Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.
Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.
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DOI：
10.1016/j.jconrel.2021.10.031
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发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Mitchell MJ
中科院分区：
文献类型：
--
作者：
Swingle KL;Billingsley MM;Bose SK;White B;Palanki R;Dave A;Patel SK;Gong N;Hamilton AG;Alameh MG;Weissman D;Peranteau WH;Mitchell MJ
导致病理性蛋白质缺乏的先天性疾病最常在出生后用蛋白质或酶替代疗法治疗。然而，在不可逆疾病发作前在子宫内治疗这些疾病可以显著降低疾病负担、发病率和死亡率。先天性疾病的产前治疗的一种可能的策略是在子宫内递送信使RNA（mRNA）。mRNA是一种基因治疗剂，以前曾被研究用于蛋白质替代疗法和基因编辑技术。虽然已经探索了病毒载体来诱导mRNA的细胞内表达，但是由于免疫原性和基因组整合的问题，它们在临床应用中受到限制。作为病毒载体的替代方案，使用可电离脂质纳米颗粒（LNP）可以实现安全且有效的子宫内mRNA递送。虽然LNP已经证明在静脉内施用后有效的体内mRNA递送至肝脏，但是羊膜内递送具有将mRNA递送至肝脏中的细胞和组织以外的细胞和组织（例如皮肤、肺和消化道中的细胞和组织）的潜力。然而，LNP在胎儿羊水中的稳定性以及这种稳定性如何影响mRNA递送先前尚未研究。在这里，我们使用正交实验设计（DOE）设计了LNP文库，以评估LNP结构如何影响羊水中的子宫外稳定性，以及从这些稳定性测量中鉴定的先导候选物是否能够在子宫内进行羊膜内mRNA递送。我们使用的技术，包括动态光散射（DLS），透射电子显微镜（TEM），和色谱法，然后蛋白质含量定量筛选羊水中的离体LNP稳定性的组合。这些结果鉴定了多种铅LNP制剂，其在从小动物到人类的羊水中高度稳定，包括小鼠、绵羊、猪和人类羊水样品。然后，我们证明了在小鼠羊水中来自子宫外筛选的稳定的LNP能够在小鼠模型中在羊膜内注射后在体外和子宫内进行有效的mRNA递送。羊水中子宫外稳定性的这种探索证明了一种方法，通过该方法鉴定用于通过子宫内mRNA递送产前治疗先天性疾病的新型LNP制剂。
Congenital disorders resulting in pathological protein deficiencies are most often treated postnatally with protein or enzyme replacement therapies. However, treatment of these disorders in utero before irreversible disease onset could significantly minimize disease burden, morbidity, and mortality. One possible strategy for the prenatal treatment of congenital disorders is the in utero delivery of messenger RNA (mRNA). mRNA is a gene therapeutic that has previously been investigated for protein replacement therapies and gene editing technologies. While viral vectors have been explored to induce intracellular expression of mRNA, they are limited in their clinical application due to concerns of immunogenicity and genomic integration. As an alternative to viral vectors, safe and efficient in utero mRNA delivery can be achieved using ionizable lipid nanoparticles (LNPs). While LNPs have demonstrated potent in vivo mRNA delivery to the liver following intravenous administration, intra-amniotic delivery has the potential to deliver mRNA to cells and tissues beyond those in the liver, such as in the skin, lung, and digestive tract. However, LNP stability in fetal amniotic fluid and how this stability affects mRNA delivery has not been previously investigated. Here, we engineered a library of LNPs using orthogonal design of experiments (DOE) to evaluate how LNP structure affects ex utero stability in amniotic fluid, and whether a lead candidate identified from these stability measurements enables intra-amniotic mRNA delivery in utero. We used a combination of techniques including dynamic light scattering (DLS), transmission electron microscopy (TEM), and chromatography followed by protein content quantification to screen ex vivo LNP stability in amniotic fluids. These results identified multiple lead LNP formulations that are highly stable in amniotic fluids ranging from small animals to humans, including mouse, sheep, pig, and human amniotic fluid samples. We then demonstrate that stable LNPs from the ex utero screen in mouse amniotic fluid enabled potent mRNA delivery in vitro and in utero following intra-amniotic injection in a murine model. This exploration of ex utero stability in amniotic fluids demonstrates a means by which to identify novel LNP formulations for prenatal treatment of congenital disorders via in utero mRNA delivery.
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影响因子：
46.9
作者：
Gilleron, Jerome;Querbes, William;Zerial, Marino
通讯作者：
Zerial, Marino
影响因子：
2.3
作者：
Connolly B;Isaacs C;Cheng L;Asrani KH;Subramanian RR
通讯作者：
Subramanian RR
DOI：
10.1016/j.biocel.2015.11.005
发表时间：
2016-06-01
影响因子：
4
作者：
Graefe, Christine;Weidner, Andreas;Dutz, Silvio
通讯作者：
Dutz, Silvio
影响因子：
15
作者：
Chen, Delai;Love, Kevin T.;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
影响因子：
4.9
作者：
Gaspar, Diana P.;Gaspar, Maria Manuela;Remunan-Lopez, Carmen
通讯作者：
Remunan-Lopez, Carmen