Abstract
Opioid use disorder (OUD) is a public health crisis in the U.S. that causes over 50 thousand deaths annually due to overdose. Using next-generation RNA sequencing and proteomics techniques, we identified 394 differentially expressed (DE) coding and long noncoding (lnc) RNAs as well as 213 DE proteins in Brodmann Area 9 of OUD subjects. The RNA and protein changes converged on pro-angiogenic gene networks and cytokine signaling pathways. Four genes (LGALS3, SLC2A1, PCLD1, and VAMP1) were dysregulated in both RNA and protein. Dissecting these DE genes and networks, we found cell type-specific effects with enrichment in astrocyte, endothelial, and microglia correlated genes. Weighted-genome correlation network analysis (WGCNA) revealed cell-type correlated networks including an astrocytic/endothelial/microglia network involved in angiogenic cytokine signaling as well as a neuronal network involved in synaptic vesicle formation. In addition, using ex vivo magnetic resonance imaging, we identified increased vascularization in postmortem brains from a subset of subjects with OUD. This is the first study integrating dysregulation of angiogenic gene networks in OUD with qualitative imaging evidence of hypervascularization in postmortem brain. Understanding the neurovascular effects of OUD is critical in this time of widespread opioid use.
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References
Hedegaard H, Miniño AM, Warner M. Drug overdose deaths in the United States, 1999–2018. NCHS Data Brief. 2020:1–8.
National Institute on Drug Abuse (NIDA). Overdose death rates. 2019. https://www.drugabuse.gov/related-topics/trends-statistics/overdose-death-rates. Accessed 16 Sep 2019.
Opioid Overdose Crisis | National Institute on Drug Abuse (NIDA). https://www.drugabuse.gov/drug-topics/opioids/opioid-overdose-crisis. Accessed 1 Jan 2021.
Browne CJ, Godino A, Salery M, Nestler EJ. Epigenetic mechanisms of opioid addiction. Biol Psychiatry. 2020;87(Jan):22–33.
Encode Project Consortium T. Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project. Nature. 2007;447:799–816.
Carninci P, Kasukawa T, Katayama S, Gough J, Frith MC, Maeda N, et al. Molecular biology: The transcriptional landscape of the mammalian genome. Science (80-) 2005;309:1559–63.
Lee JT. Epigenetic regulation by long noncoding RNAs. Science (80-) 2012;338:1435–9.
Monoranu CM, Apfelbacher M, Grünblatt E, Puppe B, Alafuzoff I, Ferrer I, et al. pH measurement as quality control on human post mortem brain tissue: a study of the BrainNet Europe consortium HHS Public Access. Neuropathol Appl Neurobiol. 2009;35:329–37.
Bateman A. UniProt: a worldwide hub of protein knowledge. Nucleic Acids Res. 2019;47:D506–15.
Zhang X, Smits AH, Van Tilburg GBA, Ovaa H, Huber W, Vermeulen M. Proteome-wide identification of ubiquitin interactions using UbIA-MS. Nat Protoc. 2018;13:530–50.
Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, et al. Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43:e47.
McCarthy DJ, Chen Y, Smyth GK. Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Res. 2012;40:4288–97.
Robinson MD, Oshlack A. A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol. 2010;11:R25.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 2001;25:402–8.
Huang SY, Lu W, Ge D, Meng N, Li Y, Su L. et al. A new microRNA signal pathway regulated by long noncoding RNA TGFB2-OT1 in autophagy and inflammation of vascular endothelial cells. Autophagy. 2015;11:2172–83.
Steen CB, Liu CL, Alizadeh AA, Newman AM. Profiling cell type abundance and expression in bulk tissues with CIBERSORTx. Methods Mol Biol. 2020;2117:135–57.
Newman AM, Liu CL, Green MR, Gentles AJ, Feng W, Xu Y, et al. Robust enumeration of cell subsets from tissue expression profiles. Nat Methods. 2015;12:453–7.
Yu Q, He Z. Comprehensive investigation of temporal and autism-associated cell type composition-dependent and independent gene expression changes in human brains. Sci Rep. 2017;7:1–12.
Darmanis S, Sloan SA, Zhang Y, Enge M, Caneda C, Shuer LM, et al. A survey of human brain transcriptome diversity at the single cell level. Proc Natl Acad Sci USA. 2015;112:7285–90.
McKenzie AT, Wang M, Hauberg ME, Fullard JF, Kozlenkov A, Keenan A, et al. Brain cell type specific gene expression and co-expression network architectures. Sci Rep. 2018;8:1–19.
Alexa A, Rahnenfuhrer J. topGO: enrichment analysis for gene ontology. R package version 2.40.0; 2020.
Langfelder P, Horvath S. WGCNA: an R package for weighted correlation network analysis. BMC Bioinform. 2008;9:559.
Collado-Torres L, Jaffe AE, Burke EE. jaffelab: commonly used functions by the Jaffe lab. 2019. https://github.com/LieberInstitute/jaffelab.
Kolde R. pheatmap: pretty heatmaps. R package version 1.0.12; 2019. https://cran.r-project.org/package=pheatmap.
Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13:2498–504.
Chavhan GB, Babyn PS, Thomas B, Shroff MM, Haacke EM. Principles, techniques, and applications of T2*-based MR imaging and its special applications. Radiographics. 2009;29:1433–49.
Sosina O, Tran M, Maynard K, Tao R, Taub M, Martinowich K, et al. Strategies for cellular deconvolution in human brain RNA sequencing data. bioRxiv. 2020. 01.19.910976.
Harlan RE, Garcia MM. Drugs of abuse and immediate-early genes in the forebrain. Mol Neurobiol. 1998;16:221–67.
Bisagno V, Cadet JL. Expression of immediate early genes in brain reward circuitries: differential regulation by psychostimulant and opioid drugs. Neurochem Int. 2019;124:10–8.
Sosnowski DW, Jaffe AE, Tao R, Deep-Soboslay A, Kleinman JE, Hyde TM, et al. Differential expression of NPAS4 in the dorsolateral prefrontal cortex following acute opioid intoxication. bioRxiv. 2020.12.23.424239.
Healy S, Khan P, Davie JR. Immediate early response genes and cell transformation. Pharm Ther. 2013;137:64–77.
Tullai JW, Schaffer ME, Mullenbrock S, Sholder G, Kasif S, Cooper GM. Immediate-early and delayed primary response genes are distinct in function and genomic architecture. J Biol Chem. 2007;282:23981–95.
Fahmy RG, Dass CR, Sun LQ, Chesterman CN, Khachigian LM. Transcription factor Egr-1 supports FGF-dependent angiogenesis during neovascularization and tumor growth. Nat Med. 2003;9:1026–32.
Khachigian LM, Collins T. Inducible expression of Egr-1-dependent genes. Circ Res. 1997;81:457–61.
Santiago FS, Lowe HC, Day FL, Chesterman CN, Khachigian LM. Early growth response factor-1 induction by injury is triggered by release and paracrine activation by fibroblast growth factor-2. Am J Pathol. 1999;154:937–44.
Wang D, Mayo MW, Baldwin AS. Basic fibroblast growth factor transcriptional autoregulation requires EGR-1. Oncogene. 1997;14:2291–9.
Thijssen VL, Barkan B, Shoji H, Aries IM, Mathieu V, Deltour L, et al. Tumor cells secrete galectin-1 to enhance endothelial cell activity. Cancer Res. 2010;70:6216–24.
Hsieh SH, Ying NW, Wu MH, Chiang WF, Hsu CL, Wong TY. et al. Galectin-1, a novel ligand of neuropilin-1, activates VEGFR-2 signaling and modulates the migration of vascular endothelial cells. Oncogene. 2008;27:3746–53.
Nangia-Makker P, Honjo Y, Sarvis R, Akahani S, Hogan V, Pienta KJ, et al. Galectin-3 induces endothelial cell morphogenesis and angiogenesis. Am J Pathol. 2000;156:899–909.
Funasaka T, Raz A, Nangia-Makker P. Galectin-3 in angiogenesis and metastasis. Glycobiology. 2014;24:886–91.
Veys K, Fan Z, Ghobrial M, Bouché A, García-Caballero M, Vriens K, et al. Role of the GLUT1 glucose transporter in postnatal CNS angiogenesis and blood-brain barrier integrity. Circ Res. 2020;127:466–82.
Hassan HE, Myers AL, Lee IJ, Chen H, Coop A, Eddington ND. Regulation of gene expression in brain tissues of rats repeatedly treated by the highly abused opioid agonist, oxycodone: microarray profiling and gene mapping analysis. Drug Metab Dispos. 2010;38:157–67.
Ma HL, Su L, Zhang SL, Kung HF, Miao JY. Inhibition of ANXA7 GTPase activity by a small molecule promotes HMBOX1 translation of vascular endothelial cells in vitro and in vivo. Int J Biochem Cell Biol. 2016;79:33–40.
Nelson EC, Agrawal A, Heath AC, Bogdan R, Sherva R, Zhang B, et al. Evidence of CNIH3 involvement in opioid dependence. Mol Psychiatry. 2016;21:608–14.
Niu DG, Peng F, Zhang W, Guan Z, Zhao HD, Li JL. et al. Morphine promotes cancer stem cell properties, contributing to chemoresistance in breast cancer. Oncotarget. 2015;6:3963–76.
Jimenez-Gonzalez A, García-Concejo A, León-Lobera F, Rodriguez RE. Morphine delays neural stem cells differentiation by facilitating Nestin overexpression. Biochim Biophys Acta 2018;1862:474–84.
Eisenstein TK. The role of opioid receptors in immune system function. Front Immunol. 2019;10:2904.
Seney ML, Kim S-M, Glausier JR, Hildebrand MA, Xue X, Zong W, et al. Transcriptional alterations in dorsolateral prefrontal cortex and nucleus accumbens implicate neuroinflammation and synaptic remodeling in opioid use disorder. Biol Psychiatry. 2021. https://doi.org/10.1016/j.biopsych.2021.06.007.
Liu A, Dai Y, Mendez EF, Hu R, Fries GR, Najera KE, et al. Genome-wide correlation of DNA methylation and gene expression in postmortem brain tissues of opioid use disorder patients. Int J Neuropsychopharmacol. 2021 https://doi.org/10.1093/ijnp/pyab043.
Muñoz-Chápuli R, Quesada AR, Medina MÁ. Angiogenesis and signal transduction in endothelial cells. C. Cell Mol Life Sci. 2004;61:2224–43.
Liu D, Jia H, Holmes DIR, Stannard A, Zachary I. Vascular endothelial growth factor-regulated gene expression in endothelial cells: KDR-mediated induction of Egr3 and the related nuclear receptors Nur77, Nurr1, and Nor1. Arterioscler Thromb Vasc Biol. 2003;23:2002–7.
Arkenbout EK, Van Bragt M, Eldering E, Van Bree C, Grimbergen JM, Quax PHA, et al. TR3 orphan receptor is expressed in vascular endothelial cells and mediates cell cycle arrest. Arterioscler Thromb Vasc Biol. 2003;23:1535–40.
Johnson MM, Michelhaugh SK, Bouhamdan M, Schmidt CJ, Bannon MJ. The transcription factor NURR1 exerts concentration-dependent effects on target genes mediating distinct biological processes. Front Neurosci. 2011;5:135.
Chen C, Li Y, Hou S, Bourbon PM, Qin L, Zhao K, et al. Orphan nuclear receptor TR3/Nur77 biologics inhibit tumor growth by targeting angiogenesis and tumor cells. Microvasc Res. 2020;128:103934.
Zeng H, Qin L, Zhao D, Tan X, Manseau EJ, Mien VH, et al. Orphan nuclear receptor TR3/Nur77 regulates VEGF-A-induced angiogenesis through its transcriptional activity. J Exp Med. 2006;203:719–29.
Proia P, Schiera G, Mineo M, Ingrassia AMR, Santoro G, Savettieri G, et al. Astrocytes shed extracellular vesicles that contain fibroblast growth factor-2 and vascular endothelial growth factor. Int J Mol Med. 2008;21:63–7.
Vallon M, Chang J, Zhang H, Kuo CJ. Developmental and pathological angiogenesis in the central nervous system. Cell Mol Life Sci. 2014;71:3489–506.
Bernal GM, Peterson DA. Phenotypic and gene expression modification with normal brain aging in GFAP-positive astrocytes and neural stem cells. Aging Cell. 2011;10:466–82.
Hashimoto K, Noshiro M, Ohno S, Kawamoto T, Satakeda H, Akagawa Y, et al. Characterization of a cartilage-derived 66-kDa protein (RCD-CAP/βig-h3) that binds to collagen. Biochim Biophys Acta. 1997;1355:303–14.
Madry C, Kyrargyri V, Arancibia-Cárcamo IL, Jolivet R, Kohsaka S, Bryan RM. et al. Microglial ramification, surveillance, and interleukin-1β release are regulated by the two-pore domain K+ channel THIK-1. Neuron. 2018;97:299–312.e6.
Sha L, MacIntyre L, MacHell JA, Kelly MP, Porteous DJ, Brandon NJ, et al. Transcriptional regulation of neurodevelopmental and metabolic pathways by NPAS3. Mol Psychiatry. 2012;17:267–79.
Hatakeyama M, Ninomiya I, Kanazawa M. Angiogenesis and neuronal remodeling after ischemic stroke. Neural Regen Res. 2020;15:16–9.
Borne J, Riascos R, Cuellar H, Vargas D, Rojas R. Neuroimaging in drug and substance abuse part II: opioids and solvents. Top Magn Reson Imaging. 2005;16:239–45.
Geibprasert S, Gallucci M, Krings T. Addictive illegal drugs: structural neuroimaging. Am J Neuroradiol. 2010;31:803–8.
Andersen SN, Skullerud K. Hypoxic/ischaemic brain damage, especially pallidal lesions, in heroin addicts. Forensic Sci Int. 1999;102:51–9.
Brust JCM, Richter RW. Stroke associated with addiction to heroin. J Neurol Neurosurg Psychiatry. 1976;39:194–9.
Khodneva Y, Muntner P, Kertesz S, Kissela B, Safford MM. Prescription opioid use and risk of coronary heart disease, stroke, and cardiovascular death among adults from a prospective cohort (REGARDS study). Pain Med. 2016;17:444–55.
Peyravian N, Dikici E, Deo S, Toborek M, Daunert S. Opioid antagonists as potential therapeutics for ischemic stroke. Prog Neurobiol. 2019;182:101679.
Benyó Z, Wahl M. Opiate receptor-mediated mechanisms in the regulation of cerebral blood flow. Cerebrovasc Brain Metab Rev. 1996;8:326–57.
Volkow ND, Valentine A, Kulkarni M. Radiological and neurological changes in the drug abuse patient: a study with MRI. J Neuroradiol. 1988;15:288–93.
Wigmore T, Farquhar-Smith P. Opioids and cancer. Curr Opin Support Palliat Care. 2016;10:109–18.
Leo S, Nuydens R, Meert TF. Opioid-induced proliferation of vascular endothelial cells. J Pain Res. 2009;2:59–66.
Gupta K, Kshirsagar S, Chang L, Schwartz R, Law P-Y, Yee D, et al. Morphine stimulates angiogenesis by activating proangiogenic and survival-promoting signaling and promotes breast tumor growth 1. Cancer Res. 2002;62:4491–8.
Koodie L, Ramakrishnan S, Roy S. Morphine suppresses tumor angiogenesis through a HIF-1α/p38MAPK pathway. Am J Pathol. 2010;177:984–97.
Yamamizu K, Furuta S, Hamada Y, Yamashita A, Kuzumaki N, Narita M, et al. Opioids inhibit tumor angiogenesis by suppressing VEGF signaling. Sci Rep. 2013;3:3213.
Yamamizu K, Hamada Y, Narita M. κ Opioid receptor ligands regulate angiogenesis in development and in tumours. Br J Pharmol. 2015;172:268–76.
Acknowledgements
We are grateful for the invaluable donations and participation from families, as well as for the generous collaboration of the medical examiners at the Harris County Institute of Forensic Sciences. This study was supported by R01DA044859 to CWB. ZZ was supported by R01LM012806. The University of Texas System provided funding for the Neuropsychiatric Proteome Database, for which proteomics data from brain tissue was generated by the Mass Spectrometry Core at the University of Texas Medical Branch.
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EM and CWB designed and organized the study, and wrote the manuscript; EM analyzed the data, interpreted the results, and wrote the manuscript; LS, GRF, KN, and KM contributed to data acquisition; HW, RH, ZZ, and JQW contributed to analysis of data; XW, MM, and JQW performed lncRNA validation studies; CML, JK and JX contributed to MRI data acquisition and analysis; KN, TM, SS, and ALT contributed to psychological autopsies and subject diagnosis. All authors revised and approved the final manuscript.
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Mendez, E.F., Wei, H., Hu, R. et al. Angiogenic gene networks are dysregulated in opioid use disorder: evidence from multi-omics and imaging of postmortem human brain. Mol Psychiatry 26, 7803–7812 (2021). https://doi.org/10.1038/s41380-021-01259-y
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DOI: https://doi.org/10.1038/s41380-021-01259-y
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