Human mesenchymal stem/stromal cells suppress spinal inflammation in mice with contribution of pituitary adenylate cyclase-activating polypeptide (PACAP)

Journal of Neuroinflammation

Seiji Shioda³  Takashi Atsumi⁶  Hitoshi Hashimoto²  Kenji Dohi⁵  Zhifang Xu³  Tomoya Nakamachi¹  Yutaka Hiraizumi⁴  Jun Watanabe³  Atsushi Sato⁶  Dandan Song³  Hirokazu Ohtaki³  Tomomi Tsumuraya⁶ 

[1] Laboratory of Regulatory Biology, Graduate School of Science and Engineering, University of Toyama, 3190 Gofuku, Toyama 930-8555, Toyama, Japan;Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita 565-0871, Osaka, Japan;Department of Anatomy, Showa University School of Medicine, 1-5-8 Hatanodai, Shinagawa-ku 142-8555, Tokyo, Japan;Department of Orthopedic Surgery, Showa University School of Medicine, 1-5-8 Hatanodai, Shinagawa-ku 142-8555, Tokyo, Japan;Department of Emergency Medicine, Tokyo Jikei University School of Medicine, 3-25-8 Nishishinnbashi, Minato-ku 105-8461, Tokyo, Japan;Department of Orthopedic Surgery, Showa University Fujigaoka Hospital, 1-30 Fujigaoka, Aoba-ku, Yokohama 227-8501, Kanagawa, Japan

关键词: Microglia;    Macrophage;    Pituitary adenylate cyclase-activating polypeptide (PACAP);    Human mesenchymal stem/stromal cells (hMSCs);    Spinal cord injury;   

Others  :  1133036
DOI  :  10.1186/s12974-015-0252-5

 received in 2014-11-07, accepted in 2015-01-21,  发布年份 2015

【 摘 要 】

Background

Adult human mesenchymal stem/stromal cells (hMSCs) from bone marrow have been reported to exhibit beneficial effects on spinal cord injury (SCI). A neuropeptide, pituitary adenylate cyclase-activating polypeptide (PACAP) is known to decrease neuronal cell death and inflammatory response after ischemia, SCI, and other neuronal disorders. Recently, we found that expression of the gene for mouse PACAP (Adcyap1) was greater in animals receiving hMSCs with neural injury such as ischemia. However, the association of PACAP with hMSCs to protect nerve cells against neural injuries is still unclear.

Methods

Wild-type and PACAP-gene-deficient (Adcyap1^+/−) mice were subjected to spinal cord transection, and hMSCs (5 × 10⁵ cells) were injected into the intervertebral spinal cord on day 1 post-operation (p.o.). Locomotor activity, injury volume, retention of hMSCs, mouse and human cytokine genes (which contribute to macrophage (MΦ) and microglial activation), and Adcyap1 were evaluated.

Results

hMSCs injected into wild-type mice improved locomotor activity and injury volume compared with vehicle-treated mice. In contrast, non-viable hMSCs injected into wild-type mice, and viable hMSCs injected into Adcyap1^+/− mice, did not. Wild-type mice injected with hMSCs exhibited increased Adcyap1 expression, and observed PACAP immunoreaction in neuron-like cells. Gene expression levels for IL-1, tumor necrosis factor α (TNFα), interleukin-10 (IL-10), and transforming growth factor β (TGFβ) decreased, while that for interleukin-4 (IL-4) increased, in hMSC-injected wild-type mice. In contrast, IL-1, TGFβ, and IL-4 gene expression levels were all abolished in hMSC-injected Adcyap1^+/− mice on day 7 post-operation. Moreover, the mice-implanted hMSCs increased an alternative activating macrophage/microglial marker, arginase activity. The human gene profile indicated that hMSCs upregulated the gene of IL-4 and growth factors which were reported to enhance Adcyap1 expression. Finally, we demonstrated that hMSCs express human ADCYAP1 and its receptor gene after the inflammation-related interferon-γ (IFNγ) in vitro.

Conclusions

These results suggest that hMSCs attenuate the deleterious effects of SCI by reducing associated inflammatory responses and enhancing IL-4 production. This effect could be mediated in part by cell-cell cross-talk involving the neuropeptide PACAP.

