Phosphorylation of PRAS40 contributes to the activation of the PI3K/AKT/mTOR signaling pathway and the inhibition of autophagy following status epilepticus in rats.

Status epilepticus (SE) is a neurological disorder associated with high morbidity and mortality rates, and is often difficult to treat. Moreover, the underlying mechanism of SE remains unknown. The lithium-pilocarpine model is a validated animal model that can reproduce the main clinical and neuropathological features of SE. In the present study, this SE model was utilized and SE was successfully established in rats, as determined by the corresponding epileptic electroencephalogram. Histology, immunohistochemistry, western blot analysis and co-immunoprecipitation were used to detect the phosphorylation (p-) of AKT substrate of 40 kDa (PRAS40), the combination of p-PRAS40 and 14-3-3 protein and the activation of the PI3K/mTOR signaling pathway in SE. In addition, the present study analyzed the dynamics of the expression of autophagy-associated factors in the hippocampus after SE induction, and the influence of suppressing the p- of PRAS40 on the autophagy process was detected in the pathogenesis of SE. The results indicated that increased p-PRAS40 expression could activate the mTOR pathway to decrease the level of autophagy. However, inhibition of the mTOR signaling pathway promoted autophagy flux. These results may provide further understanding of p-PRAS40 functions in SE.

Expression profiling of circular RNAs and their potential role in early­stage diabetic cardiomyopathy.

Diabetic cardiomyopathy (DCM) is a severe cardiovascular complication of diabetes mellitus (DM). Detecting DCM during the early stages of the disease remains a challenge, as the molecular mechanisms underlying early­stage DCM are not clearly understood. Circular RNA (circRNA), a type of non­coding RNA, has been confirmed to be associated with numerous diseases. However, it is still unclear how circRNAs are involved in early­stage DCM. In the present study, heart tissues harvested from BKS­db/db knock­out mice were identified through high­throughput RNA sequencing technology. A total of 58 significantly differentially expressed circRNAs were identified in the db/db sample. Among these, six upregulated circRNAs and seven downregulated circRNAs were detected by reverse transcription­quantitative PCR and analyzed using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Furthermore, based on the predicted binding site with microRNAs (miRNAs) involved in DCM, five circRNAs (mmu_circ_0000652, mmu_circ_0000547, mmu_circ_0001058, mmu_circ_0000680 and novel_circ_0004285) were shown to serve as competing endogenous (ce)RNAs. The corresponding miRNAs and mRNAs of the ceRNAs were also verified, and two promising circRNA­miRNA­mRNA regulatory networks were determined. Finally, internal ribosome entry site prediction combined with open reading frame prediction indicated that it was highly possible that mmu_circ_0001160 encoded a protein. In the present study, a comprehensive analysis of the circRNA expression profile during the early phase of DCM was performed, and two promising circRNA­miRNA­mRNA regulatory networks were identified. These results lay the foundation for unravelling the underlying pathogenesis of DCM, and highlight potential biomarkers and therapeutic targets for the treatment of DCM at an early stage.

Aberrant endoplasmic reticulum stress mediates coronary artery spasm through regulating MLCK/MLC2 pathway.

Coronary artery spasm (CAS) is a pathophysiological phenomenon that may cause myocardial infarction and lead to circulatory collapse and death. Aberrant endoplasmic reticulum (ER) stress causes accumulation of misfolding proteins and has been reported to be involved in a variety of vascular diseases. The present study investigated the role of ER stress in the development of CAS and explored the possible molecular mechanisms. Initially, it was found that ER stress markers were elevated in response to drug-induced vascular smooth muscle cells (VSMCs) contraction. Pharmacologic activation of ER stress using Tunicamycin (Tm) persistently induced CAS and significantly promoted Pituitrin-induced CAS in mice as well as in a collagen gel contraction assay. On the contrary, pharmacologic inhibition of ER stress using 4-phenylacetic acid (4-PBA) completely blunted Pituitrin-induced CAS development in mice. Moreover, during the drug-induced VSMCs contraction, expression of ER stress markers were increased in parallel to those of myosin light chain kinase (MLCK) and phosphor-MLC2 (p-MLC2, at Ser19). After inhibiting MLCK activity by using its specific inhibitor ML-7, the ER stress activator Tm failed to activate the MLCK/MLC2 pathway and could neither trigger CAS in mice nor induce VSMCs contraction in vitro. Our results suggested that aberrant ER stress mediated CAS via regulating the MLCK/MLC2 pathway. ER stress activators might be more robust than the common drugs (Pituitrin or acetylcholine) as to induce vasocontraction and thus may serve as potential therapeutics against chronic bleeding, while its inhibitor might be useful for treatment of severe CAS caused by other medication.

Evaluation of the inclusion of circular RNAs in mRNA profiling in forensic body fluid identification.

The use of messenger RNA (mRNA) profiling is considered a promising method in the identification of forensically relevant body fluids which can provide crucial information for reconstructing a potential crime. However, casework samples are usually of limited quantity or have been subjected to degradation, which requires improvement of body fluid identification. Circular RNAs (circRNAs), a class of products from the backsplicing of pre-mRNAs, are shown to have high abundance, remarkable stability, and cell type-specific expression in human cells. In this study, we investigated whether the inclusion of circRNAs in mRNA profiling improve the detection of biomarkers including δ-aminolevulinate synthase 2 (ALAS2) and matrix metallopeptidase 7 (MMP7) in body fluid identification. The major circRNAs of ALAS2 and MMP7 were first identified and primer sets for the simultaneous detection of linear and circular transcripts were developed. The inclusion of circRNAs in mRNA profiling showed improved detection sensitivity and stability of biomarkers revealed by using serial dilutions, mixed samples, and menstrual bloodstains as well as degraded and aged samples. Therefore, the inclusion of circRNAs in mRNA profiling should facilitate the detection of mRNA markers in forensic body fluid identification.

Axin1 up-regulated 1 accelerates stress-induced cardiomyocytes apoptosis through activating Wnt/ß-catenin signaling.

Stress-induced cardiomyocyte apoptosis contributes to the pathogenesis of a variety of cardiovascular diseases, but how stress induces cardiomyocyte apoptosis remains largely unclear. The present study aims to investigate the effects of Axin1 up-regulated 1 (Axud1), a novel pro-apoptotic protein, on the cardiomyocyte survival and the underlying mechanisms. To this end, a rat model under restraint stress (RS) was established and in vitro stress-induced cardiomyocytes culture was achieved. Our data showed that Axud1 was upregulated in the rat myocardia after exposure to RS. Anti-apoptotic Bcl-2 was decreased, whereas pro-apoptotic Bax and Cleaved caspase-3 (Cc3) were increased in a time-dependent manner. The Wnt/ß-catenin signaling was observed to be interestingly activated in heart undergoing RS. In addition, the treatment of norepinephrine (NE) to in vitro cardiomyocytes increased Axud1 level and induced cell apoptosis. Wnt/ß-catenin signaling was consistently activated. Knockdown of Axud1 using specific siRNA blunted NE-induced cardiomyocytes apoptosis and also inactivated the Wnt/ß-catenin signaling. XAV-939, an inhibitor of Wnt/ß-catenin signaling, partially reversed the pro-apoptotic effect of NE. In conclusion, Axud1 accelerated stress-induced cardiomyocytes apoptosis through activation of Wnt/ß-catenin signaling pathway. Our data provided novel evidence that therapeutic strategies against Axud1 or Wnt/ß-catenin signaling might be promising in relation to RS-induced myocardial injury.

Demographic, clinical and pathological features of sudden deaths due to myocarditis: Results from a state-wide population-based autopsy study.

Causes of sudden cardiac deaths have been widely reported with limited data focused specifically on myocarditis. A retrospective review of cases from the Office of the Chief Medical Examiner (OCME), State of Maryland yielded a total of 103 sudden unexpected deaths (SUDs) due to myocarditis (0.17% of all SUDs and 0.70% of autopsied SUDs) from 2005 through 2014. Most deaths occurred in patients <30 years of age with a male:female ratio 1.3:1. Of the 103 cases, 45 (43.7%) patients were witnessed collapsed. Four deaths occurred during exertion, such as exercising at the gym or performing heavy physical work, and 2 deaths were associated with emotional stress. The common cardiac macroscopic findings included ventricular dilatation (39.8%), mild coronary stenosis (17.5%), mottled myocardial appearance (15.5%), and myocardial fibrosis (10.7%). The histological classification of myocarditis was based on the predominant type of inflammatory cell infiltration. In our study group, lymphocytic myocarditis was most common, accounting for 56 cases (54.4%), followed by neutrophilic (32 cases, 31.7%), eosinophilic (13 cases, 12.6%) and giant cell type (2 cases, 1.9%). Microscopic examination revealed myocyte necrosis in 69 cases (67.0%) and interstitial or perivascular fibrosis in 48 cases (46.6%). The percentage of myocyte necrosis was 75.0% (42/58 cases) in lymphocytic, 65.6% (21/31 cases) in neutrophilic, 30.8% (4/13 cases) in eosinophilic, and 100% (2/2 cases) in giant cell myocarditis. Determination of myocarditis as cause of death continues to present a major challenge to forensic pathologists, because histopathologic findings can be subtle and the diagnosis of myocarditis remains difficult.

