Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Effects of GOT1 inhibition in the pancreatic TME. (a) Schematic of glutamine metabolism. (b) Determination of half‐maximal inhibitory concentration (IC 50 ) of the GOT1 inhibitor using mono‐ and multicellular 3D cultures. (c) Cell viability of mono‐ and multicellular 3D cultures. n = 3, **** = p ≤ 0.0001, (d) live/dead staining (scale bar = 100 µm), and (e) relative metabolite quantification following GOT1 inhibition. (f) Gene ontology enrichment analysis, and (g) volcano plot showing the significantly up/downregulated matrisome proteins of treated versus non‐treated multicellular 3D cultures. Blue = upregulated, red = downregulated. FDR cut‐off = 0.05, log2(fold‐change) ≥ 2. (h) Immunoblot of multicellular 3D cultures upon GOT1 inhibition. (i) Cell metabolic activity for PDAC cells cultured in type III collagen. n = 6, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001. (j) Cytokine profiling of multicellular 3D cultures following GOT1 inhibition. GOT1, glutamic‐oxaloacetic transaminase 1. TME, tumor microenvironment. PDAC, pancreatic ductal adenocarcinoma.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Inhibition, Concentration Assay, Staining, Western Blot, Activity Assay, Cell Culture

Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Analyses of cell responses following combined GOT1 inhibition and treatment with cytotoxic drugs. (a) Schematic of experimental rationale. (b) Cell viability of mono‐ and multicellular 3D cultures grown in GelMA‐HAMA hydrogels following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. n = 3, * = p ≤ 0.05, *** = p ≤ 0.001. (c) Gene ontology enrichment analysis, (d) signaling pathway analysis, and (e) volcano plot showing significantly up/downregulated matrisome proteins of treated multicellular 3D cultures compared to non‐treated cells grown in GelMA‐HAMA hydrogels. Blue = upregulated, red = downregulated. (f) Relative metabolite quantification in multicellular 3D cultures following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. FDR cut‐off = 0.05, log2(fold‐change) ≥ 2. GOT1, glutamic‐oxaloacetic transaminase 1. GelMA, gelatin methacrylate. HAMA, hyaluronic acid methacrylate.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Inhibition

Journal: Advanced Science

Article Title: GOT1 Inhibition Induces Extracellular Matrix Remodeling in Pancreatic Cancer

doi: 10.1002/advs.202516578

Figure Lengend Snippet: Multicellular 3D cultures including patient‐derived PDAC cells and their response toward GOT1 inhibition. (a) Micrograph of multicellular 3D cultures grown in GelMA‐HAMA hydrogels (scale bar = 100 µm). (b) Cell metabolic activity and (c) DNA content of mono‐ and multicellular 3D cultures including patient‐derived PDAC cells in GelMA‐HAMA hydrogels. (d) Cell viability, and (e) DNA content of mono‐ and multicellular TKCC‐22‐LO cultures following treatment with GOT1 inhibitor ± gemcitabine and nab‐paclitaxel. n = 3, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001. (f) Live/dead staining (scale bar = 100 µm), and (g) heatmap of amino acid metabolism microarray of multicellular 3D cultures treated with the GOT1 inhibitor compared to non‐treated cells. PDAC, pancreatic ductal adenocarcinoma. GOT1, glutamic‐oxaloacetic transaminase 1. GelMA, gelatin methacrylate. HAMA, hyaluronic acid methacrylate.

Article Snippet: Mono‐ and multicellular 3D cultures were established as described above and grown for 7 days and then treated with 25 μM GOT1 inhibitor (MedchemExpress HY‐122723) alone or combined with 100 nM gemcitabine (Sigma‐Aldrich G6423) and 100 nM nab‐paclitaxel (Bristol‐Myers Squibb), which were dissolved directly into PDAC or TKCC cell media.

Techniques: Derivative Assay, Inhibition, Activity Assay, Staining, Microarray

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Scheme of the glutamate-oxaloacetate transaminases (GOTs) and their roles in cellular metabolism. OAA=oxaloacetate, Asp=aspartate, Glu=glutamine, αKG=α-ketoglutarate, Asn=asparagine. B, Scheme of alleles used for Got knockout pancreata. Got1 and/or Got2 deletion driven by epithelial pancreas-specific Cre recombinase (p48-Cre) on an immunocompetent (C57B/6) background. C, Pancreas and body weight of Ptf1a-Cre ; Got1 f/f ;Got2 f/f double knockout (n=11) mice compared to controls. Controls include 8-week old single KO (n=5) or wild type (WT) mice, as indicated by colored circles. D, Representative hematoxylin and eosin (H&E) staining of pancreata from 8-week old Got1 KO and Got1/Got2 knockout mice. Scale bar is 800µm for 1.5x and 200µm for 5x. E, Percent acinar and F, islet area calculated from H&E images of 8-week old Ptf1a-Cre ; Got1 f/f (n=3) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f mice (n=7) mice. G, Fasting glucose and H, insulin after 5 hour fast in Got1 f/f ;Got2 f/f (n=4) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f (n=3) female mice. I, Blood glucose and J, plasma insulin measurements after 1.5g/kg glucose bolus with plasma analysis at 0, 15, 30, 60, and 120 minutes for Got1 f/f ;Got2 f/f (n=4) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f (n=3) female mice. K, Glucose-insulin index and L, HOMA-IR index of Got1 f/f ;Got2 f/f (n=4) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f (n=3) female mice. M, Plasma amylase for no serum controls, cerulein-treated wildtype mice (positive controls, n=4), Cre negative Got1 f/f ;Got2 f/f mice (n=5) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f double knockout mice (n=2). N-Q, Relative metabolite abundance, reported as fold change (FC) relative to control, as determined by LC-MS/MS in pancreata of 8-week old Got1 f/f ;Got2 f/f (n=3) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f (n=4) mice. N, aspartate, O, glutamate, P , α-ketoglutarate (αKG), and Q , lactate/pyruvate..

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Knock-Out, Double Knockout, Staining, Clinical Proteomics, Control, Liquid Chromatography with Mass Spectroscopy

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Representative hematoxylin and eosin (H&E) staining of Got1 f/f ;Got2 f/f (Cre negative) and Ptf1a-Cre ; Got1 f/f ;Got2 f/f pancreata. Scale bar is 800µm. B, Unsupervised hierarchical clustering of differentially abundant metabolites (p<0.1, compared to Cre negative mice) in pancreata of 8-week old Got1 f/f ;Got2 f/f (n=3) or Ptf1a-Cre ; Got1 f/f ;Got2 f/f (n=4) mice, as measured by LC-MS/MS. C, Curated list of glycolytic, pentose phosphate, and TCA cycle intermediate metabolites from the dataset generated in B .

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Staining, Liquid Chromatography with Mass Spectroscopy, Generated

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Scheme of alleles used to study Got in pancreatic precancerous lesions. Got1 or Got2 deletion with expression of mutant Kras (LSL- Kras G12D ) driven by epithelial pancreas-specific Cre recombinase ( p48 -Cre) on an immunocompetent (C57B/6) background (KC-Got1 and KC-Got2). B, Pancreas and body weight of KC (n=4) and KC; Got1 f/f (n=3) mice aged to 6 months. C, Pancreas and body weight of KC (n=4), KC; Got2 f/+ (n=10), and KC; Got2 f/f (n=5) mice aged to 6 months. D, Pancreas and body weight of KC (n=4), KC; Got1 f/+ KC (n=3); Got1 f/f (n=8) mice aged to 12 months. E, Pancreas and body weight of KC (n=4), KC; Got2 f/+ (n=3), and KC; Got2 f/f (n=4) mice aged to 12 months. F, Histological grading of pancreata of KC (n=3), KC; Got1 f/f (n=5), and KC; Got2 f/f (n=5) mice aged to 12 months. G, Representative hematoxylin and eosin (H&E) staining of pancreata from 6- and H, 12-month aged KC mice. Scale bar is 250µm for 4x and 50µm for 20x images.

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Expressing, Mutagenesis, Staining

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Tumor and body weight of KPC (n=6), KPC; Got1 f/+ (n=11), and KPC; Got1 f/f (n=5) mice. B, Tumor and body weight of KPC (n=6), KPC; Got2 f/+ (n=12), and KPC; Got2 f/f (n=11) mice. C, Representative hematoxylin and eosin (H&E) staining of 12-month-aged KC; Got2 f/+ pancreata. Scale bar is 100µm. D, Individual immunofluorescence panels for multiplex IF. Scale bar is 250µm for 4x and 50µm for 20x images.

