
11.1
作者：
Haabeth, Ole A. W.;Blake, Timothy R.;Levy, Ronald
通讯作者：
Levy, Ronald
影响因子：
4.4
作者：
Jasiulewicz, Aleksandra;Lisowska, Katarzyna A.;Witkowski, Jacek M.
通讯作者：
Witkowski, Jacek M.