A5110778611- Immunotherapy and Immune Responses
- Cancer Immunotherapy and Biomarkers
- Advanced Breast Cancer Therapies
- Cancer-related Molecular Pathways
- Immune cells in cancer
- Microtubule and mitosis dynamics
- Lung Cancer Research Studies
Roswell Park Comprehensive Cancer Center
2021-2025
Cancer Genetics (United States)
2021-2024
Abstract CDK2 inhibitors have recently been developed and entered clinical trials. Combination approaches can help broaden the use of therapeutic agents establish more effective treatments. Here, we evaluated selective inhibitor BLU-222 for mechanisms response in context ovarian breast cancer models. Sensors cellular CDK activity indicated that sensitivity to either CDK4/6 or inhibition was related differential dependence on a single G1/S transition. Unlike inhibitors, able robustly inhibit...
Abstract Cyclin dependent kinase 2 (CDK2) regulates cell cycle and is an emerging target for cancer therapy. There are relatively small numbers of tumor models that exhibit strong dependence on CDK2 undergo G1 arrest following inhibition. The expression P16INK4A cyclin E1 determines this sensitivity to co-expression these genes occurs in breast patients highlighting their clinical significance as predictive biomarkers CDK2-targeted therapies. In genetically independent CDK2, pharmacological...
<p>Differential vulnerabilities to CDK2 versus CDK4/6 inhibition. A, Table of top identified predictive genes in breast and ovarian carcinomas for response inhibition from the publicly available DepMap dataset using CRISPR (top) RNAi (bottom) screens. B, Cell Titer Glo values normalized 0 nM treatment cancer cells treated with BLU-222 5 days. C, summarizing cell line treatment. D, COV-504 PE-O1 lines serial concentrations or palbociclib days monitored Cellcyte Incucyte live...
<p>Supplemental Figure S1-7 Legends</p>
<p>Figure S6. P16INK4A is a key determinant of response to BLU-222. A, Heatmap analysis from the cancer dependency map (DepMap) database displaying relative expression <i>CCNE1</i> and several endogenous CDK inhibitor genes in panel established ovarian breast cell lines. B, Western blot RB1-E2F pathway activity MDA-MB-157 cells following overexpression (OE) <i>CDK4</i> with without BLU-222 treatment. C, Live proliferative monitoring WT (left) overexpressing (OE,...
<p>Figure S7. Prevalence of Cyclin E1-P16INK4A<sup>high</sup> tumors that express RB. A, Kaplan Meier curves based on <i>CCNE1</i> status generated from The Cancer Genome Atlas (TCGA) serous ovarian carcinoma database. HR, hazard ratio; DFS, disease-free survival; OS, overall survival. Amp, amplification. B, Oncoprint the TCGA database for indicated cell cycle genes. C, Normalized intensity D1, E1, or P16INK4A expression within clusters 1, 3, and 4 (n = 82002...
<div>Abstract<p>Cyclin-dependent kinase 2 (CDK2) inhibitors have recently been developed and entered clinical trials. Combination approaches can help broaden the use of therapeutic agents establish more effective treatments. Here, we evaluated selective CDK2 inhibitor BLU-222 for mechanisms response in context ovarian breast cancer models. Sensors cellular CDK activity indicated that sensitivity to either CDK4/6 or inhibition was related differential dependence on a single...
<p>Figure S5. Cooperative and antagonistic combination strategies with BLU-222. A, BLISS synergistic analysis for antagonism between BLU-222 CHK1 checkpoint kinase inhibitors in MDA-MB-231 cells (n = 3 per group; scores calculated SynergyFinder v3). B, Live cell proliferative monitoring of the indicated lines treated BLU-222, palbociclib, or 3–4 one-way ANOVA; error bars represent SEM). C, Quantification mVenus-DHB mCherry-RB CDK activity sensor localization to cytosol nucleus HCC-1806...
<p>Figure S3. BLU-222 elicits potent G1 arrest in Cyclin E1 and P16INK4A-high models expressing RB. A, Cell cycle profiling from univariate flow cytometric analysis the indicated cell lines following treatment with palbociclib. B, Bivariate transfection si<i>NT</i> or si<i>CCNE1</i> constructs. C, RNA sequencing gene set enrichment plots for E2F targets interferon alpha response pathways Kuramochi MDA-MB-157 cells 125 nM 100 nM, respectively, 48 hours compared...
<p>Figure S2. Contextual biochemical response to CDK inhibition. A, Western blot of RB1-E2F pathway activity in OVSAHO and MDA-MB-157 cells following BLU-222 or palbociclib treatment. B, Schematic depicting mVenus-DHB mCherry-RB reporter constructs. C, Representative images sensor OVCAR-3 T-47D treated with 500 nM (scale bar = 100 µm). D, Quantification localization the cytosol nucleus indicated ovarian breast cancer cell lines treatment either palbociclib. E, transfected...
<p>Figure S4. Loss of RB1 shifts phase arrest and sensitivity to BLU-222. A, Heatmap RNA sequencing data comparing differential gene expression between BLU-222-treated Kuramochi (125 nM BLU-222) OVCAR-8 (250 cells normalized respective DMSO-treated controls. B, Cell cycle profiling from univariate flow cytometric analysis at baseline, after 1 5 days treatment with 500 BLU-222, drug washout. C, Western blot for abrogation G2/M checkpoint signaling in following doxycycline (Dox)...
<p>Supplemental File 1. BLU-222 combinatorial drug screen data from MDA-MB-231 (tab 1) and COV-504 cells 2) reported in Figure 5. Data as percent growth compared to sole treatment with 0.1% DMSO.</p>
Abstract Therapeutic inhibition of programmed cell death ligand (PD-L1) is linked to alterations in interferon (IFN) signaling. Since IFN-regulated intracellular signaling can control extracellular secretory programs tumors modulate immunity, we examined IFN-related changes tumor cells following resistance PD-L1 inhibition. Here report an anti-PD-L1 treatment-induced secretome (PTIS) models acquired that regulated by type I IFNs. These suppress activation T ex vivo while diminishing...
Abstract Background Interferon (IFN) pathway activation in tumors can have dual, sometimes opposing, influences on immune responses. Therapeutic inhibition of programmed cell death ligand (PD-L1) – a treatment that reverses PD-1-mediated suppression tumor-killing T-cells - is linked to alterations IFN signaling; however, less known about the role IFNs after resistance. Since IFN-regulated intracellular signaling control extracellular secretory programs modulate immunity, we examined...