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Staining, Immunofluorescence, Multiplex Assay

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Scheme of alleles used to study Got in pancreatic cancer. Got1 or Got2 deletion with expression of mutant Kras (LSL- Kras G12D ) and mutant p53 (LSL- Tp53 R17H2/+ ) driven by epithelial pancreas-specific Cre recombinase ( p48 -Cre) on an immunocompetent (C57B/6) background (KPC-Got1 and KPC-Got2). B, Kaplan-Meier survival curves for overall survival of KPC (n=6), KPC; Got1 f/+ (n=11), and KPC; Got1 f/f (n=5) mice. C, Kaplan-Meier survival curves for overall survival of KPC (n=6), KPC; Got2 f/+ (n=12), and KPC; Got2 f/f (n=11) mice. D, Representative hematoxylin and eosin (H&E) staining of pancreata from KPC mice. Scale bar is 250µm for 4x and 50µm for 20x images.

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Expressing, Mutagenesis, Staining

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Representative multiplex IF images of KPC, KPC; Got1 f/+ , KPC; Got1 f/f , KPC; Got2 f/+ , and KPC; Got2 f/f mice. Scale bar is 100µm. B, C, D, E, T cell populations identified by multiplex IF of KPC, KPC; Got1 f/+ , and KPC; Got1 f/f mice (n=5 per group). F, G, H, Macrophage populations identified by multiplex IF of KPC, KPC; Got1 f/+ , and KPC; Got1 f/f mice (n=5 per group). I, J, K, Epithelial cell populations identified by multiplex IF of KPC, KPC; Got1 f/+ , and KPC; Got1 f/f mice (n=5 per group). L, M, N, O, T cell populations identified by multiplex IF of KPC, KPC; Got2 f/+ , and KPC; Got2 f/f mice (n=5 per group). P, Q, R, Macrophage populations identified by multiplex IF of KPC, KPC; Got2 f/+ , and KPC; Got2 f/f mice (n=5 per group). S, T, U, Epithelial cell populations identified by multiplex IF of KPC, KPC; Got2 f/+ , and KPC; Got2 f/f mice (n=5 per group).

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Multiplex Assay

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: A, Western blot analysis of murine KPC pancreatic cancer cell lines (7940B and MT3) stably expressing a non-targeting sgRNA (sgNT) or with Got1 or Got2 KO. Two independent sgRNAs against GOT2 are designated sg1 and sg2. B, Cell count of 7940B and MT3 cell lines with Got1 or Got2 KO over 48-72 hours. Each clone plated with 5-10 replicates. C,D, Oxygen consumption rate (fmol/mm 2 /s) of 7940B and MT3 cell lines with Got1 or Got2 KO normalized to cell number. Each clone plated with 6 replicates. F-M , Relative intracellular metabolite abundance, reported as fold change (FC), as determined by LC-MS/MS in MT3 and 7940B murine KPC cells. F,G aspartate, H,I, glutamate, J,K , α-KG (α-ketoglutarate), L,M , lactate/pyruvate. N,O, Pancreas and body weight of mice at endpoint after orthotopic implantation of pancreatic cancer cells (n=5 per group).

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Western Blot, Stable Transfection, Expressing, Cell Counting, Liquid Chromatography with Mass Spectroscopy

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: Unsupervised hierarchical clustering of differentially abundant intracellular metabolites (p<0.1, compared to sgNT) from MT3 cells with Got1 or Got2 knockout as measured by LC-MS/MS. n=3 biologically independent samples per group.

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Knock-Out, Liquid Chromatography with Mass Spectroscopy

Journal: bioRxiv

Article Title: Aspartate Transaminases Are Dispensable for Pancreatic Development and Pancreatic Cancer Progression

doi: 10.64898/2025.12.17.694916

Figure Lengend Snippet: Unsupervised hierarchical clustering of differentially abundant extracellular metabolites (p<0.1, compared to sgNT) from 7940B cells with Got1 or Got2 knockout as measured by LC-MS/MS. n=3 biologically independent samples per group; n=2 for Got2 sg2.

Article Snippet: Floxed Got1 f/f and Got2 f/f mice were generated by Ozgene ( Got1 floxed around exon 3; Got2 floxed around exon 2) , .

Techniques: Knock-Out, Liquid Chromatography with Mass Spectroscopy