Loss of Eph-receptor expression correlates with loss of cell adhesion and chondrogenic capacity in Hoxa13 mutant limbs

Development ◽  
2001 ◽  
Vol 128 (21) ◽  
pp. 4177-4188 ◽  
Author(s):  
H. Scott Stadler ◽  
Kay M. Higgins ◽  
Mario R. Capecchi
Keyword(s):  
Cell Adhesion ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Receptor Expression ◽  
Loss Of Function ◽  
Vitro Analysis ◽  
Missing Digit ◽  
Umbilical Arteries

Mesenchymal patterning is an active process whereby genetic commands coordinate cell adhesion, sorting and condensation, and thereby direct the formation of morphological structures. In mice that lack the Hoxa13 gene, the mesenchymal condensations that form the autopod skeletal elements are poorly resolved, resulting in missing digit, carpal and tarsal elements. In addition, mesenchymal and endothelial cell layers of the umbilical arteries (UAs) are disorganized, resulting in their stenosis and in embryonic death. To further investigate the role of Hoxa13 in these phenotypes, we generated a loss-of-function allele in which the GFP gene was targeted into the Hoxa13 locus. This allele allowed FACS isolation of mesenchymal cells from Hoxa13 heterozygous and mutant homozygous limb buds. Hoxa13GFP expressing mesenchymal cells from Hoxa13 mutant homozygous embryos are defective in forming chondrogenic condensations in vitro. Analysis of pro-adhesion molecules in the autopod of Hoxa13 mutants revealed a marked reduction in EphA7 expression in affected digits, as well as in micromass cell cultures prepared from mutant mesenchymal cells. Finally, antibody blocking of the EphA7 extracellular domain severely inhibits the capacity of Hoxa13GFP heterozygous cells to condense and form chondrogenic nodules in vitro, which is consistent with the hypothesis that reduction in EphA7 expression affects the capacity of Hoxa13–/– mesenchymal cells to form chondrogenic condensations in vivo and in vitro. EphA7 and EphA4 expression were also decreased in the mesenchymal and endothelial cells that form the umbilical arteries in Hoxa13 mutant homozygous embryos. These results suggest that an important role for Hoxa13 during limb and UA development is to regulate genes whose products are required for mesenchymal cell adhesion, sorting and boundary formation.

scholarly journals Characterization in mice of the stromal niche maintaining AT2 stem cell self-renewal in homeostasis and disease

2021 ◽  
Author(s):  
Sara Taghizadeh ◽  
Monika Heiner ◽  
Jochen Wilhelm ◽  
Susane Herold ◽  
Chengshui Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
The Self ◽  
Loss Of Function ◽  
Self Renewal ◽  
Alveolar Epithelial

AbstractResident mesenchymal cells (rMCs defined as Cd31NegCd45NegEpcamNeg) control the self-renewal and differentiation of alveolar epithelial type 2 (AT2) stem cells in vitro. The identity of these rMCs is still elusive. Among them, Axin2Pos mesenchymal alveolar niche cells (MANCs), which are expressing Fgf7, have been previously described. We propose that an additional population of rMCs, expressing Fgf10 (called rMC-Sca1PosFgf10Pos) are equally important to maintain AT2 stem cell self-renewal.The alveolosphere model, based on the AT2-rMC co-culture in growth factor reduced Matrigel, was used to test the efficiency of different rMC subpopulations isolated by FACS from adult murine lung to sustain the self-renewal and differentiation of AT2 stem cells.We demonstrate that rMC-Sca1PosFgf10Pos cells are efficient to promote the self-renewal and differentiation of AT2 stem cells. Co-staining of adult lung for Fgf10 mRNA and Sftpc protein respectively, indicate that 28% of Fgf10Pos cells are located close to AT2 cells. Co-ISH for Fgf7 and Fgf10 indicate that these two populations do not significantly overlap. Gene arrays comparing rMC-Sca1PosAxin2Pos and rMC-Sca1PosFgf10Pos support that these two cell subsets express differential markers. In addition, rMC function is decreased in diabetic and obese ob/ob mutant compared to WT mice with a much stronger loss of function in males compared to females.In conclusion, rMC-Sca1PosFgf10Pos cells play important role in supporting AT2 stem cells self-renewal and differentiation. This result sheds a new light on the subpopulations of rMCs contributing to the AT2 stem cell niche in homeostasis and in the context of COVID-19 pathogenesis.Key messageWhat is already known about the subject?Resident mesenchymal cells (rMCs defined as Cd31NegCd45NegEpcamNeg) control the self-renewal and differentiation of alveolar epithelial type 2 (AT2) stem cells in vitro. The identity of these rMCs is still elusive. Among them, Axin2Pos mesenchymal alveolar niche cells (MANCs), which are expressing Fgf7, have been previously described.What does this study add?Our study shows that an additional population of rMCs, expressing Fgf10 (called rMC-Sca1PosFgf10Pos) is equally important to maintain AT2 stem cell self-renewal. rMC-Sca1PosFgf10Pos are LipidToxHigh and are located close to AT2s. In addition, rMC-Sca1PosFgf10Pos cells support AT2 stem cell self-renewal and differentiation thereby identifying these cells as bone fide functional lipofibroblasts (LIFs). We have previously reported that LIF can transdifferentiate into activated MYF in the context of bleomycin-induced fibrosis in mice [1] and that activated MYF isolated from the lungs of end stage idiopathic fibrosis human patients can respond to Metformin to undergo transdifferentiation back to the LIF phenotype [2]. We also show that the function of rMCs-Sca1Pos is negatively impacted by gender and obesity, which represent two major aggravating factors for COVID-19 pathogenesis, leading to either death or major complications after infection recovery such as lung fibrosis.How might this impact on clinical practice and future development?By establishing that rMC-Sca1PosFgf10Pos are different from the MANCs, our study opens the way for a new key mesenchymal cell population that should be targeted to either prevent or reverse fibrosis. In addition, as this population maintains the AT2 stem cells self-renewal and differentiation, such targeting will also allow to progressively recover the loss in respiratory function.

Cell adhesion and movement in relation to the developing limb pattern in normal and talpid3 mutant chick embryos

Development ◽  
1968 ◽  
Vol 20 (1) ◽  
pp. 81-100
Author(s):  
D. A. Ede ◽  
G. S. Agerbak
Keyword(s):  
Cell Adhesion ◽  
Quantitative Methods ◽  
Cell Movement ◽  
Genetic Effect ◽  
Mesenchymal Cell ◽  
Cellular Level ◽  
Lethal Mutant ◽  
Wing Bud

Descriptive studies of the talpid3 chick embryonic lethal mutant (ta3/ta3) have suggested that the multiple effects produced by this gene are of mesodermal origin, and that they arise from defective mesenchymal cell movement and condensation (Ede & Kelly, 1964 a, b). It may be argued that condensation in vivo is comparable to cell reaggregation of dissociated cells in vitro, and that defects in the former are likely to be reflected in the latter. In this case it should be possible to obtain experimental verification of this effect of the gene at the cellular level, using the quantitative methods for assessing aggregation developed by Moscona (1961 a, b) and Curtis & Greaves (1965). The experiments reported here show a clear genetic effect upon cell adhesion in the wing-bud mesenchyme of the talpid3 mutant. The wing-bud was chosen because it was hoped to establish a connexion between the effect of the gene at the cellular level and its dramatic effect on limb morphogenesis.

Nonhematopoietic/endothelial SSEA-1+ cells define the most primitive progenitors in the adult murine bone marrow mesenchymal compartment

Blood ◽  
2006 ◽  
Vol 109 (3) ◽  
pp. 1298-1306 ◽  
Author(s):  
Fernando Anjos-Afonso ◽  
Dominique Bonnet
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Cell ◽  
Cell Types ◽  
Mesenchymal Cells ◽  
Hierarchical Organization ◽  
Mesenchymal Progenitor Cells ◽  
Murine Bone Marrow

Abstract It is believed that a primitive cell type that maintains the mesenchymal compartment exists in the bone marrow. However, this putative mesenchymal stem/progenitor cell is yet to be identified and isolated. We are reporting the identification, isolation, and detailed characterization of the most primitive mesenchymal progenitor cells in the adult murine bone marrow, based on the expression of stage-specific embryonic antigen–1 (SSEA-1). This primitive subset can be identified in mesenchymal cell cultures and also directly in the bone marrow, thus ascertaining for the first time their existence in an adult organism. Characterization of SSEA-1+ mesenchymal cells revealed that upon purification these cells gave rise to SSEA-1− mesenchymal cells, whereas the reverse could not be observed. Also, these SSEA-1+ cells have a much higher capacity to differentiate than their negative counterparts, not only to several mesenchymal cell types but also to unconventional cell types such as astrocyte-, endothelial-, and hepatocyte-like cells in vitro. Most importantly, a single-cell–derived population was capable of differentiating abundantly into different mesenchymal cell types in vivo. Altogether we are proposing a hierarchical organization of the mesenchymal compartment, placing SSEA-1+ cells at the apex of this hierarchy.

scholarly journals Induction of PDGF receptor-α in rat myofibroblasts during pulmonary fibrogenesis in vivo

1998 ◽  
Vol 274 (1) ◽  
pp. L72-L80 ◽  
Author(s):  
James C. Bonner ◽  
Pamela M. Lindroos ◽  
Annette B. Rice ◽  
Cindy R. Moomaw ◽  
Daniel L. Morgan
Keyword(s):  
Interleukin 1 ◽  
Immunohistochemical Analysis ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Platelet Derived Growth Factor ◽  
Pdgf Receptor ◽  
Collagen Deposition ◽  
Cell Hyperplasia

Platelet-derived growth factor (PDGF) is a potent mitogen for mesenchymal cells. Induction of the PDGF receptor-α (PDGF-Rα) in vitro enhances PDGF-induced mitogenesis and chemotaxis. Thus we investigated whether the PDGF-Rα is induced in vivo during pulmonary fibrogenesis using a vanadium pentoxide (V2O5) model of lung injury. PDGF-Rα mRNA expression was induced 24 h postinstillation. PDGF-Rβ mRNA was constitutively expressed and did not increase. Western blotting showed upregulation of PDGF-Rα protein by 48 h, and immunohistochemical analysis localized PDGF-Rα primarily in mesenchymal cells residing within fibrotic lesions. Upregulation of PDGF-Rα in vivo preceded mesenchymal cell hyperplasia (3–7 days) and collagen deposition by day 15. Supernatants from alveolar macrophages treated with V2O5in vitro released upregulatory activity for PDGF-Rα on cultured lung myofibroblasts, and this activity was blocked by the interleukin-1-receptor antagonist. These data suggest that interleukin-1β-mediated induction of PDGF-Rα in vivo is important to lung myofibroblast hyperplasia during fibrogenesis.

scholarly journals Loss of miR-24-3p promotes epithelial cell apoptosis and impairs the recovery from intestinal inflammation

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Artin Soroosh ◽  
Kai Fang ◽  
Jill M. Hoffman ◽  
Ivy K. M. Law ◽  
Elizabeth Videlock ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Apoptosis ◽  
Intestinal Inflammation ◽  
Loss Of Function ◽  
Epithelial Cell Apoptosis ◽  
Vitro Analysis ◽  
Double Blinded ◽  
Active Inflammation

AbstractWhile apoptosis plays a significant role in intestinal homeostasis, it can also be pathogenic if overactive during recovery from inflammation. We recently reported that microRNA-24-3p (miR-24-3p) is elevated in the colonic epithelium of ulcerative colitis patients during active inflammation, and that it reduced apoptosis in vitro. However, its function during intestinal restitution following inflammation had not been examined. In this study, we tested the influence of miR-24-3p on mucosal repair by studying recovery from colitis in both novel miR-24-3p knockout and miR-24-3p-inhibited mice. We observed that knockout mice and mice treated with a miR-24-3p inhibitor had significantly worsened recovery based on weight loss, colon length, and double-blinded histological scoring. In vivo and in vitro analysis of miR-24-3p inhibition in colonic epithelial cells revealed that inhibition promotes apoptosis and increases levels of the pro-apoptotic protein BIM. Further experiments determined that silencing of BIM reversed the pro-apoptotic effects of miR-24-3p inhibition. Taken together, these data suggest that miR-24-3p restrains intestinal epithelial cell apoptosis by targeting BIM, and its loss of function is detrimental to epithelial restitution following intestinal inflammation.

scholarly journals Increased Regeneration Following Stress-Induced Lung Injury in Bleomycin-Treated Chimeric Mice with CD44 Knockout Mesenchymal Cells

Cells ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 1211
Author(s):  
Dmytro Petukhov ◽  
Mark Richter-Dayan ◽  
Zvi Fridlender ◽  
Raphael Breuer ◽  
Shulamit B. Wallach-Dayan
Keyword(s):  
Lung Injury ◽  
Cell Motility ◽  
Tissue Regeneration ◽  
Nuclear Translocation ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Alveolar Epithelial Cells ◽  
P53 Signaling

CD44, an adhesion-molecule promoting cell-migration, is shown here to increase in stress conditions following bleomycin-induced apoptosis in alveolar epithelial cells (AECs), a main target of lung injury. In vivo, it inhibits tissue regeneration and leads to fibrosis. We show that some AECs survive by the ataxia-telangiectasia mutated kinase/ATM pathway, and undergo a CD44-mediated epithelial-mesenchymal transdifferentiation (EMT) with migratory capacities in vitro, and in vivo. We assessed apoptosis vs. proliferation of AECs following bleomycin, ATM/P53 signaling pathway in AECs, and CD44 involvement in EMT, cell motility and tissue regeneration in vitro and in vivo. Expression of survival genes, CD44, and ATM/p53 pathway was elevated in AECs surviving bleomycin injury, as were the markers of EMT (downregulation of E-cadherin, upregulation of N-cadherin and vimentin, nuclear translocation of β-catenin). Inhibition of CD44 decreased AECs transdifferentiation. Bleomycin-treated chimeric CD44KO-mice had decreased EMT markers, ATM, and mesenchymal cells (α-SMA+) accumulation in lung, increased surfactant-b, diminished lung mesenchymal cell motility, and increased lung tissue regenerative capacity following bleomycin injury, as indicated by lung collagen content and semiquantitave morphological index scoring. Thus, AECs surviving lung injury are plastic and undergo ATM-mediated, CD44-dependent transdifferentiation, preventing tissue regeneration and promoting fibrosis. Synthetic or natural compounds that downregulate CD44 may improve tissue regeneration following injury.

Characterization of a Bone Marrow Derived Mesenchymal Cell Line from Nude Rat Assembling Tumor Stem/Progenitor Cell Properties In Vitro and In Vivo.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4256-4256
Author(s):  
Mengyu Wang ◽  
Jingfang Ju ◽  
Stein Saeboe-Larssen ◽  
Svein-Ole Mikalsen ◽  
Elena Olsen ◽  
...  
Keyword(s):  
Cell Line ◽  
Connexin 43 ◽  
Transforming Growth Factor ◽  
Abdominal Cavity ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Nude Rat ◽  
Biological Studies

Abstract Here we report the in vitro and in vivo properties of a spontaneously transformed nude rat mesenchymal stem cells (rTMSCs) following culture passages 23. When this cell line was compared with early passages of mesenchymal cells (rMSCs) from the same animal, no apparent differences could be seen with regard to the phenotype and Gap junctions’ connexin 43. However, the telomerase activity in rTMSCs is lower than in the rMSCs. Karyotyping of rTMSC gave a trisome 6 phenotype. Microarry analysis of rTMSC shows over expression of the transforming growth factor family including TGF-beta and BMp6. Both rMSC and rTMSC had equal capability to differentiate into adipocytic and “neural like” linage. In contrast rTMSC was unable to differentiate into bone and only this cell line form spheroids in cultures. Subcutaneously injection of rTMSC in doses down to 2x 104 into nude/nude mice formed tumor in all animals tested. Examination of the tumor assembles morphology like an immature sarcoma. When 100–200 SP + cells were administered to the animals, tumor could also be formed. After intravenous injections and direct injections into left heart ventricle all animals developed metastasis in the lung; abdominal cavity; bone and skin. The rTMSC has been stabled marked with reporter gene GFP and Leucipherase. Currently the proliferation and migration capacity of the linage-differentiated rTMSC is studied in vivo by imaging and these results will be presented. We conclude that our normal and transformed rMSCs could be potential useful for further biological studies on mesenchymal cells both in vitro and in vivo.

Role of the 807 C/T Polymorphism of the α2 Gene in Platelet GP Ia Collagen Receptor Expression and Function

1999 ◽  
Vol 81 (06) ◽  
pp. 951-956 ◽  
Author(s):  
J. Corral ◽  
R. González-Conejero ◽  
J. Rivera ◽  
F. Ortuño ◽  
P. Aparicio ◽  
...  
Keyword(s):  
Venous Thrombosis ◽  
Cell Types ◽  
Receptor Expression ◽  
Case Control Studies ◽  
Platelet Surface ◽  
Vitro Analysis ◽  
And Function ◽  
In Vitro Analysis

SummaryThe variability of the platelet GP Ia/IIa density has been associated with the 807 C/T polymorphism (Phe 224) of the GP Ia gene in American Caucasian population. We have investigated the genotype and allelic frequencies of this polymorphism in Spanish Caucasians. The T allele was found in 35% of the 284 blood donors analyzed. We confirmed in 159 healthy subjects a significant association between the 807 C/T polymorphism and the platelet GP Ia density. The T allele correlated with high number of GP Ia molecules on platelet surface. In addition, we observed a similar association of this polymorphism with the expression of this protein in other blood cell types. The platelet responsiveness to collagen was determined by “in vitro” analysis of the platelet activation and aggregation response. We found no significant differences in these functional platelet parameters according to the 807 C/T genotype. Finally, results from 3 case/control studies involving 302 consecutive patients (101 with coronary heart disease, 104 with cerebrovascular disease and 97 with deep venous thrombosis) determined that the 807 C/T polymorphism of the GP Ia gene does not represent a risk factor for arterial or venous thrombosis.

scholarly journals Functional and Transcriptional Adaptations of Blood Monocytes Recruited to the Cystic Fibrosis Airway Microenvironment In Vitro

2021 ◽  
Vol 22 (5) ◽  
pp. 2530
Author(s):  
Bijean D. Ford ◽  
Diego Moncada Giraldo ◽  
Camilla Margaroli ◽  
Vincent D. Giacalone ◽  
Milton R. Brown ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Bactericidal Activity ◽  
Scavenger Receptor ◽  
Receptor Expression ◽  
Blood Monocytes ◽  
Bacterial Killing ◽  
Blood Neutrophils ◽  
Vitro Model

Cystic fibrosis (CF) lung disease is dominated by the recruitment of myeloid cells (neutrophils and monocytes) from the blood which fail to clear the lung of colonizing microbes. In prior in vitro studies, we showed that blood neutrophils migrated through the well-differentiated lung epithelium into the CF airway fluid supernatant (ASN) mimic the dysfunction of CF airway neutrophils in vivo, including decreased bactericidal activity despite an increased metabolism. Here, we hypothesized that, in a similar manner to neutrophils, blood monocytes undergo significant adaptations upon recruitment to CFASN. To test this hypothesis, primary human blood monocytes were transmigrated in our in vitro model into the ASN from healthy control (HC) or CF subjects to mimic in vivo recruitment to normal or CF airways, respectively. Surface phenotype, metabolic and bacterial killing activities, and transcriptomic profile by RNA sequencing were quantified post-transmigration. Unlike neutrophils, monocytes were not metabolically activated, nor did they show broad differences in activation and scavenger receptor expression upon recruitment to the CFASN compared to HCASN. However, monocytes recruited to CFASN showed decreased bactericidal activity. RNASeq analysis showed strong effects of transmigration on monocyte RNA profile, with differences between CFASN and HCASN conditions, notably in immune signaling, including lower expression in the former of the antimicrobial factor ISG15, defensin-like chemokine CXCL11, and nitric oxide-producing enzyme NOS3. While monocytes undergo qualitatively different adaptations from those seen in neutrophils upon recruitment to the CF airway microenvironment, their bactericidal activity is also dysregulated, which could explain why they also fail to protect CF airways from infection.

scholarly journals TRIM27 interacts with Iκbα to promote the growth of human renal cancer cells through regulating the NF-κB pathway

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chengwu Xiao ◽  
Wei Zhang ◽  
Meimian Hua ◽  
Huan Chen ◽  
Bin Yang ◽  
...  
Keyword(s):  
Cell Lines ◽  
Prognostic Indicator ◽  
The Cancer Genome Atlas ◽  
Mrna Levels ◽  
Loss Of Function ◽  
Protein Levels ◽  
Kaplan Meier ◽  
Cancer Genome Atlas

Abstract Background The tripartite motif (TRIM) family proteins exhibit oncogenic roles in various cancers. The roles of TRIM27, a member of the TRIM super family, in renal cell carcinoma (RCC) remained unexplored. In the current study, we aimed to investigate the clinical impact and roles of TRIM27 in the development of RCC. Methods The mRNA levels of TRIM27 and Kaplan–Meier survival of RCC were analyzed from The Cancer Genome Atlas database. Real-time PCR and Western blotting were used to measure the mRNA and protein levels of TRIM27 both in vivo and in vitro. siRNA and TRIM27 were exogenously overexpressed in RCC cell lines to manipulate TRIM27 expression. Results We discovered that TRIM27 was elevated in RCC patients, and the expression of TRIM27 was closely correlated with poor prognosis. The loss of function and gain of function results illustrated that TRIM27 promotes cell proliferation and inhibits apoptosis in RCC cell lines. Furthermore, TRIM27 expression was positively associated with NF-κB expression in patients with RCC. Blocking the activity of NF-κB attenuated the TRIM27-mediated enhancement of proliferation and inhibition of apoptosis. TRIM27 directly interacted with Iκbα, an inhibitor of NF-κB, to promote its ubiquitination, and the inhibitory effects of TRIM27 on Iκbα led to NF-κB activation. Conclusions Our results suggest that TRIM27 exhibits an oncogenic role in RCC by regulating NF-κB signaling. TRIM27 serves as a specific prognostic indicator for RCC, and strategies targeting the suppression of TRIM27 function may shed light on future therapeutic approaches.

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