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Biology subjects

Taketo, M. M.

Publications and source records attributed to Taketo, M. M..

11 recordsLinked to original sources

Sharp cell type boundaries emerge from coordinated morphogen signaling

Classic models of the French flag problem depict sharp cell-type boundaries emerging from threshold responses to morphogen gradients. How discrete cell-type boundaries arise from morphogen signals that vary continuously across developing tissues is not completely understood. We use hair follicle dermal condensate formation to study a sharp developmental transition in which proliferative progenitors undergo cell-cycle exit concurrent with molecular differentiation. Using genetic and genomic approaches, we show that Wnt and Hedgehog signaling interact to coordinate the timing of these two processes. We identify a division of labor between the pathways: Wnt signaling promotes cell-cycle exit by regulating chromatin binding of the Hedgehog mediator GLI3, while Hedgehog signaling induces differentiation genes in a Wnt-dependent manner and simultaneously elevates Wnt activity. When Wnt and Hedgehog activities are temporally aligned, differentiation and cell-cycle exit occur within the same developmental window, restricting both the duration and abundance of intermediate states and producing a sharp cell-type boundary. When these signals are misaligned, intermediate states persist and expand, producing fuzzy boundaries. These findings reveal a mechanism in which interacting morphogen signals regulate the duration and abundance of intermediate states during a developmental transition, thereby controlling how continuous cell-state progression is translated into discrete tissue patterning.

developmental biology↗

Tumors accumulate expanded GATA3-dependent tissue Tregs

Targeting Tregs is a potential strategy to improve cancer therapies. However, which Tregs accumulate in response to tumoral processes, and how tumors affect their phenotype, is poorly understood. Here we show that tumor Tregs are equivalent to effector tissue Tregs in steady state organs. We used a mouse model of intestinal neoplasia to demonstrate that one early event in carcinogenesis is sufficient to induce local accumulation of Tregs resembling human tumor Tregs. Treg accumulation was driven by TCR-dependent oligoclonal expansion of tissue Tregs with an effector Treg phenotype. Treg expansion was independent of CCR8, IL33R and CD137, which were previously linked to tumor Treg. In contrast, GATA3 was required for effector tissue Tregs and for their expansion in response to neoplasia. Our findings identify GATA3-dependent clonal expansion of effector tissue Tregs as a key event in promoting tumor growth. HighlightsO_LIAn early tumorigenic event alone drives accumulation of effector tissue Tregs C_LIO_LITregs in tumors are phenotypically akin to effector tissue Tregs C_LIO_LIThe accumulation of Tregs is driven by TCR-dependent oligoclonal expansion C_LIO_LIGATA3 controls tumor-promoting effector tissue Tregs C_LI

immunology↗

Canonical Wnt signaling controls the fate and plasticity of NG2 glia in the healthy and ischemic adult mouse cortex

NG2 glia, also known as oligodendrocyte precursor cells (OPCs), exhibit unexpected plasticity in the adult brain after injury, yet the molecular cues guiding their fate remain poorly defined. Wnt signaling contributes to tissue responses following ischemic stroke, but its precise role in post-injury glial remodeling is not fully understood. Here, we define how Wnt/{beta}-catenin signaling shapes NG2 glial behavior after focal cerebral ischemia. Using genetic mouse models enabling cell-specific Wnt pathway activation or inhibition, combined with single-cell RNA sequencing, immunohistochemistry, and electrophysiological recording, we disclosed a central role for Wnt signaling in post-injury fate specification. We identified 12 transcriptionally distinct subpopulations within the oligodendroglial lineage, including a subset with an astrocyte-like transcriptional profile. Wnt signaling strongly influenced the balance between OPC proliferation and differentiation: pathway hyperactivation impaired oligodendrocyte maturation and expanded astrocyte-like NG2-derived cells, likely through concomitant activation of the Notch pathway. Remarkably, Wnt hyperactivation also induced the appearance of cholinergic neuron-like cells derived from NG2-expressing cells exclusively in the somatosensory cortex; these cells generated action potentials and exhibited sodium conductance characteristic of functional neurons. Together, these findings demonstrate that NG2 glia undergo distinct fate transitions after stroke and that their lineage plasticity is highly sensitive to Wnt pathway dynamics. Targeted fine-tuning of Wnt/{beta}-catenin signaling may enable directed redirection of NG2 glia toward specific reparative outcomes, including neuronal reprogramming, in the injured adult brain. TEASERWnt activation promotes neuronal conversion of NG2 glia, while impairing oligodendrocyte maturation.

