Journal: NPJ Precision Oncology

Article Title: Leveraging patient derived models of FGFR2 fusion positive intrahepatic cholangiocarcinoma to identify synergistic therapies

doi: 10.1038/s41698-022-00320-5

Figure Lengend Snippet: A High-throughput small molecule screen inclusive of >2100 clinically relevant anticancer drugs suggest targets synergistic with pemigatinib. Each dot represents a compound plotted against that drug’s synergy score on the y -axis. Smaller differential viability is synonymous with increased synergy. Targets of commonly synergistic pathways are specifically highlighted. B Quisinostat and pemigatinib have synergistic effects on viability in the PDC-DUC18828 model based on MacSynergy II calculation (95% confidence interval). Results are color-coded from purple (most antagonistic) to dark red (most synergistic). C Dose–response curves in the PDC-DUC18828 model shift left with increasing concentrations of pemigatinib, resulting in significant reduction in the IC 50 value of quisinostat, concordant with synergy observed in B . All curves are normalized to 0 nM quisinostat under the corresponding concentrations of pemigatinib. D Quisinostat and pemigatinib have synergistic effects on viability in the PDO-DUC18828 model based on MacSynergy II calculation (95% confidence interval). Data are represented as in B . E Dose–response curves in the PDO model shift left with increasing concentrations of pemigatinib, resulting in significant reduction in the IC 50 value of quisinostat, concordant with synergy observed in D (normalized as per C ). Viability results are normalized to DMSO control and presented as the mean +/− standard deviation (error bars) for three biologically independent replicates. F Quisinostat and pemigatinib have synergistic effects on tumor growth in vivo. Mice bearing subcutaneous xenografts (~125 mm 3 ) were randomized to control (sham/sham gavage with Ora-plus suspension) vs pemigatinib (3 mg/kg oral gavage + sham gavage with Ora-plus suspension) vs quisinostat (16 mg/kg oral gavage + sham gavage with Ora-plus suspension) vs combination treatment (pemigatinib 3 mg/kg oral gavage + quisinostat 16 mg/kg oral gavage) and were treated 5 days per week. Tumor volume was measured 3×/week until study endpoints were reached. Compared to the control cohort (sham/sham), combined treatment with pemigatinib and quisinostat impaired tumor growth by 59.4%, compared to 23.8% with pemigatinib alone and 15.1% with quisinostat alone. Solid brackets (also recognized by arrows) indicate p < 0.05 between treatment arms starting on day 10 of treatment, indicated by the asterisk (*), whereas dashed brackets were not significant. Data are represented as mean +/− standard error of the mean for each experimental condition, consisting of 12 mice each and compared using the Student’s t test.

Article Snippet: Pemigatinib (Cat# T12401), BGJ398 (infigratinib, Cat# T1975) and quisinostat (Cat# T6055) were purchased from TargetMol Chemicals (Boston, MA).

Techniques: High Throughput Screening Assay, Standard Deviation, In Vivo

Journal: NPJ Precision Oncology

Article Title: Leveraging patient derived models of FGFR2 fusion positive intrahepatic cholangiocarcinoma to identify synergistic therapies

