Senktide blocks aberrant RTN3 interactome to retard memory decline and tau pathology in social isolated Alzheimer’s disease mice

Abstract

Sporadic or late-onset Alzheimer’s disease (LOAD) accounts for more than 95% of Alzheimer’s disease (AD) cases without any family history. Although genome-wide association studies have identified associated risk genes and loci for LOAD, numerous studies suggest that many adverse environmental factors, such as social isolation, are associated with an increased risk of dementia. However, the underlying mechanisms of social isolation in AD progression remain elusive. In the current study, we found that 7 days of social isolation could trigger pattern separation impairments and presynaptic abnormalities of the mossy fibre-CA3 circuit in AD mice. We also revealed that social isolation disrupted histone acetylation and resulted in the downregulation of 2 dentate gyrus (DG)-enriched miRNAs, which simultaneously target reticulon 3 (RTN3), an endoplasmic reticulum protein that aggregates in presynaptic regions to disturb the formation of functional mossy fibre boutons (MFBs) by recruiting multiple mitochondrial and vesicle-related proteins. Interestingly, the aggregation of RTN3 also recruits the PP2A B subunits to suppress PP2A activity and induce tau hyperphosphorylation, which, in turn, further elevates RTN3 and forms a vicious cycle. Finally, using an artificial intelligence-assisted molecular docking approach, we determined that senktide, a selective agonist of neurokinin3 receptors (NK3R), could reduce the binding of RTN3 with its partners. Moreover, application of senktide in vivo effectively restored DG circuit disorders in socially isolated AD mice. Taken together, our findings not only demonstrate the epigenetic regulatory mechanism underlying mossy fibre synaptic disorders orchestrated by social isolation and tau pathology but also reveal a novel potential therapeutic strategy for AD.

Key words： Alzheimer’s disease; memory impairment; synaptic disorder; tau pathology

References

1 Andel R, Crowe M, Hahn EA et al. Work-related stress may increase the risk of vascular dementia. J Am Geriatr Soc 2012;60:60–7.

2 Armitage R, Nellums LB. COVID-19 and the consequences of isolating the elderly. Lancet Public Health 2020;5:e256.

3 Azevedo L, Calandri IL, Slachevsky A et al. Impact of social isolation on people with dementia and their family caregivers. J Alzheimers Dis 2021;81:607–17.

4 Bao WD, Zhou XT, Zhou LT et al. Targeting miR-124/ Ferroportin signaling ameliorated neuronal cell death through inhibiting apoptosis and ferroptosis in aged intracerebral hemorrhage murine model. Aging Cell 2020;19:e13235.

5 Barnes DE, Yaffe K. The projected effect of risk factor reduction on Alzheimer’s disease prevalence. Lancet Neurol 2011;10:819–28.

6 Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004;116:281–97.

7 Beermann J, Piccoli MT, Viereck J et al. Non-coding RNAs in development and disease: background, mechanisms, and therapeutic approaches. Physiol Rev 2016;96:1297–325.

8 Berron D, Schutze H, Maass A et al. Strong evidence for pattern separation in human dentate gyrus. J Neurosci 2016;36:7569–79.

9 Bot AM, Debski KJ, Lukasiuk K. Alterations in miRNA levels in the dentate gyrus in epileptic rats. PLoS One 2013;8:e76051.

10 Buzsaki G. Theta oscillations in the hippocampus. Neuron 2002;33:325–40.

11 Deng M, Zhang Q, Wu Z et al. Mossy cell synaptic dysfunction causes memory imprecision via miR-128 inhibition of STIM2 in Alzheimer’s disease mouse model. Aging Cell 2020;19:e13144.

12 De Souza Silva MA, Lenz B, Rotter A et al. Neurokinin3 receptor as a target to predict and improve learning and memory in the aged organism. Proc Natl Acad Sci U S A 2013;110:15097–102.

13 De-Yi Liu H-ZH, Li KE, Lu Y et al. EPAC2 knockout causes abnormal tau pathology through calpain-mediated CDK5 activation. Adv Neurol 2022;1:8.

14 Fakoya OA, Mccorry NK, Donnelly M. Loneliness and social isolation interventions for older adults: a scoping review of reviews. BMC Public Health 2020;20:129.

15 Ferrero H, Larrayoz IM, Martisova E et al. Increased levels of brain adrenomedullin in the neuropathology of Alzheimer’s disease. Mol Neurobiol 2018;55:5177–83.

16 Fone KC, Porkess MV. Behavioural and neurochemical effects of post-weaning social isolation in rodents-relevance to developmental neuropsychiatric disorders. Neurosci Biobehav Rev 2008;32:1087–102.

17 Fredes F, Silva MA, Koppensteiner P et al. Ventro-dorsal hippocampal pathway gates novelty-induced contextual memory formation. Curr Biol 2021;31:25–38 e5.

18 Ge QD, Tan Y, Luo Y et al. MiR-132, miR-204 and BDNF-TrkB signaling pathway may be involved in spatial learning and memory impairment of the offspring rats caused by fluorine and aluminum exposure during the embryonic stage and into adulthood. Environ Toxicol Pharmacol 2018;63:60–8.

19 Grumati P, Morozzi G, Holper S et al. Full length RTN3 regulates turnover of tubular endoplasmic reticulum via selective autophagy. Elife 2017;6:e25555.

20 Hainmueller T, Bartos M. Dentate gyrus circuits for encoding, retrieval and discrimination of episodic memories. Nat Rev Neurosci 2020;21:153–68.

21 Hatada Y, Wu F, Sun ZY et al. Presynaptic morphological changes associated with long-term synaptic facilitation are triggered by actin polymerization at preexisting varicositis. J Neurosci 2000;20:RC82.

22 He W, Lu Y, Qahwash I et al. Reticulon family members modulate BACE1 activity and amyloid-beta peptide generation. Nat Med 2004;10:959–65.

23 Hersi M, Irvine B, Gupta P et al. Risk factors associated with the onset and progression of Alzheimer’s disease: a systematic review of the evidence. Neurotoxicology 2017;61:143–87.

24 Hu X, Shi Q, Zhou X et al. Transgenic mice overexpressing reticulon 3 develop neuritic abnormalities. EMBO J 2007;26:2755–67.

25 Hu J, Huang HZ, Wang X et al. Activation of glycogen synthase kinase-3 Mediates the olfactory deficit-induced hippocampal impairments. Mol Neurobiol 2015a;52:1601–17.

26 Hu S, Wang H, Chen K et al. MicroRNA-34c downregulation ameliorates amyloid-beta-induced synaptic failure and memory deficits by targeting VAMP2. J Alzheimers Dis 2015b;48:673–86.

27 Huang H, Wang L, Cao M et al. Isolation housing exacerbates Alzheimer’s disease-like pathophysiology in aged APP/PS1 mice. Int J Neuropsychopharmacol 2015;18:pyu116.

28 Huang HZ, Wang X, Liu D. Cognition damage due to disruption of cyclic adenosine monophosphate-related signaling pathway in melatonin receptor 2 knockout mice. Advanced Neurology 2023;2:0974.

29 Hwang TJ, Rabheru K, Peisah C et al. Loneliness and social isolation during the COVID-19 pandemic. Int Psychogeriatr 2020;32:1217–20.

30 Ibi D, Takuma K, Koike H et al. Social isolation rearing- induced impairment of the hippocampal neurogenesis is associated with deficits in spatial memory and emotion-related behaviors in juvenile mice. J Neurochem 2008;105:921–32.

31 Irwin JJ, Tang KG, Young J et al. ZINC20-A free ultralarge-scale chemical database for ligand discovery. J Chem Inf Model 2020;60:6065–73.

32 Jiang N, Cupolillo D, Grosjean N et al. Impaired plasticity of intrinsic excitability in the dentate gyrus alters spike transfer in a mouse model of Alzheimer’s disease. Neurobiol Dis 2021;154:105345.

33 Jo S, Yarishkin O, Hwang YJ et al. GABA from reactive astrocytes impairs memory in mouse models of Alzheimer’s disease. Nat Med 2014;20:886–96.

34 Joyce J, Ryan J, Owen A et al. Social isolation, social support, and loneliness and their relationship with cognitive health and dementia. Int J Geriatr Psychiatry 2021;37:10.

35 Kaya-Okur HS, Wu SJ, Codomo CA et al. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun 2019;10:1930.

36 Keegan KD, Woodruff GN, Pinnock RD. The selective NK3 receptor agonist senktide excites a subpopulation of dopamine-sensitive neurones in the rat substantia nigra pars compacta in vitro. Br J Pharmacol 1992;105:3–5.

37 Kheirbek MA, Drew LJ, Burghardt NS et al. Differential control of learning and anxiety along the dorsoventral axis of the dentate gyrus. Neuron 2013;77:955–68.

38 Koca RO, Gormus ZIS, Solak H et al. Neurokinin 3 Receptor Effects on Cognitive Behaviour in a Rat Model of Alzheimer’s Disease. PREPRINT (Version 1). 2021. available at Research Square [doi.org/10.21203/rs.3.rs-517911/ v1].

39 Kohen R, Dobra A, Tracy JH et al. Transcriptome profiling of human hippocampus dentate gyrus granule cells in mental illness. Transl Psychiatry 2014;4:e366.

40 Kuiper JS, Zuidersma M, Oude Voshaar RC et al. Social relationships and risk of dementia: a systematic review and meta-analysis of longitudinal cohort studies. Ageing Res Rev 2015;22:39–57.

41 Kumamaru E, Kuo CH, Fujimoto T et al. Reticulon3 expression in rat optic and olfactory systems. Neurosci Lett 2004;356:17–20.

42 Kumar S, Reddy PH. The role of synaptic microRNAs in Alzheimer’s disease. Biochim Biophys Acta Mol Basis Dis 2020;1866:165937.

43 Lau P, Bossers K, Janky R et al. Alteration of the microRNA network during the progression of Alzheimer’s disease. EMBO Mol Med 2013;5:1613–34.

44 Lee SH, Kim KR, Ryu SY et al. Impaired short-term plasticity in mossy fiber synapses caused by mitochondrial dysfunction of dentate granule cells is the earliest synaptic deficit in a mouse model of Alzheimer’s disease. J Neurosci 2012;32:5953–63.

45 Leutgeb JK, Leutgeb S, Moser MB et al. Pattern separation in the dentate gyrus and CA3 of the hippocampus. Science 2007;315:961–6.

46 Lewis BP, Burge CB, Bartel DP. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 2005;120:15–20.

47 Li JH, Liu S, Zhou H et al. starBase v20: decoding miRNA- ceRNA, miRNA-ncRNA and protein-RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Res 2014;42:D92–7.

48 Lin JH, Lin IP, Ohyama Y et al. FAM20C directly binds to and phosphorylates Periostin. Sci Rep 2020;10:17155.

49 Liu Z, Magal P, Seydi O et al. A COVID-19 epidemic model with latency period. Infect Dis Model 2020;5:323–37.

50 Livingston G, Sommerlad A, Orgeta V et al. Dementia prevention, intervention, and care. Lancet 2017;390:2673–734.

51 Livingston G, Huntley J, Sommerlad A et al. Dementia prevention, intervention, and care: 2020 report of the Lancet Commission. Lancet 2020;396:413–46.

52 Llorens-Martin M, Blazquez-Llorca L, Benavides-Piccione R et al. Selective alterations of neurons and circuits related to early memory loss in Alzheimer’s disease. Front Neuroanat 2014;8:38.

