Proteomic Analysis of Spinal Cord Tissue in a Rat Model of Cancer-induced Bone Pain

Overview

Journal Front Mol Neurosci

Specialty Molecular Biology

Date 2022 Dec 22

PMID 36545122

Authors

Heyu Yang

Ji Wu

Shuqing Zhen

Yindi Hu

Dai Li

Min Xie

Haili Zhu

Affiliations

Soon will be listed here.

Abstract

Background: Cancer-induced bone pain (CIBP) is a moderate to severe pain and seriously affects patients' quality of life. Spinal cord plays critical roles in pain generation and maintenance. Identifying differentially expressed proteins (DEPs) in spinal cord is essential to elucidate the mechanisms of cancer pain.

Methods: CIBP rat model was established by the intratibial inoculation of MRMT-1 cells. Positron emission tomography (PET) scan and transmission electron microscopy (TEM) were used to measure the stats of spinal cord in rats. Label free Liquid Chromatography with tandem mass spectrometry (LC-MS-MS) were used to analyze the whole proteins from the lumbar spinal cord. Differentially expressed proteins (DEPs) were performed using Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, and verified using Western blot and immunofluorescence assay.

Results: In the current study, CIBP rats exhibited bone damage, spontaneous pain, mechanical hyperalgesia, and impaired motor ability. In spinal cord, an hypermetabolism and functional abnormality were revealed on CIBP rats. An increase of synaptic vesicles density in active zone and a disruption of mitochondrial structure in spinal cord of CIBP rats were observed. Meanwhile, 422 DEPs, consisting of 167 up-regulated and 255 down-regulated proteins, were identified among total 1539 proteins. GO enrichment analysis indicated that the DEPs were mainly involved in catabolic process, synaptic function, and enzymic activity. KEGG pathway enrichment analysis indicated a series of pathways, including nervous system disease, hormonal signaling pathways and amino acid metabolism, were involved. Expression change of synaptic and mitochondrial related protein, such as complexin 1 (CPLX1), synaptosomal-associated protein 25 (SNAP25), synaptotagmin 1 (SYT1), aldehyde dehydrogenase isoform 1B1 (ALDH1B1), Glycine amidinotransferase (GATM) and NADH:ubiquinone oxidoreductase subunit A11 (NDUFA11), were further validated using immunofluorescence and Western blot analysis.

Conclusion: This study provides valuable information for understanding the mechanisms of CIBP, and supplies potential therapeutic targets for cancer pain.

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References

Gokhale A, Lee C, Zlatic S, Freeman A, Shearing N, Hartwig C . Mitochondrial Proteostasis Requires Genes Encoded in a Neurodevelopmental Syndrome Locus. J Neurosci. 2021; 41(31):6596-6616. PMC: 8336702. DOI: 10.1523/JNEUROSCI.2197-20.2021. View

Lei T, Qian H, Lei P, Hu Y . Ferroptosis-related gene signature associates with immunity and predicts prognosis accurately in patients with osteosarcoma. Cancer Sci. 2021; 112(11):4785-4798. PMC: 8586685. DOI: 10.1111/cas.15131. View

Jang S, Javadov S . Elucidating the contribution of ETC complexes I and II to the respirasome formation in cardiac mitochondria. Sci Rep. 2018; 8(1):17732. PMC: 6286307. DOI: 10.1038/s41598-018-36040-9. View

Luo C, Kuner T, Kuner R . Synaptic plasticity in pathological pain. Trends Neurosci. 2014; 37(6):343-55. DOI: 10.1016/j.tins.2014.04.002. View

Dhara M, Yarzagaray A, Schwarz Y, Dutta S, Grabner C, Moghadam P . Complexin synchronizes primed vesicle exocytosis and regulates fusion pore dynamics. J Cell Biol. 2014; 204(7):1123-40. PMC: 3971750. DOI: 10.1083/jcb.201311085. View

Chen Y, Orlicky D, Matsumoto A, Singh S, Thompson D, Vasiliou V . Aldehyde dehydrogenase 1B1 (ALDH1B1) is a potential biomarker for human colon cancer. Biochem Biophys Res Commun. 2011; 405(2):173-9. PMC: 3112362. DOI: 10.1016/j.bbrc.2011.01.002. View

Wang L, Chen S, Ma H, Chen H, Hittelman W, Pan H . Regulating nociceptive transmission by VGluT2-expressing spinal dorsal horn neurons. J Neurochem. 2018; 147(4):526-540. PMC: 6263733. DOI: 10.1111/jnc.14588. View

Yanagisawa Y, Furue H, Kawamata T, Uta D, Yamamoto J, Furuse S . Bone cancer induces a unique central sensitization through synaptic changes in a wide area of the spinal cord. Mol Pain. 2010; 6:38. PMC: 3020802. DOI: 10.1186/1744-8069-6-38. View

Othman A, Winogradzki M, Lee L, Tandon M, Blank A, Pratap J . Bone Metastatic Breast Cancer: Advances in Cell Signaling and Autophagy Related Mechanisms. Cancers (Basel). 2021; 13(17). PMC: 8431094. DOI: 10.3390/cancers13174310. View

10.

Tilley D, Lietz C, Cedeno D, Kelley C, Li L, Vallejo R . Proteomic Modulation in the Dorsal Spinal Cord Following Spinal Cord Stimulation Therapy in an In Vivo Neuropathic Pain Model. Neuromodulation. 2020; 24(1):22-32. PMC: 7484326. DOI: 10.1111/ner.13103. View

11.

Doncheva N, Morris J, Gorodkin J, Jensen L . Cytoscape StringApp: Network Analysis and Visualization of Proteomics Data. J Proteome Res. 2018; 18(2):623-632. PMC: 6800166. DOI: 10.1021/acs.jproteome.8b00702. View

12.

Liu M, Cheng X, Yan H, Chen J, Liu C, Chen Z . MiR-135-5p Alleviates Bone Cancer Pain by Regulating Astrocyte-Mediated Neuroinflammation in Spinal Cord through JAK2/STAT3 Signaling Pathway. Mol Neurobiol. 2021; 58(10):4802-4815. DOI: 10.1007/s12035-021-02458-y. View

13.

Carney E . GATM mutations cause mitochondrial abnormalities and kidney failure. Nat Rev Nephrol. 2018; 14(7):414. DOI: 10.1038/s41581-018-0017-3. View

14.

Kawada K, Iwamoto M, Sakai Y . Mechanisms underlying F-fluorodeoxyglucose accumulation in colorectal cancer. World J Radiol. 2016; 8(11):880-886. PMC: 5120247. DOI: 10.4329/wjr.v8.i11.880. View

15.

Siegel R, Miller K, Fuchs H, Jemal A . Cancer statistics, 2022. CA Cancer J Clin. 2022; 72(1):7-33. DOI: 10.3322/caac.21708. View

16.

Wang K, Gu Y, Liao Y, Bang S, Donnelly C, Chen O . PD-1 blockade inhibits osteoclast formation and murine bone cancer pain. J Clin Invest. 2020; 130(7):3603-3620. PMC: 7324182. DOI: 10.1172/JCI133334. View

17.

Ban J, Fock V, Aryee D, Kovar H . Mechanisms, Diagnosis and Treatment of Bone Metastases. Cells. 2021; 10(11). PMC: 8616226. DOI: 10.3390/cells10112944. View

18.

Kapoor R, Saxena A, Vasudev P, Sundriyal D, Kumar A . Cancer induced bone pain: current management and future perspectives. Med Oncol. 2021; 38(11):134. DOI: 10.1007/s12032-021-01587-7. View

19.

Zhang X, Karim M, Hasan M, Hooper J, Wahab R, Roy S . Cancer-on-a-Chip: Models for Studying Metastasis. Cancers (Basel). 2022; 14(3). PMC: 8833392. DOI: 10.3390/cancers14030648. View

20.

Bosma R, Mojarad E, Leung L, Pukall C, Staud R, Stroman P . FMRI of spinal and supra-spinal correlates of temporal pain summation in fibromyalgia patients. Hum Brain Mapp. 2016; 37(4):1349-60. PMC: 4783193. DOI: 10.1002/hbm.23106. View