Association of rs1122608 with Coronary Artery Disease and Lipid Profile: A Meta-analysis.

BACKGROUND AND AIMS: It has been reported that rs1122608 adjacent to low-density lipoprotein cholesterol receptor (LDLR) locus is associated with the risk of coronary artery disease (CAD) and blood lipid profile in the Caucasian population. Due to the contradictory results in the Asian population, we conducted a meta-analysis to systematically summarize and clarify the association between rs1122608 with CAD risk and lipid profile. METHODS: A systematic search regarding studies on the association of rs1122608 with CAD risk and lipid profile was conducted in databases including PubMed, Embase, and Cochrane library. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were used to pool the effect size. RESULTS: A total of five case-control studies were included in this study. A statistically significant association was identified between rs1122608-G allele and CAD risk in overall analysis (OR = 2.09, 95% CI 1.48-2.97) and in both Asian (OR = 1.82, 95% CI 1.04-3.18) and Caucasian subgroups (OR = 2.31, 95% CI 1.48-3.60). The rs1122608-G allele was associated with increased triglyceride (TG) level (OR = 1.25, 95% CI 1.03-1.52), but not with total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), or LDL cholesterol level. Moreover, the rs1122608-G allele is associated with increased CAD risk in the Asian male population (OR = 3.37, 95% CI 1.51-9.86) but not in the Asian female population. CONCLUSIONS: The rs1122608 is associated with the risk of CAD and TG level. The rs1122608-G allele was a significant risk factor of CAD in the Asian male population but not in the Asian female population.

RNA degradation as described by a mathematical model for postmortem interval determination.

Precisely determining the postmortem interval (PMI) is crucial to civil, criminal and forensic cases. A technique to exploit the postmortem RNA transcript level was developed to increase the accuracy and practicality of PMI estimation. For this purpose, lung tissues and muscle tissues were removed at twelve time points (0-144 h) from rat corpses that had been stored at three different temperatures (10, 20 and 30 °C). Human tissues were collected at autopsy from twelve real cases with known PMI values and other parameters. After the RNA was extracted from all these samples, the transcript levels of nine biomarkers were analyzed by real-time quantitative PCR (RT-qPCR). With the assistance of geNorm, miR-195, miR-200c, 5S, U6 and RPS29 were selected as reference biomarkers for lung specimens; miR-1, miR-206, 5S and RPS29 were chosen as control markers for muscle tissues. On the contrary, ACTB and GAPDH were significantly correlated with the PMI. The mathematical models using these target biomarkers were constructed to describe the characteristic relationship between △Ct values (normalized to reference biomarkers) and the observed PMI for each temperature group. Following validation, the relatively low error rates (7.4% and 12.5% for rat and human samples, respectively) demonstrated the accuracy and reliability of the mathematical model. We believe these results indicate that the multi-parametric mathematical model can become a practical tool for PMI estimation.

Phosphorylated Myosin Light Chain 2 (p-MLC2) as a Molecular Marker of Antemortem Coronary Artery Spasm.

BACKGROUND It is not uncommon that only mild coronary artery stenosis is grossly revealed after a system autopsy. While coronary artery spasm (CAS) is the suspected mechanism of these deaths, no specific biomarker has been identified to suggest antemortem CAS. MATERIAL AND METHODS To evaluate the potential of using phosphorylated myosin light chain 2 (p-MLC2) as a diagnostic marker of antemortem CAS, human vascular smooth muscle cells (VSMCs) were cultured and treated with common vasoconstrictors, including prostaglandins F2α (PGF2α), acetylcholine (ACh), and 5-hydroxy tryptamine (5-HT). The p-MLC2 level was examined in the cultured cells using Western blot analysis and in a rat model of spasm provocation tests using immunohistochemistry (IHC). Effects of increased p-MLC2 level on VSMCs contractile activities were assessed in vitro using confocal immunofluorescence assay. Four fatal cases with known antemortem CAS were collected and subject to p-MLC2 detection. RESULTS The p-MLC2 was significantly increased in VSMCs after treatments with vasoconstrictors and in the spasm provocation tests. Myofilament was well-organized and densely stained in VSMCs with high p-MLC2 level, but disarrayed in VSMCs with low p-MLC2 level. Three of the 4 autopsied cases showed strongly positive staining of p-MLC2 at the stenosed coronary segment and the adjacent interstitial small arteries. The fourth case was autopsied at the 6th day after death and showed negative-to-mild positive staining of p-MLC2. CONCLUSIONS p-MLC2 might be a useful marker for diagnosis of antemortem CAS. Autopsy should be performed as soon as possible to collect coronary arteries for detection of p-MLC2.

Morphologic features of culprit lesions in sudden coronary death with family history of premature coronary artery disease.

The morphologic features of familial coronary artery disease (CAD) resulting in sudden coronary death (SCD) are poorly studied. The presence and type of culprit lesions may have important implications in the genetic basis for familial heart disease. Autopsies of SCD victims over a 5-year period from a statewide medical examiner's office were studied. Premature familial disease was defined as sudden death at ≤50 years in women and ≤45 years in men, with premature SCD or acute coronary syndrome in a first-degree relative. Culprit lesion was defined as acute plaque rupture, plaque erosion, and severe narrowing without thrombus (stable plaque). There were 174 acute plaque ruptures (age 49±10 years, 9% women), 49 plaque erosions (age 45±8 years, 37% women), and 213 stable plaques (age 53±11 years, 22% women). There were 8 plaque rupture with family history. There were 9 plaque erosions with family history. There were 7 stable plaques with family history. The rate of familial history in premature coronary disease was 18.4% in erosions, 4.6% in ruptures (p=.02 vs. erosion), and 3.3% in stable plaque (p=.002 vs. erosion). We concluded that the frequency of family history of premature sudden death due to CAD may be higher in plaque erosion as compared to patients dying with acute plaque rupture or stable plaque.

Mutation analysis of 19 autosomal short tandem repeats in Chinese Han population from Shanghai.

The mutation of short tandem repeat (STR) loci is affected by several factors, such as sex, age, and DNA architectures. Previous studies have shown a different profile of mutation rates at autosomal STR loci among populations. It is important to provide population data and reveal underlying factors influencing the evaluation of STR mutation rates. In this study, we performed a comprehensive analysis on the mutation of 19 autosomal STR loci through 124,773 parent-child allelic transfers from 5846 paternity testing cases. A total of 197 mutations were observed including 187 single-step mutations. The observed mutation rates ranged from 0.15 × 10-3 (TH01) to 4.57 × 10-3 (FGA), and the average mutation rate across all the 19 loci was 1.58 × 10-3. Furthermore, the average mutation rate of STR loci increases with the paternal conception ages and remains relatively stable in different maternal age groups, which suggest the profile of paternal conception ages as a potential factor influencing the evaluation of STR mutation rates and the ratio of paternal versus maternal mutation rate in populations. Multidimensional scaling analysis (MDS) shows a difference in the profile of mutation rates at 13 CODIS STR loci among ethnical groups. Based on our data, our results support that short alleles are biased towards expansion mutation and longer alleles favor contraction mutation. In conclusion, our results provide useful information for further investigation on STR mutation in forensic genetics and population genetics.

MicroRNA-19b Downregulates Gap Junction Protein Alpha1 and Synergizes with MicroRNA-1 in Viral Myocarditis.

Viral myocarditis (VMC) is a life-threatening disease that leads to heart failure or cardiac arrhythmia. A large number of researches have revealed that mircroRNAs (miRNAs) participate in the pathological processes of VMC. We previously reported that miR-1 repressed the expression of gap junction protein α1 (GJA1) in VMC. In this study, miR-19b was found to be significantly upregulated using the microarray analysis in a mouse model of VMC, and overexpression of miR-19b led to irregular beating pattern in human cardiomyocytes derived from the induced pluripotent stem cells (hiPSCs-CMs). The upregulation of miR-19b was associated with decreased GJA1 in vivo. Furthermore, a miR-19b inhibitor increased, while its mimics suppressed the expression of GJA1 in HL-1 cells. When GJA1 was overexpressed, the miR-19b mimics-mediated irregular beating was reversed in hiPSCs-CMs. In addition, the effect of miR-19b on GJA1 was enhanced by miR-1 in a dose-dependent manner. These data suggest miR-19b contributes to irregular beating through regulation of GJA1 by cooperating with miR-1. Based on the present and our previous studies, it could be indicated that miR-19b and miR-1 might be critically involved in cardiac arrhythmia associated with VMC.

Induction of miRNA-181a by genotoxic treatments promotes chemotherapeutic resistance and metastasis in breast cancer.

Acquired therapeutic resistance is the major drawback to effective systemic therapies for cancers. Aggressive triple-negative breast cancers (TNBC) develop resistance to chemotherapies rapidly, whereas the underlying mechanisms are not completely understood. Here we show that genotoxic treatments significantly increased the expression of miR-181a in TNBC cells, which enhanced TNBC cell survival and metastasis upon Doxorubicin treatment. Consistently, high miR-181a level associated with poor disease free survival and overall survival after treatments in breast cancer patients. The upregulation of miR-181a was orchestrated by transcription factor STAT3 whose activation depended on NF-κB-mediated IL-6 induction in TNBC cells upon genotoxic treatment. Intriguingly, activated STAT3 not only directly bound to MIR181A1 promoter to drive transcription but also facilitated the recruitment of MSK1 to the same region where MSK1 promoted a local active chromatin state by phosphorylating histone H3. We further identified BAX as a direct functional target of miR-181a, whose suppression decreased apoptosis and increased invasion of TNBC cells upon Dox treatment. These results were further confirmed by evidence that suppression of miR-181a significantly enhanced therapeutic response and reduced lung metastasis in a TNBC orthotopic model. Collectively, our data suggested that miR-181a induction had a critical role in promoting therapeutic resistance and aggressive behavior of TNBC cells upon genotoxic treatment. Antagonizing miR-181a may serve as a promising strategy to sensitize TNBC cells to chemotherapy and mitigate metastasis.

Exploration of the R code-based mathematical model for PMI estimation using profiling of RNA degradation in rat brain tissue at different temperatures.

Precise estimation of postmortem interval (PMI) is crucial in some criminal cases. This study aims to find some optimal markers for PMI estimation and build a mathematical model that could be used in various temperature conditions. Different mRNA and microRNA markers in rat brain samples were detected using real-time fluorescent quantitative PCR at 12 time points within 144 h postmortem and at temperatures of 4, 15, 25, and 35 °C. Samples from 36 other rats were used to verify the animal mathematical model. Brain-specific mir-9 and mir-125b are effective endogenous control markers that are not affected by PMI up to 144 h postmortem under these temperatures, whereas the commonly used U6 is not a suitable endogenous control in this study. Among all the candidate markers, ΔCt (ß-actin) has the best correlation coefficient with PMI and was used to build a new model using R software which can simultaneously manage both PMI and temperature parameters. This animal mathematical model is verified using samples from 36 other rats and shows increased accuracy for higher temperatures and longer PMI. In this study, ß-actin was found to be an optimal marker to estimate PMI and some other markers were found to be suitable to act as endogenous controls. Additionally, we have used R code software to build a model of PMI estimation that could be used in various temperature conditions.

Mutations of desmoglein-2 in sudden death from arrhythmogenic right ventricular cardiomyopathy and sudden unexplained death.

Desmoglein-2 (DSG2), a member of the desmosomal cadherin superfamily, has been linked to arrhythmogenic right ventricular cardiomyopathy (ARVC)which may cause life-threatening ventricular arrhythmias and sudden death. Fatal arrhythmias resulting in sudden death also occur in the absence of morphologic cardiac abnormalities at autopsy. We sequenced all 15 exons of DSG2 in DNA extracted from post-mortem heart tissues of 25 patients dying with ARVC and 25 from sudden unexplained death (SUD). The primers were designed using the Primer Express 3.0 software. Direct sequencing for both sense and antisense strands was performed with a BigDye Terminator DNA sequencing kit on a 3130 xl Genetic Analyzer. Mutation damage prediction was made using Mutation Taster, Polyphen and SIFT software. 2 DSG2 mutations (p. S1026Q fsX12, p. G678R)in two ARVC samples and 2 DSG2 mutations(p. E 896K, p. A858 V) in two SUD samples were identified, all the mutations were novel. We concluded that DSG2 mutations may not specific for ARVC and may be related to the fatal arrhythmic events even in patients with a morphological normal heart.

Deregulation of RGS17 Expression Promotes Breast Cancer Progression.

OBJECTIVE: A high level of RGS17 expression is observed in diverse human cancers and correlates with tumor progression. Herein, we aim to investigate its expression and function in breast cancer. METHODS: The expression of RGS17 was detected by immunohistochemical analysis and western blot analysis. The level of miR-32 expression was investigated by qRT-PCR. Western blot analysis was used to determine the relationship between RGS17 and miR-32. A series of loss or gain of function assays was performed to measure the effects of RGS17 or miR-32 on tumor migration, invasion, and proliferation. RESULTS: Compared to that in normal breast specimen, the expression of RGS17 had a significantly higher expression level in breast cancer tissues and cell lines. Although the potential relationship of RGS17 expression with clinicopathological features was not observed, there was a significant correlation of RGS17 expression with p63 expression. In cells, inhibition of RGS17 expression impaired cell migration, invasion, and proliferation. Further, RGS17 was identified as a direct and functional target of miR-32. Overexpression of miR-32 in cells could decrease the expression of RGS17 and inhibit cell migration, invasion, and proliferation. In contrast, ectopic expression of RGS17 could attenuate phenotypes caused by miR-32 overexpression. CONCLUSION: The expression of RGS17 was upregulated in breast cancer, which could enhance cell migration, invasion, and proliferation. Moreover, the RGS17 was identified as a target of miR-32. Our results suggest that RGS17 might play an important role in breast cancer progression and could be a potential target for human breast cancer treatment.

Magnetic Resonance Spectrum Technique in the Follow-up of an Ulnar Nerve Injured Patient.

A 49-year-old Chinese man sustained laceration of the right forearm by a dagger, with his right ulnar nerve completely transected. Four months postinjury, he underwent surgery to repair the nerve. He was examined by electromyogram, nerve conduction velocity, magnetic resonance imaging, and proton magnetic resonance spectroscopy ((1)H-MRS) 6, 12, 18, and 24 months after the injury. Before surgery, intramyocellular lipid (IMCL)/creatine (Cr) and extramyocellular lipid (EMCL)/Cr were observed to be higher than those of the uninjured side. During the recovery, IMCL/Cr and EMCL/Cr became lower and closer to the uninjured side. This case demonstrates that the change of IMCL/Cr and EMCL/Cr may be related to the recovery of peripheral nerve.

TRAF Family Member-associated NF-κB Activator (TANK) Inhibits Genotoxic Nuclear Factor κB Activation by Facilitating Deubiquitinase USP10-dependent Deubiquitination of TRAF6 Ligase.

DNA damage-induced NF-κB activation plays a critical role in regulating cellular response to genotoxic stress. However, the molecular mechanisms controlling the magnitude and duration of this genotoxic NF-κB signaling cascade are poorly understood. We recently demonstrated that genotoxic NF-κB activation is regulated by reversible ubiquitination of several essential mediators involved in this signaling pathway. Here we show that TRAF family member-associated NF-κB activator (TANK) negatively regulates NF-κB activation by DNA damage via inhibiting ubiquitination of TRAF6. Despite the lack of a deubiquitination enzyme domain, TANK has been shown to negatively regulate the ubiquitination of TRAF proteins. We found TANK formed a complex with MCPIP1 (also known as ZC3H12A) and a deubiquitinase, USP10, which was essential for the USP10-dependent deubiquitination of TRAF6 and the resolution of genotoxic NF-κB activation upon DNA damage. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated deletion of TANK in human cells significantly enhanced NF-κB activation by genotoxic treatment, resulting in enhanced cell survival and increased inflammatory cytokine production. Furthermore, we found that the TANK-MCPIP1-USP10 complex also decreased TRAF6 ubiquitination in cells treated with IL-1ß or LPS. In accordance, depletion of USP10 enhanced NF-κB activation induced by IL-1ß or LPS. Collectively, our data demonstrate that TANK serves as an important negative regulator of NF-κB signaling cascades induced by genotoxic stress and IL-1R/Toll-like receptor stimulation in a manner dependent on MCPIP1/USP10-mediated TRAF6 deubiquitination.