Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 exerts a more substantial inhibitory growth effect in PDAC spheroids than other tested compounds. (A) hF37 PDAC organoid-derived 2D cell line, (B) BxPC-3, (C) Capan-2, (D) MIA PaCa-2, and (E) PANC-1 PDAC cell lines were treated as spheroids with DMSO (vehicle) and 1 μM and 10 μM concentrations of BEZ-235, GDC-0941, TP-3654, or AUM-302 for 72 h. The figure displays representative images from a sample of N = 5 . The scale bar depicts 100 μm.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Derivative Assay

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: BEZ-235, GDC-0941, TP-3654, and AUM-302 inhibit the viability of multiple pancreatic cancer cell lines in a spheroid setting. Pancreatic cancer cell lines BxPC-3 (A) Capan-2 (B) MIA PaCa-2 (C) PANC-1 (D) and organoid-derived cell line hF37 2D (E) were treated with variable concentrations of BEZ-235 ( A small solid pink circle on a white background. ) GDC-0941( A plain blue square with a solid color. ) TP-3654( Black triangle with a solid fill. ) and AUM-302 ( A green downward-facing triangle on a white background. ) for 72 h, and cell viability was measured using 3D Cell Titer-Glo. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). The LOG -9 represents 1 nM, -8 – 10 nM, -7 – 100nM, -6 – 1 μM, -5 – 10 μM, -4 – 100 μM.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Derivative Assay, Control

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 exerts a more substantial inhibitory growth effect in hF31, hF37, hF44, hT1, and hM1a PDAC organoids than other tested compounds. hF31 (A–D) hF37 (E–H) hF44 (I–L) hT1 (M–P) and hM1a (Q–T) PDAC organoids were seeded in Matrigel and treated with DMSO (vehicle) and 10 nM, 100 nM, and 1 μM concentrations of GDC-0941 (A,E,I,M,Q) TP-3654 (B,F,J,N,R) BEZ-235 (C,G,K,O,S) or AUM-302 (D,H,L,P,T) for 72 h. The figure displays representative images from a sample of N = 5 . The scale bar depicts 100 μm.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 significantly exerts a stronger inhibitory growth effect on PDAC organoids than BEZ-235. hF31 (A) hF37 (B) hF44 (C) hT1 (D) and hM1a (E) PDAC organoids were seeded in Matrigel and treated with DMSO (vehicle) and 10 nM, 100 nM, and 1 μM concentrations of BEZ-235 and AUM-302 for 72 h. At the end of the incubation, cell viability was measured using 3D Cell Titer-Glo. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). The statistical analysis was performed using a two-way ANOVA in GraphPad Prism version 10.4.1. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Incubation, Control

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 synergizes with RMC-6236 to inhibit the growth of hT1 PDAC organoids. hT1 PDAC organoids were seeded in Matrigel and treated with various concentrations of AUM-302, RMC-6236, and a combination of both compounds. Seventy-two hours post-treatment, the cell viability was assessed using 3D Cell Titer-Glo and analyzed using a non-linear regression model of log(inhibitor) vs. response–variable slope (four parameters) equation in GraphPad Prism version 10.4.1. and Synergy Finder. (A) Cell viability of mono-and combinatorial treatment. The results are shown as mean ± SD ( N = 5 ) and normalized to the control (DMSO). (B) The HSA Synergy Score is a dose matrix inhibition response. (C) The HSA Synergy Score is shown as a plot contour. (D) The graph was generated using a Plot Barometer using viability results. The needle points to the observed growth inhibition, denoted in percentage values, for the synergy between AUM-302 0.1 μM and RMC-6236 0.33 μM. The four white lines represent the predicted inhibition values calculated by the HSA, Loewe, ZIP, and BLISS models at specific concentrations.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Control, Inhibition, Generated

Journal: Frontiers in Pharmacology

Article Title: AUM-302, a novel triple PIM/PI3K/mTOR inhibitor, synergizes with RAS inhibition and impedes the growth of pancreatic ductal adenocarcinoma spheroids and organoids

doi: 10.3389/fphar.2026.1685433

Figure Lengend Snippet: AUM-302 and RMC-6236 affected the activity of the signaling pathways in the hT1 PDAC organoids. hT1 organoids were treated with DMSO (vehicle), 300nM RMC-6236, 10 nM and 100 nM AUM-302, and combinations of thereof for 24 and 48 h. (A) Western blots were performed for mTOR, AKT, ERK, S6, phosphorylated counterparts, and β-Actin as a loading control. (B,C) Densitometry analysis was conducted to quantify relative protein expression, normalized to β-actin control and non-phosphorylated proteins in samples after 24- and 48-h post-treatment. Densitometry was measured with ImageJ and analyzed with GraphPad Prism as described in “Materials and Methods.” Data are presented as mean ± SD values ( N = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: PDAC cell lines BxPC-3 (CRL-1687), Capan-2 (HTB-80), MIA PaCa-2 (CRL-1420), and PANC-1 (CRL-1469) were purchased from ATCC (Manassas, VA).

Techniques: Activity Assay, Protein-Protein interactions, Western Blot, Control, Expressing

Journal: Nature Communications

Article Title: Combined targeted and epigenetic-based therapy enhances antitumor immunity by stabilizing GATA6-dependent MHCI expression in pancreatic ductal adenocarcinoma

doi: 10.1038/s41467-026-69013-y

Figure Lengend Snippet: a Nine primary murine PDAC cell lines were established from spontaneous murine PDAC models ( CKP , KPC , and KPCY ). Created with BioRender.com. Flow cytometry analysis was used to determine GATA6 protein expression in each line. Representative histograms show GATA6 staining (GATA6 antibody, GATA6 AB, red) versus isotype control (blue) in GATA6 high (GATA6 hi ) and low (GATA6 lo ) expressing cell lines. The cell lines were dichotomized into GATA6 high ( n = 5, orange) and GATA6 low ( n = 4, blue) groups. b The parental cells (Ctrl) of three GATA6 high cell lines (60400, 511950, 70301) and three GATA6 low cell lines (60590, 60531, 511892) were treated with increasing doses of MEKi (trametinib) until 100x of the cells’ initial IC50 to generate the corresponding long-term MEKi-treated cells (MEKi). Transcriptomic profiling by RNA sequencing of the parental ( Ctrl) and long-term MEKi-treated (MEKi) cells was performed. GSEA of KEGG and HALLMARKs revealed gene sets that were significantly ( P -adjust < 0.05) different between parental (Ctrl) and long-term MEKi-treated (MEKi) cells within GATA6 high and GATA6 low groups, respectively. c Surface MHCI (H-2Db) expression on control (Ctrl) and MEKi-treated (MEKi) cells of GATA6 high (upper panel) and GATA6 low group (lower panel) was assessed by flow cytometry ( n = 4 biological replicates for cell line 60400, 511950, 511892, 60590, and 5 biological replicates for cell line 110299, 2838c3, 60531, 6694C2). MFI mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. d Flow cytometric analysis of GATA6 expression in GATA6 knockout (KPCY1-CRISPR-GFP v2.1-gRNA-gCas9): gGATA6_KO-3 and gGATA6_KO-4, and negative control (gNT) cell lines ( n = 4 biological replicates). Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. e Surface MHCI (H-2Db) expression on GATA6 knockout cell lines and negative control upon MEKi treatment ( n = 4 biological replicates). MFI: mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by unpaired two-tailed Mann–Whitney test. f Schematic of the knock-in strategy for N-terminal AID-tagged gata6. Shown are components of the knock-in cassette, and the positions of primers for genomic PCR are marked by arrows. g Western blot showing GATA6 protein expression in 110299 WT and 110299 AID-GATA6 cells after 24 h treatment with 1 µM 5-Ph-IAA ( n = 1 biological replicates). h Surface MHCI (H-2Db) expression on 110299 WT and 110299 AID-GATA6 cells treated with or without MEKi and/or 1 µM 5-Ph-IAA ( n = 5 biological replicates). MFI: mean fluorescence intensity. Mean ± SD is shown. Statistical significance was calculated by One-way ANOVA and Kruskal–Wallis test.

Article Snippet: HAs were PCR-amplified using murine primary PDAC cell line (110299) genomic DNA as template and cloned into pJET (Thermo Fisher Scientific) flanking the Blast-P2A-V5-AID cassette to generate pJET_gata6_N-AID_HDR.

Techniques: Flow Cytometry, Expressing, Staining, Control, RNA Sequencing, Fluorescence, Two Tailed Test, MANN-WHITNEY, Knock-Out, CRISPR, Negative Control, Knock-In, Western Blot

Journal: Nature Communications

Article Title: Combined targeted and epigenetic-based therapy enhances antitumor immunity by stabilizing GATA6-dependent MHCI expression in pancreatic ductal adenocarcinoma

doi: 10.1038/s41467-026-69013-y

Figure Lengend Snippet: a Patient-derived xenografts (PDXs) from 15 PDAC patients were transplanted subcutaneously into athymic nude mice, and the mice of third generation were used for MEKi treatment. When tumor volume reached 200 mm 3 , the mice were treated with vehicle control (Ctrl) or MEKi. Tumors were harvested after 4 weeks of treatment. Created with BioRender.com. b IHC staining of GATA6 and MHCI of the xenograft tumors was performed, and the proportion of marker-positive cells out of total cells was quantified by Definiens software. Xenografts were dichotomized into GATA6 high (GATA6 hi , n = 7 independent mice) and GATA low (GATA lo, n = 8 independent mice) expression groups using median GATA6 + cells (% total) as cutoff. MHCI + cells (% total) were quantified by Definiens and compared between vehicle control and corresponding MEKi-treated mice in GATA6 high and GATA low expression groups, respectively. Statistical significance was calculated by two-tailed Wilcoxon matched-pairs signed rank test. c IHC staining showing reduction of GATA6 expression in MEKi-treated tumors in GATA6 high expression group. Signal intensities were quantified by Definiens and compared between vehicle control and MEKi-treated mice groups ( n = 7 independent mice for each group). Statistical significance was calculated by two-tailed Wilcoxon matched-pairs signed rank test. d Flow cytometry analysis of GATA6 expression in 110299 cells treated with MEKi, class I HDAC inhibitors (mocetinostat, quisinostat, domatinostat), class II HDAC inhibitors (tasquinimod, LMK235, ricolinostat), or their combinations ( n = 4 independent experiments for each cell line). Data are presented as mean values ± SD. Individual dots represent independent biological replicates. Statistical significance was calculated by One-way ANOVA, Kruskal–Wallis test. e Western blot analysis of GATA6 protein levels in four GATA6 high (orange: 110299, 511950, 2838c3, 60400) and four GATA6 low (blue: 60531, 511892, 60590, 6694c2) murine PDAC cell lines following treatment with MEKi and/or domatinostat; β-actin served as a loading control ( n = 1 independent experiment). f Surface MHCI (H-2Db) expression in the four GATA6 high (orange) and four GATA6 low (blue) murine PDAC cell lines treated with MEKi and/or domatinostat, assessed by flow cytometry ( n = 4 independent experiments for cell line 511950, 60400, 511892, 60590, and 5 independent experiments for cell line 110299, 2838c3, 60531, 6694C2). Data are presented as box plots: the center line indicates the median, the bounds of the box indicate the 25th and 75th percentiles, and whiskers extend to minima and maxima. One-way ANOVA and Kruskal–Wallis test were used. Ctrl: vehicle control; MEKi: trametinib; Moce: mocetinostat; Quis: quisinostat; Doma: domatinostat; Tas: tasquinimod; LMK: LMK235; Rico: ricolinostat. MFI mean fluorescence intensity. Scale bar: μm.

Article Snippet: HAs were PCR-amplified using murine primary PDAC cell line (110299) genomic DNA as template and cloned into pJET (Thermo Fisher Scientific) flanking the Blast-P2A-V5-AID cassette to generate pJET_gata6_N-AID_HDR.

Techniques: Derivative Assay, Control, Immunohistochemistry, Marker, Software, Expressing, Two Tailed Test, Flow Cytometry, Western Blot, Fluorescence