Targeted alveolar regeneration with Frizzled-specific agonists
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE208770 | GEO | 2023/06/16
REPOSITORIES: GEO
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Project description:The rapid regeneration of the small intestinal epithelium is sustained by crypt intestinal stem cells (ISCs). Wnt/b-catenin signaling is essential for intestinal crypt homeostasis and maintenance of Lgr5+ ISC, and yet no single or combinatorial knockout of Frizzled (FZD) genes, representing Wnt receptors, has phenocopied the severe intestinal epithelial effects of Wnt signaling blockade. The elusive identification of specific Frizzled (Fzd) receptor(s) underlying homeostatic proliferation and ISC function would greatly inform therapeutic mucosal repair strategies. In prior pharmacologic studies, bioengineered antagonists that block Wnt binding to both FZD5 and FZD8 receptors induced lethal crypt and villus loss, implicating FZD5 and/or FZD8 as essential for ISCs maintenance. Here, the potential function of Fzd5 in during intestinal homeostasis was examined by epithelial-specific Fzd5 ko, which rapidly elicited lethal pan-intestinal crypt and villus loss, and by Lgr5-specific Fzd5 cKO, which strongly reduced Lgr5+ ISC while inducing their premature differentiation. In parallel, Fzd5 cKO potently repressed Wnt target gene expression, with phenotypic rescue by constitutive activation of b-catenin in vivo and confirmation upon in vitro organoid culture. Fzd5 cKO but not Fzd8 cKO in organoids ablated responsiveness to dual specificity bioengineered FZD5/FZD8-selective Wnt surrogate agonists, which reversed DSS-induced colitis phenotypes in both wild-type and Fzd8 cKO mice. Overall, our results implicate the FZD5 receptor as an essential regulator of crypt homeostasis, Lgr5+ ISCs and intestinal response to bioengineered Wnt surrogate agonists.
2024-10-18 | GSE233260 | GEO
Project description:Frizzleds (FZD) function as receptors for secreted lipoglycoproteins of the Wingless/Int-1 (WNT) family, initiating an important signal transduction network in multicellular organisms. FZDs are classified as G protein-coupled receptors (GPCRs), which control a wide variety of physiological functions. GPCRs are well known to be regulated by phosphorylation, which can lead to specification of downstream signaling or receptor desensitization. The role and underlying mechanisms of phosphorylation of FZDs remains largely unexplored. Here, we investigated the phosphorylation of human FZD6. In order to assess FZD6 phosphorylation experimentally, we employed mass spectrometry. HEK293 cells expressing FZD6-GFP were treated with either control, WNT-3A or WNT-5A containing conditioned medium for 30 min and processed for LC-MS/MS analysis. Six phosphorylated serine residues were detected in the C-terminus of FZD6 − S592, S620, S629, S648, S653 and S656.
2018-10-18 | PXD011173 | Pride
Project description:Background and aims: Current management of inflammatory bowel disease (IBD) leaves a clear unmet need to treat the severe epithelial damage. Modulation of Wnt signaling might present an opportunity to achieve histological remission and mucosal healing when treating IBD. Exogenous R-spondin (RSPO), which amplifies Wnt signals by maintaining cell surface expression of Frizzled (FZD) and low-density lipoprotein receptor-related protein (LRP) receptors, helps repair intestine epithelial damage, but it also induces hyperplasia of normal epithelium. Wnt signaling may also be modulated with the recently developed Wnt mimetics, recombinant antibody-based molecules mimicking endogenous Wnts. Methods: We first compared the epithelial healing effects of RSPO2 and a Wnt mimetic with broad Fzd-specificity in an acute Dextran Sodium Sulfate (DSS) mouse colitis model. Guided by Fzd expression patterns in the colon epithelium, we also examined the effects of Wnt mimetics with sub-family Fzd-specificities. Results: In the DSS model, Wnt mimetics repaired damaged colon epithelium and reduced disease activity and inflammation and had no apparent effect on uninjured tissue. We further identified that the FZD5/8 and LRP6 receptor-specific Wnt mimetic, SZN-1326-p, was associated with the robust repair effect. Through a range of approaches including single-cell transcriptome analyses, we demonstrated that SZN-1326-p directly impacted epithelial cells, driving transient expansion of stem and progenitor cells, promoting differentiation of epithelial cells, histologically restoring the damaged epithelium and, secondarily to epithelial repair, reducing inflammation. Conclusion: It is feasible for treating epithelial damage in IBD to design Wnt mimetics such as SZN-1326-p that affect damaged intestine epithelium specifically and restore its physiological functions.
