A role for TGFβ signalling in medium spiny neuron differentiation of human pluripotent stem cells

Abstract

Activin A and other TGFβ family members have been shown to exhibit a certain degree of promiscuity between their family of receptors. We previously developed an efficient differentiation protocol using Activin A to obtain medium spiny neurons (MSNs) from human pluripotent stem cells (hPSCs). However, the mechanism underlying Activin A-induced MSN fate specification remains largely unknown. Here we begin to tease apart the different components of TGFβ pathways involved in MSN differentiation and demonstrate that Activin A acts exclusively via ALK4/5 receptors to induce MSN progenitor fate during differentiation. Moreover, we show that Alantolactone, an indirect activator of SMAD2/3 signalling, offers an alternative approach to differentiate hPSC-derived forebrain progenitors into MSNs. Further fine tuning of TGFβ pathway by inhibiting BMP signalling with LDN193189 achieves accelerated MSN fate specification. The present study therefore establishes an essential role for TGFβ signalling in human MSN differentiation and provides a fully defined and highly adaptable small molecule-based protocol to obtain MSNs from hPSCs.

Dysfunction of cAMP–Protein Kinase A–Calcium Signaling Axis in Striatal Medium Spiny Neurons: A Role in Schizophrenia and Huntington’s Disease Neuropathology

“…All experiments were performed in the HUES9 iCas9 hESC line and genome-edited derivatives lacking BCL11B protein (clones #4, #33, #34) ( 11 ). As in previous studies, BCL11B knockout (BCL11B KO ) and control lines were reliably differentiated into MSN, cortical glutamatergic, and mDA neuronal subtypes using established protocols ( Figure 1A ; Figure S1 ) ( 22 , 23 , 24 ).

Figure 1 BCL11B KO MSNs and CTX neurons manifest signs of mitochondrial dysfunction and abnormal intracellular Ca 2+ regulation.

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Identification of molecular signatures and pathways involved in Rett syndrome using a multi-omics approach

“…The 100 DEGs upregulated in RTT and downregulated in MDS enrich processes related to neurogenesis regulation; signalling cascades, such as Wnt, BMP, and TGFß; and the cytoskeleton [Figure 4B, Table S4c]. The Wnt, BMP, and TGFß signalling pathways participate in neurogenesis regulation, synapse formation, myelin synthesis, and neural stem cell renewal [109][110][111][112]. The BMP signalling cascade has recently been found upregulated in neural stem/precursor cells from patients with RTT, and treatment with BMP inhibitors partially rescued the abnormal development detected in RTT brain organoids [113].…”

Dysfunction of cAMP-PKA-calcium signaling axis in striatal medium spiny neurons: a role in schizophrenia and Huntington’s disease pathogenesis

“…All experiments were performed in HUES9 iCas9 hESC line and genome-edited derivatives lacking BCL11B protein (clones #4, #33, #34) (11). As in previous study, BCL11B knockout (BCL11B KO ) and control lines were reliably differentiated into MSN, cortical glutamatergic and mDA neuronal subtypes using established protocols (Figure 1A and S1) (19)(20)(21).…”