Neural Regeneration Research ›› 2020, Vol. 15 ›› Issue (12): 2335-2343.doi: 10.4103/1673-5374.285002

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Insulin-like growth factor 1 partially rescues early developmental defects caused by SHANK2 knockdown in human neurons

Shu-Ting Chen 1 , Wan-Jing Lai 1, 2 , Wei-Jia Zhang 1 , Qing-Pei Chen 1 , Li-Bing Zhou 1 , Kwok-Fai So 1 , Ling-Ling Shi 1, 3, 4   

  1. 1 Guangdong-Hong Kong-Macau Institute of CNS Regeneration, Ministry of Education CNS Regeneration Collaborative Joint Laboratory, Jinan University, Guangzhou, Guangdong Province, China
    2 Clinical Medicine, Jinan University, Guangzhou, Guangdong Province, China
    3 Department of Psychiatry, the First Affiliated Hospital of Jinan University, Guangzhou, Guangdong Province, China
    4 Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
  • Online:2020-12-15 Published:2020-08-05
  • Contact: Ling-Ling Shi, MD, PhD,tlingshi@jnu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81771222 (to LLS); the Natural Science Foundation of Guangdong Province of China, No. 2019A1515011316 (to LLS); the National Key Research and Development Program of China, Stem Cell and Translational Research, No. 2017YFA0105102 (to LLS); the Guangzhou Science and Technology Innovation Development Special Fund Project of China, No. 201804010212 (to LLS); the Program of Introducing Talents of Discipline to Universities of China, No. B14036 (to KFS).

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Abstract: SHANK2 is a scaffold protein that serves as a protein anchor at the postsynaptic density in neurons. Genetic variants of SHANK2 are strongly associated with synaptic dysfunction and the pathophysiology of autism spectrum disorder. Recent studies indicate that early neuronal developmental defects play a role in the pathogenesis of autism spectrum disorder, and that insulin-like growth factor 1 has a positive effect on neurite development. To investigate the effects of SHANK2 knockdown on early neuronal development, we generated a sparse culture system using human induced pluripotent stem cells, which then differentiated into neural progenitor cells after 3–14 days in culture, and which were dissociated into single neurons. Neurons in the experimental group were infected with shSHANK2 lentivirus car- rying a red fluorescent protein reporter (shSHANK2 group). Control neurons were infected with scrambled shControl lentivirus carrying a red fluorescent protein reporter (shControl group). Neuronal somata and neurites were reconstructed based on the lentiviral red fluo- rescent protein signal. Developmental dendritic and motility changes in VGLUT1 + glutamatergic neurons and TH + dopaminergic neurons were then evaluated in both groups. Compared with shControl VGLUT1 + neurons, the dendritic length and arborizations of shSHANK2 VGLUT1 + neurons were shorter and fewer, while cell soma speed was higher. Furthermore, dendritic length and arborization were sig- nificantly increased after insulin-like growth factor 1 treatment of shSHANK2 neurons, while cell soma speed remained unaffected. These results suggest that insulin-like growth factor 1 can rescue morphological defects, but not the change in neuronal motility. Collectively, our findings demonstrate that SHANK2 deficiency perturbs early neuronal development, and that IGF1 can partially rescue the neuronal defects caused by SHANK2 knockdown. All experimental procedures and protocols were approved by the Laboratory Animal Ethics Com- mittee of Jinan University, China (approval No. 20170228010) on February 28, 2017.

Key words: cells, factor, growth, in vitro, model, neural differentiation, neurogenesis, plasticity