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Metabolic programming of distinct cancer stem cells promotes metastasis of pancreatic ductal adenocarcinoma

Oncogene volume 40, pages 215–231 (2021)Cite this article

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Abstract

Pancreatic ductal adenocarcinoma (PDAC) metastasizes to distant organs, which is the primary cause of mortality; however, specific features mediating organ-specific metastasis remain unexplored. Emerging evidence demonstrates that cancer stem cells (CSCs) and cellular metabolism play a pivotal role in metastasis. Here we investigated the role of distinct subtypes of pancreatic CSCs and their metabolomic signatures in organ-specific metastatic colonization. We found that PDAC consists of ALDH+/CD133+ and drug-resistant (MDR1+) subtypes of CSCs with specific metabolic and stemness signatures. Human PDAC tissues with gemcitabine treatment, autochthonous mouse tumors from KrasG12D; Pdx1-Cre (KC) and KrasG12D; Trp53R172H; Pdx-1 Cre (KPC) mice, and KPC- Liver/Lung metastatic cells were used to evaluate the CSC, EMT (epithelial-to-mesenchymal transition), and metabolic profiles. A strong association was observed between distinct CSC subtypes and organ-specific colonization. The liver metastasis showed drug-resistant CSC- and EMT-like phenotype with aerobic glycolysis and fatty acid β-oxidation-mediated oxidative (glyco-oxidative) metabolism. On the contrary, lung metastasis displayed ALDH+/CD133+ and MET-like phenotype with oxidative metabolism. These results were obtained by evaluating FACS-based side population (SP), autofluorescence (AF+) and Alde-red assays for CSCs, and Seahorse-based oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and fatty acid β-oxidation (FAO)-mediated OCR assays for metabolic features along with specific gene signatures. Further, we developed in vitro human liver and lung PDAC metastasis models by using a combination of liver or lung decellularized scaffolds, a co-culture, and a sphere culture methods. PDAC cells grown in the liver-mimicking model showed the enrichment of MDR1+ and CPT1A+ populations, whereas the PDAC cells grown in the lung-mimicking environment showed the enrichment of ALDH+/CD133+ populations. In addition, we observed significantly elevated expression of ALDH1 in lung metastasis and MDR1/LDH-A expression in liver metastasis compared to human primary PDAC tumors. Our studies elucidate that distinct CSCs adapt unique metabolic signatures for organotropic metastasis, which will pave the way for the development of targeted therapy for PDAC metastasis.

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Fig. 1: Distinct subtypes of CSC populations in pancreatic cancer show specific stem cell features.
Fig. 2: Distinct subtypes of CSC populations show specific metabolic profiles.
Fig. 3: Liver and lung metastasis of PDAC show specific stem cell features.
Fig. 4: Liver and lung metastases of PDAC show specific metabolic features.
Fig. 5: Liver and lung metastases of PDAC show specific EMT/MET features.
Fig. 6: Organ-specific colonization potentials of distinct pancreatic CSCs.
Fig. 7: Human PDAC lung and liver metastases show specific stem cell and metabolic profiles relative to primary PDAC.

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Acknowledgements

We thank Craig Semerad, Victoria B. Smith, and Samantha Wall of the Flow Cytometry Research Facility, University of Nebraska Medical Center, for assisting with flow cytometry. We thank Janice A. Taylor and James R. Talaska of the Advanced Microscopy Core Facility at the University of Nebraska Medical Center for assisting with confocal microscopy. Further, we thank Dr. Kelly Stauch, Seahorse Core director at the University of Nebraska Medical Center, for assisting with Seahorse experiments. We were supported primarily by the following grants from the National Institutes of Health P01 CA217798, R01 CA210637, R01 CA183459, R01 CA195586, R01 CA201444, R01 CA228524, U01 CA200466, R01 CA228524, R01 CA247471, R50CA211462, P50CA12729 and U01 CA210240, and the Nebraska Department of Health and Human Services LB595.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, 68198-5870, USA

    Rama Krishna Nimmakayala, Frank Leon, Satyanarayana Rachagani, Sanchita Rauth, Palanisamy Nallasamy, Saravanakumar Marimuthu, Gautam K. Shailendra, Seema Chugh, Ramakanth Chirravuri, Rohitesh Gupta, Kavita Mallya, Surinder K. Batra & Moorthy P. Ponnusamy

  2. Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, 986145 Nebraska Medical Center, Omaha, NE, 68198-6145, USA

    Yashpal S. Chhonker & Daryl J. Murry

  3. Department of Pathology and Microbiology, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA

    Dipakkumar R. Prajapati & Subodh M. Lele

  4. Eppley Institute for Research in Cancer and Allied Diseases, Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA

    Thomas C. Caffrey, Jean L. Grem, Paul M. Grandgenett, Michael A. Hollingsworth, Surinder K. Batra & Moorthy P. Ponnusamy

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  1. Rama Krishna Nimmakayala
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Contributions

MPP, SKB, and RKN conceived and designed the experiments. RKN performed the experiments. FL, SR, PN, GSK, SC, and RG assisted with in vitro experiments. SR and SM assisted with in vivo experiments. YSC assisted and performed mass spectrometry experiments. MPP, SKB, RKN, DJM, MAH, SML, and PDR analyzed the data. PMG and MAH collected and provided human specimens. RKN and MPP wrote the manuscript.

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Correspondence to Surinder K. Batra or Moorthy P. Ponnusamy.

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SKB is one of the co-founders of Sanguine Diagnostics and Therapeutics, Inc. The other authors disclosed no potential conflicts of interest.

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Nimmakayala, R.K., Leon, F., Rachagani, S. et al. Metabolic programming of distinct cancer stem cells promotes metastasis of pancreatic ductal adenocarcinoma. Oncogene 40, 215–231 (2021). https://doi.org/10.1038/s41388-020-01518-2

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