2022-05-19 | GSE201723 | GEO
Project description:Pulmonary inflammation compromises lung barrier function and underlies many lung diseases including acute lung injury and acute respiratory distress syndrome (ARDS). However, mechanisms by which lung cells respond to the damage caused by the inflammatory insults are not completely understood. Here we show that Fzd6-deficiency in Foxj1+ ciliated cells reduces pulmonary permeability, lipid peroxidation, and alveolar cell death accompanied with an increase in alveolar number in lungs insulted by LPS or mouse coronavirus MHV-1. Single cell RNA sequencing of lung cells indicates that the lack of Fzd6, which is primarily expressed in Foxj1+ ciliated cells, increases expression of the aldo-keto reductase Akr1b8. Intratracheal administration of the Akr1b8 protein phenocopies Fzd6-deficient lung phenotypes. In addition, ferroptosis inhibitors also phenocopy Fzd6-deficient lung phenotypes and exert no further effects in Fzd6-deficient lungs. These results indicate that Fzd6-deficiency suppresses inflammation-induced ferroptotic death of alveolar cells via the release of Akr1b8, thus revealing a previously unknown mechanism by which Fzd6 signaling regulates ferroptosis via a paracrine pathway. In addition, this study demonstrates the yet-to-be appreciated importance of ciliated cells in protecting alveolar cells during pulmonary inflammation.
2023-01-31 | GSE221068 | GEO
Project description:Pulmonary inflammation compromises lung barrier function and underlies many lung diseases including acute lung injury and acute respiratory distress syndrome (ARDS). However, mechanisms by which lung cells respond to the damage caused by the inflammatory insults are not completely understood. Here we show that Fzd6-deficiency in Foxj1+ ciliated cells reduces pulmonary permeability, lipid peroxidation, and alveolar cell death accompanied with an increase in alveolar number in lungs insulted by LPS or mouse coronavirus MHV-1. Single cell RNA sequencing of lung cells indicates that the lack of Fzd6, which is primarily expressed in Foxj1+ ciliated cells, increases expression of the aldo-keto reductase Akr1b8. Intratracheal administration of the Akr1b8 protein phenocopies Fzd6-deficient lung phenotypes. In addition, ferroptosis inhibitors also phenocopy Fzd6-deficient lung phenotypes and exert no further effects in Fzd6-deficient lungs. These results indicate that Fzd6-deficiency suppresses inflammation-induced ferroptotic death of alveolar cells via the release of Akr1b8, thus revealing a previously unknown mechanism by which Fzd6 signaling regulates ferroptosis via a paracrine pathway. In addition, this study demonstrates the yet-to-be appreciated importance of ciliated cells in protecting alveolar cells during pulmonary inflammation.
2023-01-31 | GSE222753 | GEO
Project description:Wnt/Fzd signaling has been implicated in hematopoietic stem cell maintenance and in acute leukemia establishment. We previously described a recurrent rearrangement involving the WNT10B locus (WNT10BR), characterized by expression of the WNT10BIVS1 transcript variant, in acute myeloid leukemia. To determine the occurrence of WNT10BR in T-cell acute lymphoblastic leukemia (T-ALL), we retrospectively analyzed an italian cohort of patients (n=20) and detected the WNT10BIVS1 with a high prevalence (14/20). To address genes involved in WNT10B molecular response, we designed a Wnt targeted RNA sequencing panel. We identify Wnt agonists and antagonists, from which the expression of FZD6, PLCB4, and PROM1 stand out in WNT10BIVS1-positive patients compared to negative ones. Using MOLT4 and MUTZ-2 as cell models, characterized by the expression of WNT10BIVS1, we found that WNT10B drives major Wnt activation through receipt of ligand to the FZD6 receptor complex. The WNT10B/FZD6 interaction were interfered with by short hairpin RNAs (shRNAs)-mediated gene silencing and by small molecules-mediated inhibition of WNTs secretion. We found that WNT10BIVS1 knockdown, or pharmacological interference by the LGK974 porcupine (PORCN) inhibitor, reduces WNT10B/FZD6 protein complex formation and significantly impairs intracellular effectors and leukemic expansion. These results describe the molecular circuit induced by WNT10B and suggest WNT10B/FZD6 as new targets in the T-ALL treatment strategy.
