Professor Shandong First Medical University Jinan, China (People's Republic)
Abstract: Patient-specific induced pluripotent stem cells (iPSCs) have great therapeutic potential for neurodegenerative diseases such as Parkinson’ disease (PD). However, dopaminergic neurons (iPSC-DAN) derived from the iPSCs of PD patients with genetic mutations have decreased function and survival after these iPSC-DANs are transplanted to animal models or the PD patients. Therefore, the mutations in the iPSC-DAN have to be corrected before these cells are translated to the clinical application. we have generated three iPSC lines with mutations of GBA, VPA35 and LPR10 by electroporating the transgene-free episomal plasmids expressing OCT3/4, SOX2, KLF4, L-MYC, and LIN28 into the skin fibroblast cells from patients with PD. These three iPSC lines are carrying the mutations of GBA RecNciI mutation (c.1448T > C, c.1483G > C, and c.1497G > C), VPS13A mutation (c. 4282_4289delinsA), and LRP10 mutation (c.688C > T) respectively. We have also modified the protocol to efficiently differentiate iPSCs to iPSC-DANs with more than 60% cells expressing TUJ1/TH by adding the small molecules to different stages of differentiation by regulating the dopaminergic lineage commitment and maturation. To correct mutations of GBA, VPA35 and LPR10 in three iPSC lines, we have used the CRISPR/Cas9 editing to correct these mutations in the iPSC lines. The GBA mutation was corrected and was confirmed by sequencing analysis. We are comparing the phenotypes and mitochondrial functions of the iPSC-DANs carrying GBA mutation to that without GBA mutation-corrected. Next we are going to transplant the iPSC-DANs from normal individuals, patient carrying GBA mutation and corrected GBA mutation to the rat PD model to see whether or not iPSC-DANs with corrected GBA mutation has similar ability as that from normal individuals to improve the motor deficits of PD rats.
Funding Source: The Key Basic Research project (ZR2019ZD39). , Natural Science Foundation of Shandong Province, China.
