Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a Schematic of experimental setup and timeline of lineage tracing experiments. Created in BioRender. Garcia, C. (2026) https://BioRender.com/housa2k . Mice were given tamoxifen at ~4 weeks of age (prior to significant obesity) and analyzed at ~5 weeks of age (one-week post-tamoxifen) to ensure fidelity of labeling or ~16 weeks of age (when islet CCK is present) for lineage tracing analysis. b Average percentage (mean ± SEM) of insulin+ cells labeled with TdTomato in obese Ins1-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 4 mice (2 male, 2 female), 5 weeks; n = 8 mice (4 male, 4 female), 16 weeks), obese Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 2 mice (1 male, 1 female), 5 weeks; n = 10 mice (5 male, 5 female), 16 weeks), and lean Ngn3-Cre ERT2 ;Lep ob/+ ;Rosa26 LSL-TdTomato mice ( n = 2 mice (1 male, 1 female), 5 weeks; n = 4 mice (2 male, 2 female), 16 weeks) administered tamoxifen at 4 weeks of age. p -values of two-sided Welch’s t -test are shown. c , d Terminal weight ( c ) and random glucose ( d ) (mean ± SEM) of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. e Images of CCK immunofluorescence and lineage-traced TdTomato-labeling of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgene. DAPI labels nuclei blue. Scale bar, 100 μm. f Average percentage (mean ± SEM) of CCK+ cells labeled with TdTomato in Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ER transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. For ( c–f ), 16 week-old Lep ob/ob Gcg-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 7 mice (4 male, 3 female)), Ins-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 8 mice (4 male, 4 female)), and Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 10 mice (5 male,5 female)) mice administered tamoxifen at 4 weeks of age were analyzed. Source data are provided as a file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Labeling, Immunofluorescence

Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a AAnet-learned archetypes for WT ( n = 5,847 cells), HFD ( n = 3,887 cells), and Lep ob/ob ( n = 7602 cells) samples (top row). Color embedding of denoised expression (color scale represents min to max of normalized UMI) of Neurogenin 3 ( Ngn3 ) across different conditions (bottom row). b Heatmap for marker gene expression (color scale represents min to max of mean normalized UMI, scaled per gene within condition) for each AT in ( a ). c TrajectoryNet-learned trajectories tracing paths from cells from WT mice to Cck + Lep ob/ob cells ( n = 300 randomly sampled trajectories). Gray cells are all endocrine cells ( n = 20,294 cells). d Enrichment score of each islet cell type for cell-of-origin of Cck + Lep ob/ob β cells (left) and Cck + Lep ob/ob polyhormonal cells (right), calculated over two TrajectoryNet runs that yielded identical results. e TrajectoryNet-inferred proliferation for cells from WT condition ( n = 5847 cells), where proliferation rate > 1 implies growth. Gray cells are all β cells ( n = 17,336 cells). f Cells committed to WT AT 4 ( n = 168 cells) have a higher growth rate than other archetypes (WT AT 1 n = 703 cells, WT AT 2 n = 127 cells, WT AT 3 n = 263 cells, WT AT 5 n = 136 cells, WT AT 6 n = 117 cells, WT AT 7 n = 983 cells) or uncommitted cells ( n = 3350 cells). Box plots display 25, 50, and 75th percentiles ± 1.5 interquartile range (IQR). g Enrichment scores (mean over n = 2 TrajectoryNet runs) for each WT archetype within cells-of-origin for Cck -hi Lep ob/ob AT 2 shows high enrichment of WT AT 4. h TrajectoryNet-learned trajectories tracing paths from cells from WT AT 4 to Lep ob/ob AT 2 ( n = 133 trajectories). Gray cells are all β cells ( n = 17,336 cells). i Enrichment scores (mean over n = 2 TrajectoryNet runs) for each archetype within cells on the trajectory from WT AT 4 to Lep ob/ob AT 2, demonstrating archetypes (HFD AT 4 and Lep ob/ob AT 3) that represent potential intermediate states. j Heatmap of Cck expression (color scale represents min to max of mean normalized UMI per AT) for WT AT 4, HFD AT 4, Lep ob/ob AT 3, and Lep ob/ob AT 2. Source data are provided as a Source Data file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Expressing, Marker, Gene Expression

Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a Schematic of experimental setup and timeline of lineage tracing experiments. Created in BioRender. Garcia, C. (2026) https://BioRender.com/housa2k . Mice were given tamoxifen at ~4 weeks of age (prior to significant obesity) and analyzed at ~5 weeks of age (one-week post-tamoxifen) to ensure fidelity of labeling or ~16 weeks of age (when islet CCK is present) for lineage tracing analysis. b Average percentage (mean ± SEM) of insulin+ cells labeled with TdTomato in obese Ins1-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 4 mice (2 male, 2 female), 5 weeks; n = 8 mice (4 male, 4 female), 16 weeks), obese Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 2 mice (1 male, 1 female), 5 weeks; n = 10 mice (5 male, 5 female), 16 weeks), and lean Ngn3-Cre ERT2 ;Lep ob/+ ;Rosa26 LSL-TdTomato mice ( n = 2 mice (1 male, 1 female), 5 weeks; n = 4 mice (2 male, 2 female), 16 weeks) administered tamoxifen at 4 weeks of age. p -values of two-sided Welch’s t -test are shown. c , d Terminal weight ( c ) and random glucose ( d ) (mean ± SEM) of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. e Images of CCK immunofluorescence and lineage-traced TdTomato-labeling of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgene. DAPI labels nuclei blue. Scale bar, 100 μm. f Average percentage (mean ± SEM) of CCK+ cells labeled with TdTomato in Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ER transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. For ( c–f ), 16 week-old Lep ob/ob Gcg-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 7 mice (4 male, 3 female)), Ins-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 8 mice (4 male, 4 female)), and Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 10 mice (5 male,5 female)) mice administered tamoxifen at 4 weeks of age were analyzed. Source data are provided as a file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Labeling, Immunofluorescence

Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a Gene clusters derived from gene trends (mean scaled expression ± SD) to Lep ob/ob AT 2, grouped into highly variable genes decreasing ( n = 1137 genes) or increasing ( n = 1141 genes). b Gene set enrichment analysis shows significantly ( q < 0.05) enriched gene sets for each gene cluster. Red is upregulated. Blue is downregulated. c Gene set overlap with primary human islet cells treated with ER stress inducers ( p = 1.21 × 10 −48 , hypergeometric test). d Gene set overlap with primary human islet cells exposed to inflammatory cytokines ( p = 7.88 × 10 −5 , hypergeometric test). e Serial random glucose (mean ± SEM, n = 4 male mice per group) of C57/B6 male mice treated with STZ or PBS (vehicle). p -values of two-sided repeated measures ANOVA with Tukey’s post-hoc test are shown. f Final pancreatic Ins1 and Cck expression (qRT-PCR, mean ± SEM, n = 4 male mice per group) normalized to islet marker synaptophysin ( Syn ) of mice in ( e ). p -values of two-sided Welch’s t -test are shown. g Immunofluorescence of islet insulin and CCK expression of mice in ( e ). Images are representative of n = 4 mice per group. h Embedding of β cells ( n = 6760 cells) from vehicle and STZ-treated conditions, colored by treatment, cluster annotation from original work , and marker gene expression (color scale represents min to max of normalized UMI for Ins1 and Ins2 and 1-5.5 for Cck ; gray denotes <1 normalized UMI). i Heatmap of marker gene expression (color scale represents min to max of mean normalized UMI, scaled per gene) for three β cell subclusters in ( h ). j Visualization of β cells ( n = 6760 cells) in ( h ) mapped to all archetypes. k Enrichment scores (mean over n = 2 scMMGAN runs) for each WT archetype for β2 cells and each Lep ob/ob archetype for Cck -hi mSTZ β cells. l TrajectoryNet-inferred proliferation for cells from vehicle condition ( n = 5600 cells), where proliferation rate > 1 implies growth. Gray cells are all β cells ( n = 6760 cells). TrajectoryNet-inferred proliferation rates calculated for β1 ( n = 5531 cells) and β2 ( n = 263 cells) cells in vehicle-treated mice. Box plots display 25, 50, and 75th percentiles ± 1.5 interquartile range (IQR). m Left: Visualization of trajectories from vehicle to β-mSTZ cells mapped to Lep ob/ob AT 2 ( n = 200 randomly sampled trajectories). All β cells visualized ( n = 6760 cells), colored by annotation assignment. Middle: The cell-of-origin enrichment score (mean over n = 2 TrajectoryNet runs) is higher for β2 than β1. Right: TrajectoryNet-learned trajectories tracing paths from cells mapped to WT AT 4 to cells mapped to Lep ob/ob AT 2 ( n = 195 trajectories). Gray cells are all β cells ( n = 6760 cells). Source data are provided as a file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Marker, Immunofluorescence, Gene Expression

Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a PHATE visualization of k -means clustering of acinar cell clusters (n = 260 cells). b Differential proportions of acinar cell clusters in ( a ) in lean (WT) vs. obese (HFD/ Lep ob/ob ) conditions. p < 0.0001, chi-square test. c Heatmap of Tff2 , protease, and Reg gene expression (color scale represents min to max of mean normalized UMI, scaled per gene) for each cluster in ( a ). d PHATE visualization (n = 260 cells) of Tff2 gene expression (color scale represents min to max of normalized UMI) and distribution (violin plots showing min/max with 25 th , 50 th , and 75 th percentiles delineated by lines) in acinar cell clusters (cluster 1 n = 121 cells, cluster 2 n = 124 cells, cluster 3 n = 15 cells). e Violin plots of Reg gene expression distribution (min/max with 25 th , 50 th , and 75 th percentiles delineated by lines) in acinar cell clusters. Cell numbers same as ( d ). f Violin plot of enrichment score distribution (min/max with 25 th , 50 th , and 75 th percentiles delineated by lines) of the human peri-islet acinar-REG+ differential gene expression signature derived from Tosti et al. for each murine acinar cell cluster. Cell numbers same as ( d ). g Violin plots of pancreatic Reg gene expression (RNA-seq) distribution (min/max with 25 th , 50 th , and 75 th percentiles delineated by lines) of 12-week-old WT ( n = 5 mice), Lep ob/ob ( n = 7 mice), and Lep ob/ob mice treated with CCK receptor antagonists ( n = 4 mice ( n = 2 each with proglumide or lorglumide)) for 6 weeks. q -values of Wilcoxon rank sum test with two-stage Benjamini, Krieger, Yekutieli step-up are shown. h IHC images demonstrate increased peri-islet (synaptophysin-positive) acinar cell expression of Reg2 in 3-month-old β cell CCK-expressing tumor models ( KC;Lep ob/+ ;Ins1-Cck/+ and KCO ) compared to controls ( KC;Lep ob/+ ). Images are representative of n = 10 islets analyzed for n = 2 mice per genotype. Scale bar, 100 µm. Source data are provided as a Source Data file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Gene Expression, Derivative Assay, RNA Sequencing, Expressing

Journal: Nature Communications

Article Title: Beta cell-derived cholecystokinin drives obesity-associated pancreatic adenocarcinoma development

doi: 10.1038/s41467-026-69821-2

Figure Lengend Snippet: a Schematic of experimental setup and timeline of lineage tracing experiments. Created in BioRender. Garcia, C. (2026) https://BioRender.com/housa2k . Mice were given tamoxifen at ~4 weeks of age (prior to significant obesity) and analyzed at ~5 weeks of age (one-week post-tamoxifen) to ensure fidelity of labeling or ~16 weeks of age (when islet CCK is present) for lineage tracing analysis. b Average percentage (mean ± SEM) of insulin+ cells labeled with TdTomato in obese Ins1-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 4 mice (2 male, 2 female), 5 weeks; n = 8 mice (4 male, 4 female), 16 weeks), obese Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 2 mice (1 male, 1 female), 5 weeks; n = 10 mice (5 male, 5 female), 16 weeks), and lean Ngn3-Cre ERT2 ;Lep ob/+ ;Rosa26 LSL-TdTomato mice ( n = 2 mice (1 male, 1 female), 5 weeks; n = 4 mice (2 male, 2 female), 16 weeks) administered tamoxifen at 4 weeks of age. p -values of two-sided Welch’s t -test are shown. c , d Terminal weight ( c ) and random glucose ( d ) (mean ± SEM) of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. e Images of CCK immunofluorescence and lineage-traced TdTomato-labeling of Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ERT transgene. DAPI labels nuclei blue. Scale bar, 100 μm. f Average percentage (mean ± SEM) of CCK+ cells labeled with TdTomato in Lep ob/ob ; Rosa26 LSL-TdTomato mice harboring designated Cre ER transgenes. p -values of Kruskal-Wallis with Dunn’s post-hoc test are shown. For ( c–f ), 16 week-old Lep ob/ob Gcg-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 7 mice (4 male, 3 female)), Ins-Cre ERT ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 8 mice (4 male, 4 female)), and Ngn3-Cre ERT2 ;Lep ob/ob ;Rosa26 LSL-TdTomato ( n = 10 mice (5 male,5 female)) mice administered tamoxifen at 4 weeks of age were analyzed. Source data are provided as a file.

Article Snippet: Akita (Stock #003548), Kras LSL-G12D (Stock #008179), Lep ob/ob (Stock #000632), Ins1-Cre ERT (Stock #024709), Gcg-Cre ERT2 (Stock #042277), Ngn3-Cre ERT2 (Stock #028365), Pdx1-Cre (Stock #014647), Rosa26 LSL-TdTomato (Stock #007914), Pgk1-FlpO (Stock #011065), and Trp53 LSL-R172H (Stock #008652) mice were obtained from the Jackson Laboratory (JAX).

Techniques: Labeling, Immunofluorescence