Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Direct evidence for cancer-cell-autonomous extracellular protein catabolism in pancreatic tumors

Nature Medicine volume 23, pages 235–241 (2017)Cite this article

Subjects

Abstract

Mammalian tissues rely on a variety of nutrients to support their physiological functions1. It is known that altered metabolism is involved in the pathogenesis of cancer, but which nutrients support the inappropriate growth of intact malignant tumors is incompletely understood2,3. Amino acids are essential nutrients for many cancer cells4,5 that can be obtained through the scavenging and catabolism of extracellular protein via macropinocytosis6,7. In particular, macropinocytosis can be a nutrient source for pancreatic cancer cells, but it is not fully understood how the tumor environment influences metabolic phenotypes8 and whether macropinocytosis supports the maintenance of amino acid levels within pancreatic tumors. Here we utilize miniaturized plasma exchange to deliver labeled albumin to tissues in live mice, and we demonstrate that breakdown of albumin contributes to the supply of free amino acids in pancreatic tumors. We also deliver albumin directly into tumors using an implantable microdevice, which was adapted and modified from ref. 9. Following implantation, we directly observe protein catabolism and macropinocytosis in situ by pancreatic cancer cells, but not by adjacent, non-cancerous pancreatic tissue. In addition, we find that intratumoral inhibition of macropinocytosis decreases amino acid levels. Taken together, these data suggest that pancreatic cancer cells consume extracellular protein, including albumin, and that this consumption serves as an important source of amino acids for pancreatic cancer cells in vivo.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Albumin-derived amino acids are found in pancreatic tumors.
Figure 2: Direct assessment of macropinocytosis and albumin catabolism in tumors.
Figure 3: Albumin catabolism and fibronectin internalization in autochthonous pancreatic tumors.
Figure 4: Local depletion of amino acids following inhibition of macropinocytosis in KrasG12D-driven pancreatic tumors in vivo.

Similar content being viewed by others

References

  1. Metallo, C.M. & Vander Heiden, M.G. Understanding metabolic regulation and its influence on cell physiology. Mol. Cell 49, 388–398 (2013).

    Article  CAS  Google Scholar 

  2. Mayers, J.R. & Vander Heiden, M.G. Famine versus feast: understanding the metabolism of tumors in vivo. Trends Biochem. Sci. 40, 130–140 (2015).

    Article  CAS  Google Scholar 

  3. White, E. Exploiting the bad eating habits of Ras-driven cancers. Genes Dev. 27, 2065–2071 (2013).

    Article  CAS  Google Scholar 

  4. Locasale, J.W. Serine, glycine and one-carbon units: cancer metabolism in full circle. Nat. Rev. Cancer 13, 572–583 (2013).

    Article  CAS  Google Scholar 

  5. Wise, D.R. & Thompson, C.B. Glutamine addiction: a new therapeutic target in cancer. Trends Biochem. Sci. 35, 427–433 (2010).

    Article  CAS  Google Scholar 

  6. Kamphorst, J.J. et al. Human pancreatic cancer tumors are nutrient poor and tumor cells actively scavenge extracellular protein. Cancer Res. 75, 544–553 (2015).

    Article  CAS  Google Scholar 

  7. Commisso, C. et al. Macropinocytosis of protein is an amino acid supply route in Ras-transformed cells. Nature 497, 633–637 (2013).

    Article  CAS  Google Scholar 

  8. Davidson, S.M. et al. Environment impacts the metabolic dependencies of Ras-driven non–small cell lung cancer. Cell Metab. 23, 517–528 (2016).

    Article  CAS  Google Scholar 

  9. Jonas, O. et al. An implantable microdevice to perform high-throughput in vivo drug sensitivity testing in tumors. Sci. Transl. Med. 7, 284ra57 (2015).

    Article  Google Scholar 

  10. Schilling, U. et al. Design of compounds having enhanced tumour uptake, using serum albumin as a carrier. Part II. In vivo studies. Int. J. Rad. Appl. Instrum. B 19, 685–695 (1992).

    Article  CAS  Google Scholar 

  11. Steinfeld, J.L. I131 albumin degradation in patients with neoplastic diseases. Cancer 13, 974–984 (1960).

    Article  CAS  Google Scholar 

  12. Rothschild, M.A., Oratz, M. & Schreiber, S.S. Regulation of albumin metabolism. Annu. Rev. Med. 26, 91–104 (1975).

    Article  CAS  Google Scholar 

  13. Gupta, D. & Lis, C.G. Pretreatment serum albumin as a predictor of cancer survival: a systematic review of the epidemiological literature. Nutr. J. 9, 69 (2010).

    Article  Google Scholar 

  14. Andersson, C., Iresjö, B.M. & Lundholm, K. Identification of tissue sites for increased albumin degradation in sarcoma-bearing mice. J. Surg. Res. 50, 156–162 (1991).

    Article  CAS  Google Scholar 

  15. Brenner, D.A., Buck, M., Feitelberg, S.P. & Chojkier, M. Tumor necrosis factor-α inhibits albumin gene expression in a murine model of cachexia. J. Clin. Invest. 85, 248–255 (1990).

    Article  CAS  Google Scholar 

  16. Fearon, K.C. et al. Albumin synthesis rates are not decreased in hypoalbuminemic cachectic cancer patients with an ongoing acute-phase protein response. Ann. Surg. 227, 249–254 (1998).

    Article  CAS  Google Scholar 

  17. Jewell, W.R., Krishnan, E.C. & Schloerb, P.R. Apparent cellular ingress of albumin in Walker 256 tumor and rat muscle. Cancer Res. 35, 405–408 (1975).

    CAS  PubMed  Google Scholar 

  18. Swanson, J.A. & Watts, C. Macropinocytosis. Trends Cell Biol. 5, 424–428 (1995).

    Article  CAS  Google Scholar 

  19. Daly, R. & Hearn, M.T.W. Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production. J. Mol. Recognit. 18, 119–138 (2005).

    Article  CAS  Google Scholar 

  20. Tolner, B., Smith, L., Begent, R.H.J. & Chester, K.A. Production of recombinant protein in Pichia pastoris by fermentation. Nat. Protoc. 1, 1006–1021 (2006).

    Article  CAS  Google Scholar 

  21. Merlot, A.M., Kalinowski, D.S. & Richardson, D.R. Unraveling the mysteries of serum albumin—more than just a serum protein. Front. Physiol. 5, 299 (2014).

    Article  Google Scholar 

  22. Hou, H.W. et al. Deformability based cell margination—a simple microfluidic design for malaria-infected erythrocyte separation. Lab Chip 10, 2605–2613 (2010).

    Article  CAS  Google Scholar 

  23. Hou, H.W. et al. A microfluidics approach towards high-throughput pathogen removal from blood using margination. Biomicrofluidics 6, 024115 (2012).

    Article  Google Scholar 

  24. Mayers, J.R. et al. Elevation of circulating branched-chain amino acids is an early event in human pancreatic adenocarcinoma development. Nat. Med. 20, 1193–1198 (2014).

    Article  CAS  Google Scholar 

  25. Racoosin, E.L. & Swanson, J.A. Macropinosome maturation and fusion with tubular lysosomes in macrophages. J. Cell Biol. 121, 1011–1020 (1993).

    Article  CAS  Google Scholar 

  26. Klionsky, D.J. Autophagy in mammalian systems, Part B. Preface. Methods Enzymol. 452, xxi–xxii (2009).

    Article  Google Scholar 

  27. Koivusalo, M. et al. Amiloride inhibits macropinocytosis by lowering submembranous pH and preventing Rac1 and Cdc42 signaling. J. Cell Biol. 188, 547–563 (2010).

    Article  CAS  Google Scholar 

  28. Commisso, C., Flinn, R.J. & Bar-Sagi, D. Determining the macropinocytic index of cells through a quantitative image-based assay. Nat. Protoc. 9, 182–192 (2014).

    Article  CAS  Google Scholar 

  29. Bardeesy, N. et al. Both p16Ink4a and the p19Arf–p53 pathway constrain progression of pancreatic adenocarcinoma in the mouse. Proc. Natl. Acad. Sci. USA 103, 5947–5952 (2006).

    Article  CAS  Google Scholar 

  30. Whatcott, C.J. et al. Desmoplasia in primary tumors and metastatic lesions of pancreatic cancer. Clin. Cancer Res. 21, 3561–3568 (2015).

    Article  CAS  Google Scholar 

  31. Mahadevan, D. & Von Hoff, D.D. Tumor–stroma interactions in pancreatic ductal adenocarcinoma. Mol. Cancer Ther. 6, 1186–1197 (2007).

    Article  CAS  Google Scholar 

  32. Manic, G., Obrist, F., Kroemer, G., Vitale, I. & Galluzzi, L. Chloroquine and hydroxychloroquine for cancer therapy. Molecular & Cell. Oncol. 1, e29911–e29912 (2014).

    Article  Google Scholar 

  33. Palm, W. et al. The utilization of extracellular proteins as nutrients is suppressed by mTORC1. Cell 162, 259–270 (2015).

    Article  CAS  Google Scholar 

  34. Andersson, C., Lönnroth, C., Moldawer, L.L., Ternell, M. & Lundholm, K. Increased degradation of albumin in cancer is not due to conformational or chemical modifications in the albumin molecule. J. Surg. Res. 49, 23–29 (1990).

    Article  CAS  Google Scholar 

  35. Von Hoff, D.D. et al. Increased survival in pancreatic cancer with nab–paclitaxel plus gemcitabine. N. Engl. J. Med. 369, 1691–1703 (2013).

    Article  CAS  Google Scholar 

  36. Wosikowski, K. et al. In vitro and in vivo antitumor activity of methotrexate conjugated to human serum albumin in human cancer cells. Clin. Cancer Res. 9, 1917–1926 (2003).

    CAS  PubMed  Google Scholar 

  37. Cregg, J.M., Cereghino, J.L., Shi, J. & Higgins, D.R. Recombinant protein expression in Pichia pastoris. Mol. Biotechnol. 16, 23–52 (2000).

    Article  CAS  Google Scholar 

  38. Wyckoff, J., Gligorijevic, B., Entenberg, D., Segall, J. & Condeelis, J. High-resolution multiphoton imaging of tumors in vivo. Cold Spring Harb. Protoc. 2011, 1167–1184 (2011).

    PubMed  PubMed Central  Google Scholar 

  39. Sahai, E. et al. Simultaneous imaging of GFP, CFP and collagen in tumors in vivo using multiphoton microscopy. BMC Biotechnol. 5, 14 (2005).

    Article  Google Scholar 

Download references

Acknowledgements

The authors wish to dedicate this paper to the memory of Katherine Kellersberger. We thank the MIT Department of Chemical Engineering and J. Francois-Hamel for access to bioreactor equipment, R. Bronson for pathological grading of tumors and A. Lau for providing tumor-bearing mice. S.M.D. and A.L. received support from a National Science Foundation Graduate Research Award Fellowship, and support from T32 GM007287 is also acknowledged. O.J. received support from the Koch Institute Frontier Grant and the Prostate Cancer Foundation. J.R.M. acknowledges support from grant F30CA183474 from the NCI and grant T32 GM007753 from NIGMS. M.A.K. and G.S. acknowledge support from NIH grants 1R01 DK075850-01 and 1R01 CA160458-01A1. M.G.V.H. acknowledges support from the Lustgarten Foundation, the Ludwig Center at MIT, the Broad Institute SPARC program, the Burroughs Wellcome Fund, SU2C and the NIH (P30 CA1405141, R01 CA168653). H.W.H. and J.H. acknowledge support by DARPA's Dialysis-Like Therapy (DLT) program under SSC Pacific grant N66001-11-1-4182. This work is also supported by the use of MIT's Microsystems Technology Laboratories.

Author information

Author notes

  1. Shawn M Davidson and Oliver Jonas: These authors contributed equally to this work.

Authors and Affiliations

  1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Shawn M Davidson, Oliver Jonas, Alba Luengo, Jared R Mayers, Jeffrey Wyckoff, Amanda M Del Rosario, Matthew Whitman, Christopher R Chin, Alex Lammers, Michael J Cima, Robert Langer & Matthew G Vander Heiden

  2. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Shawn M Davidson, Alba Luengo, Jared R Mayers, Kendall J Condon & Matthew G Vander Heiden

  3. Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA

    Shawn M Davidson & Matthew G Vander Heiden

  4. Department of Radiology, Brigham and Women's Hospital and Harvard Medical School, Cambridge, Massachusetts, USA

    Oliver Jonas

  5. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Mark A Keibler, Gregory Stephanopoulos & Robert Langer

  6. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Han Wei Hou & Jongyoon Han

  7. Bruker Daltronics, Inc., Billerica, Massachusetts, USA

    Katherine A Kellersberger & Brian K Stall

  8. New York University School of Medicine, New York University, New York, New York, USA

    Dafna Bar-Sagi

  9. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Jongyoon Han

  10. Department of Chemistry and Integrative Genomics, Princeton University, Princeton, New Jersey, USA

    Joshua D Rabinowitz

  11. Department of Materials Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Michael J Cima

  12. Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, Massachusetts, USA

    Matthew G Vander Heiden

Authors
  1. Shawn M Davidson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Oliver Jonas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mark A Keibler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Han Wei Hou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alba Luengo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jared R Mayers
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jeffrey Wyckoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Amanda M Del Rosario
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Matthew Whitman
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Christopher R Chin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Kendall J Condon
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Alex Lammers
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Katherine A Kellersberger
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Brian K Stall
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Gregory Stephanopoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Dafna Bar-Sagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Jongyoon Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Joshua D Rabinowitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Michael J Cima
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Robert Langer
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Matthew G Vander Heiden
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization: S.M.D., O.J. and M.G.V.H. Methodology: S.M.D., O.J., M.A.K., H.W.H., A.L., J.R.M., J.W., A.M.D., M.W., C.R.C., K.J.C., A.L., K.A.K., B.K.S., G.S., J.H. and D.B.-S. Formal analysis: S.M.D., O.J. and A.M.D. Investigation: S.M.D. and O.J. Writing original draft: S.M.D., O.J. and M.G.V.H. Visualization: S.M.D., O.J. and M.G.V.H. Supervision: M.J.C., R.L. and M.G.V.H. Funding acquisition: O.J., J.D.R., R.L. and M.G.V.H.

Corresponding author

Correspondence to Matthew G Vander Heiden.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

Supplementary Figures 1–4 (PDF 452 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Davidson, S., Jonas, O., Keibler, M. et al. Direct evidence for cancer-cell-autonomous extracellular protein catabolism in pancreatic tumors. Nat Med 23, 235–241 (2017). https://doi.org/10.1038/nm.4256

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.4256

This article is cited by

Search

Advanced search

Quick links

Nature Briefing: Cancer

Sign up for the Nature Briefing: Cancer newsletter — what matters in cancer research, free to your inbox weekly.

Get what matters in cancer research, free to your inbox weekly. Sign up for Nature Briefing: Cancer