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.

  • Matters Arising
  • Published:

Apolipoprotein C3 induces inflammasome activation only in its delipidated form

Nature Immunology volume 24, pages 408–411 (2023)Cite this article

Subjects

Matters Arising to this article was published on 13 February 2023

The Original Article was published on 09 December 2019

arising from S. Zewinger et al. Nature Immunology https://doi.org/10.1038/s41590-019-0548-1 (2020)

Increased levels of apolipoprotein C3 (APOC3) in the blood predict cardiovascular disease risk in people with and without diabetes1,2,3,4. Conversely, APOC3 loss-of-function variants are associated with cardioprotection in humans5,6,7. The mechanisms whereby APOC3 promotes atherosclerotic cardiovascular disease risk have been the topic of intense studies. It is known that APOC3 slows the clearance of triglyceride (TG)-rich lipoproteins (TRLs) and their remnant lipoprotein particles (RLPs) by inhibiting the activity of lipoprotein lipase and by preventing hepatic uptake of TRLs and RLPs, leading to increased levels of TRLs and their remnants in circulation8. The atherogenic effects of APOC3 are likely mediated, at least in part, by its effects on TRL metabolism, but recent findings that APOC3 predicts future coronary artery disease events in individuals with type 1 diabetes (T1D), who often have normal plasma lipid profiles1,2, suggest that other mechanisms might be at play. As cardiovascular disease is accompanied by heightened inflammation, and the recent Canakinumab Anti-inflammatory Thrombosis Outcome Study highlighted a causal role for the cytokine interleukin-1β (IL-1β) in incident cardiovascular disease9, a possible proinflammatory role for APOC3 has generated much interest.

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

Fig. 1: Delipidated APOC3, but not lipid-bound APOC3, induces IL-1β release from human and mouse monocytes.
Fig. 2: Endogenous APOC3 does not alter plasma IL-18.

Data availability

All original data are available from the corresponding author upon request. Source data are provided with this paper.

References

  1. Kanter, J. E. et al. Increased apolipoprotein C3 drives cardiovascular risk in type 1 diabetes. J. Clin. Invest. 129, 4165–4179 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  2. Basu, A. et al. Serum apolipoproteins and apolipoprotein-defined lipoprotein subclasses: a hypothesis-generating prospective study of cardiovascular events in T1D. J. Lipid Res. 60, 1432–1439 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Jansson Sigfrids, F. et al. Apolipoprotein C-III predicts cardiovascular events and mortality in individuals with type 1 diabetes and albuminuria. J. Intern. Med. 291, 338–349 (2022).

    Article  CAS  PubMed  Google Scholar 

  4. Scheffer, P. G. et al. Increased plasma apolipoprotein C-III concentration independently predicts cardiovascular mortality: the Hoorn Study. Clin. Chem. 54, 1325–1330 (2008).

    Article  CAS  PubMed  Google Scholar 

  5. TG and HDL Working Group of the Exome Sequencing Project, National Heart, Lung, and Blood Institute, et al. Loss-of-function mutations in APOC3, triglycerides, and coronary disease. N. Engl. J. Med. 371, 22–31 (2014).

  6. Jorgensen, A. B., Frikke-Schmidt, R., Nordestgaard, B. G. & Tybjaerg-Hansen, A. Loss-of-function mutations in APOC3 and risk of ischemic vascular disease. N. Engl. J. Med. 371, 32–41 (2014).

    Article  PubMed  Google Scholar 

  7. Pollin, T. I. et al. A null mutation in human APOC3 confers a favorable plasma lipid profile and apparent cardioprotection. Science 322, 1702–1705 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Boren, J., Packard, C. J. & Taskinen, M. R. The roles of ApoC-III on the metabolism of triglyceride-rich lipoproteins in humans. Front. Endocrinol. 11, 474 (2020).

    Article  Google Scholar 

  9. Ridker, P. M. et al. Antiinflammatory therapy with canakinumab for atherosclerotic disease. N. Engl. J. Med. 377, 1119–1131 (2017).

    Article  CAS  PubMed  Google Scholar 

  10. Zewinger, S. et al. Apolipoprotein C3 induces inflammation and organ damage by alternative inflammasome activation. Nat. Immunol. https://doi.org/10.1038/s41590-019-0548-1 (2020).

  11. Segrest, J. P. et al. The amphipathic helix in the exchangeable apolipoproteins: a review of secondary structure and function. J. Lipid Res. 33, 141–166 (1992).

    Article  CAS  PubMed  Google Scholar 

  12. Davidson, W. S. et al. The biotin-capture lipid affinity assay: a rapid method for determining lipid binding parameters for apolipoproteins. J. Lipid Res. 47, 440–449 (2006).

    Article  CAS  PubMed  Google Scholar 

  13. Netea, M. G. et al. Differential requirement for the activation of the inflammasome for processing and release of IL-1beta in monocytes and macrophages. Blood 113, 2324–2335 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Schlotter, F. et al. ApoC-III is a novel inducer of calcification in human aortic valves. J. Biol. Chem. 296, 100193 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Liu, Z. & Zhang, Z. Mapping cell types across human tissues. Science 376, 695–696 (2022).

    Article  CAS  PubMed  Google Scholar 

  16. Mund, A., Brunner, A. D. & Mann, M. Unbiased spatial proteomics with single-cell resolution in tissues. Mol. Cell. 82, 2335–2349 (2022).

    Article  CAS  PubMed  Google Scholar 

  17. Dinarello, C. A., Novick, D., Kim, S. & Kaplanski, G. Interleukin-18 and IL-18 binding protein. Front. Immunol. 4, 289 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  18. Qamar, A. et al. Plasma apolipoprotein C-III levels, triglycerides, and coronary artery calcification in type 2 diabetics. Arterioscler. Thromb. Vasc. Biol. 35, 1880–1888 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Renard, C. B. et al. Diabetes and diabetes-associated lipid abnormalities have distinct effects on initiation and progression of atherosclerotic lesions. J. Clin. Invest. 114, 659–668 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Prakash, T. P. et al. Comprehensive structure-activity relationship of triantennary N-acetylgalactosamine conjugated antisense oligonucleotides for targeted delivery to hepatocytes. J. Med. Chem. 59, 2718–2733 (2016).

    Article  CAS  PubMed  Google Scholar 

  21. Reaven, G. M., Mondon, C. E., Chen, Y. D. & Breslow, J. L. Hypertriglyceridemic mice transgenic for the human apolipoprotein C-III gene are neither insulin resistant nor hyperinsulinemic. J. Lipid Res. 35, 820–824 (1994).

    Article  CAS  PubMed  Google Scholar 

  22. Vaisar, T. et al. High concentration of medium-sized HDL particles and enrichment in HDL paraoxonase 1 associate with protection from vascular complications in people with long-standing type 1 diabetes. Diabetes Care 43, 178–186 (2020).

    Article  CAS  PubMed  Google Scholar 

  23. Shao, B. et al. Pulmonary surfactant protein B carried by HDL predicts incident CVD in patients with type 1 diabetes. J. Lipid Res. 63, 100196 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. MacLean, B. et al. Skyline: an open source document editor for creating and analyzing targeted proteomics experiments. Bioinformatics 26, 966–968 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Sudlow, C. et al. UK biobank: an open access resource for identifying the causes of a wide range of complex diseases of middle and old age. PLoS Med. 12, e1001779 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  26. Das, S. et al. Next-generation genotype imputation service and methods. Nat. Genet. 48, 1284–1287 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Taliun, D. et al. Sequencing of 53,831 diverse genomes from the NHLBI TOPMed Program. Nature 590, 290–299 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Mbatchou, J. et al. Computationally efficient whole-genome regression for quantitative and binary traits. Nat. Genet. 53, 1097–1103 (2021).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by NIH (grant nos. R35HL150754, P01HL151328 and R01HL161829 to K.E.B.) and the American Heart Association (Predoctoral Fellowship Award no. 828090 to C.-C.H.). UK Biobank analyses were conducted using the UK Biobank Resource (Application no. 34229). We also acknowledge the assistance from the University of Washington Diabetes Research Center (P30DK017047) and Nutrition and Obesity Research Center (P30DK035816) core facilities.

Author information

Authors and Affiliations

  1. Division of Metabolism, Endocrinology and Nutrition, Department of Medicine, University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA, USA

    Cheng-Chieh Hsu, Baohai Shao, Jenny E. Kanter, Yi He, Tomas Vaisar & Karin E. Bornfeldt

  2. Department of Medicine, University of California San Diego School of Medicine, La Jolla, CA, USA

    Joseph L. Witztum

  3. Barbara Davis Center for Diabetes, University of Colorado Denver, Aurora, CO, USA

    Janet Snell-Bergeon

  4. Empirico Inc, San Diego, CA, USA

    Gregory McInnes, Shannon Bruse & Omri Gottesman

  5. Ionis Pharmaceuticals, Carlsbad, CA, USA

    Adam E. Mullick

Authors
  1. Cheng-Chieh Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Baohai Shao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jenny E. Kanter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yi He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tomas Vaisar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Joseph L. Witztum
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Janet Snell-Bergeon
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Gregory McInnes
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shannon Bruse
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Omri Gottesman
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Adam E. Mullick
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Karin E. Bornfeldt
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

C.-C.H. performed the experiments, analyzed data and wrote the paper. K.E.B. designed and directed the study. B.S., Y.H., T.V., J.E.K., O.G., S.B. and G.M. performed a subset of experiments and analyzed corresponding data. J.L.W. provided advice. A.E.M. provided advice and reagents and J.S.-B. provided clinical samples. All authors reviewed the paper and provided final approval for submission.

Corresponding author

Correspondence to Karin E. Bornfeldt.

Ethics declarations

Competing interests

A.E.M. is employed by Ionis Pharmaceuticals. O.G., S.B. and G.M. are employed by Empirico Inc. K.E.B. serves on the scientific advisory board of Esperion Therapeutics, Inc. J.L.W. receives royalties from US patents 9,075,050 B2, 6716410B1, 9,347,959, 11,008,381 B2, 11,008 82 B2 and 11,168,148 B2 on oxidation-specific antibodies and biomarkers related to oxidized lipoproteins held by UCSD. J.L.W. is a cofounder of Oxitope, Inc. and Kleanthi Diagnostics, LLC, and is a consultant for Ionis Pharmaceuticals. The other authors declare no competing interests.

Peer review

Peer review information

Nature Immunology thanks Marit Westerterp and Laurent Yvan-Charvet for their contribution to the peer review of this work.

Additional information

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

Extended data

Extended Data Fig. 1 Characterization of small unilamellar vesicles (SUVs) and APOC3.

a, IL-1β release from mouse bone marrow-derived monocytes incubated in indicated APOC3 concentrations. IL-1β measurements were conducted by ELISA and the data were normalized to total cellular protein determined by a BCA protein assay. Data show means ± SEM (n = 5, 5, 5, 5, 5, 8 replicates/group, respectively). b, Endotoxin levels in the final concentrations of APOC3 used in panel a, measured by the amebocyte lysate assay (Pierce™ Chromogenic Endotoxin Quant Kit). Data show means ± SEM (n = 3 replicates/group). In a and b, statistical analyses were performed by Kruskal-Wallis tests and Dunn’s multiple comparisons tests. c, 1-palmitoyl-2-oleoyl-glycerol-3-phosphocholine (POPC) in PBS was sonicated on ice to allow the formation of small unilamellar vesicles (SUVs). The SUVs were purified on a Superose 6 column before the incorporation of APOC3. SUV-APOC3 was isolated with various centrifugation columns and was sterile filtered. The molar concentrations of APOC3 and SUVs were measured by ELISA, BCA protein assay, and phospholipid assay. d, FPLC profile of synthesized SUVs and fractions 16 and 17 (shown in red) were purified and used for the lipidation of APOC3. e, Ion mobility analysis on differential mobility analyzer (DMA) of SUVs from fractions 16 and 17, demonstrating a homogenized particle population around 27 nm. Panel c created with BioRender.com.

Source data

Extended Data Fig. 2 Distribution and density of serum triglyceride and high sensitivity C-reactive protein (hsCRP) values by APOC3 rs138326449 genotype in the UK Biobank.

Unadjusted serum levels of a, triglycerides (n = 430,833) and b, hsCRP (n = 426,774) among UK Biobank carriers of 0, 1 or 2 alternative (A) alleles of the APOC3 loss-of-function variant rs138326449 (IVS2 + 1G-A). Specifically, 0/0 indicates homozygosity of the reference (G) allele, 0/1 indicates heterozygosity and 1/1 indicates homozygosity of the alternative (A) allele. The data are shown in box-and-whisker plots (the bound of the box extends from the 25th to 75th percentiles and the whisker covers the minima and maxima) with the median marked by a center line, remaining distribution and kernel density estimations of each trait (outlier values ± 5 standard deviations from the mean were removed). Compared with rs138326449-G homozygotes, rs138326449-A heterozygotes demonstrate a 0.62 mmol/L decrease in median triglycerides (p = <2.00 ×10−16 by one-way ANOVA) and a 0.16 mg/L increase in median hsCRP (p = 1.54 × 10−5 by one-way ANOVA).

Source data

Extended Data Table 1 Associations between APOC3 rs138326449 and serum triglycerides, high-sensitivity C-reactive protein, and myocardial infarction were evaluated in UK Biobank participants
Full size table
Extended Data Table 2 Conditioned APOC3 rs138326449 high-sensitivity C-reactive protein (hsCRP) associations in the UK Biobank
Full size table

Supplementary information

Source data

Source Data Fig. 1

Unprocessed statistical source data.

Source Data Fig. 2

Unprocessed statistical source data.

Source Data Extended Data Fig. 1

Unprocessed statistical source data and images.

Source Data Extended Data Fig. 2

Unprocessed statistical source data.

Source Data Extended Data Table 1

Unprocessed statistical source data.

Source Data Extended Data Table 2

Unprocessed statistical source data.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hsu, CC., Shao, B., Kanter, J.E. et al. Apolipoprotein C3 induces inflammasome activation only in its delipidated form. Nat Immunol 24, 408–411 (2023). https://doi.org/10.1038/s41590-023-01423-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41590-023-01423-2

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing