Human autologous iPSC–derived dopaminergic progenitors restore motor function in Parkinson’s disease models
Bin Song, … , Jeffrey S. Schweitzer, Kwang-Soo Kim
Bin Song, … , Jeffrey S. Schweitzer, Kwang-Soo Kim
Published November 12, 2019
Citation Information: J Clin Invest. 2020;130(2):904-920. https://doi.org/10.1172/JCI130767.
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Human autologous iPSC–derived dopaminergic progenitors restore motor function in Parkinson’s disease models

Abstract

Parkinson’s disease (PD) is a neurodegenerative disorder associated with loss of striatal dopamine, secondary to degeneration of midbrain dopamine (mDA) neurons in the substantia nigra, rendering cell transplantation a promising therapeutic strategy. To establish human induced pluripotent stem cell–based (hiPSC-based) autologous cell therapy, we report a platform of core techniques for the production of mDA progenitors as a safe and effective therapeutic product. First, by combining metabolism-regulating microRNAs with reprogramming factors, we developed a method to more efficiently generate clinical-grade iPSCs, as evidenced by genomic integrity and unbiased pluripotent potential. Second, we established a “spotting”-based in vitro differentiation methodology to generate functional and healthy mDA cells in a scalable manner. Third, we developed a chemical method that safely eliminates undifferentiated cells from the final product. Dopaminergic cells thus express high levels of characteristic mDA markers, produce and secrete dopamine, and exhibit electrophysiological features typical of mDA cells. Transplantation of these cells into rodent models of PD robustly restores motor function and reinnervates host brain, while showing no evidence of tumor formation or redistribution of the implanted cells. We propose that this platform is suitable for the successful implementation of human personalized autologous cell therapy for PD.

Authors

Bin Song, Young Cha, Sanghyeok Ko, Jeha Jeon, Nayeon Lee, Hyemyung Seo, Kyung-Joon Park, In-Hee Lee, Claudia Lopes, Melissa Feitosa, María José Luna, Jin Hyuk Jung, Jisun Kim, Dabin Hwang, Bruce M. Cohen, Martin H. Teicher, Pierre Leblanc, Bob S. Carter, Jeffrey H. Kordower, Vadim Y. Bolshakov, Sek Won Kong, Jeffrey S. Schweitzer, Kwang-Soo Kim

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Figure 7

In vivo safety of C4-derived mDA cells in NOD SCID mice.

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In vivo safety of C4-derived mDA cells in NOD SCID mice. (A) H&E sta...
(A) H&E staining of NOD SCID mouse brain after striatal transplantation of C4 iPS cells (d0, left), or of C4-derived mDAPs at d14 (middle) or d28 (right). The white circle in the d14 group identifies rosette-like structures. (B) Quantification of teratoma formation percentage at d0 (n = 4) and d14 without quercetin (n = 4) and at d14 (n = 19) and d28 (n = 23) with quercetin treatment groups. (C) Quantification of rosette formation at d14 of differentiation without quercetin and at d14 and d28 with quercetin treatment. (D) Immunohistochemistry of vimentin in d14 and d28 groups. (E and F) Immunofluorescence staining of SOX1, PAX6, and KI67 in d14 (E) and d28 groups (F). (G) Quantification of SOX1+, KI67+, SOX1+KI67+, SOX1+PAX6+, SOX1+PAX6+KI67+ populations in d14 and d28 groups. Data are presented as mean ± SEM. n = 4. ***P < 0.001, Student’s t test. (H) Biodistribution assay. RT-PCR of human- or mouse-specific gene expression in “brain mix” (mixture of olfactory bulb and cerebellum), spinal cord, lung, heart, spleen, kidney, and liver of the NOD SCID mice that had received intrastriatal hiPSC-derived d28 dopaminergic progenitor grafts 6 months previously. hiPSC serves as a positive control. The human-specific gene is located on chromosome 10 at 29125650 to 29125967. The mouse-specific gene is part of mouse TNF-α. Scale bars: 100 μm (unless otherwise specified).