scholarly journals p53 mutants cooperate with HIF-1 in transcriptional regulation of extracellular matrix components to promote tumor progression

2018 ◽  
Vol 115 (46) ◽  
pp. E10869-E10878 ◽  
Author(s):  
Ivano Amelio ◽  
Mara Mancini ◽  
Varvara Petrova ◽  
Rob A. Cairns ◽  
Polina Vikhreva ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cancer Progression ◽  
Treatment Options ◽  
Hypoxia Inducible Factor ◽  
Gene Expression Signature ◽  
Gene Signature ◽  
Cancer Pathogenesis ◽  
Extracellular Matrix Components

Mutations in the TP53 gene and microenvironmentally driven activation of hypoxia-inducible factor-1 (HIF-1) typically occur in later stages of tumorigenesis. An ongoing challenge is the identification of molecular determinants of advanced cancer pathogenesis to design alternative last-line therapeutic options. Here, we report that p53 mutants influence the tumor microenvironment by cooperating with HIF-1 to promote cancer progression. We demonstrate that in non-small cell lung cancer (NSCLC), p53 mutants exert a gain-of-function (GOF) effect on HIF-1, thus regulating a selective gene expression signature involved in protumorigenic functions. Hypoxia-mediated activation of HIF-1 leads to the formation of a p53 mutant/HIF-1 complex that physically binds the SWI/SNF chromatin remodeling complex, promoting expression of a selective subset of hypoxia-responsive genes. Depletion of p53 mutants impairs the HIF-mediated up-regulation of extracellular matrix (ECM) components, including type VIIa1 collagen and laminin-γ2, thus affecting tumorigenic potential of NSCLC cells in vitro and in mouse models in vivo. Analysis of surgically resected human NSCLC revealed that expression of this ECM gene signature was highly correlated with hypoxic tumors exclusively in patients carrying p53 mutations and was associated with poor prognosis. Our data reveal a GOF effect of p53 mutants in hypoxic tumors and suggest synergistic activities of p53 and HIF-1. These findings have important implications for cancer progression and might provide innovative last-line treatment options for advanced NSCLC.

scholarly journals Selection of Effective Therapies Using Three-Dimensional in vitro Modeling of Chondrosarcoma

2020 ◽  
Vol 7 ◽  
Author(s):  
Ieva Palubeckaitė ◽  
Sanne Venneker ◽  
Inge H. Briaire-de Bruijn ◽  
Brendy E. van den Akker ◽  
Augustinus D. Krol ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Large Fraction ◽  
Treatment Options ◽  
Malignant Neoplasms ◽  
Animal Testing ◽  
Long Term Treatment ◽  
Immunohistochemical Assessment ◽  
Inhibitor Treatment

Purpose: Chondrosarcomas are a group of cartilaginous malignant neoplasms characterized by the deposition of chondrogenic extracellular matrix. Surgical resection is currently the only curative treatment option, due to their high resistance to conventional chemotherapy and radiotherapy. Novel therapeutic treatment options may improve outcome. Predominantly used cell line monolayer in vitro models lack in vivo complexity, such as the presence of extracellular matrix, and differing oxygen access. Hence, we aimed to improve pre-clinical chondrosarcoma research by developing an alginate-based 3D cell culture model.Method: An alginate scaffold was applied to generate spheroids of three chondrosarcoma cell lines (CH2879, JJ012, SW1353). Morphological, histological and immunohistochemical assessment of the spheroids were used to characterize the chondrosarcoma model. Presto blue assay, morphological and immunohistochemical assessment were applied to assess spheroid response to a panel of chemotherapeutics and targeted therapies, which was compared to conventional 2D monolayer models. Synergistic effect of doxorubicin and ABT-737 (Bcl-2 inhibitor) was compared between monolayer and spheroid models using excess over Bliss. A 3D colony formation assay was developed for assessment of radiotherapy response.Results: Chondrosarcoma spheroids produced chondrogenic matrix and remained proliferative after 2 weeks of culture. When treated with chemotherapeutics, the spheroids were more resistant than their monolayer counterparts, in line with animal models and clinical data. Moreover, for sapanisertib (mTOR inhibitor) treatment, a recovery in chondrosarcoma growth, previously observed in mice models, was also observed using long-term treatment. Morphological assessment was useful in the case of YM-155 (survivin inhibitor) treatment where a fraction of the spheroids underwent cell death, however a large fraction remained proliferative and unaffected. Synergy was less pronounced in 3D compared to 2D. A 3D clonogenic assay confirmed increased resistance to radiotherapy in 3D chondrosarcoma spheroids.Conclusion: We demonstrate that the chondrosarcoma alginate spheroid model is more representative of chondrosarcoma in vivo and should be used instead of the monolayer model for therapy testing. Improved selection at in vitro stage of therapeutic testing will increase the amount of information available for experimental design of in vivo animal testing and later, clinical stages. This can potentially lead to increased likelihood of approval and success at clinical trials.

scholarly journals Laminin promotes neuritic regeneration from cultured peripheral and central neurons.

1983 ◽  
Vol 97 (6) ◽  
pp. 1882-1890 ◽  
Author(s):  
M Manthorpe ◽  
E Engvall ◽  
E Ruoslahti ◽  
F M Longo ◽  
G E Davis ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Neurite Growth ◽  
Sensory Ganglia ◽  
Culture Methods ◽  
Fetal Rat ◽  
Extracellular Matrix Components ◽  
Ciliary Ganglion Neurons ◽  
Ganglion Neurons

The ability of axons to grow through tissue in vivo during development or regeneration may be regulated by the availability of specific neurite-promoting macromolecules located within the extracellular matrix. We have used tissue culture methods to examine the relative ability of various extracellular matrix components to elicit neurite outgrowth from dissociated chick embryo parasympathetic (ciliary ganglion) neurons in serum-free monolayer culture. Purified laminin from both mouse and rat sources, as well as a partially purified polyornithine-binding neurite promoting factor (PNPF-1) from rat Schwannoma cells all stimulate neurite production from these neurons. Laminin and PNPF-1 are also potent stimulators of neurite growth from cultured neurons obtained from other peripheral as well as central neural tissues, specifically avian sympathetic and sensory ganglia and spinal cord, optic tectum, neural retina, and telencephalon, as well as from sensory ganglia of the neonatal mouse and hippocampal, septal, and striatal tissues of the fetal rat. A quantitative in vitro bioassay method using ciliary neurons was used to (a) measure and compare the specific neurite-promoting activities of these agents, (b) confirm that during the purification of laminin, the neurite-promoting activity co-purifies with the laminin protein, and (c) compare the influences of antilaminin antibodies on the neurite-promoting activity of laminin and PNPF-1. We conclude that laminin and PNPF-1 are distinct macromolecules capable of expressing their neurite-promoting activities even when presented in nanogram amounts. This neurite-promoting bioassay currently represents the most sensitive test for the biological activity of laminin.

scholarly journals Regulation of Tissue Fibrosis by the Biomechanical Environment

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Wayne Carver ◽  
Edie C. Goldsmith
Keyword(s):  
Extracellular Matrix ◽  
Intracellular Signaling ◽  
Mechanical Load ◽  
Mechanical Force ◽  
Mechanical Signals ◽  
Organ Systems ◽  
Extracellular Matrix Components ◽  
Matrix Components

The biomechanical environment plays a fundamental role in embryonic development, tissue maintenance, and pathogenesis. Mechanical forces play particularly important roles in the regulation of connective tissues including not only bone and cartilage but also the interstitial tissues of most organs.In vivostudies have correlated changes in mechanical load to modulation of the extracellular matrix and have indicated that increased mechanical force contributes to the enhanced expression and deposition of extracellular matrix components or fibrosis. Pathological fibrosis contributes to dysfunction of many organ systems. A variety ofin vitromodels have been utilized to evaluate the effects of mechanical force on extracellular matrix-producing cells. In general, application of mechanical stretch, fluid flow, and compression results in increased expression of extracellular matrix components. More recent studies have indicated that tissue rigidity also provides profibrotic signals to cells. The mechanisms whereby cells detect mechanical signals and transduce them into biochemical responses have received considerable attention. Cell surface receptors for extracellular matrix components and intracellular signaling pathways are instrumental in the mechanotransduction process. Understanding how mechanical signals are transmitted from the microenvironment will identify novel therapeutic targets for fibrosis and other pathological conditions.

scholarly journals Salmonella Extracellular Matrix Components Influence Biofilm Formation and Gallbladder Colonization

2016 ◽  
Vol 84 (11) ◽  
pp. 3243-3251 ◽  
Author(s):  
Haley E. Adcox ◽  
Erin M. Vasicek ◽  
Varun Dwivedi ◽  
Ky V. Hoang ◽  
Joanne Turner ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Typhoid Fever ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Bacterial Survival ◽  
Content Type ◽  
Extracellular Matrix Components ◽  
Matrix Components

