Directed induction of alveolar type I cells derived from pluripotent stem cells via Wnt signaling inhibition

Kanagaki, Shuhei; Ikeo, Satoshi; Suezawa, Takahiro; Yamamoto, Yuki; Seki, Masahide; Hirai, Toyohiro; Hagiwara, Masatoshi; Suzuki, Yutaka; Gotoh, Shimpei

doi:10.1002/stem.3302

Cited by 43 publications

(51 citation statements)

References 29 publications

(80 reference statements)

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“…These findings were consistent with a recent report by Zepp et al (2021) showing that hedgehog signaling from AT1 cells was required for the induction of secondary crest myofibroblasts (SCMFs), which are described as highly contractile fibroblasts that are present during lung development. Because of the presence of AT1-like cells in our organoids (Kanagaki et al, 2021a), we assessed whether SCMF lineage markers varied depending on the presence or absence of epithelial cells. The SCMF low-expression markers, WNT2 and TCF21, were decreased while the SCMF high-expression marker, STC1, was upregulated in the presence of epithelial cells (Figure S2D).…”

Section: Resultsmentioning

confidence: 99%

Disease modeling of pulmonary fibrosis using human pluripotent stem cell-derived alveolar organoids

et al. 2021

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Summary Although alveolar epithelial cells play a critical role in the pathogenesis of pulmonary fibrosis, few practical in vitro models exist to study them. Here, we established a novel in vitro pulmonary fibrosis model using alveolar organoids consisting of human pluripotent stem cell-derived alveolar epithelial cells and primary human lung fibroblasts. In this human model, bleomycin treatment induced phenotypes such as epithelial cell-mediated fibroblast activation, cellular senescence, and presence of alveolar epithelial cells in abnormal differentiation states. Chemical screening performed to target these abnormalities showed that inhibition of ALK5 or blocking of integrin αVβ6 ameliorated the fibrogenic changes in the alveolar organoids. Furthermore, organoid contraction and extracellular matrix accumulation in the model recapitulated the pathological changes observed in pulmonary fibrosis. This human model may therefore accelerate the development of highly effective therapeutic agents for otherwise incurable pulmonary fibrosis by targeting alveolar epithelial cells and epithelial-mesenchymal interactions.

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Section: Resultsmentioning

confidence: 99%

Disease modeling of pulmonary fibrosis using human pluripotent stem cell-derived alveolar organoids

et al. 2021

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“…Taking advantage of our in vitro drug screening platform of self-renewable hNPCs, we also asked whether CHIR or XAV might be cytotoxic, by measuring cell proliferation and survival. Both drugs were used in this study at the same concentration or lower as in similar studies employing human or murine ESCs, hiPSCs and other types of stem cells ( De Kumar et al, 2017 ; Malleske et al, 2018 ; Shafa et al, 2018 ; Gomez et al, 2019 ; Hamad et al, 2019 ; Qiu et al, 2019 ; Shin et al, 2019 ; Almasoud et al, 2020 ; Bataille et al, 2020 ; Bejoy et al, 2020 ; Govarthanan et al, 2020 ; Han et al, 2020 ; Kanagaki et al, 2020 ; Leigh et al, 2020 ; Yang J. et al, 2020 ; Yang Y. et al, 2020 ; Ren et al, 2021 ; Wang et al, 2021 ). When stem cells differentiate into more mature cell types, their proliferation potential is gradually reduced, until cells reach a post-mitotic fate.…”

Section: Discussionmentioning

confidence: 99%

“…CHIR has been used to promote neural and neuronal differentiation in embryonic stem cells and induced pluripotent stem cells ( De Kumar et al, 2017 ; Shafa et al, 2018 ; Gomez et al, 2019 ; Qiu et al, 2019 ; Shin et al, 2019 ; Bejoy et al, 2020 ) and in several different types of neural precursors ( Yang Y. et al, 2020 ; Ren et al, 2021 ; Wang et al, 2021 ), as well as to induce trans- differentiation into neural lineages from non-neural cell types, such as skin ( Bataille et al, 2020 ; Yang J. et al, 2020 ) and mesenchymal cells ( Govarthanan et al, 2020 ). XAV has been used to promote differentiation of pluripotent stem cells into lung cells ( Malleske et al, 2018 ; Kanagaki et al, 2020 ), to increase osteogenic differentiation of mesenchymal stem cells ( Almasoud et al, 2020 ; Han et al, 2020 ), and to promote generation of cardiomyocytes ( Hamad et al, 2019 ; Leigh et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Pharmacological Manipulation of Wnt/β-Catenin Signaling Pathway in Human Neural Precursor Cells Alters Their Differentiation Potential and Neuronal Yield

Telias

Ben‐Yosef

2021

Front. Mol. Neurosci.

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The canonical Wnt/β-catenin pathway is a master-regulator of cell fate during embryonic and adult neurogenesis and is therefore a major pharmacological target in basic and clinical research. Chemical manipulation of Wnt signaling during in vitro neuronal differentiation of stem cells can alter both the quantity and the quality of the derived neurons. Accordingly, the use of Wnt activators and blockers has become an integral part of differentiation protocols applied to stem cells in recent years. Here, we investigated the effects of the glycogen synthase kinase-3β inhibitor CHIR99021, which upregulates β-catenin agonizing Wnt; and the tankyrase-1/2 inhibitor XAV939, which downregulates β-catenin antagonizing Wnt. Both drugs and their potential neurogenic and anti-neurogenic effects were studied using stable lines human neural precursor cells (hNPCs), derived from embryonic stem cells, which can be induced to generate mature neurons by chemically-defined conditions. We found that Wnt-agonism by CHIR99021 promotes induction of neural differentiation, while also reducing cell proliferation and survival. This effect was not synergistic with those of pro-neural growth factors during long-term neuronal differentiation. Conversely, antagonism of Wnt by XAV939 consistently prevented neuronal progression of hNPCs. We show here how these two drugs can be used to manipulate cell fate and how self-renewing hNPCs can be used as reliable human in vitro drug-screening platforms.

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“…Active WNT signaling is a feature of alveolar progenitors in homeostasis and regeneration, and blocking the WNT pathway by Ctnnb1 knockout in AT2 cells was shown to induce AT1 differentiation at postnatal stages ( Frank et al, 2016 ; Nabhan et al, 2018 ). In addition, an expansion of the AT1 cell population was observed in vitro after treatment of hiPSC-derived AT2 cells with XAV-939 ( Kanagaki et al, 2021 ). Differentiation of cultured adult human alveolar progenitors (TM4SF1 + ) also showed WNT responsiveness, with the AT1 fate induced after XAV-939 treatment.…”

Section: Resultsmentioning

confidence: 99%

Differentiation of mouse fetal lung alveolar progenitors in serum-free organotypic cultures

Gkatzis

Panza

Peruzzo

et al. 2021

eLife

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Lung epithelial progenitors differentiate into alveolar type 1 (AT1) and type 2 (AT2) cells. These cells form the air-blood interface and secrete surfactant, respectively, and are essential for lung maturation and function. Current protocols to derive and culture alveolar cells do not faithfully recapitulate the architecture of the distal lung, which influences cell fate patterns in vivo. Here, we report serum-free conditions that allow for growth and differentiation of mouse distal lung epithelial progenitors. We find that Collagen I promotes the differentiation of flattened, polarized AT1 cells. Using these organoids, we performed a chemical screen to investigate WNT signaling in epithelial differentiation. We identify an association between Casein Kinase activity and maintenance of an AT2 expression signature; Casein Kinase inhibition leads to an increase in AT1/progenitor cell ratio. These organoids provide a simplified model of alveolar differentiation and constitute a scalable screening platform to identify and analyze cell differentiation mechanisms.

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Directed induction of alveolar type I cells derived from pluripotent stem cells via Wnt signaling inhibition

Cited by 43 publications

References 29 publications

Disease modeling of pulmonary fibrosis using human pluripotent stem cell-derived alveolar organoids

Disease modeling of pulmonary fibrosis using human pluripotent stem cell-derived alveolar organoids

Pharmacological Manipulation of Wnt/β-Catenin Signaling Pathway in Human Neural Precursor Cells Alters Their Differentiation Potential and Neuronal Yield

Differentiation of mouse fetal lung alveolar progenitors in serum-free organotypic cultures

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