Experimentally Induced Epithelial–Mesenchymal Transition of Human Hair as a Model of Scarring Ex Vivo

2020 ◽  
pp. 143-152
Author(s):  
Ilaria Piccini ◽  
Marta Bertolini
Keyword(s):  
Human Hair ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Mesenchymal Transition ◽  
Experimentally Induced

scholarly journals Mechanophenotyping of 3D multicellular clusters using displacement arrays of rendered tractions

2020 ◽  
Vol 117 (11) ◽  
pp. 5655-5663 ◽  
Author(s):  
Susan E. Leggett ◽  
Mohak Patel ◽  
Thomas M. Valentin ◽  
Lena Gamboa ◽  
Amanda S. Khoo ◽  
...  
Keyword(s):  
Tumor Invasion ◽  
Wound Repair ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Human Patient ◽  
Mesenchymal Transition ◽  
Successive Loss ◽  
Surrounding Extracellular Matrix

Epithelial tissues mechanically deform the surrounding extracellular matrix during embryonic development, wound repair, and tumor invasion. Ex vivo measurements of such multicellular tractions within three-dimensional (3D) biomaterials could elucidate collective dissemination during disease progression and enable preclinical testing of targeted antimigration therapies. However, past 3D traction measurements have been low throughput due to the challenges of imaging and analyzing information-rich 3D material deformations. Here, we demonstrate a method to profile multicellular clusters in a 96-well-plate format based on spatially heterogeneous contractile, protrusive, and circumferential tractions. As a case study, we profile multicellular clusters across varying states of the epithelial–mesenchymal transition, revealing a successive loss of protrusive and circumferential tractions, as well as the formation of localized contractile tractions with elongated cluster morphologies. These cluster phenotypes were biochemically perturbed by using drugs, biasing toward traction signatures of different epithelial or mesenchymal states. This higher-throughput analysis is promising to systematically interrogate and perturb aberrant mechanobiology, which could be utilized with human-patient samples to guide personalized therapies.

scholarly journals Role of Epithelial–Mesenchymal Transition in Repair of the Lacrimal Gland after Experimentally Induced Injury

2012 ◽  
Vol 53 (1) ◽  
pp. 126 ◽  
Author(s):  
Samantha You ◽  
Orna Avidan ◽  
Ayesha Tariq ◽  
Ivy Ahluwalia ◽  
Paul C. Stark ◽  
...  
Keyword(s):  
Lacrimal Gland ◽  
Epithelial Mesenchymal Transition ◽  
Mesenchymal Transition ◽  
Experimentally Induced

scholarly journals Explanting Is anEx VivoModel of Renal Epithelial-Mesenchymal Transition

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Catherine E. Winbanks ◽  
Ian A. Darby ◽  
Kristen J. Kelynack ◽  
Dodie Pouniotis ◽  
Gavin J. Becker ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
De Novo ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Rat Kidney ◽  
Smooth Muscle Actin ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Mesenchymal Transition ◽  
Renal Tubulointerstitial Fibrosis

Recognised by theirde novoexpression of alpha-smooth muscle actin (SMA), recruitment of myofibroblasts is key to the pathogenesis of fibrosis in chronic kidney disease. Increasingly, we realise that epithelial-mesenchymal transition (EMT) may be an important source of these cells. In this study we describe a novel model of renal EMT. Rat kidney explants were finely diced on gelatin-coated Petri dishes and cultured in serum-supplemented media. Morphology and immunocytochemistry were used to identify mesenchymal (vimentin+, α-smooth muscle actin (SMA)+, desmin+), epithelial (cytokeratin+), and endothelial (RECA+) cells at various time points. Cell outgrowths were all epithelial in origin (cytokeratin+) at day 3. By day 10, 50 ± 12% (mean ± SE) of cytokeratin+ cells double-labelled for SMA, indicating EMT. Lectin staining established a proximal tubule origin. By day 17, cultures consisted only of myofibroblasts (SMA+/cytokeratin−). Explanting is a reproducibleex vivomodel of EMT. The ability to modify this change in phenotype provides a useful tool to study the regulation and mechanisms of renal tubulointerstitial fibrosis.

