scholarly journals TRIP6 promotes inflammatory damage via the activation of TRAF6 signaling in a murine model of DSS-induced colitis

2022 ◽  
Vol 19 (1) ◽  
Author(s):  
Yun Yang ◽  
Xiu-Ming Li ◽  
Jing-Ru Wang ◽  
Yan Li ◽  
Wen-Long Ye ◽  
...  
Keyword(s):  
Adaptor Protein ◽  
Biological Processes ◽  
Wild Type ◽  
Colonic Inflammation ◽  
Murine Colitis ◽  
Inflammatory Damage ◽  
Disease Induction ◽  
Severe Colitis ◽  
Pro Inflammatory Cytokines

Abstract Background TRIP6 is a zyxin family member that serves as an adaptor protein to regulate diverse biological processes. In prior reports, TRIP6 was shown to play a role in regulating inflammation. However, its in vivo roles and mechanistic importance in colitis remain largely elusive. Herein, we therefore employed TRIP6-deficient (TRIP6−/−) mice in order to explore the mechanistic importance of TRIP6 in a dextran sodium sulfate (DSS)-induced model of murine colitis. Findings Wild-type (TRIP6+/+) mice developed more severe colitis following DSS-mediated disease induction relative to TRIP6−/− mice, as evidenced by more severe colonic inflammation and associated crypt damage. TRIP6 expression in wild-type mice was significantly elevated following DSS treatment. Mechanistically, TRIP6 binds to TRAF6 and enhances oligomerization and autoubiquitination of TRAF6. This leads to the activation of NF-κB signaling and the expression of pro-inflammatory cytokines such as TNFα and IL-6, in the in vivo mouse model of colitis. Conclusions These in vivo data demonstrate that TRIP6 serves as a positive regulator of DSS-induced colitis through interactions with TRAF6 resulting in the activation of inflammatory TRAF6 signaling, highlighting its therapeutic promise as a protein that theoretically can be targeted to prevent or treat colitis.

scholarly journals TRIP6 deficiency inhibits inflammatory damage in a murine model of DSS-induced colitis

2021 ◽  
Author(s):  
Yun Yang ◽  
Xiu-Ming Li ◽  
Yan Li ◽  
Jing-Ru Wang ◽  
Wen-Long Ye ◽  
...  
Keyword(s):  
Adaptor Protein ◽  
Nuclear Factor Κb ◽  
Nuclear Factor ◽  
Biological Processes ◽  
Wild Type ◽  
Murine Colitis ◽  
Inflammatory Damage ◽  
Disease Induction ◽  
Severe Colitis

Abstract BackgroundTRIP6 is a zyxin family member that serves as an adaptor protein to regulate diverse biological processes. In prior reports, TRIP6 was shown to play a role in regulating inflammation by inducing the activation of the nuclear factor-κB (NF-κB) signaling cascade, though its specific role in vivo in inflammatory contexts remains to be clarified. Herein, we therefore employed TRIP6-deficient (TRIP6 −/− ) mice in order to explore the mechanistic importance of TRIP6 in a dextran sodium sulfate (DSS)-induced model of murine colitis. Findings Wild-type (TRIP6 +/+ ) mice developed more severe colitis following DSS-mediated disease induction relative to TRIP6 −/− mice, as evidenced by more severe colonic inflammation and associated crypt damage. From a mechanistic perspective, TRIP6 expression in wild-type mice was significantly elevated in the period of DSS treatment; TRIP6 deficiency was associated with the inhibition of IκBα degradation, thus suppressing NF-κB signaling activity and impairing the secretion of proinflammatory cytokines such as TNFα and IL-6. Conclusions Together, these in vivo data demonstrate that TRIP6 serves as a positive regulator of DSS-induced colitis by regulating inflammatory NF-κB signaling, highlighting its therapeutic promise as a protein that theoretically can be targeted to prevent or treat colitis.

scholarly journals Molecular Interplay between AURKA and SPOP Dictates CRPC Pathogenesis via Androgen Receptor

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3247
Author(s):  
Kumar Nikhil ◽  
Mohini Kamra ◽  
Asif Raza ◽  
Hanan S. Haymour ◽  
Kavita Shah
Keyword(s):  
Androgen Receptor ◽  
Feedback Loop ◽  
Adaptor Protein ◽  
Castration Resistant Prostate Cancer ◽  
Wild Type ◽  
Protein Levels ◽  
Upstream Regulator ◽  
Castration Resistant ◽  
Tumor Enhancer

SPOP, an adaptor protein for E3 ubiquitin ligase can function as a tumor-suppressor or a tumor-enhancer. In castration-resistant prostate cancer (CRPC), it inhibits tumorigenesis by degrading many oncogenic targets, including androgen receptor (AR). Expectedly, SPOP is the most commonly mutated gene in CRPC (15%), which closely correlates with poor prognosis. Importantly, 85% of tumors that retain wild-type SPOP show reduced protein levels, indicating that SPOP downregulation is an essential step in CRPC progression. However, the underlying molecular mechanism remains unknown. This study uncovered the first mechanism of SPOP regulation in any type of cancer. We identified SPOP as a direct substrate of Aurora A (AURKA) using an innovative technique. AURKA directly phosphorylates SPOP at three sites, causing its ubiquitylation. SPOP degradation drives highly aggressive oncogenic phenotypes in cells and in vivo including stabilizing AR, ARv7 and c-Myc. Further, SPOP degrades AURKA via a feedback loop. SPOP upregulation is one of the mechanisms by which enzalutamide exerts its efficacy. Consequently, phospho-resistant SPOP fully abrogates tumorigenesis and EMT in vivo, and renders CRPC cells sensitive to enzalutamide. While genomic mutations of SPOP can be treated with gene therapy, identification of AURKA as an upstream regulator of SPOP provides a powerful opportunity for retaining WT-SPOP in a vast majority of CRPC patients using AURKA inhibitors ± enzalutamide, thereby treating the disease and inhibiting its progression.

scholarly journals Knock out of sHSP genes determines some modifications in the probiotic attitude of Lactiplantibacillus plantarum

2020 ◽  
Author(s):  
Angela Longo ◽  
Pasquale Russo ◽  
Vittorio Capozzi ◽  
Giuseppe Spano ◽  
Daniela Fiocco
Keyword(s):  
Small Heat Shock Protein ◽  
Cellular Adhesion ◽  
Probiotic Properties ◽  
Wild Type ◽  
Knock Out ◽  
Human Enterocyte ◽  
Transcriptional Induction ◽  
Pro Inflammatory Cytokines

Abstract Objective We investigated whether the knock out of small heat shock protein (sHSP) genes (hsp1, hsp2 and hsp3) impact on probiotic features of Lactiplantibacillus plantarum WCFS1, aiming to find specific microbial effectors involved in microbe-host interplay. Results The probiotic properties of L. plantarum WCFS1 wild type, hsp1, hsp2 and hsp3 mutant clones were evaluated and compared through in vitro trials. Oro-gastro-intestinal assays pointed to significantly lower survival for hsp1 and hsp2 mutants under stomach-like conditions, and for hsp3 mutant under intestinal stress. Adhesion to human enterocyte-like cells was similar for all clones, though the hsp2 mutant exhibited higher adhesiveness. L. plantarum cells attenuated the transcriptional induction of pro-inflammatory cytokines on lipopolysaccharide-treated human macrophages, with some exception for the hsp1 mutant. Intriguingly, this clone also induced a higher IL10/IL12 ratio, which is assumed to indicate the anti-inflammatory potential of probiotics. Conclusions sHSP genes deletion determined some differences in gut stress resistance, cellular adhesion and immuno-modulation, also implying effects on in vivo interaction with the host. HSP1 might contribute to immunomodulatory mechanisms, though additional experiments are necessary to test this feature.

scholarly journals Aggravated Lyme Carditis in CD11a−/− and CD11c−/− Mice

2005 ◽  
Vol 73 (11) ◽  
pp. 7637-7643 ◽  
Author(s):  
Mireia Guerau-de-Arellano ◽  
Joseph Alroy ◽  
Daniel Bullard ◽  
Brigitte T. Huber
Keyword(s):  
Wild Type ◽  
Lyme Carditis ◽  
Disease Induction ◽  
In Vivo Analysis ◽  
The Common ◽  
Vitro Analysis ◽  
Β2 Integrins ◽  
In Vitro Analysis

ABSTRACT CD18 hypomorph mice expressing reduced levels of the common β2 integrin chain develop aggravated Lyme carditis, compared to that developed by wild-type (WT) mice, upon infection with the spirochete Borrelia burgdorferi. The enhancement of Lyme carditis in these mice is characterized by increased macrophage infiltration, correlating with augmented expression of the monocyte/macrophage chemoattractant protein 1 (MCP-1). The lack of CD18 results in the deficiency of all β2 integrins, i.e., CD11a/CD18 (LFA-1), CD11b/CD18 (Mac-1/CR3), CD11c/CD18 (p150,95/CR4), and CD11d/CD18. To determine the roles of the various β2 integrins in controlling the development of aggravated Lyme carditis, disease induction was analyzed in CD11a−/−, CD11b−/−, and CD11c−/− mice. CD11a−/− and CD11c−/− mice, but not CD11b−/− mice, developed aggravated Lyme carditis after exposure to B. burgdorferi. Similarly to CD18 hypomorph mice, CD11c−/− mice expressed higher levels of MCP-1, compared to both WT and CD11a−/− mice, as determined by in vitro analysis of MCP-1 secretion by bone marrow-derived dendritic cells and in vivo analysis of MCP-1 mRNA expression in B. burgdorferi-infected hearts. On the other hand, CD11a deficiency was associated with heightened heart B. burgdorferi burden relative to that of WT mice. Overall, our results suggest that the increased severity of Lyme carditis in CD18 hypomorph mice is caused by deficiency in CD11a or CD11c, possibly via different mechanisms.

scholarly journals Genetic Targeting of Relaxin and Insulin-Like Factor 3 Receptors in Mice

Endocrinology ◽  
2004 ◽  
Vol 145 (10) ◽  
pp. 4712-4720 ◽  
Author(s):  
Aparna A. Kamat ◽  
Shu Feng ◽  
Natalia V. Bogatcheva ◽  
Anne Truong ◽  
Colin E. Bishop ◽  
...  
Keyword(s):  
Knockout Mice ◽  
Peptide Hormone ◽  
Vascular Bundles ◽  
Biological Processes ◽  
Wild Type ◽  
Putative Receptor ◽  
Collagen Accumulation ◽  
Genetic Targeting ◽  
Deficient Mice

Abstract Relaxin (RLN) is a small peptide hormone that affects a variety of biological processes. Rln1 knockout mice exhibit abnormal nipple development, prolonged parturition, agerelated pulmonary fibrosis, and abnormalities in the testes and prostate. We describe here RLN receptor Lgr7-deficient mice. Mutant females have grossly underdeveloped nipples and are unable to feed their progeny. Some Lgr7−/− females were unable to deliver their pups. Histological analysis of Lgr7 mutant lung tissues demonstrates increased collagen accumulation and fibrosis surrounding the bronchioles and the vascular bundles, absent in wild-type animals. However, Lgr7-deficient males do not exhibit abnormalities in the testes or prostate as seen in Rln1 knockout mice. Lgr7-deficient females with additional deletion of Lgr8 (Great), another putative receptor for RLN, are fertile and have normal-sized litters. Double mutant males have normal-sized prostate and testes, suggesting that Lgr8 does not account for differences in Rln1−/− and Lgr7−/− phenotypes. Transgenic overexpression of Insl3, the cognate ligand for Lgr8, does not rescue the mutant phenotype of Lgr7-deficient female mice indicating nonoverlapping functions of the two receptors. Our data indicate that neither Insl3 nor Lgr8 contribute to the RLN signaling pathway. We conclude that the Insl3/Lgr8 and Rln1/Lgr7 actions do not overlap in vivo.

scholarly journals Impact of the Listeria monocytogenes Protein InlC on Infection in Mice

2013 ◽  
Vol 81 (4) ◽  
pp. 1334-1340 ◽  
Author(s):  
Nelly Leung ◽  
Antonella Gianfelice ◽  
Scott D. Gray-Owen ◽  
Keith Ireton
Keyword(s):  
Listeria Monocytogenes ◽  
Mutant Strain ◽  
Type Strain ◽  
Wild Type Strain ◽  
Bacterial Pathogen ◽  
Adaptor Protein ◽  
Single Amino Acid ◽  
Wild Type ◽  
Content Type

ABSTRACTThe bacterial pathogenListeria monocytogenescauses serious food-borne illnesses in pregnant women and the immunocompromised.L. monocytogenespromotes its internalization into host epithelial cells and then uses an F-actin-dependent motility process to spread from infected cells to surrounding healthy cells. In cultured enterocytes, efficient spread ofL. monocytogenesrequires the secreted bacterial protein InlC. InlC promotes dissemination by physically interacting with and antagonizing the function of the human adaptor protein Tuba. Here we examine the role of InlC and its interaction with host Tuba during infection in mice. The study took advantage of a single-amino-acid substitution (K173A) in InlC that impairs binding to human Tuba but does not affect InlC-mediated inhibition of the NF-κB pathway. Mice were inoculated intravenously with the wild-typeL. monocytogenesstrain EGD, an isogenic strain deleted for theinlCgene (ΔinlC), or a strain expressing K173A mutant InlC (inlC.K173A). The 50% lethal doses (LD50) for the ΔinlCorinlC.K173Amutant strain were approximately 4- or 6-fold greater than that for the wild-type strain, indicating a role forinlCin virulence. Compared to the wild-type strain, theinlC.K173Amutant strain exhibited lower bacterial loads in the liver. Histological analysis of livers indicated that the twoinlCmutant strains produced smaller foci of infection than did the wild-type strain. These smaller foci are consistent with a role for InlC in cell-to-cell spreadin vivo. Taken together, these results provide evidence that interaction of InlC with host Tuba is important for full virulence.

scholarly journals Involvement of CD14, Toll-Like Receptors 2 and 4, and MyD88 in the Host Response to the Fungal Pathogen Cryptococcus neoformans In Vivo

2004 ◽  
Vol 72 (9) ◽  
pp. 5373-5382 ◽  
Author(s):  
Lauren E. Yauch ◽  
Michael K. Mansour ◽  
Shmuel Shoham ◽  
James B. Rottman ◽  
Stuart M. Levitz
Keyword(s):  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Adaptor Protein ◽  
Interleukin 4 ◽  
Wild Type ◽  
Necrosis Factor Alpha ◽  
Factor Alpha ◽  
Histopathologic Analysis ◽  
And Control

ABSTRACT The major capsular polysaccharide of Cryptococcus neoformans, glucuronoxylomannan (GXM), is recognized by Toll-like receptor 2 (TLR2), TLR4, and CD14. In these studies, mice deficient in CD14, TLR2, TLR4, and the TLR-associated adaptor protein, MyD88, were utilized to investigate the contribution of TLRs and CD14 to in vivo host defenses against C. neoformans. MyD88−/− mice had significantly reduced survival compared with wild-type C57BL/6 mice after intranasal (i.n.) and intravenous (i.v.) infection with live C. neoformans. CD14−/− mice had reduced survival when infected i.v., while TLR2−/− mice died significantly earlier after i.n. infection. Mortality was similar comparing TLR4 mutant C3H/HeJ mice and control C3H/HeOuJ mice following i.v. or i.n. challenge with C. neoformans. The course of pulmonary cryptococcosis was studied in more detail in the CD14−/−, TLR2−/−, and MyD88−/− mice. MyD88−/− mice infected i.n. had higher numbers of CFU in the lungs as well as higher GXM levels in the sera and lungs 7 days after infection than wild-type mice did. Surprisingly, there were no major differences in the levels of tumor necrosis factor alpha, interleukin-4 (IL-4), IL-10, IL-12p70, or gamma interferon in the lungs of C. neoformans-infected knockout mice compared with wild-type mice. Histopathologic analysis of the lungs on day 7 postinfection revealed minimal inflammation in all mouse groups. These studies demonstrate a major role for MyD88 and relatively minor roles for CD14 and TLR2 in the response to cryptococcal infection, with the decreased survival of MyD88−/− mice correlating with increased numbers of lung CFU and serum and lung GXM levels.

scholarly journals RNA m6A methylation orchestrates cancer growth and metastasis via macrophage reprogramming

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Huilong Yin ◽  
Xiang Zhang ◽  
Pengyuan Yang ◽  
Xiaofang Zhang ◽  
Yingran Peng ◽  
...  
Keyword(s):  
Regulatory T Cell ◽  
Tumour Growth ◽  
Cancer Growth ◽  
Erk Pathway ◽  
Biological Processes ◽  
Wild Type ◽  
Potential Therapeutic Target ◽  
T Cell Infiltration ◽  
Deficient Mice

AbstractN6-methyladenosine (m6A) is a reversible mRNA modification that has been shown to play important roles in various biological processes. However, the roles of m6A modification in macrophages are still unknown. Here, we discover that ablation of Mettl3 in myeloid cells promotes tumour growth and metastasis in vivo. In contrast to wild-type mice, Mettl3-deficient mice show increased M1/M2-like tumour-associated macrophage and regulatory T cell infiltration into tumours. m6A sequencing reveals that loss of METTL3 impairs the YTHDF1-mediated translation of SPRED2, which enhances the activation of NF-kB and STAT3 through the ERK pathway, leading to increased tumour growth and metastasis. Furthermore, the therapeutic efficacy of PD-1 checkpoint blockade is attenuated in Mettl3-deficient mice, identifying METTL3 as a potential therapeutic target for tumour immunotherapy.

scholarly journals Neuroinflammatory processes are augmented in mice overexpressing human heat-shock protein B1 following ethanol-induced brain injury

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Brigitta Dukay ◽  
Fruzsina R. Walter ◽  
Judit P. Vigh ◽  
Beáta Barabási ◽  
Petra Hajdu ◽  
...  
Keyword(s):  
Brain Injury ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Glial Cell ◽  
Inflammatory Cytokines ◽  
Cell Activation ◽  
Mrna Level ◽  
Wild Type ◽  
Pro Inflammatory Cytokines

