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:

The fungal mycobiome promotes pancreatic oncogenesis via activation of MBL

Matters Arising to this article was published on 02 August 2023

Abstract

Bacterial dysbiosis accompanies carcinogenesis in malignancies such as colon and liver cancer, and has recently been implicated in the pathogenesis of pancreatic ductal adenocarcinoma (PDA)1. However, the mycobiome has not been clearly implicated in tumorigenesis. Here we show that fungi migrate from the gut lumen to the pancreas, and that this is implicated in the pathogenesis of PDA. PDA tumours in humans and mouse models of this cancer displayed an increase in fungi of about 3,000-fold compared to normal pancreatic tissue. The composition of the mycobiome of PDA tumours was distinct from that of the gut or normal pancreas on the basis of alpha- and beta-diversity indices. Specifically, the fungal community that infiltrated PDA tumours was markedly enriched for Malassezia spp. in both mice and humans. Ablation of the mycobiome was protective against tumour growth in slowly progressive and invasive models of PDA, and repopulation with a Malassezia species—but not species in the genera Candida, Saccharomyces or Aspergillus—accelerated oncogenesis. We also discovered that ligation of mannose-binding lectin (MBL), which binds to glycans of the fungal wall to activate the complement cascade, was required for oncogenic progression, whereas deletion of MBL or C3 in the extratumoral compartment—or knockdown of C3aR in tumour cells—were both protective against tumour growth. In addition, reprogramming of the mycobiome did not alter the progression of PDA in Mbl- (also known as Mbl2) or C3-deficient mice. Collectively, our work shows that pathogenic fungi promote PDA by driving the complement cascade through the activation of MBL.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Fig. 1: PDA is characterized by a distinctive intratumoral and gut mycobiome.
Fig. 2: PDA in humans is associated with a distinct mycobiome.
Fig. 3: Fungal dysbiosis promotes pancreatic oncogenesis.
Fig. 4: Fungi promote progression of PDA via the MBL–C3 axis.

Similar content being viewed by others

Data availability

The sequence datasets analysed in this article are publicly available in the NCBI BioProject database, under the accession number PRJNA557226. Raw data for all experiments are available as Source Data to the relevant figures. Any other relevant data are available from the corresponding authors upon reasonable request.

References

  1. Pushalkar, S. et al. The pancreatic cancer microbiome promotes oncogenesis by induction of innate and adaptive immune suppression. Cancer Discov. 8, 403–416 (2018).

    Article  CAS  Google Scholar 

  2. Hingorani, S. R. et al. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 4, 437–450 (2003).

    Article  CAS  Google Scholar 

  3. Hingorani, S. R. et al. Trp53 R172H and Kras G12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. Cancer Cell 7, 469–483 (2005).

    Article  CAS  Google Scholar 

  4. van Asbeck, E. C., Hoepelman, A. I., Scharringa, J., Herpers, B. L. & Verhoef, J. Mannose binding lectin plays a crucial role in innate immunity against yeast by enhanced complement activation and enhanced uptake of polymorphonuclear cells. BMC Microbiol. 8, 229 (2008).

    Article  Google Scholar 

  5. Ishikawa, T. et al. Identification of distinct ligands for the C-type lectin receptors mincle and dectin-2 in the pathogenic fungus Malassezia. Cell Host Microbe 13, 477–488 (2013).

    Article  CAS  Google Scholar 

  6. Afshar-Kharghan, V. The role of the complement system in cancer. J. Clin. Invest. 127, 780–789 (2017).

    Article  Google Scholar 

  7. Cho, M. S. et al. Autocrine effects of tumor-derived complement. Cell Reports 6, 1085–1095 (2014).

    Article  CAS  Google Scholar 

  8. Sam, Q. H., Chang, M. W. & Chai, L. Y. The fungal mycobiome and its interaction with gut bacteria in the host. Int. J. Mol. Sci. 18, 330 (2017).

