oci aml2  (Procell Inc)

Journal: Cell Reports Medicine

Article Title: CDK4/6 inhibition overcomes venetoclax resistance mechanisms with enhanced combination activity in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102526

Figure Lengend Snippet: Venetoclax, in combination with the CDK4/6 inhibitor palbociclib, inhibits growth, stalls cell cycle progression, and reduces tumor burden in AML models (A–C) OCI-AML2 cells assessed after a 5-day treatment with palbociclib (1 μM), venetoclax (200 nM), or the combination (equimolar to single agents) to determine levels of apoptosis (A), cell cycle progression (B), and percentage of viable OCI-AML2 cells/mL of media (C) Data represent the mean ± SD for 3 replicates (∗∗ p ≤ 0.01). (D) Schematic depicting two independent PDX experiments. PDX model 1: evaluation of disease burden after injection of AML patient tumor cells. PDX model 2: survival studies after injection of AML patient tumor cells. The schematic was generated using BioRender. (E–G) Flow cytometry analysis at time of euthanization for PDX model 1 showing human (h)CD45 blasts in peripheral blood (E), spleen weight of animals (F), and percentage of hCD45 chimerism in spleen tissue (G). Mean values ± SEM are shown unless otherwise stated. Two-tailed Student’s t tests were used for comparisons (∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001). (H) Survival Kaplan-Meier curves for PDX model 2 (log rank [Mantel-Cox] test, ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001).

Article Snippet: Human: OCI-AML2 , DSMZ , DSMZ no:ACC 99.

Techniques: Injection, Generated, Flow Cytometry, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: CDK4/6 inhibition overcomes venetoclax resistance mechanisms with enhanced combination activity in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102526

Figure Lengend Snippet: A combination of ven+palbo leads to changes in protein synthesis rate and translational machinery (A) Bulk RNA-seq for 560 primary AML samples, showing Pearson correlations between AML cellular state eigengenes (columns) and Reactome Translation pathway eigengenes (rows). Protein synthesis pathways positively correlate (red) with a progenitor-like cell-state signature and negatively correlate (blue) with a monocyte-like cell-state signature. (B) MTS assays confirming drug responsiveness in parental/venetoclax-sensitive cells (VenS/Par) and venetoclax-resistant cells (VenR) for OCI-AML2 cell lines. Data points denote the mean normalized cell viability ± SD for 3 replicates. (C) SUnSET assay to assess protein synthesis in OCI-AML2 cells treated with drug for 24 h. Puromycin incorporation into newly synthesized proteins was measured in ven-sensitive (VenS/Par [parental]) and -resistant (VenR) cells. A representative image of n = 4 immunoblots is shown. (D) Estimation plots of intensity measurements of anti-puromycin signal from 4 separate immunoblot experiments. (E) Immunoblots of OCI-AML2 cells following 24-h or 5-day drug treatments. Vinculin is used as a loading control. (F) SUnSET assay immunoblots showing puromycin incorporation in progenitor-like or monocyte-like patient samples after drug treatment. (G) Flow-cytometry-based SUnSET assay detecting CD64 + , CD11b + , CD33 + monocytes. The percentage of puromycin-positive cells reflects active protein synthesis within this monocyte population.

Article Snippet: Human: OCI-AML2 , DSMZ , DSMZ no:ACC 99.

Techniques: RNA Sequencing, Synthesized, Western Blot, Control, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: CDK4/6 inhibition overcomes venetoclax resistance mechanisms with enhanced combination activity in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102526

Figure Lengend Snippet: Combining ven+palbo mitigates single-agent resistance due to clinically observed mutations (A) Enrichment of individual sgRNAs for RB1, BAX, and IKZF1 shown as fold change over DMSO control following a 21-day exposure to palbo, ven, or ven+palbo in OCI-AML2 Cas9 C6 cells. (B) Immunoblot showing efficiency of knockdown of RB1, BAX, and IKZF1 proteins in OCI-AML2 cell lines. A cell line expressing an NT sgRNA was used to generate a control cell line. Vinculin was used as a protein loading control. Par, parental; NT, non-targeting. (C–F) Dose-response curves for OCI-AML2 NT and KO cell lines evaluated for drug sensitivity to palbo, ven, or the combination. Data points denote the mean normalized cell viability ± SD for 3 replicates. (G) IC 50 values derived from dose-response curves of OCI-AML2 cell line drug sensitivity assays shown in (C)–(F). Data represent the mean IC 50 ± SD for 3 replicates (∗ p ≤ 0.05 and ∗∗ p ≤ 0.01 by Student’s t test). (H–J) Outgrowth of OCI-AML2 Non-targeting (H), OCI-AML2 Bax KO (I), and OCI-AML3 cell lines (J) treated with palbo, aza, and ven single agents, duplicate combinations and the triplet. Total viable cells over a 14-day drug treatment are shown. Data points denote the mean total number of viable cells ± SD for 3 replicates. One-way ANOVA with Tukey’s post-test for multiple comparisons was used for day 7 and day 14 time points as indicated. (∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001) (K) Immunoblot of apoptotic proteins in OCI-AML2 cells, drug treated for 14 days.

Article Snippet: Human: OCI-AML2 , DSMZ , DSMZ no:ACC 99.

Techniques: Control, Western Blot, Knockdown, Expressing, Derivative Assay

Journal: Cell Reports Medicine

Article Title: CDK4/6 inhibition overcomes venetoclax resistance mechanisms with enhanced combination activity in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102526

Figure Lengend Snippet: Loss of IKZF1 leads to increased expression of AXL and is increased in IKZF1 -mutated AML patient samples (A) Volcano plot highlighting genes of interest from RNA-seq in IKZF1 -KO OCI-AML2 cells, color coded based on primary known function. (B) qPCR validation of RNA-seq results showing mean fold change ± SD for 3 replicates. (C) Immunoblot shows upregulation of AXL protein with loss of IKZF1 in OCI-AML2 cells. (D) AXL mRNA is overexpressed in AML primary patient samples harboring IKZF1 mutations ( n = 9) compared to WT samples ( n = 662). (E) OCI-AML2 IKZF1 -KO cells show resistance (red dots) to palbo, ven, and ven+palbo and retain drug sensitivity to several AXL inhibitors (blue dots). Sensitivity is shown as a percentage of the maximum area under the dose response curve (AUC) derived for a 7-point concentration series ranging from 10 μM to 10 nM. (F) AUC values from ex vivo drug sensitivity assays for 4 primary AML samples, each harboring the IKZF1 hotspot mutation N159S (∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001 by Student’s t test).

Article Snippet: Human: OCI-AML2 , DSMZ , DSMZ no:ACC 99.

Techniques: Expressing, RNA Sequencing, Biomarker Discovery, Western Blot, Derivative Assay, Concentration Assay, Ex Vivo, Mutagenesis