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 3

Specification and characterization of human LGE neural progenitor and MSN subtypes in vitro.

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Specification and characterization of human LGE neural progenitor and MS...
(A) UMAP visualization of cell clusters identified from integrated scRNA-Seq data sets of day 20 and day 44 samples (n = 2 biological replicates per time point). (B) UMAP visualization the expression of marker genes for different cell types. (C) Spatial mapping of all cell clusters onto the E13 mouse brain using VoxHunt with Allen Developing Mouse Brain Atlas reference data. (D) Violin plots showing the expression of regional marker genes for the forebrain, cortex, LGE, MGE, and CGE. (E) Bar plots quantifying the proportions of cell types among total cells (left) and total neurons (right) in the day 44 data set. (F) Heatmap of the top 100 (DEGs) distinguishing MSN subtypes. (G) Volcano plot highlighting DEGs potentially driving the lineages divergence of D1-MSNs versus D2-MSNs (H) KEGG pathway enrichment analysis of DEGs across distinct cell clusters. (I) Heatmaps depicting pairwise transcriptional correlations of 3D-default XFSC–derived LGE progenitors and MSNs in vitro with cell types in the developing human brain data set (30). AUROC, area under the receiver operating characteristic curve; GLU, glutamatergic neuron; max, maximum; min, minimum.