53 Lu L, Bao G, Chen H et al. Modification of hippocampal neurogenesis and neuroplasticity by social environments. Exp Neurol 2003;183:600–9.

54 Lu SY, Fu CL, Liang L et al. miR-218-2 regulates cognitive functions in the hippocampus through complement component 3-dependent modulation of synaptic vesicle release. Proc Natl Acad Sci U S A 2021;118:e2021770118.

55 Ma M, Xiong W, Hu F et al. A novel pathway regulates social hierarchy via lncRNA AtLAS and postsynaptic synapsin IIb. Cell Res 2020;30:105–18.

56 Madronal N, Delgado-Garcia JM, Fernandez-Guizan A et al. Rapid erasure of hippocampal memory following inhibition of dentate gyrus granule cells. Nat Commun 2016;7:10923.

57 Magill ST, Cambronne XA, Luikart BW et al. microRNA-132 regulates dendritic growth and arborization of newborn neurons in the adult hippocampus. Proc Natl Acad Sci U S A 2010;107:20382–7.

58 Mchugh TJ, Jones MW, Quinn JJ et al. Dentate gyrus NMDA receptors mediate rapid pattern separation in the hippocampal network. Science 2007;317:94–9.

59 Mckhann, GM, Knopman, DS, Chertkow, H et al. The diagnosis of dementia due to Alzheimer’s disease: recommendations from the National Institute on Aging-Alzheimer’s Association workgroups on diagnostic guidelines for Alzheimer’s disease. Alzheimers Dement 2011;7:263–9.

60 Meyer RC, Giddens MM, Schaefer SA et al. GPR37 and GPR37L1 are receptors for the neuroprotective and glioprotective factors prosaptide and prosaposin. Proc Natl Acad Sci U S A 2013;110:9529–34.

61 Monday HR, Kharod SC, Yoon YJ et al. Presynaptic FMRP and local protein synthesis support structural and functional plasticity of glutamatergic axon terminals. Neuron 2022;110:2588–2606.e6.

62 Mumtaz F, Khan MI, Zubair M et al. Neurobiology and consequences of social isolation stress in animal model-A comprehensive review. Biomed Pharmacother 2018;105:1205–22.

63 Nakashiba T, Cushman JD, Pelkey KA et al. Young dentate granule cells mediate pattern separation, whereas old granule cells facilitate pattern completion. Cell 2012;149:188–201.

64 Nordquist RE, Durkin S, Jacquet A et al. The tachykinin NK3 receptor agonist senktide induces locomotor activity in male Mongolian gerbils. Eur J Pharmacol 2008;600:87–92.

65 Ozturk Z, O’kane CJ, Perez-Moreno JJ. Axonal endoplasmic reticulum dynamics and its roles in neurodegeneration. Front Neurosci 2020;14:48.

66 Palmer A, Good M. Hippocampal synaptic activity, pattern separation and episodic-like memory: implications for mouse models of Alzheimer’s disease pathology. Biochem Soc Trans 2011;39:902–9.

67 Palop JJ, Chin J, Roberson ED et al. Aberrant excitatory neuronal activity and compensatory remodeling of inhibitory hippocampal circuits in mouse models of Alzheimer’s disease. Neuron 2007;55:697–711.

68 Quignot C, Postic G, Bret H et al. InterEvDock3: a combined template-based and free docking server with increased performance through explicit modeling of complex homologs and integration of covariation-based contact maps. Nucleic Acids Res 2021;49:W277–84.

69 Rizzi S, Bianchi P, Guidi S et al. Neonatal isolation impairs neurogenesis in the dentate gyrus of the guinea pig. Hippocampus 2007;17:78–91.

70 Rollenhagen A, Lubke JH. The mossy fiber bouton: the ‘common’ or the ‘unique’ synapse? Front Synaptic Neurosci 2010;2:2.

71 Ryan B, Logan BJ, Abraham WC et al. MicroRNAs, miR-23a-3p and miR-151-3p, are regulated in dentate gyrus neuropil following induction of long-term potentiation in vivo. PLoS One 2017;12:e0170407.

72 Sahay A, Scobie KN, Hill AS et al. Increasing adult hippocampal neurogenesis is sufficient to improve pattern separation. Nature 2011;472:466–70.

73 Sanuki R, Onishi A, Koike C et al. miR-124a is required for hippocampal axogenesis and retinal cone survival through Lhx2 suppression. Nat Neurosci 2011;14:1125–34.

74 Sasaki T, Piatti VC, Hwaun E et al. Dentate network activity is necessary for spatial working memory by supporting CA3 sharp-wave ripple generation and prospective firing of CA3 neurons. Nat Neurosci 2018;21:258–69.

75 Scheff SW, Price DA. Synaptic density in the inner molecular layer of the hippocampal dentate gyrus in Alzheimer disease. J Neuropathol Exp Neurol 1998;57:1146–53.

76 Scheff SW, Price DA, Schmitt FA et al. Hippocampal synaptic loss in early Alzheimer’s disease and mild cognitive impairment. Neurobiol Aging 2006;27:1372–84.

77 Scheff SW, Sparks DL, Price DA. Quantitative assessment of synaptic density in the outer molecular layer of the hippocampal dentate gyrus in Alzheimer’s disease. Dementia 1996;7:226–32.

78 Schwarzinger M, Pollock BG, Hasan OSM et al. Contribution of alcohol use disorders to the burden of dementia in France 2008-13: a nationwide retrospective cohort study. Lancet Public Health 2018;3:e124–32.

79 Shen C, Rolls E, Cheng W et al. Associations of social isolation and loneliness with later dementia. Neurology 2022;99:e164–e175.

80 Shi Q, Hu X, Prior M et al. The occurrence of aging-dependent reticulon 3 immunoreactive dystrophic neurites decreases cognitive function. J Neurosci 2009a;29:5108–15.

81 Shi Q, Prior M, He W et al. Reduced amyloid deposition in mice overexpressing RTN3 is adversely affected by preformed dystrophic neurites. J Neurosci 2009b;29:9163–73.

82 Shi Y, Cui M, Ochs K et al. Long-term diazepam treatment enhances microglial spine engulfment and impairs cognitive performance via the mitochondrial 18 kDa translocator protein (TSPO). Nat Neurosci 2022;25:317–29.

83 Sindi S, Kareholt I, Johansson L et al. Sleep disturbances and dementia risk: a multicenter study. Alzheimers Dement 2018;14:1235–42.

84 Siuciak JA, Mccarthy SA, Martin AN et al. Disruption of the neurokinin-3 receptor (NK3) in mice leads to cognitive deficits. Psychopharmacology (Berl) 2007;194:185–95.

85 Sommerlad A, Sabia S, Singh-Manoux A et al. Association of social contact with dementia and cognition: 28-year follow-up of the Whitehall II cohort study. PLoS Med 2019;16:e1002862.

86 Song Y, Hu M, Zhang J et al. A novel mechanism of synaptic and cognitive impairments mediated via microRNA-30b in Alzheimer’s disease. EBioMedicine 2019;39:409–21.

87 Stadhouders R, Van Den Heuvel A, Kolovos P et al. Transcription regulation by distal enhancers: who’s in the loop? Transcription 2012;3:181–6.

88 Stoessl AJ, Dourish CT, Iversen SD. The NK-3 tachykinin receptor agonist senktide elicits 5-HT-mediated behaviour following central or peripheral administration in mice and rats. Br J Pharmacol 1988;94:285–7.

89 Sundstrom A, Adolfsson AN, Nordin M et al. Loneliness increases the risk of all-cause dementia and Alzheimer’s disease. J Gerontol B Psychol Sci Soc Sci 2020;75:919–26.

90 Sze CI, Troncoso JC, Kawas C et al. Loss of the presynaptic vesicle protein synaptophysin in hippocampus correlates with cognitive decline in Alzheimer disease. J Neuropathol Exp Neurol 1997;56:933–44.

91 Tayler KK, Tanaka KZ, Reijmers LG et al. Reactivation of neural ensembles during the retrieval of recent and remote memory. Curr Biol 2013;23:99–106.

92 Varadi M, Anyango S, Deshpande M et al. AlphaFold Protein Structure Database: massively expanding the structural coverage of protein-sequence space with high-accuracy models. Nucleic Acids Res 2022;50:D439–44.

93 Viana Da Silva S, zhang p, habERL MG et al. Hippocampal mossy fibers synapses in CA3 pyramidal cells are altered at an early stage in a mouse model of Alzheimer’s disease. J Neurosci 2019;39:4193–205.

94 Wang X, Liu D, Huang HZ et al. A novel MicroRNA-124/PTPN1 Signal Pathway mediates synaptic and memory deficits in Alzheimer’s disease. Biol Psychiatry 2018;83:395–405.

95 Wilhelm BG, Mandad S, Truckenbrodt S et al. Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins. Science 2014;344:1023–8.

96 Wilson RS, Krueger KR, Arnold SE et al. Loneliness and risk of Alzheimer disease. Arch Gen Psychiatry 2007;64:234–40.

97 Wu Z, Guo Z, Gearing M et al. Tonic inhibition in dentate gyrus impairs long-term potentiation and memory in an Alzheimer’s [corrected] disease model. Nat Commun 2014;5:4159.

98 Xiang L, Ren Y, Cai H et al. MicroRNA-132 aggravates epileptiform discharges via suppression of BDNF/TrkB signaling in cultured hippocampal neurons. Brain Res 2015;1622:484–95.

99 Xie AJ, Hou TY, Xiong W et al. Tau overexpression impairs neuronal endocytosis by decreasing the GTPase dynamin 1 through the miR-132/MeCP2 pathway. Aging Cell 2019;18:e12929.

100 Yan Y, Tao H, He J et al. The HDOCK server for integrated protein-protein docking. Nat Protoc 2020;15:1829–52.

101 Yang W, Liu M, Zhang Q et al. Knockdown of miR-124 reduces depression-like behavior by targeting CREB1 and BDNF. Curr Neurovasc Res 2020;17:196–203.

102 Yassa MA, Stark SM, Bakker A et al. High-resolution structural and functional MRI of hippocampal CA3 and dentate gyrus in patients with amnestic Mild Cognitive Impairment. Neuroimage 2010;51:1242–52.

103 Zelikowsky M, Hui M, Karigo T et al. The neuropeptide Tac2 controls a distributed brain state induced by chronic social isolation stress. Cell 2018;173:1265–1279.e19.

104 Zhang S, Kou Y, Hu C et al. MicroRNA profiling in the dentate gyrus in epileptic rats: the role of miR-187-3p. Medicine (Baltim) 2017;96:e6744.

105 Zhang J, Liu D, Fu P et al. Social isolation reinforces aging-related behavioral inflexibility by promoting neuronal necroptosis in basolateral amygdala. Mol Psychiatry 2022;27:4050.

106 Zheng K, Hu F, Zhou Y et al. miR-135a-5p mediates memory and synaptic impairments via the Rock2/Adducin1 signaling pathway in a mouse model of Alzheimer’s disease. Nat Commun 2021;12:1–16.

107 Zheng R, Wan C, Mei S et al. Cistrome Data Browser: expanded datasets and new tools for gene regulatory analysis. Nucleic Acids Res. 2019;47(D1):D729–D735.

108 Zlomuzica A, Dere E, Huston JP et al. NK(3) receptor agonism promotes episodic-like memory in mice. Neurobiol Learn Mem 2008;90:420–5.