doi: 10.1038/s41698-022-00320-5

Figure Lengend Snippet: A Quisinostat and pemigatinib synergistically lower the proliferative index (Ki-67) in the PDC-DUC18828 model. The percentage of Ki-67-positive cells was determined and flow cytometry data are shown on the left with corresponding Ki-67 index for each condition shown in the bar graph on the right. Compared to DMSO control, 50 nM pemigatinib decreases the Ki-67 index by ~30%, 100 nM quisinostat decreases the Ki-67 index by ~24%, while 50 nM pemigatinib + 100 nM quisinostat (Pemi + Qui) synergistically decrease the Ki-67 index by ~53% (** p < 0.01 and *** p < 0.001). B Quisinostat and pemigatinib synergistically lower the Ki-67 in the PDO-DUC18828 model. Data are presented as in A . Compared to DMSO control, 10 nM pemigatinib decreases the Ki-67 by ~36%, 25 nM quisinostat decreases the Ki-67 by ~24%, while 10 nM pemigatinib + 25 nM quisinostat (Pemi + Qui) synergistically decreases the Ki-67 index by ~50% (* p < 0.05 and ** p < 0.01). C , D Quisinostat and pemigatinib synergistically enhance apoptosis in the PDC- and PDO-DUC18828 models, respectively. In both models, under the conditions indicated, combined inhibition of HDAC and FGFR increases the proportion of apoptotic cells, especially late-stage apoptosis, but also the early apoptotic population, compared to monotherapy or DMSO control. Q1, dead cells; Q2, late apoptotic/dead cells; Q3, early-apoptotic cells; Q4, healthy cells. E Western blot analysis of PDC-DUC18828 with pemigatinib (2P = 2 nM) monotherapy, quisinostat (50Q = 50 nM) monotherapy, and combination therapy (P + Q = 2 nM pemigatinib + 50 nM quisinostat). Compared to DMSO control, quisinostat monotherapy increases FGFR2, FRS2, MEK, and ERK activation, while pemigatinib monotherapy inhibits FGFR2 signaling, including downstream nodes, FRS2, MEK, and ERK. Combination therapy similarly impairs FGFR2, FRS2, MEK, and ERK activity. Neither monotherapy nor combination therapy alter AKT activity.

Article Snippet: Pemigatinib (Cat# T12401), BGJ398 (infigratinib, Cat# T1975) and quisinostat (Cat# T6055) were purchased from TargetMol Chemicals (Boston, MA).

Techniques: Flow Cytometry, Inhibition, Western Blot, Activation Assay, Activity Assay

Journal: Blood Advances

Article Title: Utilizing epigenetic regulators to improve HSC-based lentiviral gene therapy

doi: 10.1182/bloodadvances.2024013047

Figure Lengend Snippet: CPI203 and quisinostat support ex vivo expansion of HSCs. (A) Schematic experimental design. CD34 + cells were enriched from cord blood and cultured in a cytokine-supplemented medium with quisinostat (0.5 nM) or CPI203 (150 nM) for 4 days. (B) Representative gating strategy of expanded HSCs (CD34 + CD38 – CD90 + CD45RA – CD201 + ) and MPPs (CD34 + CD38 – CD90 – CD45RA – ) in the indicated culture conditions. (C) The graph indicates the frequency of HSC and MPP after 4 days of culture in the indicated culture conditions (n = 9; mean ± standard deviation [SD]). (D) HSC and MPP count after 4 days of culture in the indicated culture conditions (n = 9; mean ± SD). (E) Stacked bar plot showing the composition of HSPCs expanded with only cytokines and in the presence of quisinostat and CPI203. (F) Frequency of cell cycle stages in the expanded HSCs (n = 3; mean ± SD). Statistical significance was determined by a 2-way analysis of variance (ANOVA) test for multiple comparisons: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. CMP, common myeloid progenitor; GMP, granulocyte monocyte progenitor.

Article Snippet: A total of 100 000 to 250 000 enriched CD34 + cells per mL (>90% purity) were cultured in X-vivo15 medium (Lonza) supplemented with recombinant human stem cell factor (SCF; 300 ng/mL), human thrombopoietin (TPO; 100 ng/mL), human FMS-like tyrosine kinase 3 (FLT3; 100 ng/mL) (Miltenyi Biotec), and small molecules of CPI203 (150 nM) (Selleckchem) and quisinostat (0.1, 0.5, and 1 nM) (MedChemExpress).

Techniques: Ex Vivo, Cell Culture, Standard Deviation

Journal: Blood Advances

Article Title: Utilizing epigenetic regulators to improve HSC-based lentiviral gene therapy

