Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells

Sun, Yubing; Yong, Koh Meng Aw; Villa‐Diaz, Luis G.; Zhang, Xiaoli; Chen, Weiqiang; Philson, Renee; Weng, Shinuo; Xu, Haoxing; Krebsbach, Paul H.; Fu, Jianping

doi:10.1038/nmat3945

Cited by 245 publications

(264 citation statements)

References 30 publications

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“…YAP acts with TEAD transcription factors to drive cell cycle progression (18,19), and YAP depletion or inhibition of YAP-TEAD interactions can promote neuronal differentiation (19,20). Fu and coworkers (21) reported that polydimethylsiloxane micropost arrays that inhibit Hippo/YAP signaling can improve neuronal differentiation of hPS cells induced by soluble neurogenic factors. We postulated that the mechanical properties of the substrate alone would be powerful enough to poise cells for neuronal differentiation.…”

mentioning

confidence: 99%

Substratum-induced differentiation of human pluripotent stem cells reveals the coactivator YAP is a potent regulator of neuronal specification

Musah¹,

Wrighton

Zaltsman

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

159

174

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Physical stimuli can act in either a synergistic or antagonistic manner to regulate cell fate decisions, but it is less clear whether insoluble signals alone can direct human pluripotent stem (hPS) cell differentiation into specialized cell types. We previously reported that stiff materials promote nuclear localization of the Yes-associated protein (YAP) transcriptional coactivator and support long-term self-renewal of hPS cells. Here, we show that even in the presence of soluble pluripotency factors, compliant substrata inhibit the nuclear localization of YAP and promote highly efficient differentiation of hPS cells into postmitotic neurons. In the absence of neurogenic factors, the effective substrata produce neurons rapidly (2 wk) and more efficiently (>75%) than conventional differentiation methods. The neurons derived from substrate induction express mature markers and possess action potentials. The hPS differentiation observed on compliant surfaces could be recapitulated on stiff surfaces by adding small-molecule inhibitors of F-actin polymerization or by depleting YAP. These studies reveal that the matrix alone can mediate differentiation of hPS cells into a mature cell type, independent of soluble inductive factors. That mechanical cues can override soluble signals suggests that their contributions to early tissue development and lineage commitment are profound.H uman pluripotent stem (hPS) cells, which include human embryonic (hES) and human induced pluripotent stem cells, possess the remarkable capacity to self-renew indefinitely and differentiate into almost any specialized cell type (1, 2). They represent a potentially unlimited supply of cells for regenerative medicine, drug screening, and studies of human development. These applications require efficient and reproducible conditions to direct hPS cell differentiation into specialized cell types, including neuronal cells. To date, the focus has been on identifying soluble factors, such as growth factors and small molecules, that can influence hPS cell differentiation. The ability of insoluble signals to promote hPS cell-lineage specification remains less clear.Studies in murine ES cells (3, 4) and tissue-specific stem cells (5-10) indicate that the adhesive and mechanical properties of the substratum used can influence cell fate decisions (11). For example, human mesenchymal stem (hMS) cells are sensitive to changes in substrate elasticity and respond by differentiating toward distinct cell lineages depending on the stiffness of the matrix (5). These hMS cells, however, tend to exist in heterogeneous cell populations and lack a specific and unique cell characterization marker (12). Their differentiation capacity is restricted to a few tissues that arise from the mesoderm lineage, such as bone, fat, and cartilage. Indeed, there are questions about whether these cells undergo transdifferentiation to cell types, such as neurons (12)(13)(14). With the unique ability to differentiate into almost any cell type, hPS cells serve as an excellent model for und...

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mentioning

confidence: 99%

Substratum-induced differentiation of human pluripotent stem cells reveals the coactivator YAP is a potent regulator of neuronal specification

Musah¹,

Wrighton

Zaltsman

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

159

174

View full text Add to dashboard Cite

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“…A similar study has been performed in Caenorhabditis elegans zygotes in which actomyosin meshwork was ablated and the initial outward velocity of adjacent cortex was proportional to cortical tension (39). In separate studies, we know cytoskeleton tension is tightly connected to the Hippo-Yes-associated protein pathway that controls cell proliferation and organ growth (40)(41)(42). With the advance in genetically encoded, ratiometric biosensors for cell signaling proteins (43), there is some exciting potential in investigating how the distribution of tensile load by optical ablation could modulate spatial signaling.…”

Section: Optical Ablation Of Intracellular Structuresmentioning

confidence: 80%

Biophysical Tools for Cellular and Subcellular Mechanical Actuation of Cell Signaling

Liu

2016

Biophysical Journal

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The ability to spatially control cell signaling can help resolve fundamental biological questions. Optogenetic and chemical dimerization techniques along with fluorescent biosensors to report cell signaling activities have enabled researchers to both visualize and perturb biochemistry in living cells. A number of approaches based on mechanical actuation using forcefield gradients have emerged as complementary technologies to manipulate cell signaling in real time. This review covers several technologies, including optical, magnetic, and acoustic control of cell signaling and behavior and highlights some studies that have led to novel insights. I will also discuss some future direction on repurposing mechanosensitive channel for mechanical actuation of spatial cell signaling.

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“…However, coculturing different types of neural tissues is still difficult because culture conditions such as medium, growth factors, and extracellular matrix (ECM) differ according to the type of neural tissues. [27][28][29][30][31][32][33] Neurons also have the characteristic of axon overproduction and, consequently, axons extend over a wide area during early development. [34][35][36] Therefore, selective connectivity between specific, spatially separated neural tissues is technically difficult.…”

Section: Rod-shaped Neural Units For Aligned 3d Neural Network Connecmentioning

confidence: 99%

“…For example, differentiation induction of motor neurons from neural stem cells requires special medium conditions. [32] The composition of the culture medium and cell density is also different in cortical cells, [28] hippocampal cells, [29] and Purkinje cells of the cerebellar. [27,31] Using the proposed assembly of rodshaped neural units releases the construction of in vitro neural connective tissue from these culture condition limitations.…”

mentioning

confidence: 99%

Rod‐Shaped Neural Units for Aligned 3D Neural Network Connection

Kato-Negishi

Onoe

Ito

et al. 2017

Adv Healthcare Materials

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COMMUNICATION (1 of 7)© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Rod-Shaped Neural Units for Aligned 3D Neural Network ConnectionMidori Kato-Negishi, Hiroaki Onoe, Akane Ito, and Shoji Takeuchi* DOI: 10.1002/adhm.201700143 guidance [12,26] have also been developed as alternative means of controlling cell patterning with high cell density and/or aligned nerve fibers. However, coculturing different types of neural tissues is still difficult because culture conditions such as medium, growth factors, and extracellular matrix (ECM) differ according to the type of neural tissues. [27][28][29][30][31][32][33] Neurons also have the characteristic of axon overproduction and, consequently, axons extend over a wide area during early development. [34][35][36] Therefore, selective connectivity between specific, spatially separated neural tissues is technically difficult.This paper proposes neural tissue units with aligned nerve fibers, called "rodshaped neural units," that can connect spatially separate neural regions (Figure 1). To fabricate these units, we employ a meterlong microfiber-shaped neural tissue [22,23] as starting material, in which stem-cell-derived or primary neural cells are encapsulated and aligned within an alginate tubular hydrogel. This tissue is cut into millimeter-sized sections and, after further cell culturing, both edges of the sections bulge out and form spherical ends (Figure 1b). The proposed rod-shaped neural units have the following characteristics: (1) nerve fibers are aligned in the longitudinal direction of the rod, (2) each unit has an insulated region covered with a thin alginate hydrogel layer to prevent the encapsulated neural tissues from connecting to surrounding cells within the regions, and (3) the edges of the unit are connectable, with the spherical ends functioning as glue to connect with other neural tissues or units. In this study, we designed and fabricated the rod-shaped neural units and characterized their connectivity in terms of neuronal morphologies and network formation. In addition, we constructed a neural network from different types of neural tissues in different culture conditions, with connections to specific, spatially neural regions. Therefore, our neural units assembly can coculture different types of neural tissues with aligned nerve fibers that recapitulate the topological complexity of in vivo neural networks with high cell density and cell population.Rod-shaped neural units were prepared by cutting microfibershaped neural tissues, as reported previously. [22,23,37,38] We used three types of neural cells to fabricate microfiber-shaped neural tissues: primary mouse neural stem cells (mNSCs) dissected from the midbrain of mice (mNSC units), primary cortical cells (cortical units), and primary hippocampal cells (hippocampal units). To prepare the rod-shaped neural units in an accurate and reproducible manner, we developed a fiber-cutting device that can fix the microfiber-shaped neural tissues in a culture In the brain, 3D neural networks such as the hippoc...

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Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells

Cited by 245 publications

References 30 publications

Substratum-induced differentiation of human pluripotent stem cells reveals the coactivator YAP is a potent regulator of neuronal specification

Substratum-induced differentiation of human pluripotent stem cells reveals the coactivator YAP is a potent regulator of neuronal specification

Biophysical Tools for Cellular and Subcellular Mechanical Actuation of Cell Signaling

Rod‐Shaped Neural Units for Aligned 3D Neural Network Connection

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