Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use

Kim, Tae Wan; Piao, Jinghua; Koo, So Yeon; Kriks, Sonja; Chung, Sung Tae; Betel, Doron; Socci, Nicholas D.; Choi, Se Joon; Zabierowski, Susan E.; Dubose, Brittany N.; Hill, Ellen J.; Mosharov, Eugene V.; Irion, Stefan; Tomishima, Mark; Tabar, Viviane; Studer, Lorenz

doi:10.1016/j.stem.2021.01.005

Cited by 138 publications

(130 citation statements)

References 53 publications

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“…When using current DA neuron differentiation protocols, DA neurons account for only a small percentage of cells of the entire graft when mesDA progenitors are grafted in vivo . A protocol using FGF8 to induce midbrain caudalization was shown to result in the production of approximately 3,700 TH-positive DA cells per 100,000 grafted cells 14 , and a more recently reported protocol using a delayed boost of WNT was found to induce the generation of 9,173 TH-positive cells per 6.22 mm 3 graft following transplantation of 450,000 cells 13 . We investigated how our 4X LR-USCs behave in vivo when transplanted into a rodent model of Parkinson’s disease.…”

Section: Resultsmentioning

confidence: 97%

“…High concentrations result in the production of hindbrain cell types, and lower concentrations result in an anterior midbrain or diencephalic identity. Timed delivery of FGF8 or sequential exposure to high levels of WNT has also been shown to improve specification to a caudal midbrain identity 12,13 . Despite these advances in stem cell differentiation protocols, the overall yield of mesDA neurons after transplantation is extremely low, and increasing the yield would significantly improve cell purity and the reliability of graft function, which is essential for the use of these cells in the clinic 14,15 .…”

Section: Introductionmentioning

confidence: 99%

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Enhanced Production of Mesencephalic Dopaminergic Neurons from Lineage-Restricted Human Undifferentiated Stem Cells

Maimaitili

Chen

Febbraro³

et al. 2021

Preprint

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The differentiation of human pluripotent stem cells (hPSCs) into mesencephalic dopaminergic (mesDA) neurons requires a precise combination of extrinsic factors that recapitulates the in vivo environment and timing. Current methods are capable of generating authentic mesDA neurons after long-term culture in vitro; however, when mesDA progenitors are transplanted in vivo, the resulting mesDA neurons are only minor components of the graft. This low yield hampers the broad use of these cells in the clinic. In this study, we genetically modified pluripotent stem cells to generate a novel type of stem cells called lineage-restricted undifferentiated stem cells (LR-USCs), which robustly generate mesDA neurons. LR-USCs are prevented from differentiating into a broad range of nondopaminergic cell types by knocking out genes that are critical for the specification of cells of alternate lineages. Specifically, we target transcription factors involved in the production of spinal cord and posterior hindbrain cell types. When LR-USCs are differentiated under caudalizing condition, which normally give rise to hindbrain cell types, a large proportion adopt a midbrain identity and develop into authentic mesDA neurons. We show that the mesDA neurons are electrophysiologically active, and due to their higher purity, are capable of restoring motor behavior eight weeks after transplantation into 6-hydroxydopamine (6-OHDA)-lesioned rats. This novel strategy improves the reliability and scalability of mesDA neuron generation for clinical use.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Enhanced Production of Mesencephalic Dopaminergic Neurons from Lineage-Restricted Human Undifferentiated Stem Cells

Maimaitili

Chen

Febbraro³

et al. 2021

Preprint

View full text Add to dashboard Cite

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“…Transplanted DA neurons anatomically and functionally integrate in the SNpc of the host rodent brain, projecting over long distances and restoring motor function by re-establishing axonal connectivity, and subsequently, the DA transmission. Several studies provided compelling evidence that both A9-and A10-like neurons are formed in the graft with the capacity for long-distance target-specific innervation [19,57,58].…”

Section: Cell Transplantationmentioning

confidence: 99%

Dopamine Neuron Diversity: Recent Advances and Current Challenges in Human Stem Cell Models and Single Cell Sequencing