neuroscience↗

Interaction of Wnt and SHH gradients synchronizes cell cycle 1 exit and differentiation

Classic models of the French flag problem depict sharp cell-type boundaries emerging from threshold responses to morphogen gradients. Yet, how such boundaries arise during dynamic cell-state transitions remains unclear. We use hair follicle dermal condensates to study a sharp cell-type transition in which proliferative progenitors synchronously undergo cell-cycle exit and molecular differentiation. Using genetic and genomic approaches, we show that Wnt and Hedgehog signaling interact to coordinate the timing of these processes. When their activities are temporally aligned, intermediate transitional states are compressed through cell-cycle exit, producing a sharp cell-type boundary; when misaligned, transitional states expand, yielding fuzzy borders. Mechanistically, elevated Wnt activity promotes cell-cycle exit by regulating chromatin binding of the Hedgehog mediator GLI3. Hedgehog signaling induces differentiation genes in a Wnt-dependent manner and simultaneously elevates Wnt activity, aligning arrest and differentiation in time. These findings reveal a mechanism wherein morphogen interactions coordinate transitions to generate precise cell-type boundaries.

developmental biology↗

Stabilization of epithelial β-catenin compromises mammary cell fate acquisition and branching morphogenesis

The Wnt/{beta}-catenin pathway plays a critical role in cell fate specification, morphogenesis, and stem cell activation across diverse tissues, including the skin. In mammals, the embryonic surface epithelium gives rise to the epidermis, as well as the associated appendages including hair follicles and mammary glands, both of which depend on epithelial Wnt/{beta}-catenin activity for initiation of their development. Later on, Wnts are thought to enhance mammary gland growth and branching while in hair follicles, they are essential for hair shaft formation. Here we report a strong downregulation of epithelial Wnt/{beta}-catenin activity as the mammary bud progresses to branching. We show that forced activation of epithelial {beta}-catenin severely compromises embryonic mammary gland branching. However, the phenotype of conditional Lef1 deficient embryos implies that a low level of Wnt/{beta}-catenin activity is necessary for mammary cell survival. Transcriptomic profiling suggests that sustained high {beta}-catenin activity leads to maintenance of mammary bud gene signature at the expense of outgrowth/branching signature. In addition, it leads to upregulation of epidermal differentiation genes. Strikingly, we find a partial switch to hair follicle fate early on upon stabilization of {beta}-catenin suggesting that the level of epithelial Wnt/{beta}-catenin signaling activity may contribute to the choice between skin appendage identities.

developmental biology↗

Targeted engagement of β-catenin-Ikaros complexes in refractory B-cell malignancies

In most cell types, nuclear {beta}-catenin functions as prominent oncogenic driver and pairs with TCF7-family factors for transcriptional activation of MYC. Surprisingly, B-lymphoid malignancies not only lacked expression and activating lesions of {beta}-catenin but critically depended on GSK3{beta} for effective {beta}-catenin degradation. Our interactome studies in B-lymphoid tumors revealed that {beta}-catenin formed repressive complexes with lymphoid-specific Ikaros factors at the expense of TCF7. Instead of MYC-activation, {beta}-catenin was essential to enable Ikaros-mediated recruitment of nucleosome remodeling and deacetylation (NuRD) complexes for transcriptional repression of MYC. To leverage this previously unrecognized vulnerability of B-cell-specific repressive {beta}-catenin-Ikaros-complexes in refractory B-cell malignancies, we examined GSK3{beta} small molecule inhibitors to subvert {beta}-catenin degradation. Clinically approved GSK3{beta}-inhibitors that achieved favorable safety prof les at micromolar concentrations in clinical trials for neurological disorders and solid tumors were effective at low nanomolar concentrations in B-cell malignancies, induced massive accumulation of {beta}-catenin, repression of MYC and acute cell death. Preclinical in vivo treatment experiments in patient-derived xenografts validated small molecule GSK3{beta}-inhibitors for targeted engagement of lymphoid-specific {beta}-catenin-Ikaros complexes as a novel strategy to overcome conventional mechanisms of drug-resistance in refractory malignancies. HIGHLIGHTSO_LIUnlike other cell lineages, B-cells express nuclear {beta}-catenin protein at low baseline levels and depend on GSK3{beta} for its degradation. C_LIO_LIIn B-cells, {beta}-catenin forms unique complexes with lymphoid-specific Ikaros factors and is required for Ikaros-mediated tumor suppression and assembly of repressive NuRD complexes. C_LIO_LICRISPR-based knockin mutation of a single Ikaros-binding motif in a lymphoid MYC superenhancer region reversed {beta}-catenin-dependent Myc repression and induction of cell death. C_LIO_LIThe discovery of GSK3{beta}-dependent degradation of {beta}-catenin as unique B-lymphoid vulnerability provides a rationale to repurpose clinically approved GSK3{beta}-inhibitors for the treatment of refractory B-cell malignancies. C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/532152v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@1b70e4dorg.highwire.dtl.DTLVardef@10bfcc8org.highwire.dtl.DTLVardef@b10eorg.highwire.dtl.DTLVardef@1683a3e_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LIAbundant nuclear {beta}-catenin C_LIO_LI{beta}-catenin pairs with TCF7 factors for transcriptional activation of MYC C_LIO_LIB-cells rely on efficient degradation of {beta}-catenin by GSK3{beta} C_LIO_LIB-cell-specific expression of Ikaros factors C_LI Unique vulnerability in B-cell tumors: O_LIGSK3{beta}-inhibitors induce nuclear accumulation of {beta}-catenin. C_LIO_LI{beta}-catenin pairs with B-cell-specific Ikaros factors for transcriptional repression of MYC C_LI

cancer biology↗

Accumulation of the GSK3 target protein β-catenin is lethal for B cell precursors and malignant B cells

Glycogen synthase kinase 3 (GSK3) is a ubiquitously expressed kinase involved in a myriad of biological processes. Although GSK3 mediated phosphorylation has been shown to induce the degradation of many pro-survival and pro-proliferation factors, cancer cells of different origin show reduced proliferation or survival after GSK3 inhibition. Our current understanding of the role GSK3 plays in normal mature B cells, B cell precursors and transformed B cells is incomplete and does not allow to assess whether GSK3 inhibitors can be used to treat B cell derived malignancies. Here we identify {beta}-catenin as the major factor driving GSK3-inhibition induced changes in B cells. We show that {beta}-catenin accumulation has opposing effects on cell metabolism and survival in mature B cells and B cell precursors. Moreover, we demonstrate that {beta}-catenin destabilizes the commitment to the B cell lineage. In summary, our study identifies {beta}-catenin induced signaling as a factor that can be exploited to limit the survival of malignant B cells.

immunology↗

Fine-tuning of β-catenin in thymic epithelial cells is required for postnatal T cell development

In the thymus, the thymic epithelium provides a microenvironment essential for the development of functionally competent and self-tolerant T cells. Previous findings showed that modulation of Wnt/{beta}-catenin signaling in thymic epithelial cells (TECs) disrupts embryonic thymus organogenesis. However, the role of {beta}-catenin in TECs for postnatal T cell development remains to be elucidated. Here, we analyzed gain-of-function (GOF) and loss-of-function (LOF) of {beta}-catenin highly specific in TECs. We found that GOF of {beta}-catenin in TECs results in severe thymic dysplasia and T cell deficiency beginning from the embryonic period. By contrast, LOF of {beta}-catenin in TECs reduces the number of cortical TECs and thymocytes modestly and only postnatally. These results indicate that fine-tuning of {beta}-catenin expression within a permissive range is required for TECs to generate an optimal microenvironment to support postnatal T cell development.

immunology↗

Canonical Wnt signaling exerts bidirectional control on choroid plexus epithelial development

The choroid plexus (ChP) secretes cerebrospinal fluid and is critical for the development and function of the brain. In the telencephalon, the ChP epithelium (ChPe) arises from the Wnt-expressing cortical hem. Embryonic mouse and human ChPe both express nuclear {beta}-CATENIN, a canonical Wnt signaling pathway effector, indicating that this pathway is active during ChPe development. Point mutations in human {beta}-CATENIN result in the constitutive activation of canonical Wnt signaling. In a mouse model that recapitulates this perturbation, we report a loss of ChPe identity and an apparent transformation of the ChPe to a neuronal identity. Aspects of this phenomenon are recapitulated in human embryonic stem cell (hESC)-derived organoids. The ChPe is also disrupted when {beta}-Catenin is conditionally inactivated in the mouse. Together, our results indicate that canonical Wnt signaling is required in a precise and regulated manner for normal ChPe development in the mammalian brain.

developmental biology↗

APC mutant cells exploit compensatory chromosome alterations to restore tumour cell fitness

Cancer cells tolerate copy number alterations (CNAs) of genomic regions that are lethal to non-cancer cells. Certain CNAs are preferentially associated with specific cancer types and lineages, but the mechanisms underlying the emergence and selection of specific CNAs remain unclear. Adenomatous polyposis coli (APC) mutations induce mitotic errors, but their impact on tumor evolution remains elusive. We investigated APC function in cultured cells and tumors and found that its loss led to {beta}-catenin accumulation at centrosomes, suppressing its maturation through inhibition of key centrosome regulators, including Aurora kinase A (AURKA) that promotes tumor growth. These defects collectively reduced cellular fitness, leading to impaired mitotic fidelity and delayed cell cycle progression. However, in APC-mutant tumors, AURKA activity was maintained, at least in part, through the amplification of chromosomes harboring AURKA and its activator genes, yet this alone was insufficient to fully restore proliferation: aberrant chromosomal reorganization also emerged and contributed to the adaptive fitness of APC-mutant cells. Such a process of adaptive CNA selection provides a framework for understanding how specific CNAs are selected to counteract disadvantages imposed by genetic alterations during tumor progression, providing one key insight into how specific CNAs are selected in this context.

cell biology↗

Canonical Wnt signaling regulates patterning, differentiation and nucleogenesis in mouse hypothalamus and prethalamus.

The hypothalamus is a small, but anatomically and functionally complex, region of the brain whose development is poorly understood. In this study, we have explored its development by studying the canonical Wntsignalling pathway, generating gain and loss of function mutations of betacaten in(Ctnnb1) in both hypothalamic and prethalamic neuroepithelium. Deletion of Ctnnb1 resulted in an anteriorized and hypoplastic hypothalamus. Posterior structures were lost or reduced, and anterior structures were expanded. In contrast, over expression of a constitutively active mutant form of Ctnnb1 resulted in severe hyperplasia of prethalamus and hypothalamus, and expanded expression of a subset of posterior and premamillary hypothalamic markers. Moderate defects in differentiation of Arx-positive GABAergic neural precursors were observed in both prethalamus and hypothalamus of Ctnnb1 loss of function mutants, while in gain of function mutants, their differentiation was completely suppressed, although markers of prethalamic progenitors were preserved. Multiple other region-specific markers, including several specific posterior hypothalamic structures, were also suppressed in Ctnnb1 gain of function mutations. Severe, region-specific defects in hypothalamic nucleogenesis were also observed in both gain and loss of function mutations of Ctnnb1. Finally, both gain and loss of function of Ctnnb1 also produced severe, cell nonautonomous disruptions of pituitary development. These findings demonstrate acentral and multifaceted role for canonical Wnt signalling in regulating growth, patterning, differentiation and nucleogenesis in multiple diencephalic regions.\n\nHighlightsO_LICanonical Wnt signalling regulates anteroposterior patterning in the hypothalamus.\nC_LIO_LICanonical Wnt signalling regulates differentiation of GABAergic neurons in both prethalamus and hypothalamus.\nC_LIO_LICanonical Wnt signalling regulates differentiation and nucleogenesis of multiple hypothalamic neuronal subtypes.\nC_LIO_LICanonical Wnt signalling in hypothalamic neuroepithelium regulates pituitary morphogenesis and differentiation.\nC_LI

developmental biology↗