Unique photoaffinity probes to study TGFβ signaling and receptor fates

2019 ◽  
Vol 55 (30) ◽  
pp. 4323-4326 ◽  
Author(s):  
D. Längle ◽  
F. Wesseler ◽  
D. Flötgen ◽  
H. Leek ◽  
A. T. Plowright ◽  
...  
Keyword(s):  
Synthetic Approach ◽  
Tgfβ Signaling ◽  
Photoaffinity Probes ◽  
Tgfβ Receptor ◽  
Photoaffinity Labels

A novel synthetic approach is used to prepare a diverse set of “first-in-class” dihydropyridine-based TGFβ receptor degraders bearing photoaffinity labels.

scholarly journals Skeletal Deformities in Osterix-Cre;Tgfbr2f/f Mice May Cause Postnatal Death

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 975
Author(s):  
Kara Corps ◽  
Monica Stanwick ◽  
Juliann Rectenwald ◽  
Andrew Kruggel ◽  
Sarah B. Peters
Keyword(s):  
Transforming Growth Factor ◽  
Skeletal Development ◽  
Transforming Growth Factor Β ◽  
Tgfβ Signaling ◽  
Skeletal Deformities ◽  
Tooth Abnormalities ◽  
Pathological Analysis ◽  
Diaphragmatic Muscle ◽  
Proliferation And Differentiation ◽  
Tgfβ Receptor

Transforming growth factor β (TGFβ) signaling plays an important role in skeletal development. We previously demonstrated that the loss of TGFβ receptor II (Tgfbr2) in Osterix-Cre-expressing mesenchyme results in defects in bones and teeth due to reduced proliferation and differentiation in pre-osteoblasts and pre-odontoblasts. These Osterix-Cre;Tgfbr2f/f mice typically die within approximately four weeks for unknown reasons. To investigate the cause of death, we performed extensive pathological analysis on Osterix-Cre- (Cre-), Osterix-Cre+;Tgfbr2f/wt (HET), and Osterix-Cre+;Tgfbr2f/f (CKO) mice. We also crossed Osterix-Cre mice with the ROSA26mTmG reporter line to identify potential off-target Cre expression. The findings recapitulated published skeletal and tooth abnormalities and revealed previously unreported osteochondral dysplasia throughout both the appendicular and axial skeletons in the CKO mice, including the calvaria. Alterations to the nasal area and teeth suggest a potentially reduced capacity to sense and process food, while off-target Cre expression in the gastrointestinal tract may indicate an inability to absorb nutrients. Additionally, altered nasal passages and unexplained changes in diaphragmatic muscle support the possibility of hypoxia. We conclude that these mice likely died due to a combination of breathing difficulties, malnutrition, and starvation resulting primarily from skeletal deformities that decreased their ability to sense, gather, and process food.

scholarly journals Cholesterol deprivation induces TGFβ signaling to promote basal differentiation in pancreatic cancer

2019 ◽  
Author(s):  
Linara G. Cornell ◽  
Suraj Peri ◽  
Diana Restifo ◽  
Alena Klochkova ◽  
Tiffiney R. Hartman ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cholesterol Metabolism ◽  
Cholesterol Biosynthesis ◽  
Ductal Adenocarcinoma ◽  
Tgfβ Signaling ◽  
Pancreatic Development ◽  
Mesenchymal Phenotype ◽  
Tgfβ Receptor ◽  
Growth Factor Beta

SummaryOncogenic transformation alters the metabolism of cellular nutrients to sustain tumor growth. We here define a mechanism by which modifications in cholesterol metabolism control the formation of pancreatic ductal adenocarcinoma (PDAC). Disruption of distal cholesterol biosynthesis by means of conditional inactivation ofNsdhlin mice bearing a tumor-inducingKrasmutation (KrasG12D) prevented PDAC formation in the context of a heterozygousTrp53f/+genotype without impairing normal pancreatic development. In mice with pancreaticNsdhlablation and homozygous loss ofTrp53, the emerging tumors presented with the aggressive basal (mesenchymal) phenotype as opposed to the classic (glandular) PDAC. This paralleled significantly reduced expression of cholesterol metabolic pathway genes in human basal PDAC subtype. Mechanistically, we demonstrate that genetic or metabolic cholesterol deprivation stabilizes the transforming growth factor beta (TGFβ) receptor to activate pro-mesenchymal effectors in human and murine PDAC, providing a direct mechanism by which cholesterol metabolism can condition tumor differentiation.

Abstract 547: TGFß Response Coupled to Stress Signaling by TGRL Lipolysis in Vascular Endothelial Cells

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Hnin Aung ◽  
Larissa Eiselein ◽  
Michael W Lamé ◽  
Kit Ng ◽  
Dennis W Wilson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Transcription Factor ◽  
Endothelial Cell ◽  
Map Kinases ◽  
Gene Array ◽  
Receptor Activation ◽  
Type I ◽  
Tgfβ Signaling ◽  
Tgfβ Receptor ◽  
Activating Transcription Factor

Elevation of circulating triglyceride-rich lipoproteins (TGRL) in the postprandial state is associated with increased endothelial cell inflammation and dysfunction potentially contributing to atherosclerosis. Previous studies from our laboratory showed that lipolysis products of TGRL induced endothelial cell barrier permeability, up-regulation of cytokines and adhesion molecules, and apoptosis. Gene array studies implicated the Stress Associated Protein Kinase and c-Jun N-terminal kinase (SAPK/JNK) pathway and identified the transcription factor activating transcription factor 3 (ATF3) as highly induced in this process. Additional studies demonstrated activation of second messengers for transforming growth factor beta (TGFβ) signaling (SMAD2/SMAD4) in response to lipolysis product treatment. In the present study, we asked whether TGFβ signaling was implicated in the induction of apoptosis by lipolysis products and whether there was a linkage between TGFβ receptor activation and pro-inflammatory responses. siRNA knockdown of ATF3 transcription abrogated both IL-8 and E-selectin up-regulation in lipolysis product-treated human aortic endothelial cells (HAEC). In addition, ATF3 knockdown prevented activation of caspase 3/7 and induction of p53, markers of apoptosis. Inhibition of the TGFβ Type I activin receptor-like kinase (ALK) receptors (ALKs 4, 5 and 7) by SB 431542 (10 μM) also inhibited inflammatory up-regulation, ATF3 induction and apoptosis. Both ATF3 knockdown and TGFβ receptor blockade prevented c-Jun phosphorylation, a co-activating transcription factor with ATF3 of AP-1 transcription sites. Our findings implicate TGFβ signaling in the inflammatory and apoptotic responses to TGRL lipolysis and suggest a complex interaction between TGFβ receptors and the SAPK subset of MAP Kinases.

scholarly journals Restoration of Transforming Growth Factor-β Receptor II Expression in Colon Cancer Cells with Microsatellite Instability Increases Metastatic Potential in Vivo

2011 ◽  
Vol 286 (18) ◽  
pp. 16082-16090 ◽  
Author(s):  
Xiao-Qiong Liu ◽  
Ashwani Rajput ◽  
Liying Geng ◽  
Melanie Ongchin ◽  
Anathbandhu Chaudhuri ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Growth Factor ◽  
Microsatellite Instability ◽  
Cell Survival ◽  
Cancer Cells ◽  
Colon Cancer Cells ◽  
Tgfβ Signaling ◽  
Tgfβ Receptor

Microsatellite instability (MSI), which occurs in 15% of colorectal cancer, has been shown to have a lower incidence of metastasis and better patient survival rates compared with microsatellite stable colorectal cancer. However, a mechanistic understanding of the basis for this difference is very limited. Here, we show that restoration of TGFβ signaling by re-expression of TGFβ receptor II in MSI colon cancer cells increased PI3K/AKT activation, conferred resistance to growth factor deprivation stress-induced apoptosis, and promoted cell motility in vitro. Treatment with a potent PI3K inhibitor (LY294002) blocked the prosurvival and promotility effects of TGFβ, indicating that TGFβ-mediated promotion of cell survival and motility is dependent upon activation of the PI3K/AKT pathway. Analysis of apoptotic effectors that are affected by TGFβ signaling indicated that Bim is an effector of TGFβ-mediated survival. In addition, TGFβ-induced down-regulation of E-cadherin contributed to the prosurvival effect of TGFβ, and restoration of TGFβ signaling in MSI colon cancer cells increased liver metastasis in an orthotopic model in vivo. Taken together, our results demonstrate that restoration of TGFβ signaling promotes cell survival, motility, and metastatic progression in MSI colon cancer cells and indicate that TGFβ receptor II mutations contribute to the favorable outcomes in colon cancer patients with MSI.

scholarly journals SCIDOT-27. TGFβ SIGNALING-INDUCED miRNA PARTICIPATES IN AUTOPHAGIC REGULATION BY TARGETING PRAS40 IN MESENCHYMAL SUBTYPE OF GLIOBLASTOMA

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi277-vi277
Author(s):  
Yingbin Xie ◽  
Luyue Chen ◽  
Junhu Zhou ◽  
Qixue Wang ◽  
Yunfei Wang ◽  
...  
Keyword(s):  
Cell Fate ◽  
Therapeutic Interventions ◽  
Functional Verification ◽  
Tgfβ Signaling ◽  
Signature Genes ◽  
Dismal Prognosis ◽  
Tgfβ Receptor ◽  
Transcriptomic Signature

Abstract Mesenchymal subtype of glioblastoma (mesGBM) is a refractory disease condition characterized by therapeutic failure and tumor recurrence, and for the past decade, no treatment option has improved the dismal prognosis of this disease. In Cancer Genome Atlas (TCGA) GBM datasets, two of the 840 transcriptomic signature genes for GBM subtyping, TGFβ1 and TGFβ receptor type II (TβRII), were exclusively upregulated in mesGBM, indicating hyperactive TGFβ signaling. In the present study, dysregulated microRNAs (miRNAs) were identified after LY2109761 (a TβRI/II inhibitor) treatment in a mesGBM-derived cell line. Importantly, several novel miRNAs that may be missing pieces of the puzzle of TGFβ signaling-mediated pathogenesis in mesGBM were identified. Novel_miR26/56/93/97/119 were validated by quantitative PCR to correlate with TGFβ signaling activity, and novel_miR56 was selected as a promising candidate for further functional verification. A proline-rich akt substrate of 40 kDa (PRAS40), which regulates autophagy initiation by targeting mTORC1, was confirmed to be a functional target of novel_miR56. Autophagy plays dichotomous roles in the determination of GBM cell fate, and in this study, TGFβ signaling-induced novel_miR56 seemed to inhibit autophagic activity in GBM and its mesenchymal subtype, resulting in the maintenance of proliferative capability. The pro-survival effect of novel_miR56 can be impeded by intentionally downregulating its expression both in vivo and in vitro. In summary, we provide novel insight into TGFβ signaling-mediated pathogenesis and targets for the development of novel therapeutic interventions for mesGBM.

scholarly journals Fibrillin-1 regulates the bioavailability of TGFβ1

2007 ◽  
Vol 176 (3) ◽  
pp. 355-367 ◽  
Author(s):  
Shazia S. Chaudhry ◽  
Stuart A. Cain ◽  
Amanda Morgan ◽  
Sarah L. Dallas ◽  
C. Adrian Shuttleworth ◽  
...  
Keyword(s):  
Transforming Growth Factor ◽  
Solid Phase ◽  
Binding Studies ◽  
Tgfβ Signaling ◽  
Intact Cells ◽  
Altered Expression ◽  
Tgfβ Receptor ◽  
Fibrillin 1 ◽  
Cell Layers ◽  
Tgf Β1