Fiorenzano

Sozzi

Parmar

et al. 2021

Cells

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Human midbrain dopamine (DA) neurons are a heterogeneous group of cells that share a common neurotransmitter phenotype and are in close anatomical proximity but display different functions, sensitivity to degeneration, and axonal innervation targets. The A9 DA neuron subtype controls motor function and is primarily degenerated in Parkinson’s disease (PD), whereas A10 neurons are largely unaffected by the condition, and their dysfunction is associated with neuropsychiatric disorders. Currently, DA neurons can only be reliably classified on the basis of topographical features, including anatomical location in the midbrain and projection targets in the forebrain. No systematic molecular classification at the genome-wide level has been proposed to date. Although many years of scientific efforts in embryonic and adult mouse brain have positioned us to better understand the complexity of DA neuron biology, many biological phenomena specific to humans are not amenable to being reproduced in animal models. The establishment of human cell-based systems combined with advanced computational single-cell transcriptomics holds great promise for decoding the mechanisms underlying maturation and diversification of human DA neurons, and linking their molecular heterogeneity to functions in the midbrain. Human pluripotent stem cells have emerged as a useful tool to recapitulate key molecular features of mature DA neuron subtypes. Here, we review some of the most recent advances and discuss the current challenges in using stem cells, to model human DA biology. We also describe how single cell RNA sequencing may provide key insights into the molecular programs driving DA progenitor specification into mature DA neuron subtypes. Exploiting the state-of-the-art approaches will lead to a better understanding of stem cell-derived DA neurons and their use in disease modeling and regenerative medicine.

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“…Hence, this signaling has been proposed as a potential therapeutic target against neurodegeneration [ 34 , 35 ]. It has also been reported that appropriate levels of Wnt signaling are critical for increasing the quantity and quality of mDA neurons derived from stem cells or reprogrammed cells, with clear implications for their use in cell replacement strategies, currently under investigation as a potential PD therapy [ 36 – 38 ]. Indeed, since the main pathological feature of PD is neuron death or deactivation in the midbrain, a promising therapeutic strategy for curing this disease is DA neuron regeneration [ 37 – 39 ], by transplanting neurons differentiated from neural stem cells in vitro or, even better, by promoting the neurogenic regeneration of neural stem cells in situ through their proliferation and differentiation into functional neurons [ 40 ].…”

Section: Introductionmentioning

confidence: 99%

Natriuretic peptides are neuroprotective on in vitro models of PD and promote dopaminergic differentiation of hiPSCs-derived neurons via the Wnt/β-catenin signaling

et al. 2021

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Over the last 20 years, the efforts to develop new therapies for Parkinson’s disease (PD) have focused not only on the improvement of symptomatic therapy for motor and non-motor symptoms but also on the discovering of the potential causes of PD, in order to develop disease-modifying treatments. The emerging role of dysregulation of the Wnt/β-catenin signaling in the onset and progression of PD, as well as of other neurodegenerative diseases (NDs), renders the targeting of this signaling an attractive therapeutic opportunity for curing this brain disorder. The natriuretic peptides (NPs) atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP), are cardiac and vascular-derived hormones also widely expressed in mammalian CNS, where they seem to participate in numerous brain functions including neural development/differentiation and neuroprotection. We recently demonstrated that ANP affects the Wnt/β-catenin pathway possibly through a Frizzled receptor-mediated mechanism and that it acts as a neuroprotective agent in in vitro models of PD by upregulating this signaling. Here we provide further evidence supporting the therapeutic potential of this class of natriuretic hormones. Specifically, we demonstrate that all the three natriuretic peptides are neuroprotective for SHSY5Y cells and primary cultures of DA neurons from mouse brain, subjected to neurotoxin insult with 6-hydroxydopamine (6-OHDA) for mimicking the neurodegeneration of PD, and these effects are associated with the activation of the Wnt/β-catenin pathway. Moreover, ANP, BNP, CNP are able to improve and accelerate the dopaminergic differentiation and maturation of hiPSCs-derived neural population obtained from two differed healthy donors, concomitantly affecting the canonical Wnt signaling. Our results support the relevance of exogenous ANP, BNP, and CNP as attractive molecules for both neuroprotection and neurorepair in PD, and more in general, in NDs for which aberrant Wnt signaling seems to be the leading pathogenetic mechanism.

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Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use

Cited by 138 publications

References 53 publications

Enhanced Production of Mesencephalic Dopaminergic Neurons from Lineage-Restricted Human Undifferentiated Stem Cells

Enhanced Production of Mesencephalic Dopaminergic Neurons from Lineage-Restricted Human Undifferentiated Stem Cells

Dopamine Neuron Diversity: Recent Advances and Current Challenges in Human Stem Cell Models and Single Cell Sequencing

Natriuretic peptides are neuroprotective on in vitro models of PD and promote dopaminergic differentiation of hiPSCs-derived neurons via the Wnt/β-catenin signaling

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