doi: 10.1182/bloodadvances.2024013047

Figure Lengend Snippet: Quisinostat increases LV transduction efficiency. (A) Schematic experimental design. CD34 + cells were transduced with LV-GFP 24 hours after stimulation in cytokine-supplemented medium alone or the presence of quisinostat (0.5 nM) or CPI203 (150 nM). Cells were harvested for flow cytometry analysis, DNA isolation, and reverse transcription polymerase chain reaction (RT-PCR), 4 days after transduction. (B) Frequency of GFP + 4 days after transduction in live cells (n = 9; mean ± SD). (C) VCN was determined by RT-PCR (n = 3; mean ± SD; ∗ P < .05; unpaired t test). (D) Frequency of transduced HSCs and MPPs in the indicated culture conditions are shown (n = 9; mean ± SD). (E) Cell counts of transduced HSCs and MPPs (n = 9; mean ± SD). (F) The plot shows the mean fluorescence intensity of H3K27ac in different populations of expanded cells in the indicated culture conditions (n = 3; mean ± SD). Statistical significance was determined by 2-way ANOVA test with multiple comparisons: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001. H3K27ac, acetylation level of lysine 27 on histone 3.

Article Snippet: A total of 100 000 to 250 000 enriched CD34 + cells per mL (>90% purity) were cultured in X-vivo15 medium (Lonza) supplemented with recombinant human stem cell factor (SCF; 300 ng/mL), human thrombopoietin (TPO; 100 ng/mL), human FMS-like tyrosine kinase 3 (FLT3; 100 ng/mL) (Miltenyi Biotec), and small molecules of CPI203 (150 nM) (Selleckchem) and quisinostat (0.1, 0.5, and 1 nM) (MedChemExpress).

Techniques: Transduction, Flow Cytometry, DNA Extraction, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Fluorescence

Journal: Blood Advances

Article Title: Utilizing epigenetic regulators to improve HSC-based lentiviral gene therapy

doi: 10.1182/bloodadvances.2024013047

Figure Lengend Snippet: CPI203 and quisinostat support expansion of functional HSCs. (A) Schematic overview of primary transplantation. A total of 25 000 and 50 000 nucleated expanded cells in the indicated culture conditions were transplanted per mouse. (B) Human engraftment in the BM of NSG mice at week 20 after transplantation (n = 5 mice per group; mean ± standard error of the mean [SEM]; ∗ P < .05). Statistical significance was calculated by 1-way ANOVA, with multiple comparisons. (C) Representative fluorescence-activated cell sorter (FACS) plots of huCD45 in NSG BM 20 weeks after transplantation. (D) Kinetics of chimerism in PB over time (n = 5 mice per group; significance of STF vs STF + CPI203 is shown in the plot). (E) Human engraftment in different organs 20 weeks after transplantation (n = 5 mice per group). (F) Frequency of different populations in BM (n = 5 mice per group). (G) Frequency of different populations in the Thy (n = 5 mice per group; mean ± SEM). Statistical significance was calculated by 2-way ANOVA with multiple comparisons: ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001.

Article Snippet: A total of 100 000 to 250 000 enriched CD34 + cells per mL (>90% purity) were cultured in X-vivo15 medium (Lonza) supplemented with recombinant human stem cell factor (SCF; 300 ng/mL), human thrombopoietin (TPO; 100 ng/mL), human FMS-like tyrosine kinase 3 (FLT3; 100 ng/mL) (Miltenyi Biotec), and small molecules of CPI203 (150 nM) (Selleckchem) and quisinostat (0.1, 0.5, and 1 nM) (MedChemExpress).

Techniques: Functional Assay, Transplantation Assay, Fluorescence

Journal: Blood Advances

Article Title: Utilizing epigenetic regulators to improve HSC-based lentiviral gene therapy

doi: 10.1182/bloodadvances.2024013047

Figure Lengend Snippet: Quisinostat supports better engraftment of transduced cells in vivo. (A) Representative FACS plots of BM at 20 weeks after transplantation in primary mice. (B) Frequency of GFP + cells in different organs (n = 5 mice per group; mean ± SEM; ∗∗ P < .01). Statistical significance was calculated using 2-way ANOVA and multiple comparisons. (C) Frequency of transduced populations within huCD45 + in the BM at 20 weeks after transplantation in primary NSG mice (n = 5 mice per group; mean ± SEM; ∗ P < .05, ∗∗ P < .01). Statistical significance was calculated by 2-way ANOVA and multiple comparisons. (D) Schematic design of the secondary transplantation into NSG mice. (E) Human chimerism in the BM at 20 weeks after transplantation (n = 5 mice per group; mean ± SEM). Statistical significance was calculated using the Mann-Whitney test. (F) Frequency of transduced cells (GFP + ) in the BM (n = 5 mice per group; mean ± SEM; ∗ P < .05, ∗∗ P < .01). Statistical significance was calculated using the Mann-Whitney test. CLP, common lymphoid progenitor.

