Partial rescue of mucopolysaccharidosis type VII mice with a lifelong engraftment of allogeneic stem cells in utero.
Partial rescue of mucopolysaccharidosis type VII mice with a lifelong engraftment of allogeneic stem cells in utero.
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DOI：
10.1111/cga.12099
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发表时间：
2015-02
影响因子：
1.3
通讯作者：
Mizutani S
中科院分区：
文献类型：
--
作者：
Ihara N;Akihiro U;Onami N;Tsumura H;Inoue E;Hayashi S;Sago H;Mizutani S
子宫内造血细胞移植（IUHCT）已在粘多糖沉积症VII型（MPSVII）小鼠中进行，但同种异体供体细胞的终身植入尚未实现。在这项研究中，我们试图确认与母亲免疫匹配的同种异体供体细胞的终身植入，并通过静脉注射IUHCT实现原始MPSVII菌株中表型的部分拯救。我们在MPSVII小鼠模型中进行了体外受精，并在接受IUHCT的胎儿均为纯合子的情况下将受影响的胚胎转移至ICR/B6-GFP代孕母亲。在E14.5静脉内注射来自ICR/B6-GFP小鼠的谱系耗尽的细胞。在出生后4周时通过流式细胞术确认嵌合体，并在8周龄或更晚时评估血清中的β-葡萄糖醛酸酶活性和几种表型。在死亡时检测到来自ICR/B6-GFP母体的嵌合小鼠中的供体细胞，并在包括处死的嵌合小鼠的脑的几个组织中确认。虽然嵌合小鼠的血清酶活性极低，但供体细胞的植入率与酶活性相关。此外，在我们有限的临床前研究中证实了骨结构的改善和生殖能力的挽救。我们使用静脉内IUHCT证实了供体细胞在原始免疫活性MPSVII小鼠模型中的终身植入，其中细胞与母亲免疫匹配，无需骨髓消融，并且多种表型得到改善。
In utero hematopoietic cell transplantation (IUHCT) has been performed in Mucopolysaccharidosis Type VII (MPSVII) mice, but a lifelong engraftment of allogeneic donor cells has not been achieved. In this study, we sought to confirm a lifelong engraftment of allogeneic donor cells immunologically matched to the mother and to achieve partial rescue of phenotypes in the original MPSVII strain through IUHCT by intravenous injection. We performed in vitro fertilization in a MPSVII murine model and transferred affected embryos to ICR/B6-GFP surrogate mothers in cases where fetuses receiving IUHCT were all homozygous. Lineage-depleted cells from ICR/B6-GFP mice were injected intravenously at E14.5. Chimerism was confirmed by flow cytometry at 4 weeks after birth, and β-glucuronidase activity in serum and several phenotypes were assessed at 8 weeks of age or later. Donor cells in chimeric mice from ICR/B6-GFP mothers were detected at death, and were confirmed in several tissues including the brains of sacrificed chimeric mice. Although the serum enzyme activity of chimeric mice was extremely low, the engraftment rate of donor cells correlated with enzyme activity. Furthermore, improvement of bone structure and rescue of reproductive ability were confirmed in our limited preclinical study. We confirmed the lifelong engraftment of donor cells in an original immunocompetent MPSVII murine model using intravenous IUHCT with cells immunologically matched to the mother without myeloablation, and the improvement of several phenotypes.