Moskowitzova Kamila, Scire Emily M, Dang Tanya T, Bechara Beatriz S, Shroff Yash V, Zacharakis Eva, Fitzgerald Isabela D, Couto Aaliyah S, Zurakowski David, Fauza Dario O
Department of Surgery Boston Children's Hospital/Harvard Medical School Boston Massachusetts USA.
FASEB Bioadv. 2025 Sep 3;7(8):e70047. doi: 10.1096/fba.2025-00200. eCollection 2025 Aug.
Hemophilia A is an X-linked monogenic disease resulting in insufficient pro-coagulant factor VIII (FVIII) levels. Hemophiliac infants are at risk for life-threatening hemorrhage, especially during birth. No perinatal treatment for Hemophilia A is currently available. It has been previously shown that the transamniotic route is a viable option to deliver exogenous mRNA to the fetus. We sought to determine whether FVIII mRNA so delivered could be translated by the fetus, leading to the presence of FVIII in the fetal circulation. Time-dated pregnant Sprague Dawley dams underwent volume-matched intra-amniotic injections in all their fetuses ( = 166) of either a human FVIII (hFVIII) mRNA encapsulated by lipopolyplex (mRNA; = 115) or of the same lipopolyplex without mRNA (control; = 51) on gestational day 17 (E17; term = E21-22). Fetal liver and serum samples were procured daily until term and screened for hFVIII protein by ELISA. There was no maternal mortality. Overall survival was 90% (149/166). Controlled by the mRNA-free injections, fetal serum levels of hFVIII were statistically significantly higher overall in the mRNA group ( = 0.002), peaking at E20 (24.4 ± 2.4 ng/mL in the mRNA group vs. 10.5 ± 1.9 ng/mL for control; < 0.001). In the fetal liver, there was variability in statistically significant differences between the groups, with the shorter time point showing significance ( = 0.003). Encapsulated exogenous mRNA encoding for factor VIII can be incorporated and translated by the fetus following simple intra-amniotic injection in a rat model. Transamniotic mRNA delivery could become a novel strategy for the perinatal management of Hemophilia A.
甲型血友病是一种X连锁单基因疾病，会导致促凝血因子VIII（FVIII）水平不足。血友病婴儿有发生危及生命的出血的风险，尤其是在出生期间。目前尚无针对甲型血友病的围产期治疗方法。先前已表明，经羊膜途径是向胎儿递送外源性mRNA的可行选择。我们试图确定如此递送的FVIII mRNA是否能被胎儿翻译，从而使胎儿循环中出现FVIII。在妊娠第17天（E17；足月为E21 - 22），对处于相应孕周的怀孕斯普拉格 - 道利母鼠的所有胎儿（n = 166）进行体积匹配的羊膜腔内注射，其中115只胎儿注射由脂质多聚体包裹的人FVIII（hFVIII）mRNA（mRNA组），51只胎儿注射不含mRNA的相同脂质多聚体（对照组）。每天采集胎儿肝脏和血清样本直至足月，并通过酶联免疫吸附测定（ELISA）检测hFVIII蛋白。没有母鼠死亡。总体存活率为90%（149/166）。在无mRNA注射的对照下，mRNA组胎儿血清中的hFVIII水平总体在统计学上显著更高（P = 0.002），在E20达到峰值（mRNA组为24.4 ± 2.4 ng/mL，对照组为10.5 ± 1.9 ng/mL；P < 0.001）。在胎儿肝脏中，两组之间的统计学显著差异存在变异性，较短时间点显示出显著性（P = 0.003）。在大鼠模型中，经简单的羊膜腔内注射后，编码因子VIII的包裹外源性mRNA可被胎儿摄取并翻译。经羊膜递送mRNA可能成为甲型血友病围产期管理的一种新策略。
