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

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.

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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.

References

  1. 1

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

    CAS  Article  Google Scholar 

  2. 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).

    CAS  Article  Google Scholar 

  3. 3

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

    CAS  Article  Google Scholar 

  4. 4

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

    CAS  Article  Google Scholar 

  5. 5

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

    CAS  Article  Google Scholar 

  6. 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).

    CAS  Article  Google Scholar 

  7. 7

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

    CAS  Article  Google Scholar 

  8. 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).

    CAS  Article  Google Scholar 

  9. 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. 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).

    CAS  Article  Google Scholar 

  11. 11

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

    CAS  Article  Google Scholar 

  12. 12

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

    CAS  Article  Google Scholar 

  13. 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. 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).

    CAS  Article  Google Scholar 

  15. 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).

    CAS  Article  Google Scholar 

  16. 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).

    CAS  Article  Google Scholar 

  17. 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. 18

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

    CAS  Article  Google Scholar 

  19. 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).

    CAS  Article  Google Scholar 

  20. 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).

    CAS  Article  Google Scholar 

  21. 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. 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).

    CAS  Article  Google Scholar 

  23. 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. 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).

    CAS  Article  Google Scholar 

  25. 25

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

    CAS  Article  Google Scholar 

  26. 26

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

    Article  Google Scholar 

  27. 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).

    CAS  Article  Google Scholar 

  28. 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).

    CAS  Article  Google Scholar 

  29. 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).

    CAS  Article  Google Scholar 

  30. 30

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

    CAS  Article  Google Scholar 

  31. 31

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

    CAS  Article  Google Scholar 

  32. 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. 33

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

    CAS  Article  Google Scholar 

  34. 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).

    CAS  Article  Google Scholar 

  35. 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).

    CAS  Article  Google Scholar 

  36. 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. 37

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

    CAS  Article  Google Scholar 

  38. 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. 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.

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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.

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Correspondence to Matthew G Vander Heiden.

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The authors declare no competing financial interests.

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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

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