Journal: bioRxiv

Article Title: Neonatal hyperoxia inhibits proliferation of atrial cardiomyocytes by suppressing fatty acid synthesis

doi: 10.1101/2020.06.01.127621

Figure Lengend Snippet: Graph shows the increase in the relative numbers of HL-1 cells grown for 72 hours in media containing DMSO (white circles), the FASN inhibitor G28UCM at 10 mM (gray squares), or the SCD1 inhibitor A939572 at 10 nM (gray diamonds). Values are normalized to number of cells in each well just prior to adding inhibitor (0hrs). The p-values shown are the results of two-way ANOVA with Dunnett’s multiple comparison tests.

Article Snippet: To confirm that Fasn and Scd1 are required for HL-1 cell proliferation, HL-1 cells were treated with DMSO as a vehicle control, the FASN inhibitor G28UCM at 10μM or the SCD1 inhibitor A939572 at 10nM (Tocris Biosciences) and grown in room air ( ).

Techniques: Comparison

Journal: bioRxiv

Article Title: Stearoyl-CoA Desaturases regulate stem and progenitor cell metabolism and function in response to nutrient abundance

doi: 10.1101/2025.09.17.676943

Figure Lengend Snippet: The requirement for SCD1 and SCD2 activity in intestinal stem cells was investigated by using mouse organoid models. (A) Established WT small intestinal organoids were treated with either vehicle (DMSO), or 250 nM of SCD inhibitors A939572 and CAY10566, or ACC1 inhibitor ND-646 for 96 hrs. (N=4). (B) The organoid forming capacity of stem cells and (C) the expression levels of intestinal stem cell markers were significantly decreased in inhibitor treated organoids which was investigated by RT-qPCR. (D) Organoids were collected and processed for EdU staining which fluorescently labeled proliferative cells. (E-F) Exogenous oleic acid addition (50 μM) rescued SCD inhibition (50 nM) phenotype, resulting in increased number of proliferative and differentiated organoids ex vivo (N=3-4). While SCD inhibition decreased the number of proliferative cells, addition of oleic acid exogenously rescued the Scd inhibition phenotype. (G) Small intestinal organoids were treated with either vehicle (DMSO) or SCD inhibitors A939572 or CAY10566 for four days. RT-qPCR was performed with organoids cDNA to investigate the expression levels of ER stress marker genes. (H) RT-qPCR was used to investigate the expression levels of Chop in small intestinal organoids under both oleic acid-treated and non-treated conditions, where Scd enzymes were inhibited. (I) WT small intestinal organoids were treated with either a DMSO vehicle, or 50 nM of the Scd inhibitor, A939572, for 24 hrs for lipidomics (N=3). Scale bar is 250μm. Graphs and images are representative of 3-4 mice, multiple wells and fields from each biological replicate are counted. Data are mean ± SD. *p<0.05, ***p<0.001, and ****p<0.0005.

Article Snippet: After desired size and budding morphology was maintained, organoids were harvested and incubated with 50 nM SCD inhibitor A939572 (Tocris) for 4 days.

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Staining, Labeling, Inhibition, Ex Vivo, Marker

Journal: Cancer Science

Article Title: SCD1 inhibition enhances the effector functions of CD8 + T cells via ACAT1 ‐dependent reduction of esterified cholesterol

doi: 10.1111/cas.15999

Figure Lengend Snippet: SCD1 inhibitors enhance the activation of human CD8 + T cells. CD8 + T cells were isolated from human PBMCs using MACS and activated with an anti‐CD3 monoclonal antibody (mAb), anti‐CD28 mAb, and IL‐2. CD8 + T cells were treated with dimethylsulfoxide (DMSO), SCD1 inhibitors (A939572 and CAY10566), SCD1 inhibitors + oleic acid (OA), or SCD1 inhibitors + palmitoleic acid (POA). (A) Intracellular oleic acid levels in human CD8 + T cells. (B) Intracellular palmitoleic acid levels in human CD8 + T cells. (C) IFN‐γ levels in culture supernatants of CD8 + T cells were measured using ELISA. (D) Cell proliferation of CD8 + T cells was evaluated by WST‐1 assay. Data are expressed as means ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n.s., not significant.

Article Snippet: SCD1 inhibitors A939572 and CAY10566 were purchased from APExBIO and Cayman, respectively.

Techniques: Activation Assay, Isolation, Enzyme-linked Immunosorbent Assay, WST-1 Assay

Journal: Cancer Science

Article Title: SCD1 inhibition enhances the effector functions of CD8 + T cells via ACAT1 ‐dependent reduction of esterified cholesterol

doi: 10.1111/cas.15999

Figure Lengend Snippet: Inhibition of SCD1 enhances the effector function of human CD8 + T cells via suppression of esterified cholesterol synthesis by ACAT1. CD8 + T cells were isolated from human PBMCs using MACS and activated by the anti‐CD3 mAb, anti‐CD28 mAb, and IL‐2. (A–C) CD8 + T cells were cultured in AIM‐V medium with DMSO + γCD, 1 μM ACAT1 inhibitors (Avasimibe and CP113818) + γCD, or 1 μM ACAT1 inhibitors + γCD + cholesteryl oleate (ChO). IFN‐γ (A), TNF‐α (B), and GzmB (C) levels in culture supernatants were measured by ELISA. (D) CD8 + T cells were treated with DMSO, SCD1 inhibitors (A939572 and CAY10566), or SCD1 inhibitors + oleic acid (OA). The ratio of esterified cholesterol/cholesterol (left panel) and the amounts of cholesterol and esterified cholesterol (right panel) are shown. (E) CD8 + T cells were cultured in AIM‐V medium with DMSO + γ‐cyclodextrin (γCD), SCD1 inhibitors (A939572 and CAY10566) + γCD, or SCD1 inhibitors + γCD + cholesteryl oleate (ChO). IFN‐γ levels in culture supernatants were measured by ELISA. Data are expressed as means ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n.s., not significant.

Article Snippet: SCD1 inhibitors A939572 and CAY10566 were purchased from APExBIO and Cayman, respectively.

Techniques: Inhibition, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: SCD1 inhibition enhances the effector functions of CD8 + T cells via ACAT1 ‐dependent reduction of esterified cholesterol

doi: 10.1111/cas.15999

Figure Lengend Snippet: Systemic administration of SCD1 inhibitor enhances IFN‐γ production by CD8 + T cells in vivo. (A–D) C57BL/6 mice bearing MCA205 tumors were treated with an SCD1 inhibitor (A939572; 10 mg/kg), an ACAT1 inhibitor (Avasimibe; 15 mg/kg) or the vehicle (mock). Tumor‐infiltrating CD8 + T cells (CD8 + TILs) were sorted using MACS. (A) Oleic acid levels in tumor tissues and CD8 + TILs. (B) The ratio of esterified cholesterol/cholesterol and the amount of cholesterol and esterified cholesterol in CD8 + TILs are shown. (C, D) CD8 + TILs were cultured ex vivo after SCD1 inhibitor (C) or ACAT1 inhibitor (D) treatment. The next day, IFN‐γ levels in culture supernatants were measured by ELISA. Data are expressed as means ± SD ( n = 5). (E) Mice bearing MCA205 tumors were treated with A939572 or the vehicle and anti‐PD‐1 antibody (200 μg/mouse) or isotype‐matched antibody. (F) Mice bearing MCA205 tumors were treated with an ACAT1 inhibitor (Avasimibe; 15 mg/kg) or the vehicle and anti‐PD‐1 antibody (200 μg/mouse) or isotype‐matched antibody. Tumor growth curves for average tumor volumes are shown (means ± SD; n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n.s., not significant.

Article Snippet: SCD1 inhibitors A939572 and CAY10566 were purchased from APExBIO and Cayman, respectively.

Techniques: In Vivo, Cell Culture, Ex Vivo, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: SCD1 inhibition enhances the effector functions of CD8 + T cells via ACAT1 ‐dependent reduction of esterified cholesterol

doi: 10.1111/cas.15999

Figure Lengend Snippet: SCD1 is a potential target for enhancing the antitumor effects of CAR‐T cell therapy. GPC1‐specific murine CAR (GPC1‐mCAR)‐T cells and murine control T (mCont‐T) cells were generated as described in Section and then treated with DMSO or A939572. (A) Intracellular oleic acid levels in GPC1‐mCAR‐T cells. (B) The ratio of esterified cholesterol/cholesterol and the amount of cholesterol and esterified cholesterol in GPC1‐mCAR‐T cells are shown. (C, D) GPC1‐mCAR‐T cells or mCont‐T cells and mGPC1‐overexpressing MCA205 tumor cells (MCA205‐mGPC1) were co‐cultured in vitro. (C) IFN‐γ secretion was evaluated by ELISA. (D) Cytotoxic activity was evaluated using a standard Cr 51 ‐releasing assay. Data are expressed as means ± SD ( n = 3). (E) Mice bearing MCA205‐mGPC1 were treated with 5 × 10 6 cells of GPC1‐mCAR‐T cells or mCont‐T cells on day 2 and SCD1 inhibitor was administered by oral gavage twice daily for 16 days starting on day 4. Tumor growth curves for average tumor volumes are shown (means ± SD; n = 5). * p < 0.05, ** p < 0.01, **** p < 0.0001. E/T ratio, effector cell (GPC1‐mCAR‐T cells)/target cell (MCA205‐mGPC1 cell) ratio.

Article Snippet: SCD1 inhibitors A939572 and CAY10566 were purchased from APExBIO and Cayman, respectively.

Techniques: Generated, Cell Culture, In Vitro, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Cancer Science

Article Title: SCD1 inhibition enhances the effector functions of CD8 + T cells via ACAT1 ‐dependent reduction of esterified cholesterol

doi: 10.1111/cas.15999

Figure Lengend Snippet: Schematic of the study. Inhibition of SCD1 enhances the antitumor activity of CD8 + T cells by reducing the intracellular oleic acid level, ACAT1‐dependent cholesterol esterification reaction, and esterified cholesterol level.

Article Snippet: SCD1 inhibitors A939572 and CAY10566 were purchased from APExBIO and Cayman, respectively.

Techniques: Inhibition, Activity Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Small molecule inhibitors of fungal Δ(9) fatty acid desaturase as antifungal agents against Candida auris

doi: 10.3389/fcimb.2024.1434939

Figure Lengend Snippet: Haploinsufficient profiling and RNA-seq assays uncover the Δ(9) fatty acid desaturase Ole1 as a potential target of SPB00525. (A) GO biological process term enrichment of C. albicans heterozygous mutants depleted or enriched upon exposure to SPB00525 in both SC and YPD media. (B) Graphical representation of results from the SPB00525-induced haploinsufficiency profiling. Mutants were plotted according to their fitness defect score from lowest to highest. The fitness scores reflect the differential abundance of each mutant strain in the SPB00525-treated condition relative to the DMSO control. (C, D) Comparison of HIP profiles in both SC and YPD growth media. Venn diagrams indicate shared depleted (C) and enriched (D) mutants in the presence of SPB00525 in SC and YPD media. SPB00525- enriched and depleted mutants were identified using a fitness score cutoff of 2.5 (Log 2 ) and a false discovery rate of 5%. (E) Validation of the SPB00525-induced haploinsufficiency assay. WT (CAI4) and heterozygous mutants ( ole1 , orf19.5285 and pep12 ) cells were grown in SC medium and exposed or not to 6 µg/ml SPB00525. Results represent growth inhibition (%) relative to the DMSO control. (F) Increased dosage of OLE1 and orf19.5285 led to decreased SPB00525 sensitivity of C. albicans . WT (WT-Cip-Act) and strains overexpressing both OLE1 and orf19.5285 were exposed to 15 and 30 µg/ml SPB00525 and OD 600 reading were acquired at 24h. (G, H) Transcriptomic analysis of C. albicans response to SPB00525. GO enrichment of upregulated (G) and downregulated (H) transcripts of C. albicans cells exposed to 6 µg/ml SPB00525 for 15 and 60 min. (I) Gene set enrichment analysis (GSEA) of the C. albicans SPB00525-modulated transcriptome at 15- and 60-min. NES (normalized enrichment score) and nominal q -value obtained from the GSEA are shown for each correlation. The complete GSEA correlations are listed in Supplementary Table S4 . False-Discovery Rate ( q -value) of 1%.

Article Snippet: The SPB00525 chemical analogs A939572 (MolPort-003-983-382), PluriSln1 (MolPort-000-564-458), MK-8245 (MolPort-023-293-543), HTS06170 (MolPort-002-901-421), HTS06154 (MolPort-002-901-407), HTS05029 (MolPort-002-901-093), HTS05668 (MolPort-002-901-311), G514 (MolPort-000-882-466) and new batches of SPB00525 (MolPort-002-923-508) were obtained from MolPort.

Techniques: RNA Sequencing, Mutagenesis, Control, Comparison, Biomarker Discovery, Inhibition

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Small molecule inhibitors of fungal Δ(9) fatty acid desaturase as antifungal agents against Candida auris

doi: 10.3389/fcimb.2024.1434939

Figure Lengend Snippet: Library screen data, hit validation, and antifungal activity of SPB00525. (A) Screen of the yeast bioactive library against molecules with antifungal activity against C. auris . Relative inhibition of each compound tested at 100 µM was determined on C. auris 381 isolate grown at 30°C in SC medium for 24 h. (B) Chemical structure of the aryl-carbohydrazide hit SPB00525. (C) Dose-response assay of SPB00525 on C. auris . The C. auris 381 strain was grown in SC medium with different concentration of SPB00525 and OD reading was taken after 24h of incubation. Results represent growth inhibition (%) relative to the DMSO control. (D) Heat map representing the dose-response assay of SPB00525 on different azole resistant (R) and sensitive (S) clinical isolates of C. auris . The minimal inhibitory concentrations (MIC) were determined following Clinical and Laboratory Standards Institute (CLSI) recommendations using RPMI medium. (E) Heat map of the dose-response assay of SPB00525 on sensitive and, azole- (Flu R ) and caspofungin-resistant (Caspo R ) clinical isolates of C. albicans . (F) Time-kill curve showing the fungicidal activity of SPB00525 against both C. auris and C. albicans . C. auris 381 and C. albicans SC5314 strains were exposed to different concentrations (3-15 µg/ml) at 24h. CFUs were calculated as described in the method section.

Article Snippet: The SPB00525 chemical analogs A939572 (MolPort-003-983-382), PluriSln1 (MolPort-000-564-458), MK-8245 (MolPort-023-293-543), HTS06170 (MolPort-002-901-421), HTS06154 (MolPort-002-901-407), HTS05029 (MolPort-002-901-093), HTS05668 (MolPort-002-901-311), G514 (MolPort-000-882-466) and new batches of SPB00525 (MolPort-002-923-508) were obtained from MolPort.

Techniques: Biomarker Discovery, Activity Assay, Inhibition, Concentration Assay, Incubation, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Small molecule inhibitors of fungal Δ(9) fatty acid desaturase as antifungal agents against Candida auris

doi: 10.3389/fcimb.2024.1434939

Figure Lengend Snippet: Inhibition of Ole1 desaturase activity by SPB00525. (A) Desaturation of fatty acyl CoAs by fungal Δ(9) fatty acid desaturase. Ole1 converts CoA-linked saturated (palmitoyl-CoA and stearyl-CoA) to monounsaturated fatty acids of 16 (palmitoleic acid) or 18 (oleic acid) carbons in length. (B, C) SPB00525 inhibits fatty acid desaturation. SPB00525 reduced both C16 and C18 desaturation (B) . (C) Fatty acid desaturation index (unsaturated/saturated ratio) of C16, C18 and total fatty acid of C. albicans SC5314 strain exposed to 9 and 30 µg/ml SPB00525 for 2.5 hours. Data are average of five experiments, and error bars are SD. UFA: Unsaturated fatty acids; SFA: Saturated fatty acids. (D, E) Supplementation of exogenous unsaturated fatty acids alleviates growth inhibition by SPB00525. C. albicans SC5314 (D) and C. auris 381 (E) cells were exposed to SPB00525 in SC medium supplemented with either saturated (palmitic + stearic acids) or unsaturated (palmitoleic and oleic acids) fatty acid mixture. (F) OLE1 essentiality was reverted by supplementation of an equimolar mixture of palmitoleic and oleic acids. The conditional shut-off mutant strain of OLE1 ( ole1 /pTet- OLE1 ) was grown in SC medium with tetracycline (SC+tet) supplemented with saturated or unsaturated fatty acids. Cells were grown at 30°C, and OD 600 readings were taken every 15 min for 2 days.

Article Snippet: The SPB00525 chemical analogs A939572 (MolPort-003-983-382), PluriSln1 (MolPort-000-564-458), MK-8245 (MolPort-023-293-543), HTS06170 (MolPort-002-901-421), HTS06154 (MolPort-002-901-407), HTS05029 (MolPort-002-901-093), HTS05668 (MolPort-002-901-311), G514 (MolPort-000-882-466) and new batches of SPB00525 (MolPort-002-923-508) were obtained from MolPort.

Techniques: Inhibition, Activity Assay, Mutagenesis

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Small molecule inhibitors of fungal Δ(9) fatty acid desaturase as antifungal agents against Candida auris

doi: 10.3389/fcimb.2024.1434939

Figure Lengend Snippet: HTS06170 chemical analog of SPB00525 exhibit potent antifungal activity in vitro . (A-F) Chemical structure of commercially available analogs of SPB00525. (G-I) Chemical structure of known human stearoyl-CoA desaturase 1 inhibitors. (J) Antifungal activity of SPB00525 analogs tested at 15 and 30 µg/ml. C. albicans SC5314 and C. auris 381 strains were grown in SC medium and exposed to the corresponding compound for 24h at 30°C. Data reported as percentage growth relative to the untreated condition (DMSO). (K) Dose-response assay of HTS06170 on C. albicans and C. auris . The C. auris 381 and the C. albicans SC5314 strains were grown in SC medium with different concentration of HTS06170 and OD reading was taken after 24h of incubation at 30°C. Results represent growth inhibition (%) relative to the DMSO control. MICs of HTS06170 are indicated in parentheses for each fungal strain.

Article Snippet: The SPB00525 chemical analogs A939572 (MolPort-003-983-382), PluriSln1 (MolPort-000-564-458), MK-8245 (MolPort-023-293-543), HTS06170 (MolPort-002-901-421), HTS06154 (MolPort-002-901-407), HTS05029 (MolPort-002-901-093), HTS05668 (MolPort-002-901-311), G514 (MolPort-000-882-466) and new batches of SPB00525 (MolPort-002-923-508) were obtained from MolPort.

Techniques: Activity Assay, In Vitro, Concentration Assay, Incubation, Inhibition, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Small molecule inhibitors of fungal Δ(9) fatty acid desaturase as antifungal agents against Candida auris

doi: 10.3389/fcimb.2024.1434939

Figure Lengend Snippet: SPB00525 and HTS06170 exhibit in vivo activity against C. albicans and C. auris . (A) SPB00525 and HTS06170 attenuate candidiasis in Galleria mellonella . Larvae were first injected with C. albicans SC5314 strain followed by a second injection of either DMSO, 15 µg/ml of SPB00525 or HTS06170 and survival was monitored at the indicated time for a period of 4 days. (B, C) SPB00525 and HTS06170 attenuate damage of enterocytes cells caused by C. albicans (B) and C. auris (C) . Damage of the human epithelial intestinal cells HT-29 infected by C. albicans SC5314 or C. auris 381 strain was assessed using LDH release assay. Cell damage was calculated as percentage of LDH activity of each treatment to that of the control experiment (HT-29 damaged by 1% Triton X-100). Results are represented as the mean of three independent replicates. (D) Evaluation of SPB00525 and HTS06170 cytotoxicity using the LDH release assay. HT-29 cells were exposed to 2x, 4x and 6x MIC C. albicans or 5x, 10x and 15x MIC C. auris of both SPB00525 and HTS06170 for 24h. (E) SPB00525 and HTS06170 inhibit biofilm formation. C. albicans SC5314 cell suspension was seeded into 96-well polystyrene plates and incubated at 37°C for 2 hours to initiate biofilm formation. Biofilms were then exposed or not to either SPB00525 or HTS06170 and incubated for 24 hours prior to biomass assessment using the crystal violet assay. Results are represented as the mean of six independent replicates. (F) SPB00525 and HTS06170 inhibit filamentation of C. albicans in response to different stimuli. C. albicans SC5315 yeast cells was grown under hyphae-promoting conditions (FBS and Spider, RPMI or Lee’s media) at 37°C for 6 hours in the absence or the presence of aryl-carbohydrazides. Filament lengths were measured for at least 100 cells using ImageJ software.

Article Snippet: The SPB00525 chemical analogs A939572 (MolPort-003-983-382), PluriSln1 (MolPort-000-564-458), MK-8245 (MolPort-023-293-543), HTS06170 (MolPort-002-901-421), HTS06154 (MolPort-002-901-407), HTS05029 (MolPort-002-901-093), HTS05668 (MolPort-002-901-311), G514 (MolPort-000-882-466) and new batches of SPB00525 (MolPort-002-923-508) were obtained from MolPort.

Techniques: In Vivo, Activity Assay, Injection, Infection, Lactate Dehydrogenase Assay, Control, Suspension, Incubation, Crystal Violet Assay, Software