    Article  Google Scholar 

  9. Zambirinis, C. P. et al. TLR9 ligation in pancreatic stellate cells promotes tumorigenesis. J. Exp. Med. 212, 2077–2094 (2015).

    Article  CAS  Google Scholar 

  10. Reikvam, D. H. et al. Depletion of murine intestinal microbiota: effects on gut mucosa and epithelial gene expression. PLoS ONE 6, e17996 (2011).

    Article  ADS  CAS  Google Scholar 

  11. Skalski, J. H. et al. Expansion of commensal fungus Wallemia mellicola in the gastrointestinal mycobiota enhances the severity of allergic airway disease in mice. PLoS Pathog. 14, e1007260 (2018).

    Article  Google Scholar 

  12. Hruban, R. H. et al. Pancreatic intraepithelial neoplasia: a new nomenclature and classification system for pancreatic duct lesions. Am. J. Surg. Pathol. 25, 579–586 (2001).

    Article  CAS  Google Scholar 

  13. Seifert, L. et al. The necrosome promotes pancreatic oncogenesis via CXCL1 and Mincle-induced immune suppression. Nature 532, 245–249 (2016).

    Article  ADS  CAS  Google Scholar 

  14. Walters, W. et al. Improved bacterial 16S rRNA gene (V4 and V4-5) and fungal internal transcribed spacer marker gene primers for microbial community surveys. mSystems 1, e00009-15 (2015).

    PubMed  PubMed Central  Google Scholar 

  15. Navas-Molina, J. A. et al. Advancing our understanding of the human microbiome using QIIME. Methods Enzymol. 531, 371–444 (2013).

    Article  CAS  Google Scholar 

  16. Caporaso, J. G. et al. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods 7, 335–336 (2010).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We acknowledge the use of the Experimental Pathology and Microscopy core facilities at NYU School of Medicine. These shared resources are partially supported by the Cancer Center Support Grant, P30CA016087, at the Laura and Isaac Perlmutter Cancer Center. This work was supported by NIH grants CA168611 (G.M.), CA206105 (G.M. and D.S.), CA215471 (G.M.), CA19311 (G.M.), DK106025 (G.M.), DE025992 (D.S. and X.L.) and UL1TR001445 (J.I.K.), Department of Defense grant CA170450 (G.M.) and Deutsche Forschungsgemeinschaft Grant AY 126/1-1 (B.A.).

Author information

Authors and Affiliations

Authors

Contributions

B.A. carried out in vivo and in vitro experiments, study design, PCR, analysis and interpretation, manuscript preparation and statistical analysis; S.P. carried out fungal DNA sequencing, analysis and interpretation, manuscript preparation, microbiology study design and statistical analysis; R.C. carried out in vivo experiments, histological analysis and manuscript preparation; Q.L. performed computational analyses and provided critical review; R.A. carried out in vivo experiments and provided technical assistance; J.I.K. carried out in vivo experiments and provided critical review; S.A.S. carried out mouse breeding and histology; D.W. performed tissue culture and cell-line generation; P.P. provided technical assistance and carried out in vivo experiments; N.V. carried out knockdown experiments; Y.G. carried out PCR; A.S. performed FISH and provided critical review; M.V. carried out DNA extraction and contributed to computational analysis; B.D. carried out in vivo experiments and critical review; W.W. provided technical assistance; J.L. provided critical review and contributed to study design; E.K. carried out in vivo experiments and contributed to study design; J.A.K.R. provided technical assistance and contributed to study design; M.H. carried out in vivo experiments; C.Z. carried out human-sample collection; X.L. provided technical assistance; D.S. and G.M. conceived, designed, supervised, analysed and interpreted the study, prepared the manuscript and provided critical review.

Corresponding authors

Correspondence to Deepak Saxena or George Miller.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Peer review information Nature thanks Marina Pasca di Magliano and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Extended data figures and tables

Extended Data Fig. 1 Fungal infiltration of the pancreas in benign disease.

