Intra-Amniotic Delivery of CRMP4 siRNA Improves Mesenchymal Stem Cell Therapy in a Rat Spina Bifida Model

Xiaowei Wei; Songying Cao; Wei Ma; Chaonan Zhang; Hui Gu; Dan Liu; Wenting Luo; Yuzuo Bai; Weilin Wang; Zhengwei Yuan

doi:10.1016/j.omtn.2020.03.007

Intra-Amniotic Delivery of CRMP4 siRNA Improves Mesenchymal Stem Cell Therapy in a Rat Spina Bifida Model

Mol Ther Nucleic Acids. 2020 Jun 5:20:502-517. doi: 10.1016/j.omtn.2020.03.007. Epub 2020 Mar 19.

Authors

Xiaowei Wei¹, Songying Cao², Wei Ma¹, Chaonan Zhang¹, Hui Gu¹, Dan Liu¹, Wenting Luo¹, Yuzuo Bai³, Weilin Wang³, Zhengwei Yuan⁴

Affiliations

¹ Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China.
² Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China. Electronic address: csymn01@163.com.
³ Department of Pediatric Surgery, Shengjing Hospital, China Medical University, Shenyang, P.R. China.
⁴ Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang 110004, P.R. China. Electronic address: yuanzw@hotmail.com.

Abstract

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.

Keywords: CRMP4 siRNA; gene transfer; intra-amniotic injection; mesenchymal stem cell transplantation; neural differentiatio; neural tube defects; neurological repair; prenatal therapy.