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
View: Text | PDF
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

×

Figure 6

Neuromodulation of striatal MSN by DA in transgenic mice and human graft in HD mice.

Options: View larger image (or click on image) Download as PowerPoint
Neuromodulation of striatal MSN by DA in transgenic mice and human graft...
(A and B) The strategy for electrophysiological recording in transgenic mice (A) and in HD mice with human graft (B). (C) Current-induced single and multiple AP traces from D1-MSNs and D2-MSNs in transgenic mice under baseline conditions and after DA application. (D) Current-induced single and multiple AP traces in human D1-like MSNs and D2-like MSNs from grafted neurons 5MPT, under baseline conditions and after DA application. (E and H) Quantification of DA-induced changes in spike numbers evoked by depolarizing pulses: for transgenic mice, 240 pA for D1-MSNs (n = 9 cells from 5 mice), 200 pA for D2-MSNs (n = 10 cells from 4 mice); for grafted neurons, 100 pA for D1-like MSN (n = 12 cells from 6 mice), 120 pA for D2-like MSN (n = 8 cells from 6 mice). (F and G) Traces of current-induced single and multiple APs for both D1-MSNs and D2-MSNs from transgenic mice (F), and human D1-like MSNs and human D2-like MSNs from grafts 5MPT (G). (I–L) Traces from selected step current injections showing near-threshold stimuli elicited delayed spiking in D1-/D2-MSNs in mice, and D1-like/D2-like MSNs in 5-month human grafts. (M) Electrophysiological recorded neurons were stained by biocytin, mCherry, MEIS2, and GABA. Scale bar: 50 μm. (N–Q) Statistical analyses of the RMP (N) and rheobase (O) in D1-/D2-MSNs from transgenic mice, or human D1-like/D2-like MSNs in 5-month human grafts (P and Q), before and after DA application (n = 10 cells from 5 mice for mouse D1-MSN; n = 10 cells from 4 mice for mouse D2-MSN; n = 12 cells from 6 mice for human D1-like MSN; n = 8 cells from 6 mice for human D2-like MSN). Paired Student’s t tests compared before and after DA treatments; unpaired Student’s t tests analyzed between-group differences. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. transpl., transplantation.