Search

XLS file - 204K, Genomic regions enriched in MLLN, AF4C, H3K79me2 and the common of the three ChIP-seq in RS4;11 cells

XLS file - 330K, Genes included in BCR-ABL1, E2A-PBX1 or MLLr DNA methylation signatures vs. normal pre-B cells

XLS file - 60K, Characteristics of the 215 E2993 B-ALL patients and the 12 normal bone marrow samples (excel file)

PDF file - 839K

XLS file - 74K, DNA methylation and gene expression signature of BCR-ABL1-positive B-ALL

Canonically, EZH2 serves as the catalytic subunit of PRC2, which mediates H3K27me3 deposition and transcriptional repression. Here, we report that in acute leukaemias, EZH2 has additional noncanonical functions by binding cMyc at non-PRC2 targets and uses a hidden transactivation domain (TAD) for (co)activator recruitment and gene activation. Both canonical (EZH2-PRC2) and noncanonical (EZH2-TAD-cMyc-coactivators) activities of EZH2 promote oncogenesis, which explains the slow and ineffective antitumour effect of inhibitors of the catalytic function of EZH2. To suppress the multifaceted activities of EZH2, we used proteolysis-targeting chimera (PROTAC) to develop a degrader, MS177, which achieved effective, on-target depletion of EZH2 and interacting partners (that is, both canonical EZH2-PRC2 and noncanonical EZH2-cMyc complexes). Compared with inhibitors of the enzymatic function of EZH2, MS177 is fast-acting and more potent in suppressing cancer growth. This study reveals noncanonical oncogenic roles of EZH2, reports a PROTAC for targeting the multifaceted tumorigenic functions of EZH2 and presents an attractive strategy for treating EZH2-dependent cancers.

Although we have gained insight into the biological and biochemical effects of natural sunlight exposure on prokaryotes, little is known about sunlight exposure on natural virus communities. To fully understand the mechanics of microbial food webs, it is essential to understand the dynamics of viral infection of bacteria and its role in bacteria-mediated processes when sunlight is present or not. An investigation of the effects of sunlight and darkness treatments on viral communities, viral production and decay rates in Kenting coastal waters was conducted. A linear increase in viral abundance was observed in the darkness treatment as expected in the viral production experiment, the average rate of net viral production in the darkness treatment was 0.018 × 106 viruses mL–1 h− 1. However, within the first 12 hours after exposure to sunlight, viral abundance increased non-significantly in the viral production experiments. Further, averaged value for viral decay in the sunlight treatment was nearly 0.038×106 virus mL− 1h− 1. The rate of viral decay almost equaled the rate of gross viral production under natural sunlight treatment, resulting in gross viral production of nearly 0.038×106 virus mL− 1h− 1. Furthermore, we estimated that the gross viral production was nearly 0.018×106 virus mL− 1h− 1 in the darkness treatment. As a result, sunlight damages a large portion of the natural viral community, affecting the role viruses play in food webs. We understanding how viruses change under significant sunlight pressures suggests considering the temporal variations of viral production in relation to diel variations.

Recent evidence suggests that melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs), a neuronal class regulating non-image forming (NIF) vision and generally thought to be injury resistant, are dysfunctional in certain neurodegenerative diseases. Although disrupted NIF visual functions have been reported in patients and animals with diabetes, it remains controversial whether ipRGCs exhibit remodeling during diabetes and if so, whether such remodeling is variable among ipRGC subtypes. Here, we demonstrate that survival, soma-dendritic profiles, and melanopsin-based functional activity of M1 ipRGCs were unaltered in streptozotocin-induced 3-month diabetic mice. Such resistance remained at 6 months after streptozotocin administration. In contrast, M2/M3 ipRGCs underwent significant remodeling in diabetic mice, manifested by enlarged somata and increased dendritic branching complexity. Consistent with the unaltered melanopsin levels, the sensitivity of melanopsin-based activity was unchanged in surviving M2 cells, but their response gain displayed a compensatory enhancement. Meanwhile, the pupillary light reflex, a NIF visual function controlled by M2 cells, was found to be impaired in diabetic animals. The resistance of M1 cells might be attributed to the adjacency of their dendrites to capillaries, which makes them less disturbed by the impaired retinal blood supply at the early stage of diabetes.

The coronavirus disease 2019 (COVID-19) pandemic caused by 2019 novel coronavirus (2019-nCoV) has been a crisis of global health, whereas the effective vaccines against 2019-nCoV are still under development. Alternatively, utilization of old drugs or available medicine that can suppress the viral activity or replication may provide an urgent solution to suppress the rapid spread of 2019-nCoV. Andrographolide is a highly abundant natural product of the medicinal plant, Andrographis paniculata, which has been clinically used for inflammatory diseases and anti-viral therapy. We herein demonstrate that both andrographolide and its fluorescent derivative, the nitrobenzoxadiazole-conjugated andrographolide (Andro- NBD), suppressed the main protease (Mpro) activities of 2019-nCoV and severe acute respiratory syndrome coronavirus (SARS-CoV). Moreover, Andro-NBD was shown to covalently link its fluorescence to these proteases. Further mass spectrometry (MS) analysis suggests that andrographolide formed a covalent bond with the active site Cys145 of either 2019-nCoV Mpro or SARS-CoV Mpro. Consistently, molecular modeling analysis supported the docking of andrographolide within the catalytic pockets of both viral Mpros. Considering that andrographolide is used in clinical practice with acceptable safety and its diverse pharmacological activities that could be beneficial for attenuating COVID-19 symptoms, extensive investigation of andrographolide on the suppression of 2019-nCoV as well as its application in COVID-19 therapy is suggested., Highlights • 2019-nCoV and SARS-CoV main proteases exhibit similar enzyme kinetics. • Andrographolide inhibits enzyme activity of 2019-nCoV and SARS-CoV main proteases. • Andrographolide is covalently linked to active cysteine of 2019-nCoV main protease.

Bioallethrin belongs to the family of pyrethroid insecticides. Previous studies have shown that bioallethrin affected the function of muscarinic receptor and subsequently induced neurotoxicity in different brain models. Reactive oxygen species (ROS) are generated in the metabolic course of the human body, which can cause human damage when overactivated. However, whether bioallethrin evokes cytotoxicity through ROS signaling and whether the antioxidant Vitamin E (VE) protects these cytotoxic responses in human glial cell model are still elusive. This study investigated the effect of bioallethrin on cytotoxicity through ROS signaling and evaluated the protective effect of the antioxidant VE in DBTRG-05MG human glioblastoma cells. The cell counting kit-8 (CCK-8) was used to measure cell viability. Intracellular ROS and glutathione (GSH) levels were measured by a cellular assay kit. The levels of apoptosis- and antioxidant-related protein were analyzed by Western blotting. In DBTRG-05MG cells, bioallethrin (25-75 μM) concentration-dependently induced cytotoxicity by increasing ROS productions, decreasing GSH contents, and regulating protein expressions related to apoptosis or antioxidation. Furthermore, these cytotoxic effects were partially reversed by VE (20 μM) pretreatment. Together, VE partially lessened bioallethrin-induced apoptosis through oxidative stress in DBTRG-05MG cells. The data assist us in identifying the toxicological mechanism of bioallethrin and offer future development of the antioxidant VE to reduce brain damage caused by bioallethrin.

In spite of the fact that the interactions between environmental parameters and prokaryotic and viral abundance have been explored in various aquatic environments, only a few independent estimates of viral production and decay in the estuarine region have been explored. In this study, data were analyzed for viral and prokaryotic abundance, viral production, and viral decay in a subtropical Danshui estuary in summer 2021. Prokaryotic abundance varied from 2.4 ± 0.6 × 105 to 12 ± 2.3 × 105 cells mL−1, and viral abundance ranged from 2.3 ± 0.9 × 105 to 6.9 ± 1.3 × 105 viruses mL−1 during the study period. Viral abundance was significantly correlated with prokaryotic abundance and chlorophyll a concentration. Furthermore, studies of changes in viral to prokaryotic abundance ratio (VPR) ranged from 0.42 ± 0.11 to 2.0 ± 0.25. Viral decay values were 2.1 ± 0.5 and 2.1 ± 0.3 × 104 virus mL−1h−1, and non-significant differences were observed between the inner estuary and coastal water region. Viral decay almost balanced gross viral production in this study. The dilution experiments revealed non-significant net viral production in July; thus, a lower VPR might be explained in this estuarine environment.

Trimethylated histone H3 lysine 27 (H3K27me3) regulates gene repression, cell-fate determination and differentiation. We report that a conserved bromo-adjacent homology (BAH) module of BAHCC1 (BAHCC1BAH) 'recognizes' H3K27me3 specifically and enforces silencing of H3K27me3-demarcated genes in mammalian cells. Biochemical, structural and integrated chromatin immunoprecipitation-sequencing-based analyses demonstrate that direct readout of H3K27me3 by BAHCC1 is achieved through a hydrophobic trimethyl-L-lysine-binding 'cage' formed by BAHCC1BAH, mediating colocalization of BAHCC1 and H3K27me3-marked genes. BAHCC1 is highly expressed in human acute leukemia and interacts with transcriptional corepressors. In leukemia, depletion of BAHCC1, or disruption of the BAHCC1BAH-H3K27me3 interaction, causes derepression of H3K27me3-targeted genes that are involved in tumor suppression and cell differentiation, leading to suppression of oncogenesis. In mice, introduction of a germline mutation at Bahcc1 to disrupt its H3K27me3 engagement causes partial postnatal lethality, supporting a role in development. This study identifies an H3K27me3-directed transduction pathway in mammals that relies on a conserved BAH 'reader'.

The experiment of flow past a freely suspended circular cylinder in the wake of an upstream stationary cylinder was carried out in a re-circulating water channel using the Particle Image Velocimetry (PIV) technique. The upstream cylinder was fixed, while the downstream cylinder was suspended from a platform and allowed to move freely in the horizontal plane. The centre-to-centre spacing ratio between two tandem cylinders was initially kept at a constant value of 3.0. The instantaneous flow field and the orbital trajectories were analyzed to reveal the effect of the presence of the upstream cylinder and flow velocity on the dynamic response of the downstream suspended cylinder. The results showed that the upstream stationary cylinder has significant effect on modifying the flow patterns behind two tandem cylinders. Different trajectories of the downstream suspended cylinder with variation of flow velocity U were observed, such as: (1) depicting a figure-8 type motion at U = 0.27 m/s; (2) undergoing intermittent oscillations as it travels downstream at 0.3 m/s ⩽ U ⩽ 0.37 m/s; (3) successive moving downstream, no obvious streamwise oscillation observed at U = 0.43 m/s.

E2A, a basic helix-loop-helix transcription factor, plays a crucial role in determining tissue-specific cell fate, including differentiation of B-cell lineages. In 5% of childhood acute lymphoblastic leukemia (ALL), the t(1,19) chromosomal translocation specifically targets the E2A gene and produces an oncogenic E2A-PBX1 fusion protein. Although previous studies have shown the oncogenic functions of E2A-PBX1 in cell and animal models, the E2A-PBX1–enforced cistrome, the E2A-PBX1 interactome, and related mechanisms underlying leukemogenesis remain unclear. Here, by unbiased genomic profiling approaches, we identify the direct target sites of E2A-PBX1 in t(1,19)–positive pre-B ALL cells and show that, compared with normal E2A, E2A-PBX1 preferentially binds to a subset of gene loci cobound by RUNX1 and gene-activating machineries (p300, MED1, and H3K27 acetylation). Using biochemical analyses, we further document a direct interaction of E2A-PBX1, through a region spanning the PBX1 homeodomain, with RUNX1. Our results also show that E2A-PBX1 binding to gene enhancers is dependent on the RUNX1 interaction but not the DNA-binding activity harbored within the PBX1 homeodomain of E2A-PBX1. Transcriptome analyses and cell transformation assays further establish a significant RUNX1 requirement for E2A-PBX1–mediated target gene activation and leukemogenesis. Notably, the RUNX1 locus itself is also directly activated by E2A-PBX1, indicating a multilayered interplay between E2A-PBX1 and RUNX1. Collectively, our study provides the first unbiased profiling of the E2A-PBX1 cistrome in pre-B ALL cells and reveals a previously unappreciated pathway in which E2A-PBX1 acts in concert with RUNX1 to enforce transcriptome alterations for the development of pre-B ALL.

Significance Acute myeloid leukemias (AMLs) with NUP98-NSD1 or MLL abnormality are generally aggressive, demanding a better understanding of the underlying oncogenic mechanisms. We show that these AMLs rely on a regulatory axis involving PRC2 – | Kdm5b – |stemness genes for sustaining an oncogenic program. The H3K27 methylase activity of polycomb repressive complex 2 (PRC2) is crucial for repressing Kdm5b, a corepressor carrying a H3K4me3 reader domain, that antagonizes the AML oncoproteins by directly binding to and down-regulating the AML stemness genes, thereby suppressing acute leukemogenesis. Such an AML-suppressing role of Kdm5b is not dependent on its intrinsic demethylase activity but requires its scaffold and/or chromatin association functions. The findings of this study shall aid in potential therapeutics of the affected AML patients.

This study used DNA methyltransferase 3b (DNMT3b) knockout cells and the functional loss of DNMT3b mutation in immunodeficiency-centromeric instability-facial anomalies syndrome (ICF) cells to understand how DNMT3b dysfunction causes genome instability. We demonstrated that R-loops contribute to DNA damages in DNMT3b knockout and ICF cells. More prominent DNA damage signal in DNMT3b knockout cells was due to the loss of DNMT3b expression and the acquirement of p53 mutation. Genome-wide ChIP-sequencing mapped DNA damage sites at satellite repetitive DNA sequences including (peri-)centromere regions. However, the steady-state levels of (peri-)centromeric R-loops were reduced in DNMT3b knockout and ICF cells. Our analysis indicates that XPG and XPF endonucleases-mediated cleavages remove (peri-)centromeric R-loops to generate DNA beaks, causing chromosome instability. DNMT3b dysfunctions clearly increase R-loops susceptibility to the cleavage process. Finally, we showed that DNA double-strand breaks (DSBs) in centromere are probably repaired by error-prone end-joining pathway in ICF cells. Thus, DNMT3 dysfunctions undermine the integrity of centromere by R-loop-mediated DNA damages and repair.

Coronaviral papain-like proteases (PLpros) are essential enzymes that mediate not only the proteolytic processes of viral polyproteins during virus replication but also the deubiquitination and deISGylation of cellular proteins that attenuate host innate immune responses. Therefore, PLpros are attractive targets for antiviral drug development. Here, we report the crystal structure of papain-like protease 2 (PLP2) of porcine epidemic diarrhea virus (PEDV) in complex with ubiquitin (Ub). The X-ray structural analyses reveal that PEDV PLP2 interacts with the Ub substrate mainly through the Ub core region and C-terminal tail. Mutations of Ub-interacting residues resulted in a moderately or completely abolished deubiquitinylating function of PEDV PLP2. In addition, our analyses also indicate that 2-residue-extended blocking loop 2 at the S4 subsite contributes to the substrate selectivity and binding affinity of PEDV PLP2. Furthermore, the PEDV PLP2 Glu99 residue, conserved in alphacoronavirus PLpros, was found to govern the preference of a positively charged P4 residue of peptidyl substrates. Collectively, our data provided structure-based information for the substrate binding and selectivity of PEDV PLP2. These findings may help us gain insights into the deubiquitinating (DUB) and proteolytic functions of PEDV PLP2 from a structural perspective. IMPORTANCE Current challenges in coronaviruses (CoVs) include a comprehensive understanding of the mechanistic effects of associated enzymes, including the 3C-like and papain-like proteases. We have previously reported that the PEDV PLP2 exhibits a broader substrate preference, superior DUB function, and inferior peptidase activity. However, the structural basis for these functions remains largely unclear. Here, we show the high-resolution X-ray crystal structure of PEDV PLP2 in complex with Ub. Integrated structural and biochemical analyses revealed that (i) three Ub core-interacting residues are essential for DUB function, (ii) 2-residue-elongated blocking loop 2 regulates substrate selectivity, and (iii) a conserved glutamate residue governs the substrate specificity of PEDV PLP2. Collectively, our findings provide not only structural insights into the catalytic mechanism of PEDV PLP2 but also a model for developing antiviral strategies.

Viral production (VP) and bacterial mortality by viral lysis critically influence the production and mortality of aquatic bacteria. Although bacterial production, mortality by viral lysis, and viral density have been found to exhibit diel variations, the diel change in viral production has rarely been investigated. In this study, we conducted two diel dilution incubation experiments in a semi-enclosed, nutrient-rich coastal region in northeastern Taiwan to estimate the diel viral production and the mortality by viral lysis. We also compared two methods (linear regression between viral density and time versus arithmetic mean of VP during incubation) of estimating viral production. We found that viral production estimated by linear regression and bacterial mortality by viral lysis were higher during the daytime than during the nighttime. A possible explanation for the high viral production at daytime is that the bacterial community was composed of cell types with higher burst sizes at daytime. We further argued that the classical linear regression method can be used only when viral density significantly linearly increases with time, which does not always occur in dilution incubations. This study offered observations of diel variation in viral dynamics and discussed the methods estimating viral production in a marine environment.

Background Frailty may increase the risk of complications and mortality in patients undergoing cardiac surgery. Few studies on frailty and its associated factors have been conducted in these patients. Objective The aim of this study was to explore frailty and related factors in patients undergoing cardiac surgery. Methods A total of 154 patients undergoing cardiac surgery in northern Taiwan were recruited using a longitudinal study design and interviewed using structured questionnaires assessing physical activity, anxiety and depression, and social support before surgery and at 1 month and 3 months after surgery. Results The prevalence of frailty in patients undergoing cardiac surgery was 16.2%, 20.5%, and 16.6% before surgery and at 1 month and 3 months after surgery, respectively. Frail and prefrail patients undergoing cardiac surgery were more likely to be unemployed, have gout, have a higher New York Heart Association class, have preoperative dysrhythmia, undergo cardiopulmonary bypass, have a lower functional ability, have a higher European System for Cardiac Operative Risk Evaluation score, have a longer anesthesia time, have longer endotracheal tube and extracorporeal circulation times, have longer intensive care unit and hospital stays, have lower hemoglobin and albumin levels, have higher anxiety and depression levels, and have lower Mini-Mental State Examination scores. The significant predictors of prefrailty and frailty included unemployment, the presence of gout, higher New York Heart Association classes, less independence in activities of daily living, lower hemoglobin levels, and higher levels of depression. Conclusions Frailty was associated with patients' functional status, perioperative conditions and psychosocial factors. Preoperative assessments of frailty and appropriate interventions are needed to improve frailty in patients undergoing cardiac surgery.

This study applied an in vitro fermentation model, whereby the catabolism of Porphyra haitanensis polysaccharides (PHP) was monitored coupled with the variations of microbiota composition and the concentration of short-chain fatty acids (SCFAs), so as to assess the effects of PHP on human intestinal microbiota. After 24 h anaerobic incubation, the level of microflora diversity increased significantly, as the microbiome structure was reshaped through promotion of intestinal probiotics proliferation and inhibition of pathogens growth. Besides, the final concentration of total SCFAs increased to 32.32 ± 1.81 mmol/L, and contained high amounts of acetic, propionic and butyric acid. Furthermore, the molecular weight of PHP decreased from 2.623 × 105 g/mol to 2.308 × 104 g/mol. The degree of polymerization of oligosaccharide products ranged from 2 to 9, with the main linkage patterns being 1 → 3 and 1 → 4 linked Galp. The current study provides new insight on the probiotic activity of PHP within the human gastrointestinal tract.

Reliability of an electronic device, concerning if it can function reliably over its designated lifetime in the field (such as 10 or 15 years), has become more and more important in today's safety-critical applications such as automotive electronics. Traditionally, the ageing has been performed in an offline setting where stress test has been applied to accelerate the ageing process and then a model is established to make the futuristic prediction. This kind of offline method has a drawback of not being able to take into account the factor of the unique operating condition and environment that a device could have experienced in the field. In this work, we present the first cloud-based ageing monitoring system to the best of our knowledge, for the Internet-of-Things (IoT) devices. It has many advantages. First of all, one can know of the ageing status of an IoT device remotely and continuously. Secondly, through data analysis in a cloud server, more accurate prediction can be achieved. Thirdly, an ageing hazard can be alarmed in advance before it actually strikes, and thereby pre-caution actions (such as online repair, or even call-for-maintenance request) can be taken in advance to avoid unnecessary system fatal failure. A prototype system using test chips with built-in design-for-ageing-monitoring circuitry will be demonstrated with measurement data collected through a cloud server.

To develop a zebrafish cataract model for screening potential anti-cataract compounds.Living zebrafish were anesthetized and exposed to ultraviolet-C (UV-C) irradiation at a dosage of 3250 mJ/cm2/d until they developed severe cataracts. These cataracts were graded based on photographs analyzed with ImageQuant TL version 7.0. Fish with severe cataracts were used to evaluate a range of compounds for cataract treatment, including the previously demonstrated hit compound lanosterol. For the initial evaluation, fish were divided into four groups: no treatment, balanced salt solution, β-cyclodextrin (β-CD), and lanosterol dissolved in β-CD. The treatments were performed for 10 days, and the clarity of lenses was evaluated. To assess the persistence of treatment, fish were treated with β-CD and lanosterol dissolved in β-CD for seven consecutive days followed by monitoring for three days without treatment.The average time for zebrafish to develop severe cataracts using the present UV-C irradiation protocol was 7.8 days (range 4-15 days). Both study designs required only another 10 days to determine the effect of hit compounds. The total experimental period could be completed within one month, and the entire experiment was economical.We could assay a large number of hit compounds at a reasonable cost and within a short time using this newly developed zebrafish cataract model. These assays may allow development of an efficient platform for screening potential anti-cataract compounds.The results may facilitate the development of ani-cataract medication for humans after further experiments and investigations.

Background Postoperative delirium (POD) is a common complication that may occur from 24 to 72 h after cardiac surgery. Frailty is a chronic syndrome that leads to a decline in physiological reserve and to disability. The associations between frailty and POD are unclear. Aims To investigate associations between POD and frailty in patients undergoing cardiac surgery and to analyse predictors of POD. Methods and results Convenience sampling was used to recruit 152 patients who underwent cardiac surgery in two medical centres in northern Taiwan. Preoperative frailty in these patients was evaluated using Fried’s frailty phenotype. Delirium in patients was assessed from postoperative day 1 to day 5 using the confusion assessment method for intensive care units. A total of 152 patients who underwent cardiac surgery included 68 (44.74%) prefrail patients and 21 (13.81%) patients with frailty after the surgery. Ten patients (6.58%) developed delirium after cardiac surgery. The occurrence of delirium peaked at postoperative day 2, and the average duration of delirium was 3 days. A case–control comparison revealed a significant correlation between preoperative frailty and POD. Significant predictors of POD in patients undergoing cardiac surgery included the European System for Cardiac Operative Risk Evaluation II, preoperative arrhythmia, and preoperative anxiety and depression. Conclusion Preoperative frailty was correlated with POD. Preoperative arrhythmia, anxiety, and depression are predictors of POD. Nurses should perform preoperative assessments of surgical risk and physiological and psychological conditions of patients undergoing cardiac surgery and monitor the occurrence of POD.

Recent evidence suggests that melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs), a neuronal class regulating non-image forming (NIF) vision and generally thought to be injury-resistant, are dysfunctional in certain neurodegenerative diseases. Although disrupted NIF visual functions have been reported in patients and animals with diabetes, it remains controversial whether ipRGCs exhibit remodeling during diabetes and if so, whether such remodeling is variable among ipRGC subtypes. Here we demonstrate that survival, soma-dendritic profiles and melanopsin-based functional activity of M1 ipRGCs were unaltered in streptozotocin-induced 3-month diabetic mice. Such resistance remained at 6 months after streptozotocin administration. In contrast, M2/M3 ipRGCs underwent significant remodeling in diabetic mice, manifested by enlarged somata and increased dendritic branching complexity. Consistent with the unaltered melanopsin levels, the sensitivity of melanopsin-based activity was unchanged in surviving M2 cells, but their response gain displayed a compensatory enhancement. Meanwhile, the pupillary light reflex, a NIF visual function controlled by M2 cells, was found to be impaired in diabetic animals. The resistance of M1 cells might be attributed to the adjacency of their dendrites to capillaries, which makes them less disturbed by the impaired retinal blood supply at the early stage of diabetes.

Disulfiram is a promising repurposed drug that, combining with radiation and chemotherapy, exhibits effective anticancer activities in several preclinical models. The cellular metabolites of disulfiram have been established, however, the intracellular targets of disulfiram remain largely unexplored. We have previously reported that disulfiram suppresses the coronaviral papain-like proteases through attacking their zinc-finger domains, suggesting an inhibitory function potentially on other proteases with similar catalytic structures. Ubiquitin-specific proteases (USPs) share a highly-conserved zinc-finger subdomain that structurally similar to the papain-like proteases and are attractive anticancer targets as upregulated USPs levels are found in a variety of tumors. Here, we report that disulfiram functions as a competitive inhibitor for both USP2 and USP21, two tumor-related deubiquitinases. In addition, we also observed a synergistic inhibition of USP2 and USP21 by disulfiram and 6-Thioguanine (6TG), a clinical drug for acute myeloid leukemia. Kinetic analyses revealed that both drugs exhibited a slow-binding mechanism, moderate inhibitory parameters, and a synergistically inhibitory effect on USP2 and USP21, suggesting the potential combinatory use of these two drugs for USPs-related tumors. Taken together, our study provides biochemical evidence for repurposing disulfiram and 6TG as a combinatory treatment in clinical applications.