Journal: bioRxiv

Article Title: Oncodevelopmental plasticity of the skeleton in myeloid neoplasms

doi: 10.64898/2026.03.19.712939

Figure Lengend Snippet: a, Experimental design of the in vitro BM organoid treatment assay. Mature human iPSC-derived bone marrow organoids were competitively engrafted with JAK2V617F;RFP⁺ and isogenic JAK2WT;GFP⁺ differentiated hematopoietic stem and progenitor cells (iHSPCs) at a 1:1 ratio and treated statically with the THBS1 antagonist LSKL or vehicle control for 7 days. b, Flow cytometric analysis of donor-derived cells from BM organoids treated with vehicle or LSKL, showing frequencies of HSPCs (CD45+CD34+) and erythroid cells (CD45-CD235a+) derived from mutant and WT competitors. Two-way-ANOVA with post hoc Tukey’s was used. c, qRT-PCR of BM-derived cells following LSKL or vehicle control treatment. Statistical analysis was performed using two-way ANOVA with Sidak’s correction. d, Schematic overview of treatment effect on patient-derived primary cells engrafted into BM organoids. Peripheral blood mononuclear cell-derived CD34+ fraction from a patient diagnosed with post-ET MF were labelled with CellVue dye and engrafted into mature iPSC-derived BM organoids. BM organoids with patient cells were treated statically with the THBS1 antagonist LSKL or vehicle control for 7 days. e, Clinical hematological parameters showing blood counts of a post-ET MF patient at the time of PBMC isolation. f, Flow cytometric analysis of CellVue-labeled patient-derived cells following vehicle or LSKL treatment. g, Schematic overview of the preclinical organ-on-a-chip platform enabling dynamic drug perfusion. BM organoids engrafted with JAK2V617F;RFP and JAK2WT;GFP CD34+ cells were cultured under continuous flow, with LSKLor vehicle delivered exclusively through the microfluidic circulation channel. h, Flow cytometric analysis of the preclinical organ-on-a-chip model with vehicle or LSKL treatment groups, showing frequencies of immature erythrocytes (CD45+CD235a+) and monocytes (CD45+CD14+) derived from mutant and WT competitors. Two-way-ANOVA with post hoc Tukey’s was used. i, Experimental design of in vivo therapeutic intervention. Lethally irradiated mice were competitively transplanted with JAK2V617F/JAK2WT and WT competitor bone marrow cells at a ratio 1:1. Blood counts were performed once in 4 weeks starting from 4 weeks post transplant. JAK2V617F mice started receiving the treatment at 16 weeks post-transplantation (time point 0) with vehicle (n=6), Fedratinib (n=4), LSKL (n=5), or combined Fedratinib + LSKL (n=4) until 20 and 24 weeks post transplant (4 and 8 weeks of treatment, respectively). j, Blood counts over time showing white blood cells (WBC), red blood cells (RBCs) and platelets. Each dot represents the value from one mouse. Two-way-ANOVA with post hoc Tukey’s was used. k, Representative images of H&E and reticulin staining on tibial sections from mice treated for 8 weeks. Scale bars, 50µm. l, Reticulin staining and grading of bone marrow fibrosis from mice treated for 8 weeks with vehicle (n=6), Fedratinib (n=4), LSKL (n=5), or combined Fedratinib + LSKL (n=4). Kruskal-Wallis H test was used. m-n, Reconstruction of 3D µCT images of femoral cortical (M) and trabecular (N) bones from mice treated for 8 weeks. o, Quantification of trabecular (Tb.) thickness from the proximal part of the femur. Mice treated for 8 weeks with vehicle (n=6), Fedratinib (n=4), LSKL (n=5), or combined Fedratinib + LSKL (n=4). One-way-ANOVA with Holm-Sidak’s correction was used. p, Reconstruction of 3D µCT images of maxillary bones from mice treated for 8 weeks.

Article Snippet: For pharmacological treatments, fedratinib (120 mg/kg, HY-10409A, MCE) was dissolved in vehicle solution (0.5% methylcellulose + 0.05% Tween-80 in water) and administered to mice via oral gavage daily, 5 days/week.

Techniques: In Vitro, Derivative Assay, Control, Mutagenesis, Quantitative RT-PCR, Isolation, Labeling, Cell Culture, In Vivo, Irradiation, Transplantation Assay, Staining

Journal: bioRxiv

Article Title: Targeting the energy metabolism of melanoma cells: FX-11 acts as a mitochondrial uncoupler

doi: 10.1101/2024.11.04.621801

Figure Lengend Snippet: (A) Graphs showing the effect of different concentrations of FX-11 on cell viability in 624Mel and Wm3248 cells. After 48h of treatment, cells were stained with Hoechst 33342 and Sytox Orange for 30min and imaged on a Cytation 5. Mean and standard deviation of one representative biological replicate out of 3 are shown, each performed in 3 technical replicates, (B) Fold induction of apoptosis following 16h drug incubation as measured by Caspase 3 activity assay. Positive controls (Staurosporine (Stauro.) and TG101348 (TG)) are shown in green. Encorafenib (Encora) which leads to growth inhibition but does not induce apoptosis is also shown. Mean and standard deviation of one representative biological replicate out of 3 are shown, each performed in 3 technical replicates, (C) Relative ROS induction following 4h drug incubation of 624Mel cells. Cells were stained with CMH2DCFDA and analysed by flow cytometry in the FITC channel. Menadione serves as positive control. Means ± SD; n=3 independent experiments except for Menadione (n=2).

Article Snippet: 2μM Staurosporine (Santa Cruz Biotechnology) or 30μM TG101348 (Selleck chemicals) were included as positive controls.

Techniques: Staining, Standard Deviation, Incubation, Caspase-3 Activity Assay, Inhibition, Flow Cytometry, Positive Control

Journal: Gene

Article Title: A novel cancer-germline gene DAZL promotes progression and cisplatin resistance of non-small cell lung cancer by upregulating JAK2 and MCM8.

doi: 10.1016/j.gene.2024.148449

Figure Lengend Snippet: Fig. 6. Inhibition of JAK2 counteracts the effects of DAZL overexpression on NSCLC cell viability, colony formation, migration, and invasion. (A-E) Cells overexpressing empty vector (Vec) or DAZL cDNA (DAZL) were treated with vehicle (Veh) or fedratinib (Fed) for 24 h, then Western blot assay (A), MTT assay (B), colony information assay (C), transwell invasion assay (D) and wound healing assay (E) were performed. (F-H) The statistical analysis results of triplicate colony information assay (F), transwell invasion assay (G), and wound healing assay (H). ns, p ≥0.05, *, p < 0.05 and **, p < 0.01.

Article Snippet: Fedratinib (TG101348) was purchased from Selleck (Houston, TX, USA).

Techniques: Inhibition, Over Expression, Migration, Plasmid Preparation, Western Blot, MTT Assay, Transwell Invasion Assay, Wound Healing Assay

Journal: Frontiers in Immunology

Article Title: Induction of T cell exhaustion by JAK1/3 inhibition in the treatment of alopecia areata

