hdac dual inhibitor corin  (MedChemExpress)

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Immunoblot analysis of WCL from P3HR-1 or MUTU I Burkitt cells treated with the indicated corin concentrations for two days. (B) FACS analysis of PM gp350 levels in corin-treated P3HR-1 or MUTU I cells. (C) Immunoblot analysis of WCL from EBV+ KEM III, Jijoye, Farage, AGS, YCCEL1, C666-1 cells treated with corin (0, 1, 2, or 5 μM) for two days. ( D ) Immunoblot analysis of WCL from EBV+/KSHV+ JSC-1 primary effusion lymphoma cells treated with corin (0, 1, 2, or 5 μM) for two days. ( E ) Workflow of corin and ganciclovir (GCV) treatment. Cells were seeded into fresh media on days of corin treatment. GCV (10 μg/ml) was added twice daily where indicated. ( F ) EBV+ Burkitt B-cells were treated as described in (E), with the following corin concentrations: P3HR-1 (0.5 μM), MUTU I (0.5 μM), Rael (0.25 μM) vs vehicle. Shown are mean ± SD live cell number relative to DMSO-treated controls on day 6 from n=3 replicates. Values of DMSO treated control cells were normalized to 1. * p <0.05, *** p < 0.001. ( G ) MUTU I murine xenograft experiment workflow. Two weeks post MUTU I Burkitt xenograft implantation, mice were treated with vehicle or corin (30 mg/ml), as indicated. ( H ) Immunohistochemical analysis of BMRF1 expression in xenograft tumors following treatment with vehicle vs. corin as in Figure 3G . Scale bar = 100 μm. ( I ) Dosing strategy for analysis of corin, GCV or corin+GCV effects on murine MUTU I xenograft tumor size. Once xenograft tumors reached a volume of 50mm 3 , n=6 or 7 mice were treated with vehicle, corin (30mg/kg), GCV (50mg/kg) or corin (30mg/kg) + GCV (50mg/kg). ( J ) Area under the curve (AUC) analysis of tumor size up to 21 days post the initial treatment. AUC integrates tumor size measurements per group across all timepoints. All blots shown are representative images of n=3 replicates. * p <0.05, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Inhibition, Western Blot, Control, Immunohistochemical staining, Expressing

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Workflow of human genome-wide CRISPR-Cas9 screen for host factors that support EBV lytic reactivation. Cas9+ P3HR-1 with stable ZHT/RHT conditional immediate early allele expression were transduced with the Brunello sgRNA library. Transduced cells were selected and then reactivated by 4-hydroxytamoxifen (4HT, 0.4 μM) together with NaB (0.5 mM) for 48 hours. The 5% of cells with the lowest gp350 expression were sorted. sgRNA abundance in input vs sorted cells was quantitated to identify hits. ( B ) Volcano plot analysis of screen hits, which are highlighted by category. ( C ) Immunoblot analysis of WCL from P3HR-1 ZHT/RHT cells expressing control or KMT2D sgRNAs and mock induced or induced by 4HT (0.4 μM) and NaB (0.5 mM) for 24 hours. ( D-E ) FACS analysis of PM gp350 levels (D) and of PM gp350 MFI ± SD from n=3 replicates (E) in Propidium Iodide (PI) negatively stained P3HR-1 ZHT/RHT cells with control vs KMT2D sgRNAs and mock induced or induced for lytic replication by 4HT and NaB for 24 hours. ( F ) qPCR of intracellular EBV genome copy number in P3HR-1 ZHT/RHT cells expressing control or KMT2D sgRNA that were treated with or without 4HT (0.4 μM) and NaB (0.5 mM) for 24 hours. ( G ) Immunoblot analysis of WCL from Cas9+ P3HR-1 ZHT/RHT cells expressing control or KMT2D sgRNA and that were treated with or without corin (2.5 μM) for 24 hours. ( H ) Immunoblot analysis of WCL from Akata cells that stably expressed cDNAs encoding control GFP or the KMT2D SET domain, which writes H3K4 methyl marks. The schematic at bottom indicates SET domain residues. All blots are representative of n=3 replicates. Bar graphs are presented as mean ± SD values from three replicates. *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Genome Wide, CRISPR, Expressing, Transduction, Western Blot, Control, Staining, Stable Transfection

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Schematic diagram highlighting chromatin conformation capture (3C) assay BZLF1p anchor primer (red box) and 12 test (T) primers locations along the linear EBV genome, including at oriLyt R (primer T6). For reference, EBV genomic terminal repeat ( TR ) and origin of plasmid replication ( oriP ) are shown. ( B ) 3C assay analysis of DNA looping between the BZLF1 anchor and 12 test primer regions. Shown are the mean ± SD 3C assay signals relative to EBV BACmid negative control from Akata cells expressing control, LSD1, ZNF217, or CoREST sgRNAs. ( C ) Mean ± SD 3C assay signals from Akata cells 24 hours post-treatment with vehicle, C12 (2.5 μM), C12 (2.5 μM) + NaB (0.5 mM), or corin (2.5 μM). * p < 0.05; ** p < 0.01; *** p < 0.001 by Student’s t-test. ( D ) Schematic model. In latency, LSD1/ZNF217/CoREST/HDAC complexes co-occupy EBV oriLyt enhancer regions, where LSD1 and HDAC erase activating H3K4 methylation and H3K4 acetylation marks, respectively. Perturbation of LSD1 and HDAC activity enables KMT2D to deposit activating H3K4 epigenetic marks at both oriLyt enhancers and supports their looping to the immediate early BZLF1 promoter. Newly synthesized BZLF1 drives the lytic cycle by binding to early gene promoters and also to oriLyt enhancers to increase their strength, which further upregulates BZLF1 in a positive feedback loop.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Plasmid Preparation, Negative Control, Expressing, Control, Methylation, Activity Assay, Synthesized, Binding Assay

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Schematic diagram depicting bifunctional small molecule antagonist corin inhibition of LSD1 and HDAC activity. ( B ) Immunoblot analysis of WCL from P3HR-1, Akata or MUTU I cells that were treated with the indicated concentration of corin for 6 hours and then grown in corin-free RPMI for the remainder of 48 hours. ( C ) Mean ± SD intracellular EBV genome copy number from n=3 replicates of P3HR-1 or MUTU I cells treated with corin for 48 hours. ( D ) Immunoblot analysis of WCL from Akata cells with stable GFP versus MYC cDNA overexpression that were treated with corin for 48 hours, as indicated. ( E ) Immunoblot analysis of WCL from control or BZLF1 knockout (KO) Akata cells treated with corin for 48 hours, suggesting that corin requires BZLF1 to induce early and late protein expression. ( F ) Immunoblot analysis of WCL from Akata, KEM I, EB3, Rael Burkitt cells, GM15892 LCLs or SNU719 gastric carcinoma cells treated with corin for 48 hours. ( G ) Immunoblot analysis of WCL from EBV+/KSHV+ BC-1 or KSHV+ BCBL-1 primary effusion lymphoma cells treated with corin for 48 hours. ( H ) KSHV+ iSLK.219 epithelial cells with conditional doxycycline (Dox) inducible KSHV immediate early RTA expression was treated with DMSO, Dox (0.5 ug/ml) or corin (5 μM) for 24 hours, and then maintained in growth media without Dox or corin. 48 hours later, immunofluorescence analysis was performed for GFP (stably expressed from the KSHV genome) versus for red fluorescence protein (RFP, controlled by lytic PAN RNA promoter, which reports KSHV lytic gene expression). Scale bar = 650 μm. Blots and immunofluorescence images are representative of n=3 independent replicates. ** p <0.01, *** p < 0.001 by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Inhibition, Activity Assay, Western Blot, Concentration Assay, Over Expression, Control, Knock-Out, Expressing, Immunofluorescence, Stable Transfection, Fluorescence, Gene Expression

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Corin and ganciclovir co-treatment effects on EBV-negative Burkitt and EBV+ LCL cells. Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment of EBV-negative MUTU I (left) and of EBV+ GM15892 LCLs (right). Cells were treated as described in Fig. 3E with DMSO or corin (0.5 μM). ( B ) Corin and ganciclovir co-treatment effects on KEM III LCLs. KEM III LCLs were treated with a single dose on day 1 of or with multiple drug doses as described in Fig. 3E . Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment. ( C ) P3HR-1 Burkitt cells were treated with a single dose on day 1 or with multiple doses of DMSO vehicle, LSD1 inhibitor C12 (0.5 μM), the HDAC inhibitor NaB (0.1 mM), C12 + NaB or with the dual LSD1/HDAC inhibitor corin (0.5 μM), using the regimen described in Fig. 3E . Shown are mean ± SD fold changes of live cell numbers, relative to DMSO-treated controls, from n=3 replicates on day 6 of treatment. ( D ) Immunoblot analysis of WCL prepared from MUTU I xenografts harvested from mice treated with DMSO vehicle vs corin as described in Fig. 3G . ( E ) Immunohistochemical analysis of BZLF1 expression in MUTU I xenografts harvested from mice treated with DMSO vs corin, as described in Fig. 3G . Scale bar = 100 μm. ( F ) Quantification of BZLF1+ (left panel) versus BMRF1+ (right panel) cell numbers in MUTUI xenograft tumors as in Fig. 3H and Extended Data Fig. 6E . Shown are mean ± SEM numbers of BZLF1+ or BMRF1+ cells from four randomly selected fields per mouse, quantitated by Image J using the Cell Counter plugin. ( G ) qPCR analysis of BZLF1, early BMRF1 and late BLLF1 (encodes gp350) mRNA abundances in xenograft tumors from tumors shown in Fig. 3H and Extended Data 6E . ( H ) qPCR defined intracellular EBV genome copy number from the xenograft tumors shown in Fig. 3H . Box heights indicate mean qPCR values. ( I ) MUTU I xenograft tumor sizes from the vehicle control, corin, GCV or corin and GCV treated mice shown in Fig. 3J . Significance was determined by cross-comparison of mean tumor sizes from dual drug treated versus vehicle or single drug treated mice. Blots shown are representative images of n=3 replicates. * p <0.05, ** p <0.01, *** p < 0.001, NS, not significant by Students t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Transformation Assay, In Vivo, Western Blot, Immunohistochemical staining, Expressing, Control, Comparison

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) ChIP-qPCR analysis of LSD1 and ZNF217 occupancy in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM), in the presence of acyclovir (100 μg/ml) to block EBV lytic DNA synthesis. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( B ) ChIP-qPCR analysis of H3K4me1, H3K4me2 and H3K4me3 abundances at the BZLF1 promoter or oriLyt regions in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( C ) ChIP-qPCR analysis of H3K9Ac and H3K27Ac abundances at the BZLF1 promoter or oriLyt regions in Akata cells 24 hours after treatment with vehicle, C12 (2.5 μM) or corin (2.5 μM). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( D ) ChIP-qPCR analysis of H3K9 and H3K27 acetylation levels in Akata cells expressing control versus independent LSD1 targeting sgRNAs. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( E ) ChIP-qPCR analysis of H3K9me2 and H3K9me3 abundances at the BZLF1 promoter or oriLyt regions in Akata cells treated with vehicle control, C12 (2.5 μM) or corin (2.5 μM) for 24 hours. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. Student’s t-test was performed to cross-compare ChIP-qPCR values from C12 or corin treated cells or LSD1 KO cells with corresponding values from control cells: * p <0.05, ** p <0.01, *** p < 0.001; NS: not significant.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: ChIP-qPCR, Blocking Assay, DNA Synthesis, Expressing, Control

Journal: Nature microbiology

Article Title: Lysine-specific histone demethylase complex restricts Epstein-Barr virus lytic reactivation

doi: 10.1038/s41564-025-02165-7

Figure Lengend Snippet: ( A ) Immunoblot analysis of WCL from Akata cells expressing control or KMT2D sgRNAs and that were mock-stimulated or stimulated for lytic reactivation by α-human IgG (10 μg/ml) cross-linking for 24 hours. ( B ) FACS analysis of PM gp350 levels on live Akata cells (as defined by exclusion of the vital dye PI and by forward and side scatter analysis) that expressed control or KMT2D sgRNA following 24 hours of mock-stimulation or αIgG crosslinking as in ( A ). ( C ) Mean ± SD MFI PM gp350 values from n=3 replicates, as in ( B ). ( D ) qPCR analysis of intracellular EBV genome copy number in Akata cells expressing control or KMT2D sgRNA, treated with or without α-human IgG (10 μg/ml) for 24 hours. ( E ) Immunoblot analysis of WCL from Akata cells expressing control or KMT2D sgRNA and treated with vehicle or corin (2.5 μM) for 24 hours. ( F ) Immunoblot analysis of WCL from KEM III LCLs expressing control or KMT2D sgRNA and treated with vehicle or corin (2.5 μM) for 24 hours. ( G ) ChIP-qPCR analysis of H3K4me1 abundances at the BZLF1 promoter or oriLyt regions in Akata cells expressing control or KMT2D sgRNA 24 hours after treatment with vehicle or corin (2.5 μM). Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( H ) ChIP-qPCR analysis of H3K4me1 abundances at the BZLF1 promoter or oriLyt regions in Akata cells expressing GFP or KMT2D SET cDNA. Shown are mean ± SD ChIP-qPCR % input values from n=3 replicates. ( I-J ) Immunoblot analysis of WCL from Akata cells expressing GFP or KMT2D SET cDNA, treated with or without α-human IgG (10 μg/ml) (I) or corin (2.5 μM) (J) for 24 hours. Blots are representative images of n = 3 replicates. ** p < 0.01; *** p < 0.001 as defined by Student’s t-test.

Article Snippet: LSD1 inhibitor C12 (also known as SP-2509, 0–10 μM, MedChemExpress, #HY-12635), LSD1 and HDAC dual inhibitor corin (0–10 μM, MedChemExpress, #HY-111048), HDAC inhibitor sodium butyrate (NaB, 0.5 mM), 4HT (0.4 μM), doxycycline (Dox, 0.5 μg/ml, Sigma-Aldrich, #D9891-1G), acyclovir (100 μg/ml, Sigma-Aldrich, #114798-25MG), ganciclovir (GCV, 10 μg/ml, MedChemExpress, #HY-13637), and anti-human IgG (10 μg/ml, Aligent, #A042301-2), were all purchased commercially and are listed in Supplementary Table 4 .

Techniques: Western Blot, Expressing, Control, ChIP-qPCR