Impaired neuronal macroautophagy in the prelimbic cortex contributes to comorbid anxiety-like behaviors in rats with chronic neuropathic pain

Su Fu; Haojie Sun; Jiaxin Wang; Shuaixin Gao; Liu Zhu; Kun Cui; Shimeng Liu; Xuetao Qi; Rui Guan; Xiaocen Fan; Qingying Liu; Wen Chen; Li Su; Shuang Cui; Feifei Liao; Fengyu Liu; Catherine C L Wong; Ming Yi; You Wan

doi:10.1080/15548627.2024.2330038

Impaired neuronal macroautophagy in the prelimbic cortex contributes to comorbid anxiety-like behaviors in rats with chronic neuropathic pain

Autophagy. 2024 Mar 24:1-18. doi: 10.1080/15548627.2024.2330038. Online ahead of print.

Authors

Su Fu¹, Haojie Sun^{1

2

3}, Jiaxin Wang^{1

2}, Shuaixin Gao^{4

5}, Liu Zhu⁶, Kun Cui^{1

2}, Shimeng Liu⁷, Xuetao Qi^{1

2}, Rui Guan^{1

2}, Xiaocen Fan^{1

2}, Qingying Liu^{1

2}, Wen Chen^{1

2}, Li Su^{1

2}, Shuang Cui^{1

2}, Feifei Liao^{1

2}, Fengyu Liu^{1

2}, Catherine C L Wong⁴, Ming Yi^{1

2}, You Wan^{1

2

8}

Affiliations

¹ Neuroscience Research Institute and Department of Neurobiology, School of Basic Medical Sciences, Peking University, Beijing, China.
² Key Laboratory for Neuroscience, Ministry of Education and National Health Commission, Peking University, Beijing, China.
³ UCL School of Pharmacy, University College London, London, UK.
⁴ Department of Medical Research Center, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing, China.
⁵ Human Nutrition Program, Department of Human Sciences & James Comprehensive Cancer Center, 309 Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA.
⁶ Department of Biochemistry and Biophysics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
⁷ Laboratory of Cardiovascular Bioactive Molecule, School of Basic Medical Sciences, Peking University, Beijing, China.
⁸ Co-innovation Center of Neuroregeneration, Nantong University, Nantong 226001, P.R. China.

PMID: 38522078
DOI: 10.1080/15548627.2024.2330038

Abstract

A large proportion of patients with chronic pain experience co-morbid anxiety. The medial prefrontal cortex (mPFC) is proposed to underlie this comorbidity, but the molecular and neuronal mechanisms are not fully understood. Here, we reported that impaired neuronal macroautophagy in the prelimbic cortical (PrL) subregion of the mPFC paralleled the occurrence of anxiety-like behaviors in rats with chronic spared nerve injury (SNI). Intriguingly, such macroautophagy impairment was mainly observed in a FOS/c-Fos⁺ neuronal subpopulation in the PrL. Chemogenetic inactivation of this comorbid anxiety-related neuronal ensemble relieved pain-induced anxiety-like behaviors. Rescuing macroautophagy impairment in this neuronal ensemble relieved chronic pain-associated anxiety and mechanical allodynia and restored synaptic homeostasis at the molecular level. By contrast, artificial disruption of macroautophagy induced early-onset co-morbid anxiety in neuropathic rats, but not general anxiety in normal rats. Taken together, our work identifies causal linkage between PrL neuronal macroautophagy dysfunction and comorbid anxiety in neuropathic pain and provides novel insights into the role of PrL by differentiating its contribution in pain-induced comorbid anxiety from its modulation over general anxiety-like behaviors.Abbreviation: AAV: adeno-associated viruses; ACC: anterior cingulate cortex; ATG5: autophagy related 5; ATG7: autophagy related 7; ATG12: autophagy related 12; CAMK2/CaMKII: calcium/calmodulin-dependent protein kinase II; CNO: clozapine-N-oxide; CQ: chloroquine; DIA: data independent acquisition; DIO: double floxed inverse orf; DLG4/PSD-95: discs large MAGUK scaffold protein 4; Dox: doxycycline; GABA: γ-aminobutyric acid; GFP: green fluorescent protein; GO: gene ontology; Gi: inhibitory guanine nucleotide-binding proteins; HsCHRM4/M4D: human cholinergic receptor muscarinic 4; HsSYN: human synapsin; KEGG: Kyoto encyclopedia of genes and genomes; LAMP1: lysosomal-associated membrane protein 1; LC3-II: PE conjugated microtubule-associated protein 1 light chain3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; mPFC: medial prefrontal cortex; P2A: 2A self-cleaving peptide; PPI: protein-protein interaction networks; PrL: prelimbic cortex; RBFOX3/NeuN: RNA binding protein, fox-1 homolog (C. elegans) 3; rtTA: reverse tetracycline-transactivator; SDS-PAGE: sodium dodecylsulfate-polyacrylamide gel electrophoresis; SHANK3: SH3 and multiple ankyrin repeat domains 3; SLC1A1/EAAC1: solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, systemXag), member 1; SNAP23: synaptosomal-associated protein 23; SNI:spared nerve injury; SQSTM1/p62: sequestosome 1; SYT3: synaptotagmin 3; TRE: tetracycline-responsive element; TRE3G: third-generation tetracycline-responsive element.

Keywords: Autophagy; chronic neuropathic pain; comorbid anxiety; neuronal ensemble; prelimbic cortex; synaptic homeostasis.