doi: 10.3389/fimmu.2022.955038

Figure Lengend Snippet: γc cytokines regulated effector T cell exhaustion. (A) to (J) T cells from C3H/HeJ mice without AA were stimulated with 500 ng/ml anti-CD3 in the presence of indicated regents in vitro for 4 (d) (A) and (B) The expression of PD-1 on CD8 + T cells was measured by FACS after treated with increasing dose of Ifidancitinib. *P < 0.05, ***P < 0.001 (one-way ANOVA). (C) and (D) The expression of PD-1 on CD8 + T cells was measured by FACS after treated with increasing dose of STAT5 inhibitor. *P < 0.05, ***P < 0.001 (one-way ANOVA). (E) and (F) The expression of PD-1 on CD8 + T cells was measured by FACS after treated with 1 µM of JAK1i (Itacitinib), JAK2i (Fedratinib), JAK3i (Ritlecitinib), JAK1/2-selective inhibitor Ruxolitinib (Ruxo), or pan-JAK inhibitor Tofacitinib (Tofa). ns indicates not significant, *P < 0.05, ***P < 0.001. P values were determined using one-way ANOVA followed by Brown-Forsythe test. (G) and (H) The expression of PD-1 on CD8 + T cells was measured by FACS after treated with 20 µg/ml IL-2 neutralizing mAbs. **P < 0.01. (Unpaired Student t test). (I) The expression of Eomes and TOX in CD8 + T cells was measured by FACS after treated with 1µM of JAK1i (Itacitinib), JAK2i (Fedratinib), JAK3i (Ritlecitinib), JAK1/2-selective inhibitor Ruxolitinib (Ruxo), Ifidancitinib, or pan-JAK inhibitor Tofacitinib (Tofa). (J) The expression of Eomes and TOX in CD8 + T cells was measured by FACS after treated with 20 µg/ml IL-2 neutralizing mAbs and isotype control mAbs. (K) to (M) C3H/HeJ mice with AA were treated with a combination of IL-2 neutralizing mAbs, IL-9 neutralizing mAbs and IL-15 neutralizing mAbs or isotype for 8 weeks. (K) Representative images of anti-IL2/9/15 or isotype treated C3H/HeJ mice before or after 8 weeks treatment. (L) Percentage of skin hair loss or regrowth is shown before and after treatment. **P < 0.01, ***P < 0.001 (Unpaired Student t test). The expression of PD-1 on CD8+ T cells was measured by FACS after treated with 20 µg/ml IL-2/9/15 neutralizing mAbs. **P < 0.01. (Unpaired Student t test). The results are representative of two separate experiments. (N) to (P) C3H/HeJ mice with AA were treated with Ritlecitinib or vehicle systemically for 8 weeks. (N) Representative images of Ritlecitinib or vehicle treated C3H/HeJ mice before or after 8 weeks treatment. (O) Percentage of skin hair loss or regrowth is shown before and after treatment. ***P < 0.001 (Unpaired Student t test). (P) The frequency of PD-1+TOX+CD44+CD8+ T cells within SDLNs were measured with mice that were systemically treated with Ritlecitinib or vehicle for 8 weeks. *P < 0.05. (Unpaired Student t test). The results are representative of two separate experiments. The results are representative of two separate experiments.

Article Snippet: Itacitinib (catalog HY-16997, MedChemExpress), Fedratinib (catalog 202893, Medkoo), Ritlecitinib (catalog PZ0316, MilliporeSigma), Ruxolitinib (catalog S1378, Selleck), Tofacitinib (catalog 200811, Medkoo)

Techniques: In Vitro, Expressing

Journal: Informatics in Medicine Unlocked

Article Title: Contribution of machine learning approaches in response to SARS-CoV-2 infection

doi: 10.1016/j.imu.2021.100526

Figure Lengend Snippet: Repurposed drugs which their efficiency in inhibition of SARS-CoV-2 infection is predicted by artificial intelligence.

Article Snippet: Fedratinib , Janus kinase (JAK) , BenevolentAI , - , Myeloproliferative diseases , [ , ] .

Techniques: Inhibition, Infection, In Vitro

Journal: eLife

Article Title: Early and delayed STAT1-dependent responses drive local trained immunity of macrophages in the spleen

doi: 10.7554/eLife.100922

Figure Lengend Snippet: ( A ) Mouse model to detect training markers in BCG-vaccinated WT and STAT1-KO mice. ( B ) CXCL9 and Sca-1 expression in MPs between control and BCG mice in WT or STAT1-KO background. ( C ) CD11b + and RPM populations between control and BCG mice in WT or STAT1-KO background. ( D ) Mouse model of BCG vaccination with early interferon inhibition using the small molecule inhibitor Fedratinib targeting Type-II IFN, or Deucravacitinib targeting Type-I IFN. ( E ) Heatmap of log2 fold change for DEGs across inhibitor conditions. ( F ) CXCL9 expression in MPs across inhibitor conditions. ( G ) Flow cytometry gating strategy used for identification of BM LSK HSCs. Gating is representative of a BCG inoculated mouse 2 weeks post injection. Lineage staining combines CD4, CD8, B220, Ter119, Gr1, B220, and CD11b for separation of T, B, erythroid, granulocyte, and myeloid cells respectively. ( H ) Flow cytometry plot of LSK expansion within the BM across inhibitor conditions. ( I ) Two-dimensional spleen area from across experimental conditions including training, Fedratinib inhibitor, and S.Tm infection (n=3–6). Significance between inhibitor and infection conditions is indicated. ( J ) PCA projection of all samples onto the space of the two leading principal components based on all DEGs across training, inhibitor, and infection conditions (two-sample t-test, 5% FDR). The percentage of variance explained by each PC is indicated at the PC axes. Color is indicative of control or BCG mice before and after S.Tm challenge. Circle or diamond shape represent treatment with DMSO or Fedratinib. ( K–L ) Heatmap of STAT-1 signature genes ( K ) and their mean log2 expression across training conditions and antibody treatment ( L ). ( M ) Total BCG CFU from spleens from isotype and α-IFNγ treated mice 2 weeks post vaccination (n=4). Data in ( I ) and ( M ) are presented as mean ± SEM. Heatmap rows in ( E ) and ( K ) indicate biological replicates. Two-tailed t -test used for data in ( I ) and ( L ) (*p≥0.05, **p≥0.01, ***p≥0.005, ****p≥0.001).

Article Snippet: JAK-STAT inhibitors Fedratinib (cat #202893), and Deucravacitinib (cat #555349) (MedKoo Biosciences) were resuspended in DMSO, aliquoted, and stored at –80 C for later use.

Techniques: Expressing, Control, Inhibition, Flow Cytometry, Injection, Staining, Infection, Two Tailed Test

Journal: eLife

Article Title: Early and delayed STAT1-dependent responses drive local trained immunity of macrophages in the spleen

doi: 10.7554/eLife.100922

Figure Lengend Snippet: ( A ) Mouse model of BCG vaccination with early interferon inhibition using the Fedratinib inhibitor. ( B ) Splenic S. Tm CFU for control and BCG mice, with and without Fedratinib inhibitor, 24 hr post infection (n=2–6).( C ) MP populations from control (gray) and BCG (black) mice, with or without Fedratinib inhibition. Percentage of CM and NCM cells calculated from CD11b + population. Percentage of RPM calculated from Lin - population. ( D ) Percentage of CXCL9 + CM-T, NCM, and RPM populations from control (gray) and BCG (black) mice, with or without Fedratinib inhibition (control: n=3, BCG: n=4, control +Fedratinib: n=3, BCG +Fedratinib: n=6). ( E ) Heatmap of normalized log2 expression of DEGs across naive and training conditions. ( F ) Gene set enrichment analysis of DEGs from E. Data in bar graphs are presented as mean ± SEM. For bar graph ( B ) each individual point is a biological repeat. Heatmap rows in E indicate biological replicates. Two-tailed t -test used for data in ( B ), ( C ), and ( D ) (*p<0.05, **p<0.01, ***p<0.005, ****p<0.001).

Article Snippet: JAK-STAT inhibitors Fedratinib (cat #202893), and Deucravacitinib (cat #555349) (MedKoo Biosciences) were resuspended in DMSO, aliquoted, and stored at –80 C for later use.

Techniques: Inhibition, Control, Infection, Expressing, Two Tailed Test

Journal: Blood Advances

Article Title: Fedratinib in myelofibrosis

doi: 10.1182/bloodadvances.2019000954

Figure Lengend Snippet: Chemical structure of TG101348.

Article Snippet: Commercial development of fedratinib: TargeGen, Sanofi, Impact, Celgene Fedratinib development took a tortuous relay path requiring multiple partners.

Techniques:

Journal: Blood Advances

Article Title: Fedratinib in myelofibrosis

doi: 10.1182/bloodadvances.2019000954

Figure Lengend Snippet: Kinase profiles of current JAK2 inhibitors

Article Snippet: Commercial development of fedratinib: TargeGen, Sanofi, Impact, Celgene Fedratinib development took a tortuous relay path requiring multiple partners.

Techniques:

Journal: Blood Advances

Article Title: Fedratinib in myelofibrosis

doi: 10.1182/bloodadvances.2019000954

Figure Lengend Snippet: Key milestones in the development of fedratinib. P, phosphorylation; Ph 1, phase 1; Ph 2, phase 2; Ph 3, phase 3; PI3K, phosphatidylinositol 3-kinase.

Article Snippet: Commercial development of fedratinib: TargeGen, Sanofi, Impact, Celgene Fedratinib development took a tortuous relay path requiring multiple partners.

Techniques:

Journal: Blood Advances

Article Title: Fedratinib in myelofibrosis

doi: 10.1182/bloodadvances.2019000954

Figure Lengend Snippet: Clinical utilization of fedratinib. Hep B, hepatitis B; JAKi, JAK inhibitor.

Article Snippet: Commercial development of fedratinib: TargeGen, Sanofi, Impact, Celgene Fedratinib development took a tortuous relay path requiring multiple partners.

Techniques: