Identifying the most effective types of integration-free human iPS cell-derived neural stem/progenitor cells in the treatment of spinal cord injury

Identifying the most effective types of integration-free human iPS cell-derived neural stem/progenitor cells in the treatment of spinal cord injury

Authors: T. IIDA, N. NAGOSHI, J. KOHYAMA, O. TSUJI, M. MATSUMOTO, M. NAKAMURA, H. OKANO
Lab Abstract:
INTRODUCTION: We have previously demonstrated the therapeutic potential of transplanting human iPS cell-derived neural stem/progenitor cells (hiPSC-NS/PCs) in the treatment of spinal cord injury (SCI) models. Recently, we have produced integration-free hiPSCs using episomal vectors which is safer for clinical use. The purpose of this study is to assess the efficacy of integration-free hiPSC-NS/PCs, and to investigate their genetic profiles in order to evaluate factors related to therapeutic efficacy. METHODS: Two integration-free human iPS cell lines were prepared (836B3-, 414C2-hiPSCs), and were induced to hiPSC-NS/PCs (836B3-, 414C2-NS/PCs). ES cells were also used for analysis as a target for comparison of hiPSCs. hiPSC-NS/PCs were transplanted into the injured spinal cord of NOD-SCID mice, and phosphate buffered saline (PBS) was injected to the control group (414C2-NS/PCs; n=27, 836B3-NS/PCs; n=23, control; n=15). Transplanted cells were monitored using bio-imaging and evaluated histologically; and motor function was evaluated by basso mouse scale (BMS) score. HumanHT-12 was used to evaluate gene-expression analyses and single-cell-RNA-sequence was performed using Illumina-Hiseq2500. RESULTS: In the in-vivo study, better motor functional recovery was observed in the 414C2-NS/PCs group compared with the control group (p< 0.001). In contrast, 836B3-NS/PCs group showed no improvement in motor function, and formed undifferentiated tissues. The gene-expression profile of 414C2-hiPSCs resembled that of ES cells with clustering analysis, and 12 genes which included genome-stabilization gene such as DPPA3 and differentiation related genes such as IRX2 were highly expressed in 414C2-hiPSCs, similar to those found in ES cells. None of these findings, however, were observed in 836B3-hiPSCs. In single-cell-RNA-sequence, Delta-Notch signal positive cells which are important for neural differentiation were more abundant in 414C2-NS/PCs, whereas 836B3-NS/PCs only contained a small population (12% and 5%, respectively).

DISCUSSION: In order to pursue our mission of conducting a clinical trial for SCI patients within the next several years, it is critical to build guidelines for selecting “effective” hiPSC-NS/PCs, such as 414C2-NS/PCs. Herein we showed that the key to good motor function recovery is to transplant integration-free hiPSC-NS/PCs that differentiate well within the spinal cord tissue. Our results suggest that examining hiPSCs quality with 12-gene markers and establishing hiPSC-NS/PCs that contain Delta-Notch (+) cells for more than 10% could be the criteria to select “effective” hiPSC-NS/PCs for SCI treatment.

Abstract Citation
*T. IIDA1, N. NAGOSHI1, J. KOHYAMA2, O. TSUJI1, M. MATSUMOTO1, M. NAKAMURA1, H. OKANO2;
1Dept of Orthop, Sch. of Med, Keio Univ., Shinjyuku-Ku Tokyo, Japan; 2Dept of Physiology, Sch. of Med, Keio Univ., Shinjyuku-Ku Tokyo, Japan. Identifying the most effective types of integration-free human iPS cell-derived neural stem/progenitor cells in the treatment of spinal cord injury. Program No. 213.06. 2018 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2018. Online.

Keio researchers plan to treat spinal cord injuries with stem cells  at The Japan Times News LINK

Keio Univ. to OK transplant of iPS-derived cells for spine injury treatment in world 1st   at The Mainichi LINK

This entry was posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, Spinal Research, Stem Cell Research and tagged , , , , . Bookmark the permalink.

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