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Disruption of the beclin 1–BCL2 autophagy regulatory complex promotes longevity in mice

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An Author Correction to this article was published on 19 June 2018

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Abstract

Autophagy increases the lifespan of model organisms; however, its role in promoting mammalian longevity is less well-established1,2. Here we report lifespan and healthspan extension in a mouse model with increased basal autophagy. To determine the effects of constitutively increased autophagy on mammalian health, we generated targeted mutant mice with a Phe121Ala mutation in beclin 1 (Becn1F121A/F121A) that decreases its interaction with the negative regulator BCL2. We demonstrate that the interaction between beclin 1 and BCL2 is disrupted in several tissues in Becn1F121A/F121A knock-in mice in association with higher levels of basal autophagic flux. Compared to wild-type littermates, the lifespan of both male and female knock-in mice is significantly increased. The healthspan of the knock-in mice also improves, as phenotypes such as age-related renal and cardiac pathological changes and spontaneous tumorigenesis are diminished. Moreover, mice deficient in the anti-ageing protein klotho3 have increased beclin 1 and BCL2 interaction and decreased autophagy. These phenotypes, along with premature lethality and infertility, are rescued by the beclin 1(F121A) mutation. Together, our data demonstrate that disruption of the beclin 1–BCL2 complex is an effective mechanism to increase autophagy, prevent premature ageing, improve healthspan and promote longevity in mammals.

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Fig. 1: Effects of beclin 1(F121A) mutation on the beclin 1–BCL2 interaction and basal autophagy.
Fig. 2: Beclin 1(F121A) knock-in mutation extends lifespan in mice.
Fig. 3: Beclin 1(F121A) knock-in mutation improves healthspan in mice.
Fig. 4: Expression of beclin 1(F121A) prevents lethality of klotho-deficient mice.

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Change history

  • 19 June 2018

    In this Letter, the graphs in Fig. 2a and c were inadvertently the same owing to a copy and paste error from the original graphs in Prism. The Source Data files containing the raw data were correct. Fig. 2c has been corrected online.

References

  1. Madeo, F., Zimmermann, A., Maiuri, M. C. & Kroemer, G. Essential role for autophagy in life span extension. J. Clin. Invest. 125, 85–93 (2015).

    Article  PubMed Central  PubMed  Google Scholar 

  2. Rubinsztein, D. C., Mariño, G. & Kroemer, G. Autophagy and aging. Cell 146, 682–695 (2011).

    Article  CAS  Google Scholar 

  3. Kuro-o, M. et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature 390, 45–51 (1997).

    Article  ADS  CAS  PubMed  Google Scholar 

  4. Levine, B. & Kroemer, G. Autophagy in the pathogenesis of disease. Cell 132, 27–42 (2008).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Meléndez, A. et al. Autophagy genes are essential for dauer development and life-span extension in C. elegans. Science 301, 1387–1391 (2003).

    Article  ADS  CAS  Google Scholar 

  6. Yamamoto, T. et al. Time-dependent dysregulation of autophagy: Implications in aging and mitochondrial homeostasis in the kidney proximal tubule. Autophagy 12, 801–813 (2016).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Eisenberg, T. et al. Cardioprotection and lifespan extension by the natural polyamine spermidine. Nat. Med. 22, 1428–1438 (2016).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Eisenberg, T. et al. Induction of autophagy by spermidine promotes longevity. Nat. Cell Biol. 11, 1305–1314 (2009).

    Article  CAS  Google Scholar 

  9. Mercken, E. M. et al. SIRT1 but not its increased expression is essential for lifespan extension in caloric-restricted mice. Aging Cell 13, 193–196 (2014).

    Article  CAS  Google Scholar 

  10. Pyo, J. O. et al. Overexpression of Atg5 in mice activates autophagy and extends lifespan. Nat. Commun. 4, 2300 (2013).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Kimmey, J. M. et al. Unique role for ATG5 in neutrophil-mediated immunopathology during M. tuberculosis infection. Nature 528, 565–569 (2015).

    Article  ADS  PubMed Central  CAS  PubMed  Google Scholar 

  12. Liang, X. H. et al. Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402, 672–676 (1999).

    Article  ADS  CAS  Google Scholar 

  13. Kihara, A., Kabeya, Y., Ohsumi, Y. & Yoshimori, T. Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network. EMBO Rep. 2, 330–335 (2001).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Levine, B., Liu, R., Dong, X. & Zhong, Q. Beclin orthologs: integrative hubs of cell signaling, membrane trafficking, and physiology. Trends Cell Biol. 25, 533–544 (2015).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Rocchi, A. et al. A Becn1 mutation mediates hyperactive autophagic sequestration of amyloid oligomers and improved cognition in Alzheimer’s disease. PLoS Genet. 13, e1006962 (2017).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Pattingre, S. et al. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 122, 927–939 (2005).

    Article  CAS  Google Scholar 

  17. Sinha, S., Colbert, C. L., Becker, N., Wei, Y. & Levine, B. Molecular basis of the regulation of Beclin 1-dependent autophagy by the γ-herpesvirus 68 Bcl-2 homolog M11. Autophagy 4, 989–997 (2008).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Mizushima, N., Yamamoto, A., Matsui, M., Yoshimori, T. & Ohsumi, Y. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol. Biol. Cell 15, 1101–1111 (2004).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Mizushima, N., Yoshimori, T. & Levine, B. Methods in mammalian autophagy research. Cell 140, 313–326 (2010).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. McKnight, N. C. et al. Beclin 1 is required for neuron viability and regulates endosome pathways via the UVRAG-VPS34 complex. PLoS Genet. 10, e1004626 (2014).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Lim, J. H. et al. Age-associated molecular changes in the kidney in aged mice. Oxid. Med. Cell. Longev. 2012, 171383 (2012).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Lapierre, L. R., Kumsta, C., Sandri, M., Ballabio, A. & Hansen, M. Transcriptional and epigenetic regulation of autophagy in aging. Autophagy 11, 867–880 (2015).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Brayton, C. in The Mouse in Biomedical Research Vol. 2 (eds Fox, J. et al.), 623–717 (Elsevier, Amsterdam, 2007).

  24. Kuro-o, M. Klotho and aging. Biochim. Biophys. Acta 1790, 1049–1058 (2009).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Tsujikawa, H., Kurotaki, Y., Fujimori, T., Fukuda, K. & Nabeshima, Y. Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system. Mol. Endocrinol. 17, 2393–2403 (2003).

    Article  CAS  Google Scholar 

  26. Kurosu, H. et al. Suppression of aging in mice by the hormone Klotho. Science 309, 1829–1833 (2005).

    Article  ADS  PubMed Central  CAS  PubMed  Google Scholar 

  27. Chen, T. H. et al. The secreted Klotho protein restores phosphate retention and suppresses accelerated aging in Klotho mutant mice. Eur. J. Pharmacol. 698, 67–73 (2013).

    Article  CAS  Google Scholar 

  28. Shi, M. et al. αKlotho mitigates progression of AKI to CKD through activation of autophagy. J. Am. Soc. Nephrol. 27, 2331–2345 (2016).

    Article  CAS  Google Scholar 

  29. Su, T. et al. Deletion of histidine triad nucleotide-binding protein 1/PKC-interacting protein in mice enhances cell growth and carcinogenesis. Proc. Natl Acad. Sci. USA 100, 7824–7829 (2003).

    Article  ADS  PubMed Central  CAS  PubMed  Google Scholar 

  30. Sebti, S. et al. BAT3 modulates p300-dependent acetylation of p53 and autophagy-related protein 7 (ATG7) during autophagy. Proc. Natl Acad. Sci. USA 111, 4115–4120 (2014).

    Article  ADS  PubMed Central  CAS  PubMed  Google Scholar 

  31. Fielding, A. B., Willox, A. K., Okeke, E. & Royle, S. J. Clathrin-mediated endocytosis is inhibited during mitosis. Proc. Natl Acad. Sci. USA 109, 6572–6577 (2012).

    Article  ADS  PubMed Central  PubMed  Google Scholar 

  32. Tacheva-Grigorova, S. K., Santos, A. J., Boucrot, E. & Kirchhausen, T. Clathrin-mediated endocytosis persists during unperturbed mitosis. Cell Reports 4, 659–668 (2013).

    Article  CAS  Google Scholar 

  33. Wang, C., Li, Q., Redden, D. T., Weindruch, R. & Allison, D. B. Statistical methods for testing effects on “maximum lifespan”. Mech. Ageing Dev. 125, 629–632 (2004).

    Article  Google Scholar 

  34. Motulsky, H. J. & Brown, R. E. Detecting outliers when fitting data with nonlinear regression — a new method based on robust nonlinear regression and the false discovery rate. BMC Bioinformatics 7, 123 (2006).

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Sciarretta for discussions; N. Mizushima for reagents; L. Nguyen for technical assistance and H. Smith for assistance with manuscript preparation. This work was supported by NIH grants RO1-CA109618 (B.L.), U19-AI199725 (B.L.), RO1-DK091392 and RO1-DK092461 (M.C.H. and O.W.M.), P30-DK07938 (O.W.M.), K99R00-DK094980 (C.H.), Cancer Prevention Research Institute of Texas grant RP120718 (B.L.) and a Fondation Leducq grant 15CBD04 (B.L., A.F.F. and S.S.).

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Nature thanks D. Harrison and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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Authors and Affiliations

Authors

Contributions

A.F.F., S.S., Y.W., C.H., T.T., Y.L., M.C.H. and B.L. designed the study. A.F.F., S.S., Y.W., Z.Z., M.S., K.L.M. and W.-C.C. performed biochemical analyses. A.F.F., S.S. and Z.Z. performed autophagy microscopic analyses. A.F.F. and S.S. performed renal and cardiac histopathological analyses. G.B. characterized malignancies. A.F.F., S.S., D.K.M., G.G.S, G.B., O.W.M., M.C.H. and B.L. discussed and analysed data. A.F.F., S.S., M.C.H. and B.L. wrote the manuscript. A.F.F. and S.S. contributed equally and the order of these authors was determined arbitrarily.

Corresponding authors

Correspondence to Ming Chang Hu or Beth Levine.

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B.L. is a Scientific Founder of Casma Therapeutics, Inc.

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Extended data figures and tables

Extended Data Fig. 1 Increased basal autophagy in tissues of beclin 1(F121A) knock-in mice.

a, Representative images and quantification of GFP–LC3 puncta (autophagosomes) in glomeruli from Becn1+/+;GFP-LC3 (wild-type, WT) and Becn1F121A/F121A;GFP-LC3 (knock-in, KI) mice with or without 50 mg kg−1 chloroquine for 6 h. Scale bars, 10 μm. Data are mean ± s.e.m. for three mice per genotype. b, Enlarged versions of the images shown in Fig. 1c. P values determined by one-sided unpaired t-test. Arrows denote representative GFP–LC3 puncta.

Source Data

Extended Data Fig. 2 Sustained increase in basal autophagy during adulthood in beclin 1(F121A) knock-in mice.

a, Co-immunoprecipitation of beclin 1 and BCL2 in representative samples of hearts and kidneys from eight-month-old Becn1+/+ (wild-type) and Becn1F121A/F121A (knock-in) animals. b, Quantification of beclin 1 co-immunoprecipitated with BCL2 in indicated tissues of eight-month-old wild-type and knock-in mice (n = 3 mice per genotype). c, Quantification of GFP–LC3 puncta in hearts and tissues from six-month-old Becn1+/+;GFP-LC3 (wild-type) and Becn1F121A/F121A;GFP-LC3 (knock-in) mice with or without chloroquine (50 mg kg−1, 6 h). d, Western blot analysis of autophagy markers in the hearts and kidneys from eight-month-old wild-type and knock-in mice. Each lane represents a different mouse. e, Quantification of p62 and total LC3 levels (normalized to β-actin), as well as LC3-II/LC3-I ratios from samples in d. Data are mean ± s.e.m. for three mice per genotype. P values were determined by one-sided unpaired t-test. For uncropped gels, see Supplementary Fig. 1.

Source Data

Extended Data Fig. 3 Increased autophagy, but not endocytosis, in beclin 1(F121A) MEFs.

a, Co-immunoprecipitation of beclin 1 and BCL2 in MEFs derived from Becn1+/+ (wild-type) and Becn1F121A/F121A (knock-in) animals. b, Representative images (top) and quantification (bottom) of GFP–LC3 puncta in wild-type and knock-in cells with or without 10 nM bafilomycin A1 (BafA1) for 3 h. Scale bars, 10 µm. c, Western blot analysis of autophagy markers in wild-type and knock-in MEFs with or without BafA1 (100 nM, 2 h). d, Representative images and quantitative electron microscopic analysis of autophagic structures in wild-type and knock-in MEFs with or without BafA1 (100 nM, 3 h). Insets show representative autophagosome (arrowhead) and autolysosome (arrow). Scale bars, 1 µm. e, Representative images and quantification of transferrin uptake kinetics in wild-type and knock-in cells. Scale bars, 20 µm. Results shown are representative of two and four independent experiments respectively for a and c. Data are mean ± s.e.m. for three replicates in b and for 50 cells per genotype and condition in d. Data points on line graph in e denote mean ± s.e.m. for cells at 7 min (n = 63, WT; n = 54, KI), 15 min (n = 58, WT; n = 52 KI), and 30 min (n = 76, WT; n = 83, KI). P values determined by unpaired one-sided (b) and two-sided (d, e) t-test. For uncropped gels, see Supplementary Fig. 1.

Source Data

Extended Data Fig. 4 Apoptosis and autophagy analyses in kidneys and hearts of aged mice.

a, b, Representative images and quantification of active caspase 3-positive cells in kidneys (a) and hearts (b) from Becn1+/+ (wild-type) and Becn1F121A/F121A (knock-in) animals. Two month-old wild-type and knock-in mouse kidneys and hearts (n = 6 per genotype) were analysed. For kidney analyses, aged (20-month-old) wild-type (n = 20) and knock-in (n = 26) mice were used. For heart analyses, aged (20-month-old) wild-type (n = 19) and knock-in (n = 26) mice were used. Scatter plot bars represent median ± interquartile ranges. P values were determined by two-sided Mann–Whitney test. c, d, Enlarged versions of the endogenous LC3 puncta (autophagosome) images shown in Fig. 3d (c) and Fig. 3h (d). Arrows denote representative LC3 puncta.

Source Data

Extended Data Fig. 5 In vitro klotho treatment disrupts the beclin 1–BCL2 interaction.

Co-immunoprecipitation of beclin 1 and BCL2 in HeLa cells treated with PBS or the indicated concentrations of recombinant full-length mouse klotho protein for 24 h. Result shown is representative of two independent experiments. For uncropped gels, see Supplementary Fig. 1.

Extended Data Table 1 Increased median lifespan and maximal lifespan in beclin 1(F121A) knock-in mice

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Fernández, Á.F., Sebti, S., Wei, Y. et al. Disruption of the beclin 1–BCL2 autophagy regulatory complex promotes longevity in mice. Nature 558, 136–140 (2018). https://doi.org/10.1038/s41586-018-0162-7

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