Fungal DNA content was tested using qPCR in pancreata from control (ctl) mice (n = 5) and mice induced to develop caerulein-induced pancreatitis (n = 5). ns, not significant. Data are mean ± s.e.m. Two-tailed Student’s t-test.

Source data

Extended Data Fig. 2 Dysbiosis of the gut mycobiome in a mouse model of PDA.

Hierarchical tree cladogram depicting changes in the taxonomic composition of the mycobiome (assigned to the genus level) in the guts of 30-week-old KC (n = 14) compared to wild-type (n = 12) mice, based on the average percentage relative abundance of genera as determined by 18S ITS sequencing.

Source data

Extended Data Fig. 3 Efficacy of antifungal treatments in pancreatic disease.

a, Wild-type mice that bear orthotopic PDA tumours were treated with vehicle (n = 7 mice) or fluconazole (n = 8 mice), and killed three weeks later. Tumours were collected and weighed. Data are representative of experiments that were performed twice. Scale bar, 1 cm. b, Germ-free wild-type mice were treated with amphotericin B (n = 6 mice) or vehicle (n = 10 mice), and orthotopic tumours from KPC mice were administered to them. Mice were killed three weeks later, and tumours were collected and weighed. Scale bar, 1 cm. ce, Wild-type mice induced to develop caerulein-induced pancreatitis were serially treated with amphotericin B (n = 5 mice) or vehicle (n = 3 mice). c, Representative H&E-stained sections of pancreata are shown, and pancreatic oedema was quantified by measuring the percentage of the area that was white space. Scale bar, 100 μm. d, CD45+ inflammatory cell infiltration was determined by immunohistochemistry. Scale bar, 20 μm. e, Serum levels of amylase were measured. n = 5 mice treated with amphotericin B, n = 3 mice treated with vehicle and n = 3 mock-treated (control) mice. f, Wild-type mice treated with amphotericin B were repopulated with C. tropicalis (n = 4 mice) or vehicle (n = 4 mice), and killed three weeks later. Tumours were collected and weighed. Scale bar, 1 cm. Data are mean ± s.e.m. P values determined by two-tailed Student’s t-test (af).

Source data

Extended Data Fig. 4 Fungal dysbiosis drives the progression of PDA via the lectin pathway.

a, Kaplan–Meier survival curve of patients with PDA, stratified by high (n = 16 patients), medium-high (n = 24 patients), medium-low (n = 26 patients) and low (n = 17 patients) expression of MBL on the basis of data from TCGA. b, Orthotopic tumours from KPC mice were administered to MBL-null mice treated with vehicle (n = 3 mice) or amphotericin B (n = 4 mice), and killed three weeks later. Tumours were collected and weighed. Data are representative of three separate experiments. c, MBL-null mice treated with amphotericin B were repopulated with M. globosa (n = 5 mice) or sham-repopulated (n = 4 mice), and killed three weeks later. Tumours were collected and weighed. Data are representative of experiments that were repeated twice. d, Kaplan–Meier survival curve of patients with PDA, stratified by high (n = 18) versus low (n = 15) expression of C3, on the basis of data from TCGA. e, Pancreata from three-month-old wild-type, KC and KC, MBL-null mice were stained using a monoclonal antibody against C3a. Representative images from two experiments are shown. Scale bar, 20 μm. f, KPC tumour cells were seeded in 96-well plates with vehicle or recombinant mouse C3a. n = 5 cells per group for each time point. Cellular proliferation was measured at serial time points using the XTT assay. Data are representative of experiments that were repeated three times. Data are mean ± s.e.m. P values determined by two-tailed log-rank test (a, d) or two-tailed Student’s t-test (bcf).

Source data

Supplementary information

Supplementary Data

Sequence match of Malassezia globosa from human samples and ATTC strain used in murine repopulation experiments.

Reporting Summary

Source data

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Aykut, B., Pushalkar, S., Chen, R. et al. The fungal mycobiome promotes pancreatic oncogenesis via activation of MBL. Nature 574, 264–267 (2019). https://doi.org/10.1038/s41586-019-1608-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41586-019-1608-2

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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