Neural tube defects (NTDs) result in prenatal mortality and lifelong morbidity, and available treatments have limited efficacy. We previously suggested that prenatal bone marrow-derived mesenchymal stem cell (BMSC) transplantation could treat neuron deficiency in NTD rats; however, BMSC-based therapy is limited by the low survival rate of BMSCs when used to treat severe NTDs. Herein, a new therapy using combined BMSC transplantation and small interfering RNA of collapsin response mediator protein 4 (CRMP4 siRNA), which was identified as a novel potential target for the NTD treatment, is proposed. The intra-amniotic CRMP4 siRNA, BMSC, and CRMP4 siRNA + BMSC injections repaired skin lesions, improved motor neural function, reduced neuronal apoptosis, and promoted expression of neural differentiation-related molecules and neurotrophic factors in the spinal cord of spina bifida rat fetuses. Therapeutic effects in the CRMP4 siRNA + BMSC injection group were superior to those of the CRMP4 siRNA only or BMSC only injection groups. CRMP4 siRNA + BMSC injection resulted in a 45.38% reduction in the skin lesion area and significantly shorter latency and higher amplitude of motor-evoked potentials (MEPs) in spina bifida fetuses. Our results suggest that intrauterine Ad-CRMP4 siRNA delivery with BMSCs is an innovative platform for developing fetal therapeutics to safely and efficaciously treat NTDs.

神经管缺陷（NTD）会导致产前死亡和终生发病，并且可用的治疗方法疗效有限。我们以前曾提出，产前骨髓间充质干细胞（BMSC）移植可以治疗NTD大鼠的神经元缺乏。但是，基于BMSC的疗法在用于治疗严重NTD时受限于BMSC的低存活率。在本文中，提出了一种新的疗法，该疗法结合了BMSC移植和胶原蛋白介导蛋白4（CRMP4 siRNA）的小干扰RNA，已被确定为NTD治疗的新靶标。羊膜内CRMP4 siRNA，BMSC和CRMP4 siRNA + BMSC注射修复了皮肤病变，改善了运动神经功能，减少了神经元凋亡，促进脊柱裂大鼠胎儿脊髓中神经分化相关分子和神经营养因子的表达。CRMP4 siRNA + BMSC注射组的治疗效果优于仅CRMP4 siRNA或仅BMSC注射组的治疗效果。CRMP4 siRNA + BMSC注射可使脊柱裂胎儿的皮肤病变面积减少45.38％，潜伏期明显缩短，运动诱发电位（MEP）幅度更高。我们的结果表明 脊柱裂胎儿的皮肤病变面积减少了38％，潜伏期明显缩短，运动诱发电位（MEP）幅度更高。我们的结果表明 脊柱裂胎儿的皮肤病变面积减少了38％，潜伏期明显缩短，运动诱发电位（MEP）幅度更高。我们的结果表明BMSCs的宫内Ad-CRMP4 siRNA递送是一种创新平台，可用于开发胎儿疗法以安全有效地治疗NTD。