We have discovered that fibrillin-1, which forms extracellular microfibrils, can regulate the bioavailability of transforming growth factor (TGF) β1, a powerful cytokine that modulates cell survival and phenotype. Altered TGFβ signaling is a major contributor to the pathology of Marfan syndrome (MFS) and related diseases. In the presence of cell layer extracellular matrix, a fibrillin-1 sequence encoded by exons 44–49 releases endogenous TGFβ1, thereby stimulating TGFβ receptor–mediated Smad2 signaling. This altered TGFβ1 bioavailability does not require intact cells, proteolysis, or the altered expression of TGFβ1 or its receptors. Mass spectrometry revealed that a fibrillin-1 fragment containing the TGFβ1-releasing sequence specifically associates with full-length fibrillin-1 in cell layers. Solid-phase and BIAcore binding studies showed that this fragment interacts strongly and specifically with N-terminal fibrillin-1, thereby inhibiting the association of C-terminal latent TGFβ-binding protein 1 (a component of the large latent complex [LLC]) with N-terminal fibrillin-1. By releasing LLC from microfibrils, the fibrillin-1 sequence encoded by exons 44–49 can contribute to MFS and related diseases.

Hsp90/Cdc37 assembly modulates TGFβ receptor-II to act as a profibrotic regulator of TGFβ signaling during cardiac hypertrophy

2015 ◽  
Vol 27 (12) ◽  
pp. 2410-2424 ◽  
Author(s):  
Ritwik Datta ◽  
Trisha Bansal ◽  
Santanu Rana ◽  
Kaberi Datta ◽  
Shiladitya Chattopadhyay ◽  
...  
Keyword(s):  
Cardiac Hypertrophy ◽  
Tgfβ Signaling ◽  
Tgfβ Receptor

scholarly journals A Key Cytoskeletal Regulator of Ubiquitination Amplifies TGFβ Signaling During Mouse Developmental Vascular Patterning

2016 ◽  
Author(s):  
Ronak Shetty ◽  
Divyesh Joshi ◽  
Mamta Jain ◽  
Madavan Vasudevan ◽  
Jasper Chrysolite Paul ◽  
...  
Keyword(s):  
Nuclear Translocation ◽  
De Novo ◽  
Yolk Sac ◽  
Dual Function ◽  
Tgfβ Signaling ◽  
Vascular Patterning ◽  
Cytoskeletal Remodeling ◽  
Tumour Metastasis ◽  
Microtubule Stability ◽  
Tgfβ Receptor

AbstractVascular development involves de novo formation of a capillary plexus, which is then pruned and remodeled by angiogenic events. Cytoskeletal remodeling and directional endothelial migration are essential for developmental and pathological angiogenesis. Smad-dependent TGFβ signaling controls vascular patterning and is negatively regulated by microtubules. Here we show that a positive regulator of TGFβ signaling is essential for developmental vascular patterning and microtubule stability. Rudhira/BCAS3 is known to bind microtubules and to play a nodal role in cytoskeletal remodeling and directional endothelial cell (EC) migration in vitro. We demonstrate that the molecular and cellular function of Rudhira is deployed at critical steps in vascular patterning. We generated the first floxed mice for rudhira and find that global or endothelial knockout of rudhira results in mid-gestation lethality due to aberrant embryonic and extra-embryonic vessel patterning and defective cardiac morphogenesis. Rudhira null yolk sac ECs show random and retarded migration. Yolk sac transcriptome analysis revealed key mediators of angiogenic processes and TGFβ receptor signaling were perturbed in rudhira null mutants. Molecular and biochemical analyses showed that rudhira depletion reduced microtubule stability but increased expression of pathway inhibitors leading to high levels of SMAD2/3 ubiquitination and reduced activation. These effects were not rescued by exogenous TGFβ. However, TGFβ treatment of wild type ECs increased Rudhira expression. Further, exogenous Rudhira, which promotes directional cell migration, caused increased SMAD2/3 nuclear translocation and reduced inhibitor levels. Therefore, we propose that Rudhira and TGFβ signaling are mutually dependent. Rudhira has a dual function in promoting TGFβ signaling, possibly by sequestering microtubules and simultaneously preventing SMAD2/3 ubiquitination to permit EC migration and vascular patterning. TGFβ signaling and aberrant human Rudhira (Breast Cancer Amplified Sequence 3, BCAS3) expression are both associated with tumour metastasis. Our study identifies a cytoskeletal, cell type-specific modulator of TGFβ signaling important in development and cancer.

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