Hyaluronic acid fuels pancreatic cancer cell growth
- Doctoral Program in Cancer Biology, University of Michigan, United States
- Department of Molecular & Integrative Physiology, University of Michigan, United States
- Program in Chemical Biology, University of Michigan, United States
- Department of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, United States
- Department of Radiology, Memorial Sloan Kettering Cancer Center, United States
- Department of Chemistry, University of Southern California, United States
- Department of Biological Sciences, University of Southern California, United States
- Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center, United States
- Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan, United States
- Rogel Cancer Center, University of Michigan, United States
Figures
Figure 1 with 1 supplement
Pancreatic ductal adenocarcinoma (PDA) requires de novo hexosamine biosynthetic pathway (HBP) fidelity in vitro but not in vivo.
(A) Schematic overview of the HBP and the nutrient inputs. Ac-CoA, acetyl-coenzyme A; GFAT1, glutamine fructose 6-phosphate amidotransferase 1; Glc, glucose; GlcNAc, N-acetyl-glucosamine; Gln, …
Figure 1—figure supplement 1
Additional characterization of GFAT1 knockout pancreatic ductal adenocarcinoma (PDA) populations and clonal lines.
(A) Proliferation kinetics of parental PDA cell lines and corresponding pooled populations of GFAT1 knockout cells supplemented with varying concentrations of N-acetyl-glucosamine (GlcNAc) (n = 3). …
Figure 2 with 1 supplement
Conditioned media (CM) from cancer-associated fibroblasts (CAFs) and wild-type pancreatic ductal adenocarcinoma (PDA) cells support proliferation of GFAT1 knockout cells.
(A) Representative wells from a colony-forming assay in parental TU8988T and GFAT1 knockout clonal line D10 in 10 mM N-acetyl-glucosamine (GlcNAc), base media (DMEM), or base media supplemented 1:3 …
Figure 2—figure supplement 1
Rescue activity of conditioned media (CM) and N-acetyl-glucosamine (GlcNAc) in GFAT1 knockout cells.
(A) Quantitation of colony-forming assay data of parental MiaPaCa2 and GFAT1 knockout clonal line M11 in base media (DMEM), positive control GlcNAc, wildtype TU8988T CM diluted 1:2 (33%) or used …
Figure 3 with 2 supplements
Hyaluronic acid rescues GFAT1 knockout pancreatic ductal adenocarcinoma (PDA) cells.
(A) Representative wells from a colony-forming assay in parental and clonally derived GFAT1 knockout TU8988T cell lines grown in base media (DMEM), positive control N-acetyl-glucosamine (GlcNAc) (10 …
Figure 3—figure supplement 1
Characterization of macropinocytosis and glycosaminoglycan rescue activity in pancreatic ductal adenocarcinoma (PDA) and GFAT1 knockout cells.
(A–F) Representative colony formation assays and their quantitation following treatment with two concentrations of heparin or necrotic cell debris that contain complete cellular contents, relative …
Figure 3—figure supplement 2
Analysis of hyaluronic acid formulation on GFAT1 rescue and composition in conditioned media (CM).
(A,B) Proliferation time course of (A) parental MiaPaCa2 and (B) GFAT1 knockout cells in base media (DMEM), positive control (N-acetyl-glucosamine [GlcNAc]), 5 kDa HA (oligo-hyaluronic acid [o-HA]), …
Figure 4
Hyaluronic acid in conditioned media (CM) rescues GFAT1 knockout.
(A) Quantification of hyaluronic acid (HA) in CM from wildtype TU8988T cells treated with varying doses of 4-methylumbelliferone (4-MU) (n = 3). (B–E) Proliferation time course of (B,C) wildtype …
Figure 5 with 1 supplement
Hyaluronic acid-derived N-acetyl-glucosamine (GlcNAc) rescues GFAT1 loss via the GlcNAc salvage pathway.
(A) Western blot of NAGK, GFAT1, and ACTIN loading control from TU8988T, MiaPaCa2, and HPAC parental (wildtype, WT) and NAGK knockout (KO) populations. NAGK was knocked out using two independent …
Figure 5—figure supplement 1
Additional characterization of hyaluronic acid (HA) rescue in GFAT1/NAGK double knockout cell lines.
(A–D) Proliferation time course of (A) parental TU8988T, (B) GFAT1 knockout cells, and (C,D) two GFAT1/NAGK double knockout cell lines in base media (DMEM), positive control (GlcNAc), 5 kDa HA (o-HA)…
Additional files
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Source data 1
Raw western blot images.
- https://cdn.elifesciences.org/articles/62645/elife-62645-supp1-v3.docx
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Transparent reporting form
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