scholarly journals YAP1 Is a Potential Predictive Molecular Biomarker for Response to SMO Inhibitor in Medulloblastoma Cells

Cancers ◽  
2021 ◽  
Vol 13 (24) ◽  
pp. 6249
Author(s):  
Gustavo Alencastro Veiga Cruzeiro ◽  
Taciani de Almeida Magalhães ◽  
Graziella Ribeiro de Sousa ◽  
Ricardo Bonfim Silva ◽  
Carlos Alberto Oliveira de Biagi Junior ◽  
...  
Keyword(s):  
Sonic Hedgehog ◽  
Cellular Response ◽  
Molecular Subtypes ◽  
In Vitro Models ◽  
Rna Seq ◽  
Preclinical Models ◽  
Molecular Biomarker ◽  
Shh Pathway ◽  
Smo Inhibitor

Advances in genomics have led to the identification of twelve relevant molecular subtypes within medulloblastoma (MB). The alpha subtype of Sonic hedgehog-driven MB is resistant to therapy (including smoothened inhibitors) due to activation of genes from the non-canonical SHH pathway, such as MYCN, YAP1, or TP53. Using retrospective cohort microarray data, we found that YAP1 is overexpressed in SHH alpha MB and patients profiled as resistant to SMO inhibitors compared to good responders. Here, we performed YAP1 depletion via CRISPR/Cas9 in two in vitro models of SHH-like MB cells and found that this protein is involved in responsiveness to the SMO inhibitor regarding proliferation, apoptosis, and colony formation. Further, considering the synergic combination of YAP1 depletion with SMO inhibition, we assessed single-cell RNA-seq data from five patients and found that SMO and YAP1 are enriched within cells of SHH MB. Importantly, our data suggest that YAP1 is not only a reliable biomarker for cellular response to SMOi but may indicate prospective testing of combination therapy using YAP1 and SMO inhibitors in preclinical models of SHH MB.

scholarly journals Effects of Platelet-Rich Plasma on Cellular Populations of the Central Nervous System: The Influence of Donor Age

2021 ◽  
Vol 22 (4) ◽  
pp. 1725
Author(s):  
Diego Delgado ◽  
Ane Miren Bilbao ◽  
Maider Beitia ◽  
Ane Garate ◽  
Pello Sánchez ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Cellular Response ◽  
Platelet Rich Plasma ◽  
Biological Response ◽  
In Vitro Models ◽  
Molecular Composition ◽  
Cellular Models ◽  
The Central Nervous System

Platelet-rich plasma (PRP) is a biologic therapy that promotes healing responses across multiple medical fields, including the central nervous system (CNS). The efficacy of this therapy depends on several factors such as the donor’s health status and age. This work aims to prove the effect of PRP on cellular models of the CNS, considering the differences between PRP from young and elderly donors. Two different PRP pools were prepared from donors 65–85 and 20–25 years old. The cellular and molecular composition of both PRPs were analyzed. Subsequently, the cellular response was evaluated in CNS in vitro models, studying proliferation, neurogenesis, synaptogenesis, and inflammation. While no differences in the cellular composition of PRPs were found, the molecular composition of the Young PRP showed lower levels of inflammatory molecules such as CCL-11, as well as the presence of other factors not found in Aged PRP (GDF-11). Although both PRPs had effects in terms of reducing neural progenitor cell apoptosis, stabilizing neuronal synapses, and decreasing inflammation in the microglia, the effect of the Young PRP was more pronounced. In conclusion, the molecular composition of the PRP, conditioned by the age of the donors, affects the magnitude of the biological response.

scholarly journals The Role of Biomimetic Hypoxia on Cancer Cell Behaviour in 3D Models: A Systematic Review

Cancers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1334
Author(s):  
Ye Liu ◽  
Zahra Mohri ◽  
Wissal Alsheikh ◽  
Umber Cheema
Keyword(s):  
Systematic Review ◽  
Cellular Response ◽  
3D Culture ◽  
3D Models ◽  
In Vitro Models ◽  
Research Findings ◽  
Tissue Models

The development of biomimetic, human tissue models is recognized as being an important step for transitioning in vitro research findings to the native in vivo response. Oftentimes, 2D models lack the necessary complexity to truly recapitulate cellular responses. The introduction of physiological features into 3D models informs us of how each component feature alters specific cellular response. We conducted a systematic review of research papers where the focus was the introduction of key biomimetic features into in vitro models of cancer, including 3D culture and hypoxia. We analysed outcomes from these and compiled our findings into distinct groupings to ascertain which biomimetic parameters correlated with specific responses. We found a number of biomimetic features which primed cancer cells to respond in a manner which matched in vivo response.

Barriers, Nephrotoxicology and Chronic Testing In Vitro

2002 ◽  
Vol 30 (2_suppl) ◽  
pp. 101-105 ◽  
Author(s):  
Pilar Prieto
Keyword(s):  
Chronic Exposure ◽  
Cellular Response ◽  
Chronic Toxicity ◽  
Intestinal Barrier ◽  
Toxicity Testing ◽  
In Vitro Models ◽  
Renal Epithelium ◽  
Acute And Chronic Exposure

In many organs of the human body, there are effective physiological barriers which contribute to regulation of the uptake, transport and secretion of endogenous and exogenous materials. ECVAM is involved in the development of several in vitro models for detecting damage to various barriers, including, the renal epithelium, the intestinal barrier, and the blood–brain barrier, after acute and chronic exposure to chemicals and products of various kinds. Long-term toxicity testing is an important issue in toxicology. At present, there are no generally accepted in vitro models available for replacing chronic testing in animals. Under chronic exposure conditions, the cellular response is greater than that which can be predicted in the standard cytotoxicity models. Therefore, the approach to predicting chronic toxicity will need to involve more-complex in vitro models. Several currently available in vitro long-term toxicity systems are under evaluation.

scholarly journals Compound Phyllanthus urinaria L Inhibits HBV-Related HCC through HBx-SHH Pathway Axis Inactivation

2019 ◽  
Vol 2019 ◽  
pp. 1-15
Author(s):  
Yun Li ◽  
Mingjie Jiang ◽  
Mingshun Li ◽  
Yingjie Chen ◽  
Chunshan Wei ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Sonic Hedgehog ◽  
Dependent Manner ◽  
Shh Pathway ◽  
Phyllanthus Urinaria ◽  
Gli 1 ◽  
Underlying Mechanisms ◽  
And Migration ◽  
Dose Dependent

Compound Phyllanthus urinaria L (CP) is a traditional formula widely used in clinical practice for hepatocellular carcinoma (HCC), especially HBV-related HCC. HBx, HBV X gene encoded X protein, has positive correlation with the abnormal SHH pathway in HBV-related HCC. So, we predicted that CP has the capability of anti-HBV-related HCC maybe via inactivating the HBx-Hedgehog pathway axis. HepG2-HBx cells, HBx overexpression, were treated with CP (70μg/ml and 35 μg/ml, respectively) for 48 hours and the mice which received the HepG2-HBx cells were treated with CP (625mg/kg and 300 mg/kg, respectively) for 17 days to evaluate the effect of CP on HBV-related HCC. HBx could accelerate HepG2 cells proliferation, clone formation, and migration in vitro and also could strengthen tumor growth in mice. However, CP could significantly decrease HepG2-HBx cells proliferation, clone formation, and migration in vitro and also could inhibit tumors growth in mice in a dose-dependent manner. Mechanism studies suggested that HBx upregulated the mRNA and proteins expression of Sonic hedgehog (SHH), transmembrane receptor patched (PTCH-1), smoothened (SMO), oncogene homolog transcription factors-1 (GLI-1), and oncogene homolog transcription factors-2 (GLI-2), which are compositions of the SHH pathway. CP could inhibit the mRNA and proteins expression of SHH, PTCH-1, GLI-1, and HBx. It may be one of the underlying mechanisms of CP to delay the HBV-related HCC development through the HBx-SHH pathway axis inactivation.

scholarly journals Arl13b regulates ciliogenesis and the dynamic localization of Shh signaling proteins

2011 ◽  
Vol 22 (23) ◽  
pp. 4694-4703 ◽  
Author(s):  
Christine E. Larkins ◽  
Gladys D. Gonzalez Aviles ◽  
Michael P. East ◽  
Richard A. Kahn ◽  
Tamara Caspary
Keyword(s):  
Sonic Hedgehog ◽  
Protein Localization ◽  
Signaling Proteins ◽  
Shh Signaling ◽  
Dynamic Localization ◽  
Shh Pathway ◽  
Ciliary Protein ◽  
Adp Ribosylation Factor

Arl13b, a ciliary protein within the ADP-ribosylation factor family and Ras superfamily of GTPases, is required for ciliary structure but has poorly defined ciliary functions. In this paper, we further characterize the role of Arl13b in cilia by examining mutant cilia in vitro and determining the localization and dynamics of Arl13b within the cilium. Previously, we showed that mice lacking Arl13b have abnormal Sonic hedgehog (Shh) signaling; in this study, we show the dynamics of Shh signaling component localization to the cilium are disrupted in the absence of Arl13b. Significantly, we found Smoothened (Smo) is enriched in Arl13b-null cilia regardless of Shh pathway stimulation, indicating Arl13b regulates the ciliary entry of Smo. Furthermore, our analysis defines a role for Arl13b in regulating the distribution of Smo within the cilium. These results suggest that abnormal Shh signaling in Arl13b mutant embryos may result from defects in protein localization and distribution within the cilium.

scholarly journals Phospholipids of APOE lipoproteins activate microglia in an isoform-specific manner in preclinical models of Alzheimer’s disease

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Nicholas F. Fitz ◽  
Kyong Nyon Nam ◽  
Cody M. Wolfe ◽  
Florent Letronne ◽  
Brittany E. Playso ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Genetic Risk ◽  
Rna Seq ◽  
Preclinical Models ◽  
Preclinical Ad ◽  
Specific Manner ◽  
Microglial Migration ◽  
Microglial Response

AbstractAPOE and Trem2 are major genetic risk factors for Alzheimer’s disease (AD), but how they affect microglia response to Aβ remains unclear. Here we report an APOE isoform-specific phospholipid signature with correlation between human APOEε3/3 and APOEε4/4 AD brain and lipoproteins from astrocyte conditioned media of APOE3 and APOE4 mice. Using preclinical AD mouse models, we show that APOE3 lipoproteins, unlike APOE4, induce faster microglial migration towards injected Aβ, facilitate Aβ uptake, and ameliorate Aβ effects on cognition. Bulk and single-cell RNA-seq demonstrate that, compared to APOE4, cortical infusion of APOE3 lipoproteins upregulates a higher proportion of genes linked to an activated microglia response, and this trend is augmented by TREM2 deficiency. In vitro, lack of TREM2 decreases Aβ uptake by APOE4-treated microglia only, suggesting TREM2-APOE interaction. Our study elucidates phenotypic and transcriptional differences in microglial response to Aβ mediated by APOE3 or APOE4 lipoproteins in preclinical models of AD.

scholarly journals C28-induced autophagy of female germline stem cells in vitro and its potential mechanisms

2019 ◽  
Author(s):  
Rong Hu ◽  
Xiuying Pei ◽  
Huchen Zhou ◽  
Ji Wu ◽  
Ping Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Er Stress ◽  
Cellular Response ◽  
Germline Stem Cells ◽  
Rna Seq ◽  
Sequestosome 1 ◽  
Expression Of Genes ◽  
Female Germline ◽  
Molecular Compounds

Abstract BackgroundThere are few studies indicating that small molecular compounds affect the proliferation, differentiation, apoptosis, and autophagy of female germline stem cells (FGSCs). However, the epigenetic regulatory mechanism of small molecular compounds that induce autophagy in FGSCs remains unknown.ResultsIn this study, we found that C28 reduced the viability and proliferation of FGSCs, respectively. Additionally, western blotting showed that the expression of autophagy marker light chain 3 beta II (LC3B-II) was significantly increased and expression of sequestosome-1 (SQSTM1) was significantly reduced in C28-treated groups. Immunofluorescence showed that, in C28-treated groups, the number of LC3B-II-positive puncta was increased significantly. These results indicated that C28 induced autophagy of FGSCs in vitro. ChIP-seq data showed that autophagy-related biological processes such as regulation of mitochondrial membrane potential, Golgi vesicle transport, and cellular response to reactive oxygen species were enriched. In addition, RNA-Seq showed that the expression of genes (Trib3, DDIT3, and ATF4) related to endoplasmic reticulum (ER) stress was enhanced by C28.ConclusionC28 could induce FGSC autophagy in vitro leading to a decrease in the number of FGSCs. H3K27ac and ER stress might play roles in C28-induced autophagy of FGSCs in vitro.

scholarly journals Targeting Sonic Hedgehog Signaling by Compounds and Derivatives from Natural Products

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Yu-Chuen Huang ◽  
K. S. Clifford Chao ◽  
Hui-Fen Liao ◽  
Yu-Jen Chen
Keyword(s):  
Alternative Medicine ◽  
Complementary And Alternative Medicine ◽  
Sonic Hedgehog ◽  
Anticancer Agents ◽  
Invasion And Metastasis ◽  
Antitumor Effects ◽  
Shh Signaling ◽  
Shh Pathway ◽  
Complementary And Alternative

Cancer stem cells (CSCs) are a major cause of cancer treatment failure, relapse, and drug resistance and are known to be responsible for cancer cell invasion and metastasis. The Sonic hedgehog (Shh) signaling pathway is crucial to embryonic development. Intriguingly, the aberrant activation of the Shh pathway plays critical roles in developing CSCs and leads to angiogenesis, migration, invasion, and metastasis. Natural compounds and chemical structure modified derivatives from complementary and alternative medicine have received increasing attention as cancer chemopreventives, and their antitumor effects have been demonstrated bothin vitroandin vivo. However, reports for their bioactivity against CSCs and specifically targeting Shh signaling remain limited. In this review, we summarize investigations of the compounds cyclopamine, curcumin, epigallocatechin-3-gallate, genistein, resveratrol, zerumbone, norcantharidin, and arsenic trioxide, with a focus on Shh signaling blockade. Given that Shh signaling antagonism has been clinically proven as effective strategy against CSCs, this review may be exploitable for development of novel anticancer agents from complementary and alternative medicine.

scholarly journals Bacteriophage inhibits murine norovirus replication after co-incubation in RAW 264.7 cells

2021 ◽  
Author(s):  
Lili Zhang ◽  
Khashayar Shahin ◽  
Ran Wang
Keyword(s):  
Cellular Response ◽  
Antimicrobial Effect ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Murine Norovirus ◽  
Rna Seq ◽  
Qpcr Analysis ◽  
Ifn Γ ◽  
Phage Treatment

AbstractBacteriophages (phages) are viruses of bacteria. Despite the growing progress in research on phage interactions with eukaryotic cells, our understanding of the roles of phages and their potential implications remains incomplete. The objective of this study was to investigate the effects of the Staphylococcus aureus phage vB_SauM_JS25 on murine norovirus (MNV) replication. Experiments were performed using the RAW 264.7 cell line. After phage treatment, MNV multiplication was significantly inhibited, as indicated by real-time quantitative PCR (RT-qPCR) analysis, western blot, and immunofluorescence assay. Furthermore, we demonstrated the transcriptional changes in phage–MNV co-incubated RAW 264.7 cells through RNA sequencing (RNA-Seq) and bioinformatic analysis. Our subsequent analyses revealed that the innate immune response may play an important role in the restriction of MNV replication, such as the cellular response to IFN-γ and response to IFN-γ. In addition, the gene expression of IL-10, Arg-1, Ccl22, GBP2, GBP3, GBP5, and GBP7 was proven to increase significantly by RT-qPCR, showing a strong correlation between RT-qPCR and RNA-Seq results. Furthermore, phage treatment activated guanylate binding proteins (GBPs), as revealed by RT-qPCR analysis, western blotting, and confocal microscopy. Taken together, these data suggest that the phage affects the innate response (such as the IFN-inducible GTPases and GBPs), reflecting their direct antimicrobial effect on the membrane structure of the MNV replication complexes, and therefore, exerts an antiviral effect in vitro. Collectively, our findings provide insights into the interactions of immune cells and phages, which improve our understanding of the actual role and potential of phages.

scholarly journals RNA-Seq Analysis Reveals the Role of Omp16 in Brucella-Infected RAW264.7 Cells

2021 ◽  
Vol 8 ◽  
Author(s):  
Dong Zhou ◽  
Feijie Zhi ◽  
Jiaoyang Fang ◽  
Weifang Zheng ◽  
Junmei Li ◽  
...  
Keyword(s):  
Proinflammatory Cytokines ◽  
Cellular Response ◽  
Outer Membrane Proteins ◽  
Raw264.7 Cells ◽  
Economic Losses ◽  
Rna Seq ◽  
Protective Antigens ◽  
Cytokines And Chemokines ◽  
Relevant Role

Brucellosis is an endemic zoonotic infectious disease in the majority of developing countries, which causes huge economic losses. As immunogenic and protective antigens at the surface of Brucella spp., outer membrane proteins (Omps) are particularly attractive for developing vaccine and could have more relevant role in host–pathogen interactions. Omp16, a homolog to peptidoglycan-associated lipoproteins (Pals), is essential for Brucella survival in vitro. At present, the functions of Omp16 have been poorly studied. Here, the gene expression profile of RAW264.7 cells infected with Brucella suis vaccine strain 2 (B. suis S2) and ΔOmp16 was analyzed by RNA-seq to investigate the cellular response immediately after Brucella entry. The RNA-sequence analysis revealed that a total of 303 genes were significantly regulated by B. suis S2 24 h post-infection. Of these, 273 differentially expressed genes (DEGs) were upregulated, and 30 DEGs were downregulated. These DEGs were mainly involved in innate immune signaling pathways, including pattern recognition receptors (PRRs), proinflammatory cytokines, and chemokines by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. In ΔOmp16-infected cells, the expression of 52 total cells genes was significantly upregulated and that of 9 total cells genes were downregulated compared to B. suis S2-infected RAW264.7 cells. The KEGG pathway analysis showed that several upregulated genes were proinflammatory cytokines and chemokines, such as interleukin (IL)-6, IL-11, IL-12β, C–C motif chemokine (CCL2), and CCL22. All together, we clearly demonstrate that ΔOmp16 can alter macrophage immune-related pathways to increase proinflammatory cytokines and chemokines, which provide insights into illuminating the Brucella pathogenic strategies.

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