Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Targeting ALDH7A1 with covalent inhibitors reveals new chemical space for prostate cancer therapy

doi: 10.1080/14756366.2026.2664708

Figure Lengend Snippet: Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of DU145 cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).

Article Snippet: To validate the in vitro and in silico findings in a cellular context, the DU145 PCa cell line was selected as a suitable model due to its high expression of ALDH7A1, as reported in the Human Protein Atlas ( www.proteinatlas.org ).

Techniques: Migration, Incubation, Concentration Assay, Activity Assay, Lysis, Bradford Assay, Standard Deviation

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Targeting ALDH7A1 with covalent inhibitors reveals new chemical space for prostate cancer therapy

doi: 10.1080/14756366.2026.2664708

Figure Lengend Snippet: Effects of compound 3b and 4b in migration, cell cycle progression, and MDA accumulation in PCa cells. (a) Representative images of DU145 cells migration upon scratch generation in a confluent monolayer. Time point 0 h (T = 0 h) or 60 h (T = 60 h) points post-incubation in presence of DMSO, compound 3b or compound 4b at 20 µM final concentration, are shown. Scale bar = 100 µm. (b) Quantification of scratch area reduction over time normalised to the scratch area at T = 0 h. Wound healing was monitored at 12, 24, 36, 48 and 60 h post-incubation with the indicated compounds. (c) Quantification of cell cycle distribution in DU145 cells treated with DMSO, compound 3b or compound 4b at 20 µM final concentration for 24 h prior to cytofluorimetric analysis. The percentage of cells in each population is shown. (d) Total intracellular ALDH activity measured in DU145 cell lysates following pre-incubation with 20 µM 3b or 4b for 24 h at 37 °C. Cells were extensively washed prior to lysis to eliminate any non-covalently bound material. (e) Quantification of malondialdehyde (MDA) levels in the same cell lysates. MDA concentration was measured and normalised to the total protein content of each sample, as determined by the Bradford assay. In all plots, error bars represent the standard deviation, and asterisks indicate statistical significance levels: p ≤ 0.05 (*), p ≤ 0.01 (**), and p ≤ 0.001 (***).

Article Snippet: The PCa cells DU145 (#HTB-81, ATCC) and LNCaP (#CRL1740, ATCC) were cultured in an RPMI-1640 culture medium, supplemented with 10% foetal bovine serum (yourSIAL-FBS-SA, S.I.A.L.

Techniques: Migration, Incubation, Concentration Assay, Activity Assay, Lysis, Bradford Assay, Standard Deviation

Journal: Cell Reports Methods

Article Title: Biobank of genetically defined murine prostate cancer tumoroids uncovers oncogenic pathways and drug vulnerabilities driven by PTEN-loss

doi: 10.1016/j.crmeth.2026.101370

Figure Lengend Snippet: The PDPK1/AKT/FLT DPI and tenovin-6 (T6) show high anti-cancer efficacy in murine tumoroids and human PCa cell lines (A) Dose-response curves for DPI (top) and T6 (bottom) for in vivo and in vitro Pten KO (left), Pten/Stat3 KO (middle), and Pten/Tp53 KO (right) tumoroids. Points represent means of technical duplicates per tumoroid line ( N = 3). Curve fitting was performed using GraphPad Prism 8.0.2. (B) Bar graphs showing means and ±SD of half-maximal inhibitory concentration (IC50) for DPI (top) and T6 (bottom) for in vivo and in vitro tumoroid lines of all genotypes ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA, Tukey’s test). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05. (C) Bar graphs depicting means and ±SD of IC50 values of DPI (left) and T6 (right) on human PCa cell lines. 22RV1: primary PCa; LNCaP: metastatic PCa; DU145, PC3: metastatic castration-resistant PCa ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05; ∗∗ p ≤ 0.01. (D) Heatmaps of synergy scores calculated with the highest single agent (HSA) model for DPI and enzalutamide (left), and T6 and enzalutamide (right) on the human LNCaP cell line. Values > 0 represent synergistic effects, and values < 0 represent antagonistic effects. IC50 concentrations of respective compounds are underlined ( N = 3). See also .

Article Snippet: Human DU145 metastatic PCa cell line , ATCC , HTB-81, RRID:CVCL_0105.

Techniques: In Vivo, In Vitro, Concentration Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Site-Specific Abiraterone Protein–Drug Conjugates via Hedgehog Autoprocessing

doi: 10.1021/acsami.5c22490

Figure Lengend Snippet: Schematic of hedgehog-mediated abirateronylation as a noncanonical PTM (ncPTM) for generating protein–drug conjugates (PDCs). An intrinsically disordered protein polymer (IDPP) serves as the model protein scaffold. The resulting hybrid biopolymers are internalized by DU145 cells and exert cytotoxicity. Created with BioRender.com

Article Snippet: DU145 human prostate cancer cells (ATCC HTB-81) were cultured in high-glucose DMEM supplemented with 10% fetal bovine serum and 0.1% penicillin–streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Polymer

Journal: ACS Applied Materials & Interfaces

Article Title: Site-Specific Abiraterone Protein–Drug Conjugates via Hedgehog Autoprocessing

doi: 10.1021/acsami.5c22490

Figure Lengend Snippet: Biophysical and biological assessment of the optimized PDC and controls. (a) Hydrodynamic radius ( R h ) of (A,40) carrier (E), E–Abi and E–Chol at 50 μM and 200 μM, measured by dynamic light scattering. E–Abi forms reversible, concentration-dependent oligomers. (b) Release kinetics of conjugated sterols (E-Abi, E–Andro and E–Chol) in PBS (pH 6.5) at 37 °C, along with the chemical structures of Abi and Andro tail groups. E–Abi and E–Andro shows sustained release, whereas E–Chol displays negligible sterol release over the same period. Shaded areas represent the 95% confidence intervals of the nonlinear regression fit to a first-order kinetics model. (c) Dose–response curves for DU145 prostate cancer cells after 72 h exposure to free drug, conjugate, and controls (measured by MTT assay; nonlinear dose–response fit). (d) Viability of 3D DU145 spheroids after 24-h treatment with 100 μM of each compound, showing superior efficiency of E–Abi. (e) Representative live/dead fluorescence images of treated spheroids. Live cells are stained green (calcein AM), and dead cells are stained red (propidium iodide). Data are mean ± s.d. ( n = 3). Statistical analysis was performed using one-way ANOVA with posthoc tests (Tukey’s for a, Holm–Sidak’s for d): * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: DU145 human prostate cancer cells (ATCC HTB-81) were cultured in high-glucose DMEM supplemented with 10% fetal bovine serum and 0.1% penicillin–streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Concentration Assay, MTT Assay, Fluorescence, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Site-Specific Abiraterone Protein–Drug Conjugates via Hedgehog Autoprocessing

doi: 10.1021/acsami.5c22490

Figure Lengend Snippet: Lipidation-dependent modulation of biopolymer penetration in 3D tumor spheroids. (a–c) Representative bivariate flow cytometry dot plots of WGA-Alexa Fluor 680 (membrane label) versus carrier-Alexa Fluor 488. The gated population represents WGA + /Carrier + cells, and the percentage of uptake-positive cells is indicated for each condition. DU145 spheroids were incubated overnight with labeled (a) E, (b) E–Abi, (c) E–Chol, dissociated to single cells, and analyzed by flow cytometry. (d) Bar graph summarizing the fraction of uptake-positive cells across treatments. (e–g) Confocal z-stack images of DU145 spheroids treated with E, E–Abi, or E–Chol for 24 h. WGA (AF680, red) marks the cell membrane, and constructs (AF488, green) indicate carrier localization. Data are mean ± s.d. ( n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test (** p < 0.01).

Article Snippet: DU145 human prostate cancer cells (ATCC HTB-81) were cultured in high-glucose DMEM supplemented with 10% fetal bovine serum and 0.1% penicillin–streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Flow Cytometry, Membrane, Incubation, Labeling, Construct