【 授权许可】

   
2015 Tsumuraya et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Nesathurai S: Steroids and spinal cord injury: revisiting the NASCIS 2 and NASCIS 3 trials. J Trauma. 1998, 45:1088-93.
• [2]Rolls A, Shechter R, Schwartz M: The bright side of the glial scar in CNS repair. Nat Rev Neurosci. 2009, 10:235-41.
• [3]Owen M, Friedenstein AJ: Stromal stem cells: marrow-derived osteogenic precursors. Ciba Found Symp. 1988, 136:42-60.
• [4]Prockop DJ: “Stemness” does not explain the repair of many tissues by mesenchymal stem/multipotent stromal cells (MSCs). Clin Pharmacol Ther. 2007, 82:241-3.
• [5]Wright KT, El Masri W, Osman A, Chowdhury J, Johnson WE: Concise review: bone marrow for the treatment of spinal cord injury: mechanisms and clinical applications. Stem Cells. 2011, 29:169-78.
• [6]Uccelli A, Milanese M, Principato MC, Morando S, Bonifacino T, Vergani L, Giunti D, Voci A, Carminati E, Giribaldi F, Caponnetto C, Bonanno G: Intravenous mesenchymal stem cells improve survival and motor function in experimental amyotrophic lateral sclerosis. Mol Med. 2012, 18:794-804.
• [7]Parr AM, Tator CH, Keating A: Bone marrow-derived mesenchymal stromal cells for the repair of central nervous system injury. Bone Marrow Transplant. 2007, 40:609-19.
• [8]Shi Y, Su J, Roberts AI, Shou P, Rabson AB, Ren G: How mesenchymal stem cells interact with tissue immune responses. Trends Immunol. 2012, 33:136-43.
• [9]Uccelli A, Moretta L, Pistoia V: Mesenchymal stem cells in health and disease. Nat Rev Immunol. 2008, 8:726-36.
• [10]Németh K, Leelahavanichkul A, Yuen PS, Mayer B, Parmelee A, Doi K, Robey PG, Leelahavanichkul K, Koller BH, Brown JM, Hu X, Jelinek I, Star RA, Mezey E: Bone marrow stromal cells attenuate sepsis via prostaglandin E(2)-dependent reprogramming of host macrophages to increase their interleukin-10 production. Nat Med. 2009, 15:42-9.
• [11]Ohtaki H, Ylostalo JH, Foraker JE, Robinson AP, Reger RL, Shioda S, Prockop DJ: Stem/progenitor cells from bone marrow decrease neuronal death in global ischemia by modulation of inflammatory/immune responses. Proc Natl Acad Sci U S A. 2008, 105:14638-43.
• [12]Gordon S: Alternative activation of macrophages. Nat Rev Immunol. 2003, 3:23-35.
• [13]Arimura A, Shioda S: Pituitary adenylate cyclase activating polypeptide (PACAP) and its receptors: neuroendocrine and endocrine interaction. Front Neuroendocrinol. 1995, 16:53-88.
• [14]Vaudry D, Gonzalez BJ, Basille M, Yon L, Fournier A, Vaudry H: Pituitary adenylate cyclase-activating polypeptide and its receptors: from structure to functions. Pharmacol Rev. 2000, 52:269-324.
• [15]Ohtaki H, Nakamachi T, Dohi K, Aizawa Y, Takaki A, Hodoyama K, Yofu S, Hashimoto H, Shintani N, Baba A, et al.: Pituitary adenylate cyclase-activating polypeptide (PACAP) decreases ischemic neuronal cell death in association with IL-6. Proc Natl Acad Sci U S A 2006, 103:7488-93.
• [16]Ohtaki H, Nakamachi T, Dohi K, Shioda S: Role of PACAP in ischemic neural death. J Mol Neurosci. 2008, 36:16-25.
• [17]Fang KM, Chen JK, Hung SC, Chen MC, Wu YT, Wu TJ, Lin HI, Chen CH, Cheng H, Yang CS, Tzeng SF: Effects of combinatorial treatment with pituitary adenylate cyclase activating peptide and human mesenchymal stem cells on spinal cord tissue repair. PLoS One. 2010, 5:e15299.
• [18]Reglodi D, Kiss P, Lubics A, Tamas A: Review on the protective effects of PACAP in models of neurodegenerative diseases in vitro and in vivo. Curr Pharm Des. 2011, 17:962-72.
• [19]Tsuchikawa D, Nakamachi T, Tsuchida M, Wada Y, Hori M, Farkas J, Yoshikawa A, Kagami N, Imai N, Shintani N, et al. Neuroprotective effect of endogenous PACAP on spinal cord injury. J Mol Neurosci. 2012; in press.
• [20]Tsuchida M, Nakamachi T, Sugiyama K, Tsuchikawa D, Watanabe J, Hori M, Yoshikawa A, Imai N, Kagami N, Matkovits A, et al.: PACAP Stimulates Functional Recovery after Spinal Cord Injury through Axonal Regeneration. J Mol Neurosci. 2014, 54:380-7.
• [21]Tan YV, Abad C, Lopez R, Dong H, Liu S, Lee A, Gomariz RP, Leceta J, Waschek JA: Pituitary adenylyl cyclase-activating polypeptide is an intrinsic regulator of Treg abundance and protects against experimental autoimmune encephalomyelitis. Proc Natl Acad Sci U S A. 2009, 106:2012-7.
• [22]Armstrong BD, Abad C, Chhith S, Cheung-Lau G, Hajji OE, Nobuta H, Waschek JA: Impaired nerve regeneration and enhanced neuroinflammatory response in mice lacking pituitary adenylyl cyclase activating peptide. Neuroscience. 2008, 151:63-73.
• [23]Hashimoto H, Shintani N, Tanaka K, Mori W, Hirose M, Matsuda T, Sakaue M, Miyazaki J, Niwa H, Tashiro F, et al.: Altered psychomotor behaviors in mice lacking pituitary adenylate cyclase-activating polypeptide (PACAP). Proc Natl Acad Sci U S A. 2001, 98:13355-60.
• [24]Sato A, Ohtaki H, Tsumuraya T, Song D, Ohara K, Asano M, Iwakura Y, Atsumi T, Shioda S: Interleukin-1 participates in the classical and alternative activation of microglia/macrophages after spinal cord injury. J Neuroinflammation. 2012, 9:65. BioMed Central Full Text
• [25]Song D, Ohtaki H, Tsumuraya T, Miyamoto K, Shibato J, Rakwal R, Xu Z, Hiraizumi Y, Inoue T, Shioda S: The anti-inflammatory property of human bone marrow-derived mesenchymal stem/stromal cells is preserved in late-passage cultures. J Neuroimmunol. 2013, 263:55-63.
• [26]Basso DM, Fisher LC, Anderson AJ, Jakeman LB, McTigue DM, Popovich PG: Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. J Neurotrauma. 2006, 23:635-59.
• [27]Suzuki R, Arata S, Nakajo S, Ikenaka K, Kikuyama S, Shioda S: Expression of the receptor for pituitary adenylate cyclase-activating polypeptide (PAC1-R) in reactive astrocytes. Brain Res Mol Brain Res. 2003, 115:10-20.
• [28]Nakamachi T, Tsuchida M, Kagami N, Yofu S, Wada Y, Hori M, Tsuchikawa D, Yoshikawa A, Imai N, Nakamura K, et al.: IL-6 and PACAP receptor expression and localization after global brain ischemia in mice. J Mol Neurosci. 2012, 48:518-25.
• [29]Choi H, Lee RH, Bazhanov N, Oh JY, Prockop DJ: Anti-inflammatory protein TSG-6 secreted by activated MSCs attenuates zymosan-induced mouse peritonitis by decreasing TLR2/NF-κB signaling in resident macrophages. Blood. 2011, 118:330-8.
• [30]Hori M, Nakamachi T, Rakwal R, Shibato J, Nakamura K, Wada Y, Tsuchikawa D, Yoshikawa A, Tamaki K, Shioda S: Unraveling the ischemic brain transcriptome in a permanent middle cerebral artery occlusion mouse model by DNA microarray analysis. Dis Model Mech. 2012, 5:270-83.
• [31]Hashimoto H, Hagihara N, Koga K, Yamamoto K, Shintani N, Tomimoto S, Mori W, Koyama Y, Matsuda T, Baba A: Synergistic induction of pituitary adenylate cyclase-activating polypeptide (PACAP) gene expression by nerve growth factor and PACAP in PC12 cells. J Neurochem. 2000, 74:501-7.
• [32]Glorioso C, Sabatini M, Unger T, Hashimoto T, Monteggia LM, Lewis DA, Mirnics K: Specificity and timing of neocortical transcriptome changes in response to BDNF gene ablation during embryogenesis or adulthood. Mol Psychiatry. 2006, 11:633-48.
• [33]Watanabe J, Shetty AK, Hattiangady B, Kim DK, Foraker JE, Nishida H, Prockop DJ: Administration of TSG-6 improves memory after traumatic brain injury in mice. Neurobiol Dis. 2013, 59:86-99.
• [34]Ha CM, Kang JH, Choi EJ, Kim MS, Park JW, Kim Y, Choi WS, Chun SY, Kwon HB, Lee BJ: Progesterone increases mRNA levels of pituitary adenylate cyclase-activating polypeptide (PACAP) and type I PACAP receptor (PAC(1)) in the rat hypothalamus. Brain Res Mol Brain Res. 2000, 78:59-68.
• [35]Munoz JR, Stoutenger BR, Robinson AP, Spees JL, Prockop DJ: Human stem/progenitor cells from bone marrow promote neurogenesis of endogenous neural stem cells in the hippocampus of mice. Proc Natl Acad Sci U S A. 2005, 102:18171-6.
• [36]Lee RH, Pulin AA, Seo MJ, Kota DJ, Ylostalo J, Larson BL, Semprun-Prieto L, Delafontaine P, Prockop DJ: Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6. Cell Stem Cell. 2009, 5:54-63.
• [37]Prockop DJ, Oh JY: Mesenchymal stem/stromal cells (MSCs): role as guardians of inflammation. Mol Ther. 2012, 20:14-20.
• [38]Martinez FO, Helming L, Gordon S: Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol. 2009, 27:451-83.
• [39]Gordon S, Martinez FO: Alternative activation of macrophages: mechanism and functions. Immunity. 2010, 32:593-604.
• [40]Foraker JE, Oh JY, Ylostalo JH, Lee RH, Watanabe J, Prockop DJ: Cross-talk between human mesenchymal stem/progenitor cells (MSCs) and rat hippocampal slices in LPS-stimulated cocultures: the MSCs are activated to secrete prostaglandin E2. J Neurochem. 2011, 119:1052-63.