Article Snippet: A total of 100 000 to 250 000 enriched CD34 + cells per mL (>90% purity) were cultured in X-vivo15 medium (Lonza) supplemented with recombinant human stem cell factor (SCF; 300 ng/mL), human thrombopoietin (TPO; 100 ng/mL), human FMS-like tyrosine kinase 3 (FLT3; 100 ng/mL) (Miltenyi Biotec), and small molecules of CPI203 (150 nM) (Selleckchem) and quisinostat (0.1, 0.5, and 1 nM) (MedChemExpress).

Techniques: In Vivo, Transplantation Assay, MANN-WHITNEY

Journal: Blood Advances

Article Title: Utilizing epigenetic regulators to improve HSC-based lentiviral gene therapy

doi: 10.1182/bloodadvances.2024013047

Figure Lengend Snippet: Single-cell transcriptome analysis reveals quisinostat and CPI203 maintain HSCs upon LV transduction in CD34 + . (A) Schematic experimental design. (B) Mock-up schematic of cell-type projection. (C) Uniform manifold approximation and projection for dimension reduction (UMAP) plots of unstimulated CD34 + cells and integrated UMAP for all conditions. (D) UMAP displaying HSCs for each condition. (E) The bar plot represents population distribution. (F) Violin plots displaying the expression levels of selected HSC surface marker genes in the indicated conditions. (G) Gene set enrichment analysis and molecular processes of various conditions interrogated by scRNA-seq. LMPP, lymphoid multipotent progenitor; MEP, megakaryocyte-Eryth progenitor; ML, myeloid-lymphoid progenitor; Mono, monocyte.

Article Snippet: A total of 100 000 to 250 000 enriched CD34 + cells per mL (>90% purity) were cultured in X-vivo15 medium (Lonza) supplemented with recombinant human stem cell factor (SCF; 300 ng/mL), human thrombopoietin (TPO; 100 ng/mL), human FMS-like tyrosine kinase 3 (FLT3; 100 ng/mL) (Miltenyi Biotec), and small molecules of CPI203 (150 nM) (Selleckchem) and quisinostat (0.1, 0.5, and 1 nM) (MedChemExpress).

Techniques: Transduction, Expressing, Marker

Journal: Cell systems

Article Title: Quantifying drug combination synergy along potency and efficacy axes.

doi: 10.1016/j.cels.2019.01.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Quisinostat (JNJ-26481585) , SelleckChem , S1096.

Techniques: Virus, Recombinant, Sample Prep, Reverse Transcription, SYBR Green Assay, Expressing, Derivative Assay, Plasmid Preparation, Software

Journal: Nucleic Acids Research

Article Title: Small-molecule compounds boost genome-editing efficiency of cytosine base editor

doi: 10.1093/nar/gkab645

Figure Lengend Snippet: Establishment of a reporter system to identify small molecules increasing gene-editing efficiency of CBE in human cells. ( A ) Schematic diagram of BFP to GFP conversion reporter system for evaluation of BE3-mediated gene-editing efficiency. ( B ) Schematic illustration of BFP to GFP sequence editing by CBE. The target sequence is shown. The special residues of His and Tyr are highlighted in blue and green, respectively. The underlined nucleotide represents the PAM. ( C ) Fluorescence images of BFP to GFP conversion. Scale bar, 10 μm. ( D ) Illustration of drug screening platform in the present study. ( E ) The summary of activity of the tested drugs. ( F ) Representative images and raw flow cytometry data of Ricolinostat treatment group. Scale bar, 10 μm. ( G ) Additional HDAC inhibitors modulating BE3 efficiency. All of the compounds were used in 5 μM, except TMP269 (2.5 μM), Panobinostat (100 nM) and Quisinostat 2HCl (100 nM). Error bars, S.E.M.; n = 3; NC, negative control; Control, DMSO treated group; **** P < 0.0001.

Article Snippet: The HDAC inhibitors Ricolinostat, Nexturastat A, Citarinostat, Droxinostat, Entinostat, Panobinostat, Quisinostat 2HCl, TMP269, Tubacin, Tucidinostat were purchased from TargetMol (Boston, MA, USA).

Techniques: Sequencing, Fluorescence, Activity Assay, Flow Cytometry, Negative Control

Journal: Colloids and surfaces. B, Biointerfaces

Article Title: pH driven precipitation of quisinostat onto PLA-PEG nanoparticles enables treatment of intracranial glioblastoma

doi: 10.1016/j.colsurfb.2018.02.048

Figure Lengend Snippet: Summary of QNP formulation parameters and characterization.

Article Snippet: Quisinostat (JNJ-26481585) was obtained from APExBio (Houston, TX USA).

Techniques: Formulation, Zeta Potential Analyzer

Journal: Colloids and surfaces. B, Biointerfaces

Article Title: pH driven precipitation of quisinostat onto PLA-PEG nanoparticles enables treatment of intracranial glioblastoma

doi: 10.1016/j.colsurfb.2018.02.048

Figure Lengend Snippet: Nanoparticle’ hydrodynamic diameter, as measured by DLS, positively correlated (Pearson coefficient = 0.9108, p < 0.0001) with the quisinostat loading for each batch. Each data point represents an individual batch.

Article Snippet: Quisinostat (JNJ-26481585) was obtained from APExBio (Houston, TX USA).

Techniques:

Journal: Colloids and surfaces. B, Biointerfaces

Article Title: pH driven precipitation of quisinostat onto PLA-PEG nanoparticles enables treatment of intracranial glioblastoma

doi: 10.1016/j.colsurfb.2018.02.048

Figure Lengend Snippet: QNPs released quisinostat into PBS at 37 °C over 48 h, with nearly 50% release occurring in the first 6 h. Free quisinostat was completely released from the cassette within 4 h. Points and error bars represent the mean ± SD of 3 samples read in triplicate at each time point.

Article Snippet: Quisinostat (JNJ-26481585) was obtained from APExBio (Houston, TX USA).

Techniques:

Journal: Colloids and surfaces. B, Biointerfaces

Article Title: pH driven precipitation of quisinostat onto PLA-PEG nanoparticles enables treatment of intracranial glioblastoma

doi: 10.1016/j.colsurfb.2018.02.048

Figure Lengend Snippet: QNP and free quisinostat exhibited equipotent growth inhibition against GL261 murine glioma cells in vitro with IC50 s of 30 and 24 nM, respectively. Points and error bars represent the mean ± SD of 3 samples read in triplicate at each dilution.

Article Snippet: Quisinostat (JNJ-26481585) was obtained from APExBio (Houston, TX USA).

Techniques: Inhibition, In Vitro

Journal: Colloids and surfaces. B, Biointerfaces

Article Title: pH driven precipitation of quisinostat onto PLA-PEG nanoparticles enables treatment of intracranial glioblastoma

doi: 10.1016/j.colsurfb.2018.02.048

Figure Lengend Snippet: (A) Tumor growth was determined by the change in tumor size (mean ± SD) from day 6, as measured by bioluminescence. (B) Survival is shown on the Kaplan-Meier plot. (C) Saline (n = 5) and Free Quisinostat (n = 5) treated tumors both doubled in size every 2.4 days and had median survival times of 22 and 19 days, respectively.

Article Snippet: Quisinostat (JNJ-26481585) was obtained from APExBio (Houston, TX USA).

Techniques: Saline

Journal: Frontiers in Oncology

Article Title: A review of the therapeutic potential of histone deacetylase inhibitors in rhabdomyosarcoma

doi: 10.3389/fonc.2023.1244035

Figure Lengend Snippet: HDAC classes and interaction complexes.

Article Snippet: Quisinostat (Janssen Pharmaceuticals, Belgium), or JNJ-26481585, activates the mitochondrial apoptosis pathway by triggering the Bax/Bak complex, which activates enzyme caspase-9 ( ).

Techniques: