Journal: The Journal of general virology

Article Title: Influenza virus entry via the GM3 ganglioside-mediated platelet-derived growth factor receptor β signalling pathway.

doi: 10.1099/jgv.0.001235

Figure Lengend Snippet: Fig. 4. Virus entry and inhibition by Ki8751 in CHO-wt, CHO-K1, and transfectant cells carrying FGFR4 or VEGFR2. (a) Nuclear import of vRNP in CHO cells incubated with influenza (A/PR/8/34) virus over 1 h at 35 C. Quantitative confocal microscopy was carried out after dual nuclear staining for viral NP and DAPI. In three separate experiments, three images (each containing ~120 cells) were taken from each condition to quantify the green fluorescent signal in the nucleus. (b) Virus binding efficiency relative to CHO-wt cells, as determined by RT-qPCR quantification of cell-bound virus after 60 min of incubation at 4 C. (c) Ki8751 and two other RTK inhibitors, crenolanib and imatinib (all at 60 µM), inhibit virus nuclear entry in CHO-wt and CHO-K1 cells. Confocal microscopy after NP-immunos- taining at 1 h p.i. Green: viral NP; blue: nuclear DAPI staining. (d) vRNA synthesis at 10 h p.i., assessed by two-step RT-qPCR. The y-axis shows the fold increase in vRNA, relative to untreated CHO-wt cells. Panels a, b and d: mean±SEM (N=3). Statistical significance by two- sided Student’s t-test: *P<0.05; ***P<0.005; NS=not significant (P>0.05).

Article Snippet: The library of 276 protein kinase inhibitors was purchased from Selleck Chemicals (catalogue no. L1200).

Techniques: Virus, Inhibition, Transfection, Incubation, Confocal Microscopy, Staining, Binding Assay, Quantitative RT-PCR

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: A kinase domain–targeted CRISPR screen identifies HASPIN as a novel dependency in AML cells. (A) Schematic of human kinase domain–targeted CRISPR screen in 2 t(8;21) AML cell lines, Kasumi-1 and SKNO-1. (B) Gene rank plots (left) and volcano plots (right) depicting significant kinase hits in the kinase domain–targeted CRISPR screens. Top candidates determined by CRISPR score as calculated by MAGeCK robust ranking aggregation (RRA) (left) and a log 2 (fold change) ≤−1.0 and false discovery rate (FDR) ≤0.05 significance cutoff (right). White diamonds indicate top 10 kinase hits in each plot. (C) Bubble plot of preranked gene set enrichment analysis results performed on top kinases identified by CRISPR screen in Kasumi-1 and SKNO-1 AML cell lines. Fill color indicates normalized enrichment score (NES). Size indicates significance by –log 10 (FDR). Facets indicate Molecular Signatures Database (MSigDB) gene set collection. (D) Density plot of all individual sgRNA log 2 (fold change) values in the kinase domain–targeted library. For selected genes, log 2 (fold change) values of corresponding sgRNAs depicted for Kasumi-1 and SKNO-1 cell lines relative to all other library sgRNAs (red, blue, and gray, respectively). (E) Competitive proliferation assay of Kasumi-1 or SKNO-1 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± standard deviation (SD) of 4 independent experiments. CGP, chemical, genetic perturbation; CP, canonical pathway.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: CRISPR, Proliferation Assay, Expressing, Negative Control, Positive Control, Derivative Assay, Control, Standard Deviation

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: HASPIN is a clinically relevant, general leukemia dependency. (A) Bar plot depicting mean log 2 (fold change) of HASPIN targeting sgRNA genome-wide CRISPR screen performed in several leukemia cell lines as reported by Wang et al. Screen data were retrieved from BIOGRID ORCS. Dotted line indicates author-specified significance cutoff. (B) Competitive proliferation assay of THP-1 or OCI-AML3 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± SD of 4 independent experiments per cell line. (C) Box plots depicting median HASPIN mRNA expression in the TCGA-LAML patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (D) Box plots depicting median HASPIN mRNA expression in the BEAT-AML (2022) patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (E) Kaplan-Meier survival curve depicting comparison of overall survival of patients with TCGA-LAML belonging to the top quartile (red) and bottom quartile (blue) of HASPIN expression. Plot and data derived from GEPIA2. (F) Forest plot of hazard ratios from multivariate Cox proportional hazard analysis of overall survival of patients with TCGA LAML incorporating HASPIN expression level and significant clinical and genetic factors. High and low HASPIN -expressing patients belong to the top and bottom expression quartiles, respectively. Clinical variables include the following: patient sex (Sex), age at first diagnosis (Diagnosis_Age), genetic risk group (Risk_Group), FLT3 mutation status (FLT3_Status), NPM1 mutation status (NPM1_Status), DNMT3A mutation status (DNMT3A_Status), TP53 mutation status (TP53_Status), and NRAS mutation status (NRAS_Status). Clinical metadata and mutation calls derived from the Genomic Data Commons TCGA LAML project patient information. N.D., not defined; NOS, not otherwise specified; NP, not profiled.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: Genome Wide, CRISPR, Proliferation Assay, Expressing, Negative Control, Positive Control, Derivative Assay, Control, Translocation Assay, Comparison, Biomarker Discovery, Mutagenesis

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: HASPIN inhibitor CHR-6494 effectively targets AML and synergizes with BCL-2 inhibition. (A) Dose-response curves (left) and IC comparison (right) of Kasumi-1 and healthy CD34 + hematopoietic progenitor cells treated with CHR-6494. IC values determined by nonlinear regression. Data on curve are mean ± SD of technical triplicates. Representative curves of 3 independent experiments revealed. Data on bar plot are mean ± SD of 3 independent experiments. Significance determined by unpaired 2-tailed Student t test. ∗∗∗∗ P < .0001. (B) Bar plots comparing CHR-6494 IC values in leukemia cell lines. IC values determined by dose-response curve with nonlinear regression for each cell line. Data are mean ± SD of 3 independent experiments. Dotted line indicates CHR-6494 IC value of healthy CD34 + hematopoietic progenitor cells determined in panel A. (C) Bar plots depicting normalized HASPIN sgRNA counts in a genome-wide CRISPR screen in MOLM-13 cells treated with either DMSO or VEN for 8 or 16 days as performed by Chen et al. Screen data were retrieved from BIOGRID ORCS. Counts were normalized to initial time point (d0). One data point was removed from DMSO (d16) as a significant outlier. Data are mean ± SD. Significance determined by 1-way ANOVA with Holm-Sidak multiple comparison correction. ∗ P < .05; ∗∗ P < .01. (D) Dose-response matrix (left) and corresponding zero interaction potency (ZIP) drug synergy contour plot (right) of Kasumi-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (E) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of THP-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (F) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of OCI-AML3 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. DMSO, dimethyl sulfoxide.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: Inhibition, Comparison, Genome Wide, CRISPR

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of a selective PI3Kα inhibitor via structure-based virtual screening for targeted colorectal cancer therapy

doi: 10.1080/14756366.2025.2468852

Figure Lengend Snippet: Validation of the docking protocol was performed through ligand extraction and redocking. The co-crystal ligand, alpelisib, originally identified in complex with the PI3Kα protein via X-ray crystallography, was extracted and re-docked into the protein’s binding site. The accuracy of this protocol was evaluated by calculating the RMSD between the superimposed poses of the ligand in the original crystal structure (cyan color) and its docked conformation (purple color).

Article Snippet: The screening protocol encompassed ∼3000 compounds sourced from the Protein Kinase Diversity Set and the Phosphatidylinositol 3-Kinase (PI3K) Targeted Library by Life Chemicals (accessed 15 May 2024).

Techniques: Biomarker Discovery, Extraction, Binding Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of a selective PI3Kα inhibitor via structure-based virtual screening for targeted colorectal cancer therapy

doi: 10.1080/14756366.2025.2468852

Figure Lengend Snippet: Inhibitory effects of candidate compounds on PI3Kα and their selectivity profile across PI3K isoforms. The ADP-Glo™ assay was employed using purified PI3Kα to assess the kinase inhibition exerted by compounds F0608-0019 (A), F2490-0372 (B), and the reference inhibitor, alpelisib (C), across a range of concentrations. The percentage inhibition of PI3K isoforms β and γ was determined at a concentration of 10 µM (D) using the ADP-Glo™ kinase activity assay. The data represents the mean ± SEM of two biological replicates.

Article Snippet: The screening protocol encompassed ∼3000 compounds sourced from the Protein Kinase Diversity Set and the Phosphatidylinositol 3-Kinase (PI3K) Targeted Library by Life Chemicals (accessed 15 May 2024).

Techniques: Glo Assay, Purification, Inhibition, Concentration Assay, Kinase Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of a selective PI3Kα inhibitor via structure-based virtual screening for targeted colorectal cancer therapy

doi: 10.1080/14756366.2025.2468852

Figure Lengend Snippet: Molecular dynamics simulation of alpelisib and F0608-0019 in complex with PI3Kα. RMSD analysis of the ligand-protein complexes, illustrating the stability of alpelisib (A) and F0608-0019 (D) bound to PI3Kα. RMSF analysis depicts the flexibility of amino acid residues in the PI3Kα complexed with alpelisib (B) and F0608-0019 (E). Hydrogen bond formation analysis indicates the interactions of alpelisib (C) and F0608-0019 (F) with PI3Kα.

Article Snippet: The screening protocol encompassed ∼3000 compounds sourced from the Protein Kinase Diversity Set and the Phosphatidylinositol 3-Kinase (PI3K) Targeted Library by Life Chemicals (accessed 15 May 2024).

Techniques:

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of a selective PI3Kα inhibitor via structure-based virtual screening for targeted colorectal cancer therapy

doi: 10.1080/14756366.2025.2468852

Figure Lengend Snippet: Two-dimensional ligand interaction map of PI3Kα inhibitor complexes extracted from the last frame of molecular dynamics simulations. This analysis provides a detailed view of the ligand-protein interactions, highlighting the binding poses of alpelisib (A) and F0608-0019 (B). Interaction bonds are represented as dashed lines connecting the relevant atoms. Distinct colours denote different types of interactions: green indicates conventional hydrogen bonds, light orange represents π-sulfur interactions, dark orange signifies π-anion interactions, and pink depicts π-alkyl interactions.

Article Snippet: The screening protocol encompassed ∼3000 compounds sourced from the Protein Kinase Diversity Set and the Phosphatidylinositol 3-Kinase (PI3K) Targeted Library by Life Chemicals (accessed 15 May 2024).

Techniques: Binding Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Discovery of a selective PI3Kα inhibitor via structure-based virtual screening for targeted colorectal cancer therapy

doi: 10.1080/14756366.2025.2468852

Figure Lengend Snippet: Total energy contributions of key residues in the binding of PI3Kα inhibitors. Analysis of the energy contributions of essential residues involved in the binding interactions of PI3Kα with the inhibitors alpelisib (A) and F0608-0019 (B).

Article Snippet: The screening protocol encompassed ∼3000 compounds sourced from the Protein Kinase Diversity Set and the Phosphatidylinositol 3-Kinase (PI3K) Targeted Library by Life Chemicals (accessed 15 May 2024).

Techniques: Binding Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Akt1/protein kinase B enhances transcriptional reprogramming of fibroblasts to functional cardiomyocytes

doi: 10.1073/pnas.1516237112

Figure Lengend Snippet: List of myristoylated kinases tested in an addition analysis to enhance GHMT-mediated cardiac reprogramming

Article Snippet: The Myristoylated Kinase Library listed in was obtained from Addgene [(Kit #1000000012) deposited by William Hahn and Jean Zhao ( 17 )]. table ft1 table-wrap mode="anchored" t5 Table S2. caption a7 AAK1 CKMT1A HK1 MVK PIP5K1B RPS6KL1 ACVR1 CKMT2 HK2 NADK PIP5K2A RPSK6A3 ADCK4 CKS1B HK3 NEK11 PIP5K3 SGK ADCK5 CKS2 IHPK2 NEK3 PKM2 SNF1LK ADPGK CLK1 IKBKE NEK6 PKN1 SPHK2 ADRBK1 CLK2 ILK NME7 PKN2 SRPK2 ADRBK2 CLK3 ITK NTRK3 PLAU STK17B AKT1 CMPK ITPK1 NUAK2 PLK1 STK3 AKT3 CSNK1A1L ITPKB OXSR1 PLK2 STK32A AMHR2 CSNK1E LCK PACSIN1 PLK3 STK32B AURKA CSNK1G1 LIMK1 PAK4 PLK4 STK32C AXL CSNK1G2 LIMK2 PAPSS1 PMVK STK33 BLK DAK MAP2K5 PBK PNKP STK38L BMX DGKG MAP2K6 PCK2 PRKAA1 STK4 BTK DGUOK MAP2K7 PCTK1 PRKACB STK40 CALM2 DLG5 MAP3K14 PCTK2 PRKACG SYK CAMK1G DYRK2 MAP3K6 PCTK3 PRKAG2 TAOK3 CAMK2B DYRK4 MAP3K7 PDIK1L PRKAR2A TBK1 CAMK2D EPHA4 MAP3K8 PDK1 PRKCD TEC CAMK4 FASTK MAPK12 PDPK1 PRKCI TESK1 CAMKK1 FGFR1 MAPK13 PDXK PRKCZ TIE1 CAMKV FGR MAPK14 PFKL PRKRA TK1 CDC2 FRK MAPK6 PFKM PTK2 TNK2 CDK2 GAK MAPK7 PI4K2B RET TSSK1B CDK4 GALK2 MAPKAP1 PIK3CB RIOK1 TSSK6 CDK5 GCK MAST1 PIK3CG RIOK2 TTK CDK7 GK MATK PIK3R3 RIOK3 TYK2 CDK9 GK2 MELK PIK3R5 RPS6KA2 UCK2 CERK GRK5 MKNK1 PIK4CA RPS6KA5 ULK4 CHEK1 GRK6 MOBKL1A PIK4CB RPS6KA6 VRK2 CKB HCK MOBKL2A PIM1 RPS6KB1 VRK3 CKM HIPK1 MPP1 PIP5K1A RPS6KB2 YES1 Open in a separate window List of myristoylated kinases tested in an addition analysis to enhance GHMT-mediated cardiac reprogramming qPCR, Western Blot Analyses, Immunocytochemistry, Flow Cytometry, Beating Cell Analyses, Calcium Transient Measurements, and Cell Size Measurements.

Techniques: