Restoring carboxypeptidase E rescues BDNF maturation and neurogenesis in aged brains

Abstract

Adult neurogenesis declines with age due to the less functional neural stem cells (NSCs) and niches, but the underlying molecular bases for this impaired condition remain unclear. Here we analyzed >55,000 single-cell transcriptomes from two discrete neurogenic niches across the mouse lifespan, and identified new features and populations in NSCs, new markers, and neurogenic regional-specific alternations during aging. Intercellular communication analysis revealed defects in brain-derived neurotrophic factor (BDNF)-TrkB signaling cascade in old NSCs. Carboxypeptidase E (CPE) was found to be highly enriched in NSCs, and played a crucial role in mature/proBDNF balance and adult neurogenesis. Diminishment of CPE with aging resulted in impaired generation of BDNF, thus limiting the neurogenesis in old neurogenic niches. Restoring CPE expression markedly rescued the adult neurogenesis by increasing the production of mature BDNF, offering an attractive therapeutic strategy for the treatment of certain disorders in regions associated with constitutive neurogenesis.

Keywords

carboxypeptidase E / adult neurogenesis / aging / BDNF

References

[1] Doetsch F, Caille I, Lim DA, et al. Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell 1999;97:703–16.

[2] Eriksson PS, Perfilieva E, Bjork-Eriksson T, et al. Neurogenesis in the adult human hippocampus. Nat Med 1998;4:1313–7.

[3] Jiang D, Li T, Guo C, et al. Small molecule modulators of chromatin remodeling: from neurodevelopment to neurodegeneration. Cell Biosci 2023;13:10.

[4] Sorrells SF, Paredes MF, Cebrian-Silla A, et al. Human hippocampal neurogenesis drops sharply in children to undetectable levels in adults. Nature 2018;555:377–81.

[5] Boldrini M, Fulmore CA, Tartt AN, et al. Human hippocampal neurogenesis persists throughout aging. Cell Stem Cell 2018;22:589–599.e5.

[6] Spalding KL, Bergmann O, Alkass K, et al. Dynamics of hippocampal neurogenesis in adult humans. Cell 2013;153:1219–27.

[7] Kempermann G, Gage FH, Aigner L, et al. Human adult neurogenesis: evidence and remaining questions. Cell Stem Cell 2018;23:25–30.

[8] Ernst A, Alkass K, Bernard S, et al. Neurogenesis in the striatum of the adult human brain. Cell 2014;156:1072–83.

[9] Tartt AN, Fulmore CA, Liu Y, et al. Considerations for assessing the extent of hippocampal neurogenesis in the adult and aging human brain. Cell Stem Cell 2018;23:782–3.

[10] Encinas JM, Michurina TV, Peunova N, et al. Division-coupled astrocytic differentiation and age-related depletion of neural stem cells in the adult hippocampus. Cell Stem Cell 2011;8:566–79.

[11] Kuhn HG, Dickinson-Anson H, Gage FH. Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci 1996;16:2027–33.

[12] Zhang H, Li J, Ren J, et al. Single-nucleus transcriptomic landscape of primate hippocampal aging. Protein Cell 2021;12:695–716.

[13] Leeman DS, Hebestreit K, Ruetz T, et al. Lysosome activation clears aggregates and enhances quiescent neural stem cell activation during aging. Science 2018;359:1277–83.

[14] Silva-Vargas V, Maldonado-Soto AR, Mizrak D, et al. Age-dependent niche signals from the choroid plexus regulate adult neural stem cells. Cell Stem Cell 2016;19:643–52.

[15] Harris L, Rigo P, Stiehl T, et al. Coordinated changes in cellular behavior ensure the lifelong maintenance of the hippocampal stem cell population. Cell Stem Cell 2021;28:863–876.e6.

[16] Artegiani B, Lyubimova A, Muraro M, et al. A single-cell RNA sequencing study reveals cellular and molecular dynamics of the hippocampal neurogenic niche. Cell Rep 2017;21:3271–84.

[17] Basak O, Krieger TG, Muraro MJ, et al. Troy+ brain stem cells cycle through quiescence and regulate their number by sensing niche occupancy. Proc Natl Acad Sci U S A 2018;115:E610–9.

[18] Dulken BW, Leeman DS, Boutet SC, et al. Single-cell transcriptomic analysis defines heterogeneity and transcriptional dynamics in the adult neural stem cell lineage. Cell Rep 2017;18:777–90.

[19] Hochgerner H, Zeisel A, Lonnerberg P, et al. Conserved properties of dentate gyrus neurogenesis across postnatal development revealed by single-cell RNA sequencing. Nat Neurosci 2018;21:290–9.

[20] Llorens-Bobadilla E, Zhao S, Baser A, et al. Single-cell transcriptomics reveals a population of dormant neural stem cells that become activated upon brain injury. Cell Stem Cell 2015;17:329–40.

[21] Luo Y, Coskun V, Liang A, et al. Single-cell transcriptome analyses reveal signals to activate dormant neural stem cells. Cell 2015;161:1175–86.

[22] Mizrak D, Levitin HM, Delgado AC, et al. Single-cell analysis of regional differences in adult V-SVZ neural stem cell lineages. Cell Rep 2019;26:394–406.e5.

[23] Shin J, Berg DA, Zhu Y, et al. Single-cell RNA-Seq with waterfall reveals molecular cascades underlying adult neurogenesis. Cell Stem Cell 2015;17:360–72.

[24] Zywitza V, Misios A, Bunatyan L, et al. Single-cell transcriptomics characterizes cell types in the subventricular zone and uncovers molecular defects impairing adult neurogenesis. Cell Rep 2018;25:2457–2469.e8.

[25] Shi Z, Geng Y, Liu J, et al. Single-cell transcriptomics reveals gene signatures and alterations associated with aging in distinct neural stem/progenitor cell subpopulations. Protein Cell 2018;9:351–64.

[26] Kalamakis G, Brune D, Ravichandran S, et al. Quiescence modulates stem cell maintenance and regenerative capacity in the aging brain. Cell 2019;176:1407–1419.e14.

[27] Dulken BW, Buckley MT, Navarro Negredo P, et al. Single-cell analysis reveals T cell infiltration in old neurogenic niches. Nature 2019;571:205–10.

[28] Buckley MT SE, George BM, Liu L, et al. Cell-type-specific aging clocks to quantify aging and rejuvenation in neurogenic regions of the brain. Nature Aging 2023;3:121–37.

[29] Lazarov O, Mattson MP, Peterson DA, et al. When neurogenesis encounters aging and disease. Trends Neurosci 2010;33:569–79.

[30] Yang KY, Zhao S, Feng H, et al. Ca(2+) homeostasis maintained by TMCO1 underlies corpus callosum development via ERK signaling. Cell Death Dis 2022;13:674.

[31] Ma X, Liu H, Li J, et al. Poleta O-GlcNAcylation governs genome integrity during translesion DNA synthesis. Nat Commun 2017;8:1941.

[32] Xie W, Wang JQ, Wang QC, et al. Adult neural progenitor cells from Huntington’s disease mouse brain exhibit increased proliferation and migration due to enhanced calcium and ROS signals. Cell Prolif 2015;48:517–31.

[33] Xiao L, Yang X, Sharma VK, et al. Cloning, gene regulation, and neuronal proliferation functions of novel N-terminal-truncated carboxypeptidase E/neurotrophic factor-alphal variants in embryonic mouse brain. FASEB J 2019;33:808–20.

[34] Pang PT, Teng HK, Zaitsev E, et al. Cleavage of proBDNF by tPA/plasmin is essential for long-term hippocampal plasticity. Science 2004;306:487–91.

[35] Wang Y, Song F, Zhu J, et al. GSA: Genome Sequence Archive. Genomics Proteomics Bioinformatics. 2017;15(1):14–18.

[36] Shi Y, Chichung Lie D, Taupin P, et al. Expression and function of orphan nuclear receptor TLX in adult neural stem cells. Nature 2004;427:78–83.

[37] Navarro Negredo P, Yeo RW, Brunet A. Aging and rejuvenation of neural stem cells and their niches. Cell Stem Cell 2020;27:202–23.

[38] Marques-Torrejon MA, Williams CAC, Southgate B, et al. LRIG1 is a gatekeeper to exit from quiescence in adult neural stem cells. Nat Commun 2021;12:2594.

[39] Fricker LD. Carboxypeptidase E. In: Neil D. Rawlings GS, ed. Handbook of Proteolytic Enzymes. Amsterdam, USA: American Academic Press, Elsevier Ltd., 2013:1342–1345.

[40] Xiao L, Yang X, Loh YP. Neurotrophic, gene regulation, and cognitive functions of carboxypeptidase e-neurotrophic factor-alpha1 and its variants. Front Neurosci 2019;13:243.

[41] Cheng Y, Rodriguiz RM, Murthy SR, et al. Neurotrophic factor-alpha1 prevents stress-induced depression through enhancement of neurogenesis and is activated by rosiglitazone. Mol Psychiatry 2015;20:744–54.

[42] Cool DR, Normant E, Shen F, et al. Carboxypeptidase E is a regulated secretory pathway sorting receptor: genetic obliteration leads to endocrine disorders in Cpe(fat) mice. Cell 1997;88:73–83.

[43] Cawley NX, Wetsel WC, Murthy SR, et al. New roles of carboxypeptidase E in endocrine and neural function and cancer. Endocr Rev 2012;33:216–53.

[44] Park JJ, Cawley NX, Loh YP. A bi-directional carboxypeptidase E-driven transport mechanism controls BDNF vesicle homeostasis in hippocampal neurons. Mol Cell Neurosci 2008;39:63–73.

[45] Lou H, Kim SK, Zaitsev E, et al. Sorting and activity-dependent secretion of BDNF require interaction of a specific motif with the sorting receptor carboxypeptidase e. Neuron 2005;45:245–55.

[46] Lee C, Hu J, Ralls S, et al. The molecular profiles of neural stem cell niche in the adult subventricular zone. PLoS One 2012;7:e50501.

[47] Zheng M, Streck RD, Scott RE, et al. The developmental expression in rat of proteases furin, PC1, PC2, and carboxypeptidase E: implications for early maturation of proteolytic processing capacity. J Neurosci 1994;14:4656–73.

[48] Selvaraj P, Xiao L, Lee C, et al. Neurotrophic factor-alpha1: a key Wnt-beta-catenin dependent anti-proliferation factor and ERK-Sox9 activated inducer of embryonic neural stem cell differentiation to astrocytes in neurodevelopment. Stem Cells 2017;35:557–71.

[49] Sharma VK, Yang X, Kim SK, et al. Novel interaction between neurotrophic factor-alpha1/carboxypeptidase E and serotonin receptor, 5-HTR1E, protects human neurons against oxidative/neuroexcitotoxic stress via beta-arrestin/ERK signaling. Cell Mol Life Sci Dec 29 2021;79:24.

[50] Armoza-Eilat S, Caspi M, Shomron O, et al. Carboxypeptidase E is efficiently secreted and internalized via lysosomes. bioRxiv 2022.

[51] Mowla SJ, Pareek S, Farhadi HF, et al. Differential sorting of nerve growth factor and brain-derived neurotrophic factor in hippocampal neurons. J Neurosci 1999;19:2069–80.

[52] Balkowiec A, Katz DM. Cellular mechanisms regulating activity-dependent release of native brain-derived neurotrophic factor from hippocampal neurons. J Neurosci 2002;22:10399–407.

[53] Lu B, Pang PT, Woo NH. The yin and yang of neurotrophin action. Nat Rev Neurosci 2005;6:603–14.

[54] Nagappan G, Zaitsev E, Senatorov VV Jr, et al. Control of extracellular cleavage of ProBDNF by high frequency neuronal activity. Proc Natl Acad Sci U S A 2009;106:1267–72.

[55] Mizoguchi H, Nakade J, Tachibana M, et al. Matrix metalloproteinase- 9 contributes to kindled seizure development in pentylenetetrazole-treated mice by converting pro-BDNF to mature BDNF in the hippocampus. J Neurosci 2011;31:12963–71.

[56] Chougule A, Kolli V, Baroi S, et al. Nonenzymatic and trophic activities of carboxypeptidase e regulate bone mass and bioenergetics of skeletal stem cells in mice. JBMR Plus 2020;4:e10392.

[57] Xiao L, Sharma VK, Toulabi L, et al. Neurotrophic factor-alpha1, a novel tropin is critical for the prevention of stress-induced hippocampal CA3 cell death and cognitive dysfunction in mice: comparison to BDNF. Transl Psychiatry 2021;11:24.

[58] Gupta VK, You Y, Gupta VB, et al. TrkB receptor signalling: implications in neurodegenerative, psychiatric and proliferative disorders. Int J Mol Sci 2013;14:10122–42.

[59] Tapia-Arancibia L, Aliaga E, Silhol M, et al. New insights into brain BDNF function in normal aging and Alzheimer disease. Brain Res Rev 2008;59:201–20.

RIGHTS & PERMISSIONS

The Author(s) 2023. Published by Oxford University Press on behalf of Higher Education Press.