Salmonella enterica serovar Typhi, the causative agent of typhoid fever in humans, forms biofilms encapsulated by an extracellular matrix (ECM). Biofilms facilitate colonization and persistent infection in gallbladders of humans and mouse models of chronic carriage. Individual roles of matrix components have not been completely elucidated in vitro or in vivo . To examine individual functions, strains of Salmonella enterica serovar Typhimurium, the murine model of S . Typhi, in which various ECM genes were deleted or added, were created to examine biofilm formation, colonization, and persistence in the gallbladder. Studies show that curli contributes most significantly to biofilm formation. Expression of Vi antigen decreased biofilm formation in vitro and virulence and bacterial survival in vivo without altering the examined gallbladder pro- or anti-inflammatory cytokines. Oppositely, loss of all ECM components (Δ wcaM Δ csgA Δ yihO Δ bcsE ) increased virulence and bacterial survival in vivo and reduced gallbladder interleukin-10 (IL-10) levels. Colanic acid and curli mutants had the largest defects in biofilm-forming ability and contributed most significantly to the virulence increase of the Δ wcaM Δ csgA Δ yihO Δ bcsE mutant strain. While the Δ wcaM Δ csgA Δ yihO Δ bcsE mutant was not altered in resistance to complement or growth in macrophages, it attached and invaded macrophages better than the wild-type (WT) strain. These data suggest that ECM components have various levels of importance in biofilm formation and gallbladder colonization and that the ECM diminishes disseminated disease in our model, perhaps by reducing cell attachment/invasion and dampening inflammation by maintaining/inducing IL-10 production. Understanding how ECM components aid acute disease and persistence could lead to improvements in therapeutic treatment of typhoid fever patients.

scholarly journals Crosstalk between invadopodia and the extracellular matrix

2020 ◽  
Author(s):  
Shinji Iizuka ◽  
Ronald P. Leon ◽  
Kyle P. Gribbin ◽  
Ying Zhang ◽  
Jose Navarro ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Complex Structure ◽  
Three Dimensional ◽  
3D Culture ◽  
Model Systems ◽  
Type I ◽  
Extracellular Matrix Components

ABSTRACTThe scaffold protein Tks5α is required for invadopodia-mediated cancer invasion both in vitro and in vivo. We have previously also revealed a role for Tks5 in tumor cell growth using three-dimensional (3D) culture model systems and mouse transplantation experiments. Here we use both 3D and high-density fibrillar collagen (HDFC) culture to demonstrate that native type I collagen, but not a form lacking the telopeptides, stimulated Tks5-dependent growth, which was dependent on the DDR collagen receptors. We used microenvironmental microarray (MEMA) technology to determine that laminin, collagen I, fibronectin and tropoelastin also stimulated invadopodia formation. A Tks5α-specific monoclonal antibody revealed its expression both on microtubules and at invadopodia. High- and super-resolution microscopy of cells in and on collagen was then used to place Tks5α at the base of invadopodia, separated from much of the actin and cortactin, but coincident with both matrix metalloprotease and cathepsin proteolytic activity. Inhibition of the Src family kinases, cathepsins or metalloproteases all reduced invadopodia length but each had distinct effects on Tks5α localization. These studies highlight the crosstalk between invadopodia and extracellular matrix components, and reveal the invadopodium to be a spatially complex structure.

Megakaryocytes Contribute To The Establishment Of The Bone Marrow Environment By Expressing Extracellular Matrix proteins

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3695-3695
Author(s):  
Alessandro Malara ◽  
Cristian Gruppi ◽  
Manuela Currao ◽  
Alessandra Balduini
Keyword(s):  
Bone Marrow ◽  
Extracellular Matrix ◽  
Type Iv Collagen ◽  
Extracellular Matrix Component ◽  
Matrix Component ◽  
Type Iv ◽  
Extracellular Matrix Components ◽  
Matrix Components

Abstract Introduction the bone marrow microenvironment consists of various types of cells and their secreted extracellular matrix components that surround capillary-venous sinusoids, and plays a key role in the regulation of hematopoiesis. In general, extracellular matrix components interact with each other to form a structural framework that supports tissue organization and positional cues that regulate cellular processes. Megakaryocytes are rare cells in the bone marrow and, besides platelet release, growing evidences attribute new functions to these cells in the generation and maintenance of the bone marrow cell niche. Recent evidences, by our group, demonstrated that megakaryocytes are involved in matrix deposition and remodeling, as demonstrated by their role in fibronectin fibrillogenesis and the expression of matrix cross-linking enzymes, such as factor XIIIa, essential in the dynamic of megakaryocyte-matrix component interactions. Interestingly, individual extracellular matrix components were demonstrated to play a role in the regulation of megakaryocytes development in vitro. Fibronectin was shown to regulate megakaryocyte maturation and proplatelet extension, while type III and type IV collagens were demonstrated to support proplatelet formation in vitro. In contrast, type I collagen is an important physiological inhibitor of platelet release in vitro. However, little is known about the exact localization as well as function of these matrix components in vivo. Results in this work we have analyzed the spatial distribution of megakaryocytes and extracellular matrix components by immunofluorescence in murine femur sections. We found that megakaryocytes were predominantly located in the femur diaphysis with only 20% of megakaryocytes within 50μm from the endosteal surface and more than 80% of megakaryocytes located less than 50 μm from a sinusoid. Correlation between megakaryocyte distance from sinusoids and dimension suggested a gradient of maturing megakaryocytes towards the vascular niche. Next, we deciphered bone marrow extracellular matrix component composition by western blotting and mapped the location in situ of different collagens (I, III, IV, VI) and glycoproteins (fibronectin, laminin). We found that all these proteins were differently located in the endosteal and sinusoidal districts supporting the concept that regulation of hemopoiesis, in the bone marrow, may also depend from matrix distribution. Further, we showed, for the first time, that megakaryocytes were surrounded by a pericellular matrix mainly composed of fibronectin, laminin and type IV collagen. Interestingly, these three proteins were also demonstrated to promote thrombopoietin-dependent megakaryocyte differentiation in in vitro cultures of bone marrow hemopoietic progenitor cells. Finally, fibronectin, laminin and type IV collagen were also demonstrated to be expressed and synthesized by differentiated megakaryocytes in vitro as demonstrated by PCR and western blotting analysis. Most importantly, megakaryocyte expression of these extracellular matrix components was up-regulated in vivo during bone marrow reconstitution upon drug induced myelosuppression and, at a lesser extent, thrombocytopenia. Conclusions all together these results suggested that megakaryocytes are important extracellular matrix component-producing bone marrow cells and that released extracellular matrix components support megakaryopoiesis and concur to the generation of bone marrow niches. Disclosures: No relevant conflicts of interest to declare.

scholarly journals In Vivo and In VitroExpression of Tenascin by Human Thymic Microenvironmental Cells

1995 ◽  
Vol 4 (2) ◽  
pp. 139-147 ◽  
Author(s):  
Claudia Sondermann Freitas ◽  
Jurandy Susana Patricia O'Campo Lyra ◽  
Sergio Ranto Dalmau ◽  
Wilson Savino
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Basement Membranes ◽  
Extracellular Matrix Protein ◽  
Thymic Epithelial Cell ◽  
Normal Infant ◽  
Cell Network ◽  
Extracellular Matrix Components

Increasing evidence reveals that extracellular matrix components can be regarded as a group of mediators in intrathymic T-cell migration and/or differentiation. Yet, little is kown about the expression and putative function of one particular extracellular matrix protein, namely, tenascin in the thymus. Herein we investigated, by means of immunocytochemistry, tenascin expression in normal infant and fetal human thymuses, as well as in cultures of thymic microenvironmental cells.In situ, tenascin distribution is restricted to the medulla and cortico-medullary regions of normal thymuses. This pattern thus differed from that of fibronectin, laminin and type IV collagen, in which subseptal basement membranes were strongly labeled. Interestingly, tenascin did not co-localize with the cytokeratin-defined thymic epithelial cell network. This was in keeping with thein vitrodata showing that tenascin-bearing cells were nonepithelial (and probably nonfibroblastic) microenvironmental elements.Studies with fetal thymuses revealed a developmentally regulated expression of tenascin, with a faint but consistent network labeling, in thymic rudiments as early as 12 weeks of gestational age, that progressed to a strong TN expression at 18 weeks of fetal development, which was similar to the distribution pattern observed thereafter, including postnatally.Our results clearly indicated that tenascin is constitutively expressed in the human thymus, since early stages of thymic ontogeny, and suggest that the cell type responsible for its secretion is a nonepithelial microenvironmental cell.

scholarly journals A specific tRNA half, 5’tiRNA-His-GTG, responds to hypoxia via the HIF1α/ANG axis and promotes colorectal cancer progression by regulating LATS2

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
En-Wei Tao ◽  
Hao-Lian Wang ◽  
Wing Yin Cheng ◽  
Qian-Qian Liu ◽  
Ying-Xuan Chen ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cancer Progression ◽  
Hypoxia Inducible Factor ◽  
Luciferase Reporter ◽  
Hippo Signaling ◽  
Large Tumor ◽  
In Vivo Experiments ◽  
Trna Half

Abstract Background Currently, tRNA-derived small RNAs (tsRNAs) are recognized as a novel and potential type of non-coding RNAs (ncRNAs), which participate in various cellular processes and play an essential role in cancer progression. However, tsRNAs involvement in colorectal cancer (CRC) progression remains unclear. Methods Sequencing analyses were performed to explore the tsRNAs with differential expression in CRC. Gain- and loss-of functions of 5’tiRNA-His-GTG were performed in CRC cells and xenograft tumor to discover its role in the progression of CRC. Hypoxia culture and hypoxia inducible factor 1 subunit alpha (HIF1α) inhibitors were performed to uncover the biogenesis of 5’tiRNA-His-GTG. The regulation of 5’tiRNA-His-GTG for large tumor suppressor kinase 2 (LATS2) were identified by luciferase reporter assay, western blot, and rescue experiments. Results Here, our study uncovered the profile of tsRNAs in human CRC tissues and confirmed a specific tRNA half, 5’tiRNA-His-GTG, is upregulated in CRC tissues. Then, in vitro and in vivo experiments revealed the oncogenic role of 5’tiRNA-His-GTG in CRC and found that targeting 5’tiRNA-His-GTG can induce cell apoptosis. Mechanistically, the generation of 5’tiRNA-His-GTG seems to be a responsive process of tumor hypoxic microenvironment, and it is regulated via the HIF1α/angiogenin (ANG) axis. Remarkably, LATS2 was found to be an important and major target of 5’tiRNA-His-GTG, which renders 5’tiRNA-His-GTG to “turn off” hippo signaling pathway and finally promotes the expression of pro-proliferation and anti-apoptosis related genes. Conclusions In summary, the findings revealed a specific 5’tiRNA-His-GTG-engaged pathway in CRC progression and provided clues to design a novel therapeutic target in CRC.

scholarly journals Hypoxia-Inducible Factor 1 Alpha Is Dispensable for Host Defense of Group B Streptococcus Colonization and Infection

2021 ◽  
pp. 1-13
Author(s):  
Gregory R. Lum ◽  
Vicki Mercado ◽  
Diede van Ens ◽  
Victor Nizet ◽  
Jacqueline M. Kimmey ◽  
...  
Keyword(s):  
Host Response ◽  
Immune Cell ◽  
Systemic Disease ◽  
Treatment Options ◽  
Hypoxia Inducible Factor ◽  
Immune Defense ◽  
Hypoxia Inducible Factor 1 ◽  
Group B

Group B <i>Streptococcus</i> (GBS) is a leading cause of neonatal morbidity and mortality, and the primary source of exposure is the maternal vagina. Intrapartum antibiotic prophylaxis for GBS-positive mothers has reduced the incidence of GBS early-onset disease, however, potential long-lasting influence of an antibiotic-altered neonatal microbiota, and the frequent clinical sequelae in survivors of invasive GBS infection, compels alternative treatment options for GBS. Here, we examined the role of transcription factor hypoxia-inducible factor 1 alpha (HIF-1α), widely recognized as a regulator of immune activation during infection, in the host response to GBS. Given the importance of endogenous HIF-1α for innate immune defense, and the potential utility of HIF-1α stabilization in promoting bacterial clearance, we hypothesized that HIF-1α could play an important role in coordinating host responses to GBS in colonization and systemic disease. Counter to our hypothesis, we found that GBS infection did not induce HIF-1α expression in vaginal epithelial cells or murine macrophages, nor did HIF-1α deficiency alter GBS colonization or pathogenesis in vivo. Furthermore, pharmacological enhancement of HIF-1α did not improve control of GBS in pathogenesis and colonization models, while displaying inhibitory effects in vaginal epithelial cytokines and immune cell killing in vitro. Taken together, we conclude that HIF-1α is not a prominent aspect of the host response to GBS colonization or invasive disease, and its pharmacological modulation is unlikely to provide significant benefit against this important neonatal pathogen.

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