3D cultured human medium spiny neurons functionally integrate and rescue motor deficits in Huntington’s disease mice
Yuting Mei, Yuan Xu, Xinyue Zhang, Ban Feng, Yingying Zhou, Qian Cheng, Yuan Li, Xingsheng Peng, Mengnan Wu, Lianshun Xie, Lei Xiao, Wenhao Zhou, Yuejun Chen, Man Xiong
Yuting Mei, Yuan Xu, Xinyue Zhang, Ban Feng, Yingying Zhou, Qian Cheng, Yuan Li, Xingsheng Peng, Mengnan Wu, Lianshun Xie, Lei Xiao, Wenhao Zhou, Yuejun Chen, Man Xiong
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Research Article Cell biology

3D cultured human medium spiny neurons functionally integrate and rescue motor deficits in Huntington’s disease mice

Abstract

Dysfunction of striatal medium spiny neurons (MSNs) is implicated in several neurological disorders, including Huntington’s disease (HD). Despite progress in characterizing MSN pathology in HD, mechanisms underlying MSN susceptibility remain unknown, driving the need for MSNs derived from human pluripotent stem cells (hPSCs), especially subtypes in research and therapy. Here, we established a scalable 3D-default culture system to produce striatal MSNs efficiently from hPSCs by activation of the endogenous sonic hedgehog (SHH) pathway. These cells expressed canonical markers of striatal progenitors and dopamine D1 (D1) and dopamine D2 (D2) MSNs and presented dynamic specification and transcriptional signatures that closely resemble endogenous MSNs at single-cell resolution, both in vitro and post-transplantation in HD mice with quinolinic acid (QA) lesions. Grafted human cells survived and matured into D1-/D2-like MSNs and projected axons to endogenous targets including globus pallidus externus, globus pallidus internus, and substantia nigra pars reticulata to reconstruct the basal ganglia pathways. Functionally, they displayed spontaneous synaptic currents, received regulation from host cortex and thalamus, and were modulated by dopamine to either enhance or reduce neuronal excitability, similar to the endogenous D1-/D2-MSNs, subsequently improving behavior in QA-lesioned HD mice. Our study presents a method for generating authentic MSNs, providing a reliable cell source for HD cell therapy, mechanistic studies, and drug screening.

Authors

Yuting Mei, Yuan Xu, Xinyue Zhang, Ban Feng, Yingying Zhou, Qian Cheng, Yuan Li, Xingsheng Peng, Mengnan Wu, Lianshun Xie, Lei Xiao, Wenhao Zhou, Yuejun Chen, Man Xiong

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Figure 5

Characterization of human LGE progenitors and MSN subtypes after transplantation in HD mice using snRNA-Seq.

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Characterization of human LGE progenitors and MSN subtypes after transpl...
(A) Schematic of the experimental design for cell transplantation in HD model mice and subsequent snRNA-Seq 5MPT. (B) UMAP visualization of all identified human cell clusters in the graft. (C) Bar plot showing the proportions of each cell type among total human cells. (D) UMAP plots displaying the expression of canonical marker genes for different cell types. (E) Dot plot showing the expression of marker genes for major cell types. (F) Dot plot showing the expression of canonical D1- and D2-MSN marker genes across the 2 main MSN subtypes. (G) Dot plot showing the expression of subtype-specific marker genes across all identified MSN subtypes. (H) Heatmap displaying the top 100 DEGs distinguishing MSN subtypes. (I) KEGG pathway enrichment analysis of DEGs in D1-MSNs versus D2-MSNs. (J) Dot plot showing the expression of marker genes associated with striosome and matrix among MSN subtypes. (K) Heatmap depicting pairwise transcriptional correlations of 3D XFSC–derived progenitors and MSNs in the grafts with those in the developing human brain data set (30). (L) Heatmap depicting pairwise transcriptional correlations of graft-derived cells and developmental stages of human brain data set (30).