Unbiased transcriptomic analyses reveal distinct effects of immune deficiency in CNS function with and without injury
Dandan Luo1,2,3, Weihong Ge4(), Xiao Hu1,2,3, Chen Li1,2,3, Chia-Ming Lee4, Liqiang Zhou3, Zhourui Wu1,2,3, Juehua Yu3, Sheng Lin3, Jing Yu3, Wei Xu1,2,3, Lei Chen1,2,3, Chong Zhang3, Kun Jiang3, Xingfei Zhu1,2,3, Haotian Li1,2,3, Xinpei Gao3, Yanan Geng3, Bo Jing3, Zhen Wang3, Changhong Zheng3, Rongrong Zhu1,2, Qiao Yan3, Quan Lin3,4, Keqiang Ye5, Yi E. Sun3,4(), Liming Cheng1,2,3()
1. Division of Spine Surgery, Department of Orthopedics, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China 2. Institute of Spine and Spine Cord Injury of Tongji University, Shanghai 200065, China 3. Stem Cell Translational Research Center, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China 4. Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90095, USA 5. Department of Pathology and Laboratory Medicine, Center for neurodegeneration disease, Emory University School of Medicine, Atlanta, GA 30322, USA
The mammalian central nervous system (CNS) is considered an immune privileged system as it is separated from the periphery by the blood brain barrier (BBB). Yet, immune functions have been postulated to heavily influence the functional state of the CNS, especially after injury or during neurodegeneration. There is controversy regarding whether adaptive immune responses are beneficial or detrimental to CNS injury repair. In this study, we utilized immunocompromised SCID mice and subjected them to spinal cord injury (SCI). We analyzed motor function, electrophysiology, histochemistry, and performed unbiased RNA-sequencing. SCID mice displayed improved CNS functional recovery compared to WT mice after SCI. Weighted gene-coexpression network analysis (WGCNA) of spinal cord transcriptomes revealed that SCID mice had reduced expression of immune function-related genes and heightened expression of neural transmission-related genes after SCI, which was confirmed by immunohistochemical analysis and was consistent with better functional recovery. Transcriptomic analyses also indicated heightened expression of neurotransmission-related genes before injury in SCID mice, suggesting that a steady state of immune-deficiency potentially led to CNS hyper-connectivity. Consequently, SCID mice without injury demonstrated worse performance in Morris water maze test. Taken together, not only reduced inflammation after injury but also dampened steady-state immune function without injury heightened the neurotransmission program, resulting in better or worse behavioral outcomes respectively. This study revealed the intricate relationship between immune and nervous systems, raising the possibility for therapeutic manipulation of neural function via immune modulation.
MA Anderson, JE Burda, Y Ren, Y Ao, TM O’Shea, R Kawaguchi, G Coppola, BS Khakh, TJ Deming, MV Sofroniew (2016) Astrocyte scar formation aids central nervous system axon regeneration. Nature 532(7598):195–200 https://doi.org/10.1038/nature17623
2
GC Bosma, RP Custer, MJ Bosma (1983) A severe combined immunodeficiency mutation in the mouse. Nature 301(5900):527–530 https://doi.org/10.1038/301527a0
3
H Duan, W Ge, A Zhang, Y Xi, Z Chen, D Luo, Y Cheng, KS Fan, S Horvath, MV Sofroniewet al. (2015) Transcriptome analyses reveal molecular mechanisms underlying functional recovery after spinal cord injury. Proc Natl Acad Sci USA 112(43):13360–13365 https://doi.org/10.1073/pnas.1510176112
AJ Filiano, Y Xu, NJ Tustison, RL Marsh, W Baker, I Smirnov, CC Overall, SP Gadani, SD Turner, Z Wenget al. (2016) Unexpected role of interferon-γ in regulating neuronal connectivity and social behaviour. Nature 535(7612):425–429 https://doi.org/10.1038/nature18626
6
AJ Filiano, SP Gadani, J Kipnis (2017) How and why do T cells and their derived cytokines affect the injured and healthy brain? Nat Rev Neurosci 18(6):375–384 https://doi.org/10.1038/nrn.2017.39
7
MJ Fogarty, LA Hammond, R Kanjhan, MC Bellingham, PG Noakes (2013) A method for the three-dimensional reconstruction of NeurobiotinTM-filled neurons and the location of their synaptic inputs. Front Neural Circuits 7:153 https://doi.org/10.3389/fncir.2013.00153
S Han, C Tai, CJ Jones, T Scheuer, WA Catterall (2014) Enhancement of inhibitory neurotransmission by GABAA receptors having α2,3-subunits ameliorates behavioral deficits in a mouse model of autism. Neuron 81(6):1282–1289 https://doi.org/10.1016/j.neuron.2014.01.016
10
V Hook, KJ Brennand, Y Kim, T Toneff, L Funkelstein, KC Lee, M Ziegler, FH Gage (2014) Human iPSC neurons display activitydependent neurotransmitter secretion: aberrant catecholamine levels in schizophrenia neurons. Stem Cell Rep 3(4):531–538 https://doi.org/10.1016/j.stemcr.2014.08.001
B Jing, C Zhang, X Liu, L Zhou, J Liu, Y Yao, J Yu, Y Weng, M Pan, J Liuet al. (2017) Glycosylation of dentin matrix protein 1 is a novel key element for astrocyte maturation and BBB integrity. Protein Cell. https://doi.org/10.1007/s13238-017-0449-8
13
KA Kigerl, VM McGaughy, PG Popovich (2006) Comparative analysis of lesion development and intraspinal inflammation in four strains of mice following spinal contusion injury. J Comp Neurol 494(4):578–594 https://doi.org/10.1002/cne.20827
14
J Kipnis, H Cohen, M Cardon, Y Ziv, M Schwartz (2004) T cell deficiency leads to cognitive dysfunction: Implications for therapeutic vaccination for schizophrenia and other psychiatric conditions. Proc Natl Acad Sci USA 101(21):8180–8185 https://doi.org/10.1073/pnas.0402268101
15
P Kivisäkk, DJ Mahad, MK Callahan, C Trebst, B Tucky, T Wei, L Wu, ES Baekkevold, H Lassmann, SM Staugaitiset al. (2003) Human cerebrospinal fluid central memory CD4+ T cells: evidence for trafficking through choroid plexus and meninges via P-selectin. Proc Natl Acad Sci USA 100(14):8389–8394 https://doi.org/10.1073/pnas.1433000100
C Liu, W Zhang, G Chen, H Tian, J Li, H Qu, L Cheng, J Zhu, C Zhuo (2017) Aberrant patterns of local and long-range functional connectivity densities in schizophrenia. Oncotarget 8 (29):48196–48203 https://doi.org/10.18632/oncotarget.18441
18
A Louveau, I Smirnov, TJ Keyes, JD Eccles, SJ Rouhani, JD Peske, NC Derecki, D Castle, JW Mandell, KS Leeet al. (2015) Structural and functional features of central nervous system lymphatic vessels. Nature 523(7560):337–341 https://doi.org/10.1038/nature14432
19
P Lu, Y Wang, L Graham, K McHale, M Gao, D Wu, J Brock, A Blesch, ES Rosenzweig, LA Havtonet al. (2012) Long-distance growth and connectivity of neural stem cells after severe spinal cord injury. Cell 150(6):1264–1273 https://doi.org/10.1016/j.cell.2012.08.020
20
S Luchetti, KD Beck, MD Galvan, R Silva, BJ Cummings, AJ Anderson (2010) Comparison of immunopathology and locomotor recovery in C57BL/6, BUB/BnJ, and NOD-SCID mice after contusion spinal cord injury. J Neurotrauma 27(2):411–421 https://doi.org/10.1089/neu.2009.0930
M Michel, MJ Schmidt, K Mirnics (2012) Immune system gene dysregulation in autism and schizophrenia. Dev Neurobiol 72(10):1277–1287 https://doi.org/10.1002/dneu.22044
23
R Nardone, Y Höller, A Thomschewski, AC Bathke, AR Ellis, SM Golaszewski, F Brigo, E Trinka (2015) Assessment of corticospinal excitability after traumatic spinal cord injury using MEP recruitment curves: a preliminary TMS study. Spinal Cord 53(7):534–538 https://doi.org/10.1038/sc.2015.12
24
A Nott, J Cheng, F Gao, YT Lin, E Gjoneska, T Ko, P Minhas, AV Zamudio, J Meng, F Zhanget al. (2016) Histone deacetylase 3 associates with MeCP2 to regulate FOXO and social behavior. Nat Neurosci 19(11):1497–1505 https://doi.org/10.1038/nn.4347
25
S Partida-Sánchez, DA Cockayne, S Monard, EL Jacobson, N Oppenheimer, B Garvy, K Kusser, S Goodrich, M Howard, A Harmsenet al. (2001) Cyclic ADP-ribose production by CD38 regulates intracellular calcium release, extracellular calcium influx and chemotaxis in neutrophils and is required for bacterial clearance in vivo. Nat Med 7(11):1209–1216 https://doi.org/10.1038/nm1101-1209
PG Popovich, TB Jones (2003) Manipulating neuroinflammatory reactions in the injured spinal cord: back to basics. Trends Pharmacol Sci 24(1):13–17 https://doi.org/10.1016/S0165-6147(02)00006-8
28
PG Popovich, EE Longbrake (2008) Can the immune system be harnessed to repair the CNS? Nat Rev Neurosci 9(6):481–493 https://doi.org/10.1038/nrn2398
29
MD Richard, NC Brahm (2012) Schizophrenia and the immune system: pathophysiology, prevention, and treatment. Am J Health-Syst Pharm AJHP 69(9):757–766 https://doi.org/10.2146/ajhp110271
K Supekar, LQ Uddin, A Khouzam, J Phillips, WD Gaillard, LE Kenworthy, BE Yerys, CJ Vaidya, V Menon (2013) Brain hyperconnectivity in children with autism and its links to social deficits. Cell Rep 5(3):738–747 https://doi.org/10.1016/j.celrep.2013.10.001
32
R van den Brand, J Heutschi, Q Barraud, J DiGiovanna, K Bartholdi, M Huerlimann, L Friedli, I Vollenweider, EM Moraud, S Duiset al. (2012) Restoring voluntary control of locomotion after paralyzing spinal cord injury. Science 336(6085):1182–1185 https://doi.org/10.1126/science.1217416
33
CV Vorhees, MT Williams (2006) Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc 1(2):848–858 https://doi.org/10.1038/nprot.2006.116
34
S Whitfield-Gabrieli, HW Thermenos, S Milanovic, MT Tsuang, SV Faraone, RW McCarley, ME Shenton, AI Green, A Nieto-Castanon, P LaVioletteet al. (2009) Hyperactivity and hyperconnectivity of the default network in schizophrenia and in firstdegree relatives of persons with schizophrenia. Proc Natl Acad Sci USA 106(4):1279–1284 https://doi.org/10.1073/pnas.0809141106
35
K Yamanaka, N Sasaki, Y Fujita, A Kawamoto, KI Hirata, Y Okita (2016) Impact of acquired and innate immunity on spinal cord ischemia and reperfusion injury. Gen Thorac Cardiovasc Surg 64(5):251–259 https://doi.org/10.1007/s11748-016-0629-0
36
Z Yang, A Zhang, H Duan, S Zhang, P Hao, K Ye, YE Sun, X Li (2015) NT3-chitosan elicits robust endogenous neurogenesis to enable functional recovery after spinal cord injury. Proc Natl Acad Sci USA 112(43):13354–13359 https://doi.org/10.1073/pnas.1510194112
37
AM Yip, S Horvath (2007) Gene network interconnectedness and the generalized topological overlap measure. BMC Bioinform 8:22 https://doi.org/10.1186/1471-2105-8-22
38
Y Ziv, N Ron, O Butovsky, G Landa, E Sudai, N Greenberg, H Cohen, J Kipnis, M Schwartz (2006) Immune cells contribute to the maintenance of neurogenesis and spatial learning abilities in adulthood. Nat Neurosci 9(2):268–275 https://doi.org/10.1038/nn1629