Abstract Background Heat-shock protein B1 (HSPB1) is among the most well-known and versatile member of the evolutionarily conserved family of small heat-shock proteins. It has been implicated to serve a neuroprotective role against various neurological disorders via its modulatory activity on inflammation, yet its exact role in neuroinflammation is poorly understood. In order to shed light on the exact mechanism of inflammation modulation by HSPB1, we investigated the effect of HSPB1 on neuroinflammatory processes in an in vivo and in vitro model of acute brain injury. Methods In this study, we used a transgenic mouse strain overexpressing the human HSPB1 protein. In the in vivo experiments, 7-day-old transgenic and wild-type mice were treated with ethanol. Apoptotic cells were detected using TUNEL assay. The mRNA and protein levels of cytokines and glial cell markers were examined using RT-PCR and immunohistochemistry in the brain. We also established primary neuronal, astrocyte, and microglial cultures which were subjected to cytokine and ethanol treatments. TNFα and hHSPB1 levels were measured from the supernates by ELISA, and intracellular hHSPB1 expression was analyzed using fluorescent immunohistochemistry. Results Following ethanol treatment, the brains of hHSPB1-overexpressing mice showed a significantly higher mRNA level of pro-inflammatory cytokines (Tnf, Il1b), microglia (Cd68, Arg1), and astrocyte (Gfap) markers compared to wild-type brains. Microglial activation, and 1 week later, reactive astrogliosis was higher in certain brain areas of ethanol-treated transgenic mice compared to those of wild-types. Despite the remarkably high expression of pro-apoptotic Tnf, hHSPB1-overexpressing mice did not exhibit higher level of apoptosis. Our data suggest that intracellular hHSPB1, showing the highest level in primary astrocytes, was responsible for the inflammation-regulating effects. Microglia cells were the main source of TNFα in our model. Microglia isolated from hHSPB1-overexpressing mice showed a significantly higher release of TNFα compared to wild-type cells under inflammatory conditions. Conclusions Our work provides novel in vivo evidence that hHSPB1 overexpression has a regulating effect on acute neuroinflammation by intensifying the expression of pro-inflammatory cytokines and enhancing glial cell activation, but not increasing neuronal apoptosis. These results suggest that hHSPB1 may play a complex role in the modulation of the ethanol-induced neuroinflammatory response.

scholarly journals The Serine Protease Autotransporter Pic Modulates Citrobacter rodentium Pathogenesis and Its Innate Recognition by the Host

2015 ◽  
Vol 83 (7) ◽  
pp. 2636-2650 ◽  
Author(s):  
Kirandeep Bhullar ◽  
Maryam Zarepour ◽  
Hongbing Yu ◽  
Hong Yang ◽  
Matthew Croxen ◽  
...  
Keyword(s):  
Serine Protease ◽  
Shigella Flexneri ◽  
Citrobacter Rodentium ◽  
Wild Type ◽  
Content Type ◽  
The Family ◽  
Toll Like Receptor 2 ◽  
Severe Colitis

Bacterial pathogens produce a number of autotransporters that possess diverse functions. These include the family of serine protease autotransporters ofEnterobacteriaceae(SPATEs) produced by enteric pathogens such asShigella flexneriand enteroaggregativeEscherichia coli. Of these SPATEs, one termed “protein involved in colonization,” or Pic, has been shown to possess mucinase activityin vitro, but to date, its role inin vivoenteric pathogenesis is unknown. Testing apicnull (ΔpicC) mutant inCitrobacter rodentium, a natural mouse pathogen, found that theC. rodentiumΔpicCstrain was impaired in its ability to degrade mucinin vitrocompared to the wild type. Upon infection of mice, the ΔpicCmutant exhibited a hypervirulent phenotype with dramatically heavier pathogen burdens found in intestinal crypts. ΔpicCmutant-infected mice suffered greater barrier disruption and more severe colitis and weight loss, necessitating their euthanization between 10 and 14 days postinfection. Notably, the virulence of the ΔpicCmutant was normalized when thepicCgene was restored; however, a PicC point mutant causing loss of mucinase activity did not replicate the ΔpicCphenotype. Exploring other aspects of PicC function, the ΔpicCmutant was found to aggregate to higher levelsin vivothan wild-typeC. rodentium. Moreover, unlike the wild type, theC. rodentiumΔpicCmutant had a red, dry, and rough (RDAR) morphologyin vitroand showed increased activation of the innate receptor Toll-like receptor 2 (TLR2). Interestingly, theC. rodentiumΔpicCmutant caused a degree of pathology similar to that of wild-typeC. rodentiumwhen infecting TLR2-deficient mice, showing that despite its mucinase activity, PicC's major rolein vivomay be to limitC. rodentium's stimulation of the host's innate immune system.

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