scholarly journals Transcription Factor AP4 Mediates Cell Fate Decisions: To Divide, Age, or Die

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 676
Author(s):  
Matthew Man-Kin Wong ◽  
Sancy Mary Joyson ◽  
Heiko Hermeking ◽  
Sung Kay Chiu
Keyword(s):  
Transcription Factor ◽  
Cell Fate ◽  
Cell Cycle Progression ◽  
Leucine Zipper ◽  
Target Genes ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Mesenchymal Transition ◽  
Multiple Tumor ◽  
Cell Fate Decisions

Activating Enhancer-Binding Protein 4 (AP4)/transcription factor AP4 (TFAP4) is a basic-helix-loop-helix-leucine-zipper transcription factor that was first identified as a protein bound to SV40 promoters more than 30 years ago. Almost 15 years later, AP4 was characterized as a target of the c-Myc transcription factor, which is the product of a prototypic oncogene that is activated in the majority of tumors. Interestingly, AP4 seems to represent a central hub downstream of c-Myc and N-Myc that mediates some of their functions, such as proliferation and epithelial-mesenchymal transition (EMT). Elevated AP4 expression is associated with progression of cancer and poor patient prognosis in multiple tumor types. Deletion of AP4 in mice points to roles of AP4 in the control of stemness, tumor initiation and adaptive immunity. Interestingly, ex vivo AP4 inactivation results in increased DNA damage, senescence, and apoptosis, which may be caused by defective cell cycle progression. Here, we will summarize the roles of AP4 as a transcriptional repressor and activator of target genes and the contribution of protein and non-coding RNAs encoded by these genes, in regulating the above mentioned processes. In addition, proteins interacting with or regulating AP4 and the cellular signaling pathways altered after AP4 dysregulation in tumor cells will be discussed.

scholarly journals Cooperativity between stromal cytokines drives the invasive migration of human breast cancer cells

2019 ◽  
Vol 374 (1779) ◽  
pp. 20180231 ◽  
Author(s):  
Yair Elisha ◽  
Yael Sagi ◽  
Georg Klein ◽  
Ravid Straussman ◽  
Benjamin Geiger
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cross Talk ◽  
Breast Cancer Cells ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Cancer Invasion ◽  
Stromal Fibroblast ◽  
Mesenchymal Transition ◽  
Interdisciplinary Approaches

The cross-talk between cancer cells and the stromal microenvironment plays a key role in regulating cancer invasion. Here, we employed an ex vivo invasion model system for exploring the regulation of breast cancer cells infiltration into a variety of stromal fibroblast monolayers. Our results revealed considerable variability in the stromal induction of invasiveness, with some lines promoting and others blocking invasion. It was shown that conditioned medium (CM), derived from invasion-promoting fibroblasts, can induce epithelial–mesenchymal transition-like process in the cancer cells, and trigger their infiltration into a monolayer of invasion-blocking fibroblasts. To identify the specific invasion-promoting molecules, we analysed the cytokines in stimulatory CM, screened a library of purified cytokines for invasion-promoting activity and tested the effect of specific inhibitors of selected cytokine receptors on the CM-induced invasion. Taken together, these experiments indicated that the invasiveness of BT-474 is induced by the combined action of IL1 and IL6 and that IL1 can induce IL6 secretion by invasion-blocking fibroblasts, thereby triggering cancer cell invasion into the stroma. This unexpected observation suggests that stromal regulation of cancer invasion may involve not only cross-talk between stromal and cancer cells, but also cooperation between different stromal subpopulations. This article is part of a discussion meeting issue ‘Forces in cancer: interdisciplinary approaches in tumour mechanobiology’.

scholarly journals Epithelial-type systemic breast carcinoma cells with a restricted mesenchymal transition are a major source of metastasis

2019 ◽  
Vol 5 (6) ◽  
pp. eaav4275 ◽  
Author(s):  
Xiao Liu ◽  
Junjian Li ◽  
Bruno Loureiro Cadilha ◽  
Anamarija Markota ◽  
Cornelia Voigt ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Lung Metastases ◽  
Ex Vivo ◽  
Surrogate Marker ◽  
Metastatic Breast ◽  
Distant Metastases ◽  
Carcinoma Cells ◽  
Clinical Samples ◽  
Mesenchymal Transition ◽  
Syngeneic Mouse Model

Carcinoma cells undergo epithelial-mesenchymal transition (EMT); however, contributions of EMT heterogeneity to disease progression remain a matter of debate. Here, we addressed the EMT status of ex vivo cultured circulating and disseminated tumor cells (CTCs/DTCs) in a syngeneic mouse model of metastatic breast cancer (MBC). Epithelial-type CTCs with a restricted mesenchymal transition had the strongest lung metastases formation ability, whereas mesenchymal-type CTCs showed limited metastatic ability. EpCAM expression served as a surrogate marker to evaluate the EMT heterogeneity of clinical samples from MBC, including metastases, CTCs, and DTCs. The proportion of epithelial-type CTCs, and especially DTCs, correlated with distant metastases and poorer outcome of patients with MBC. This study fosters our understanding of EMT in metastasis and underpins heterogeneous EMT phenotypes as important parameters for tumor prognosis and treatment. We further suggest that EpCAM-dependent CTC isolation systems will underestimate CTC numbers but will quantify clinically relevant metastatic cells.

scholarly journals Epithelial-Mesenchymal Transition in Atopy: A Mini-Review

2020 ◽  
Vol 1 ◽  
Author(s):  
Erik D. Anderson ◽  
Mohammadali E. Alishahedani ◽  
Ian A. Myles
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Topical Steroid ◽  
Atopic Diseases ◽  
Barrier Dysfunction ◽  
Mesenchymal Phenotype ◽  
Mesenchymal Transition ◽  
Common Pathogenesis

Atopic diseases, particularly atopic dermatitis (AD), asthma, and allergic rhinitis (AR) share a common pathogenesis of inflammation and barrier dysfunction. Epithelial to mesenchymal transition (EMT) is a process where epithelial cells take on a migratory mesenchymal phenotype and is essential for normal tissue repair and signal through multiple inflammatory pathways. However, while links between EMT and both asthma and AR have been demonstrated, as we outline in this mini-review, the literature investigating AD and EMT is far less well-elucidated. Furthermore, current studies on EMT and atopy are mostly animal models or ex vivo studies on cell cultures or tissue biopsies. The literature covered in this mini-review on EMT-related barrier dysfunction as a contributor to AD as well as the related (perhaps resultant) atopic diseases indicates a potential for therapeutic targeting and carry treatment implications for topical steroid use and environmental exposure assessments. Further research, particularly in vivo studies, may greatly advance the field and translate into benefit for patients and families.

scholarly journals Tracking a TGF-β activator in vivo: sensitive PET imaging of αvβ8-integrin with the Ga-68-labeled cyclic RGD octapeptide trimer Ga-68-Triveoctin

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Neil Gerard Quigley ◽  
Katja Steiger ◽  
Frauke Richter ◽  
Wilko Weichert ◽  
Sebastian Hoberück ◽  
...  
Keyword(s):  
Human Subject ◽  
Transforming Growth Factor ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Transforming Growth Factor Β ◽  
Integrin Expression ◽  
Target Tissue ◽  
Mesenchymal Transition ◽  
Positron Emission

Abstract Purpose As a major activator of transforming growth factor β (TGF-β), the RGD receptor αvβ8-integrin is involved in pathogenic processes related to TGF-β dysregulation, such as tumor growth, invasion, and radiochemoresistance, metastasis and tumor cell stemness, as well as epithelial-mesenchymal transition. The novel positron emission tomography (PET) radiopharmaceutical Ga-68-Triveoctin for in vivo mapping of αvβ8-integrin expression might enhance the prognosis of certain tumor entities, as well as support and augment TGF-β-targeted therapeutic approaches. Methods Monomeric and trimeric conjugates of cyclo(GLRGDLp(NMe)K(pent-4-ynoic amide)) were synthesized by click chemistry (CuAAC), labeled with Ga-68, and evaluated in MeWo (human melanoma) xenografted SCID mice by means of PET and ex-vivo biodistribution. αvβ8-integrin expression in murine tissues was determined by β8-IHC. A human subject received a single injection of 173 MBq of Ga-68-Triveoctin and underwent 3 subsequent PET/CT scans at 25, 45, and 90 min p.i.. Results The trimer Ga-68-Triveoctin exhibits a 6.7-fold higher αvβ8-integrin affinity than the monomer (IC50 of 5.7 vs. 38 nM, respectively). Accordingly, biodistribution showed a higher tumor uptake (1.9 vs. 1.0%IA/g, respectively) but a similar baseline upon blockade (0.25%IA/g for both). IHC showed an intermediate β8-expression in the tumor while most organs and tissues were found β8-negative. Low non-target tissue uptakes (< 0.4%IA/g) confirmed a low degree of unspecific binding. Due to its hydrophilicity (log D = − 3.1), Ga-68-Triveoctin is excreted renally and shows favorable tumor/tissue ratios in mice (t/blood: 6.7; t/liver: 6.8; t/muscle: 29). A high kidney uptake in mice (kidney-to-blood and -to-muscle ratios of 126 and 505, respectively) is not reflected by human PET (corresponding values are 15 and 30, respectively), which furthermore showed notable uptakes in coeliac and choroid plexus (SUVmean 6.1 and 9.7, respectively, 90 min p.i.). Conclusion Ga-68-Triveoctin enables sensitive in-vivo imaging αvβ8-integrin expression in murine tumor xenografts. PET in a human subject confirmed a favorable biodistribution, underscoring the potential of Ga-68-Triveoctin for mapping of αvβ8-integrin expression in a clinical setting.

scholarly journals Mechanophenotyping of 3D Multicellular Clusters using Displacement Arrays of Rendered Tractions

2019 ◽  
Author(s):  
Susan E. Leggett ◽  
Mohak Patel ◽  
Thomas M. Valentin ◽  
Lena Gamboa ◽  
Amanda S. Khoo ◽  
...  
Keyword(s):  
Tumor Invasion ◽  
Wound Repair ◽  
Epithelial Mesenchymal Transition ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Human Patient ◽  
Mesenchymal Transition ◽  
Successive Loss ◽  
Surrounding Extracellular Matrix

ABSTRACTEpithelial tissues mechanically deform the surrounding extracellular matrix during embryonic development, wound repair, and tumor invasion. Ex vivo measurements of such multicellular tractions within three-dimensional (3D) biomaterials could elucidate collective dissemination during disease progression, and enable preclinical testing of targeted anti-migration therapies. However, past 3D traction measurements have been low throughput due to the challenges of imaging and analyzing information-rich 3D material deformations. Here, we demonstrate a method to profile multicellular clusters in a 96-well plate format based on spatially heterogeneous contractile, protrusive, and circumferential tractions. As a case study, we profile multicellular clusters across varying states of the epithelial-mesenchymal transition, revealing a successive loss of protrusive and circumferential tractions, as well as the formation of localized contractile tractions with elongated cluster morphologies. These cluster phenotypes were biochemically perturbed using drugs, biasing towards traction signatures of different epithelial or mesenchymal states. This higher-throughput analysis is promising to systematically interrogate and perturb aberrant mechanobiology, which could be utilized with human patient samples to guide personalized therapies.

Sign in / Sign up

Export Citation Format

Share Document