2020-10-22 | GSE159797 | GEO
Project description:<p>Pulmonary fibrosis is a heterogenous syndrome in which fibrotic scar replaces normal lung tissue. We performed massively parallel single-cell RNA-Seq on lung tissue from eight lung transplant donors and eight patients with pulmonary fibrosis. Combined with in situ RNA hybridization, with amplification, these data provide a molecular atlas of disease pathobiology. We identified a distinct, novel population of profibrotic alveolar macrophages exclusively in patients with fibrosis. Within epithelial cells, the expression of genes involved in Wnt secretion and response was restricted to non-overlapping cells. We identified rare cell populations including airway stem cells and senescent cells emerging during pulmonary fibrosis. Analysis of a cryobiopsy specimen from a patient with early disease supports the clinical application of single-cell RNA-Seq to develop personalized approaches to therapy.</p>
| phs001750 | dbGaP
Project description:Nails protect the soft tissue of the tips of digits in humans. In birds and mammals the equivalent claws are used for purposes including capturing prey, digging, climbing, fighting and maintaining dexterity and balance. The molecular mechanism of nail (and claw) development is largely unknown, but we have recently identified a Wnt receptor gene, Frizzled6 (Fzd6), as mutated in a human autosomal-recessive nail dysplasia. To investigate the action of Fzd6 in claw development at the molecular level, we compared gene expression profiles of digit tips of wild-type and Fzd6-/- mice, and show that Fzd6 regulates the transcription of a striking number of epidermal differentiation-related genes. Sixty-three genes encoding keratins, keratin associated proteins, and transglutaminases and their substrates were significantly down-regulated in the knockout mice. Among them, four hard keratins, Krt86, Krt81, Krt34 and Krt31; two epithelial keratins, Krt6a and Krt6b; and transglutaminase1 were known to be expressed in nails and involved in nail abnormality when dysregulated. Immunohistochemical studies revealed decreased expression of Krt86, Krt6b and involucrin in the epidermal portion of the claw field in the knockout embryos. We further show that Dkk4, a Wnt antagonist, was significantly down-regulated in Fzd6-/- mice along with Wnt, Bmp and Hh family genes; and Dkk4 transgenic mice showed a subtly but appreciably modified claw phenotype. Thus, Fzd6-mediated Wnt signaling likely regulates the overall differentiation process of nail/claw formation. Heterozygous Fzd6+/- mice (kindly provided by Dr. J. Nathans) were crossed to generate Fzd6+/-, Fzd6-/- and wild-type offspring. Timed matings were set up to harvest embryos at E14.5, E16.5 and E18.5. The morning after mating was designated as E0.5. Tails and the digital tips from forelimbs of three wild-type and Fzd6-/- mice from each embryonic time-point were excised, immediately frozen on dry ice, and stored at 80 C freezer until use. Genotyping was done as described previously (15). RNA was extracted using Trizol Reagent (Life technologies). RNA from dissected forelimbs from each embryo of the three time-points was used for microarray analysis.
2013-09-13 | E-GEOD-40763 | biostudies-arrayexpress
Project description:Wnt/β-catenin signaling regulates progenitor cell fate decisions during lung development and in various adult tissues. Ectopic activation of Wnt/β-catenin signaling promotes tissue repair in emphysema, a devastating lung disease with progressive loss of parenchymal lung tissue. The identity of Wnt/β-catenin responsive progenitor cells and the potential impact of Wnt/β-catenin signaling on adult distal lung epithelial progenitor cell function in emphysema, are poorly understood. Here, we used a TCF/Lef:H2B/GFP reporter mice to investigate the role of Wnt/β-catenin signaling in lung organoid formation. We identified an organoid-forming adult distal lung epithelial progenitor cell population characterized by a low Wnt/β-catenin activity, which was enriched in club and alveolar epithelial type (AT)II cells. To further characterize the lung epithelial populations with different Wnt activities, we perform microarray analysis using freshly isolated Wnthigh/low/negative lung epithelial cells to study their transcriptome, specially the enriched genes and signaling pathways in the Wnt low population related epithelial stem cell functions.
2020-06-12 | GSE150957 | GEO
Project description:Alveologenesis is the culmination of lung development and involves the correct temporal and spatial signals to generate the delicate gas exchange interface. Using a novel Wnt signaling reporter system, we have identified a Wnt-responsive alveolar epithelial sublineage arising during alveologenesis called the axin2+ alveolar type 2 cell or AT2Aaxin2. The number of AT2Aaxin2 sublineage cells increases substantially during late lung development, revealing a wave of Wnt signaling during alveologenesis. Transcriptome analysis, in vivo clonal analysis, and ex vivo lung organoid assays reveal that AT2sAaxin2s promote enhanced AT2 cellalveolar growth during generation of the alveolus compared to the overall AT2 population. Activating Wnt signaling in the AT2 lineage results in expansion of the AT2axin2 sublineageAT2s whereas inhibition of Wnt signaling inhibits AT2 cell development and shunts alveolar epithelial development towards the AT1 cell lineage. These findings reveal a novel epithelial sublineage that coordinates Wnt-dependent alveolar growthAT2 expansion required for lung alveologenesis
2016-11-30 | GSE82154 | GEO