Journal: Molecular cancer therapeutics

Article Title: Targeting Monounsaturated Fatty Acid Metabolism for Radiosensitization of KRAS Mutant 3D Lung Cancer Models

doi: 10.1158/1535-7163.MCT-24-0213

Figure Lengend Snippet: Genotype correlation of SCD1 inhibitor effects. A , Primary sphere formation assay using limited dilution in four representative cell lines from Fig. 1B . Cells seeded at the density shown were irradiated with 4 Gy and exposed to inhibitor at “high” (1 μM) and “low” (0.1 μM) concentrations. Shown is a representative replicate from up to three biological repeats. B , Radiosensitization factors (SRF) for the same four cell lines as in Panel A using two SCD1 inhibitors in parallel (MF438 over a dose range of 100 nM - 3.3 μM and CAY10655 over 30 nM - 1 μM). For each model, median SRF, interquartile range, and range of all replicates from two biological repeats are shown. C , Comparison of pooled SRF technical repeats for 27 NSCLC cell lines grown in 3D, analogous to Fig. 1C . D , Cell viability after treatment with MF438 (D4, D3, D2, and D1 being 3, 10, 33, and 100 nM, respectively). Data points represent median viability for 8 technical replicates per drug concentration based on two biological repeats. Statistical comparison by Fisher Exact, two-sided. *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001; ns, not significant.

Article Snippet: Stearoyl-CoA Desaturase 1 (SCD1) inhibitor (MF438, Cat #569406) and CAY10566 (Cat #HY-15823) were purchased from EMD Millipore, Darmstadt, Germany and MedChemExpress, Monmouth Junction, NJ, respectively (see Tab.

Techniques: Tube Formation Assay, Irradiation, Comparison, Concentration Assay

Journal: Molecular cancer therapeutics

Article Title: Targeting Monounsaturated Fatty Acid Metabolism for Radiosensitization of KRAS Mutant 3D Lung Cancer Models

doi: 10.1158/1535-7163.MCT-24-0213

Figure Lengend Snippet: SCD1 activity and expression in KRAS mutant and wild-type NSCLC models. A , Cell viability after MF438 treatment of isogenic NCI-H1703 cells with endogenous wild-type (wt) KRAS or with exogenous expression of mutant (mut) KRAS in 3D. Data points represent mean +/− SE of 8 replicates. B , Measurement of lipid peroxidation with C11 BODIPY 581/591 in NCI-H1703 cells following treatment with negative control (Ctrl, DMSO), ferroptosis activator (RSL3, 5 μM), or SCD1 inhibitor MF438 (1 μM). Left, representative images. Right, bars represent mean + SE normalized oxidation ratio for two to three biological repeats. C , Expression levels of SCD1 in 2D versus 3D cultures of isogenic NCI-H1703 cells with/without mutant KRAS. D , Left, Representative images showing individual fatty acid peaks as determined by GC-MS. Right, Quantification of fatty acid peaks plotting the ratio of SFA to MUFAs for different genotypes and culture conditions of isogenic NCI-H1703 cells. Bars represent mean + SE based on three measurements. E , SCD1 expression levels comparing the most strongly radiosensitized (S) models in Fig. 1B (NCI-H460, NCI-H1944, NCI-H2122, NCI-H441, A427, LU65 NCI-H2023, NCI-H1651, NCI-H2126) to the remaining models (R). Data points correspond to individual cell lines with KL double mutants highlighted. F , Fast pre-ranked gene set enrichment analysis comparing radiosensitized (S) versus resistant (R) models with KL mutations. Normalized enrichment scores for Hallmark pathways with adjusted p≤0.05 are shown. See also Fig. S3 . All statistical comparisons by t-test or Mann-Whitney test, two-sided; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001.

Article Snippet: Stearoyl-CoA Desaturase 1 (SCD1) inhibitor (MF438, Cat #569406) and CAY10566 (Cat #HY-15823) were purchased from EMD Millipore, Darmstadt, Germany and MedChemExpress, Monmouth Junction, NJ, respectively (see Tab.

Techniques: Activity Assay, Expressing, Mutagenesis, Negative Control, Gas Chromatography-Mass Spectrometry, MANN-WHITNEY

Journal: Molecular cancer therapeutics

Article Title: Targeting Monounsaturated Fatty Acid Metabolism for Radiosensitization of KRAS Mutant 3D Lung Cancer Models

doi: 10.1158/1535-7163.MCT-24-0213

Figure Lengend Snippet: Targeting SCD1 for radiosensitization of isogenic NCI-H1703 spheres with mutant (mut) or wild-type (wt) KRAS. A , Radiosensitization by MF438 (0.1 μM) using 6 Gy irradiation. Bars represent mean SRF +/− SEM based on five biological repeats. B , Left, representative images of γ-H2AX foci 24 hours following irradiation with 6 Gy with and without MF438 treatment. Right, Fractions of cells with at least 20 foci per nucleus. Bars represent mean + SE based on three biological repeats. C , Left, Western blot with an anti-SCD1 antibody for cells treated with scrambled (scr) or validated siRNA. Relative SCD1 band intensities are displayed. Right, Radiosensitization by siRNA against SCD1 using 6 Gy irradiation. Bars represent mean SRF +/− SE based on two repeat experiments. D , Radiosensitization by small molecule inhibitors of the targets indicated. Data represent median, interquartile range, and range for 32 pooled replicates across four different drug concentrations and two biological repeats. MF438 data are included for reference. Statistical comparisons by t-test or Mann-Whitney test, two-sided; *p≤0.05, **p≤0.01, ***p≤0.001.

Article Snippet: Stearoyl-CoA Desaturase 1 (SCD1) inhibitor (MF438, Cat #569406) and CAY10566 (Cat #HY-15823) were purchased from EMD Millipore, Darmstadt, Germany and MedChemExpress, Monmouth Junction, NJ, respectively (see Tab.

Techniques: Mutagenesis, Irradiation, Western Blot, MANN-WHITNEY