Abstract
Intracerebral hemorrhage (ICH) is a devastating form of stroke affecting millions of people worldwide. Parenchymal hematoma triggers a series of reactions leading to primary and secondary brain injuries and permanent neurological deficits. Microglia and macrophages carry out hematoma clearance, thereby facilitating functional recovery after ICH. Here, we elucidate a pivotal role for the interleukin (IL)-4)/signal transducer and activator of transcription 6 (STAT6) axis in promoting long-term recovery in both bloodand collagenase-injection mouse models of ICH, through modulation of microglia/macrophage functions. In both ICH models, STAT6 was activated in microglia/macrophages (i.e., enhanced expression of phospho-STAT6 in Iba1+cells). Intranasal delivery of IL- 4 nanoparticles after ICH hastened STAT6 activation and facilitated hematoma resolution. IL-4 treatment improved long-term functional recovery in young and aged male and young female mice. In contrast, STAT6 knockout (KO) mice exhibited worse outcomes than WT mice in both ICH models and were less responsive to IL-4 treatment. The construction of bone marrow chimera mice demonstrated that STAT6 KO in either the CNS or periphery exacerbated ICH outcomes. STAT6 KO impaired the capacity of phagocytes to engulf red blood cells in the ICH brain and in primary cultures. Transcriptional analyses identified lower level of IL- 1 receptor-like 1 (ST2) expression in microglia/macrophages of STAT6 KO mice after ICH. ST2 KO diminished the beneficial effects of IL-4 after ICH. Collectively, these data confirm the importance of IL-4/STAT6/ST2 signaling in hematoma resolution and functional recovery after ICH. Intranasal IL-4 treatment warrants further investigation as a clinically feasible therapy for ICH.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 32679-32690 |
| Number of pages | 12 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 117 |
| Issue number | 51 |
| DOIs | |
| State | Published - Dec 22 2020 |
Keywords
- Bone marrow chimera
- Intracerebral hemorrhage
- Macrophages
- Microglia
- Phagocytosis
ASJC Scopus subject areas
- General
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IL-4/STAT6 signaling facilitates innate hematoma resolution and neurological recovery after hemorrhagic stroke in mice. / Xu, Jing; Chen, Zhouqing; Yu, Fang; Liu, Huan; Ma, Cheng; Xie, Di; Hu, Xiaoming; Leak, Rehana K.; Chou, Sherry H.Y.; R. Anne Stetler, Anne Stetler; Shi, Yejie; Chen, Jun; Bennett, Michael V.L.; Chen, Gang.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, No. 51, 22.12.2020, p. 32679-32690.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - IL-4/STAT6 signaling facilitates innate hematoma resolution and neurological recovery after hemorrhagic stroke in mice
AU - Xu, Jing
AU - Chen, Zhouqing
AU - Yu, Fang
AU - Liu, Huan
AU - Ma, Cheng
AU - Xie, Di
AU - Hu, Xiaoming
AU - Leak, Rehana K.
AU - Chou, Sherry H.Y.
AU - R. Anne Stetler, Anne Stetler
AU - Shi, Yejie
AU - Chen, Jun
AU - Bennett, Michael V.L.
AU - Chen, Gang
N1 - Funding Information: 16. T. Garton, R. F. Keep, Y. Hua, G. Xi, Brain iron overload following intracranial hae-morrhage. Stroke Vasc. Neurol. 1, 172–184 (2016). Materials and Methods Key resources that are essential to reproduce the results are in SI Appendix, Table S1. ICH was induced in adult male or female mice (10-to 12-wk old, 25 to 30 g) or aged male (18-mo old) mice. All animal procedures were approved by the University of Pittsburgh Institutional Animal Care and Use Committee and performed in accordance with the Guide for the Care and Use of Laboratory Animals (51). All efforts were made to minimize animal suffering and the number of animals used. Surgeries and all outcome assessments were performed by investigators blinded to mouse genotype and experimental group assignments. All statistics are summarized in SI Appendix, Table S2. For further methodological details, please consult SI Appendix. Data Availability. All study data are included in the article and supporting information. ACKNOWLEDGMENTS. We thank T. Kevin Hitchens and Lesley M. Foley for assistance with the MRI experiments and Patricia Strickler for administrative support. The ST2 knockout breeders were a gift from A. McKenzie (Medical Research Council as part of UK Research and Innovation). This project was supported by the University of Pittsburgh School of Medicine. J.C. is the Richard King Mellon Professor of Neurology and a recipient of a VA Senior Research Career Scientist Award. M.V.L.B. is the Sylvia and Robert S. Olnick Professor of Neuroscience. 17. A. L. A. Garton, V. P. Gupta, B. R. Christophe, E. S. Connolly, Jr, Biomarkers of func-tional outcome in intracerebral hemorrhage: Interplay between clinical metrics, CD163, and ferritin. J. Stroke Cerebrovasc. Dis. 26, 1712–1720 (2017). 18. J. Wang, S. Doré, Heme oxygenase-1 exacerbates early brain injury after intracerebral haemorrhage. Brain 130, 1643–1652 (2007). 19. M. Xue, M. R. Del Bigio, Intracerebral injection of autologous whole blood in rats: Time course of inflammation and cell death. Neurosci. Lett. 283, 230–232 (2000). 20. R. A. Taylor et al., TGF-β1 modulates microglial phenotype and promotes recovery after intracerebral hemorrhage. J. Clin. Invest. 127, 280–292 (2017). 21. R. M. Dahnovici et al., Microscopic aspects of macrophage system cells in hemorrhagic stroke in humans. Rom. J. Morphol. Embryol. 52, 1249–1253 (2011). 22. X. Zhao, J. Grotta, N. Gonzales, J. Aronowski, Hematoma resolution as a therapeutic target: The role of microglia/macrophages. Stroke 40 (suppl.(3), S92–S94 (2009). 23. X. Zhao et al., Hematoma resolution as a target for intracerebral hemorrhage treatment: Role for peroxisome proliferator-activated receptor gamma in microglia/ macrophages. Ann. Neurol. 61, 352–362 (2007). 24. K. Vaibhav et al., Remote ischemic post-conditioning promotes hematoma resolution via AMPK-dependent immune regulation. J. Exp. 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Buckley et al., Increased susceptibility of ST2-deficient mice to polymicrobial sepsis is associated with an impaired bactericidal function. J. Immunol. 187, 4293–4299 (2011). 32. C. L. MacLellan et al., Intracerebral hemorrhage models in rat: Comparing collagenase to blood infusion. J. Cereb. Blood Flow Metab. 28, 516–525 (2008). 33. A. R. Saand, F. Yu, J. Chen, S. H. Chou, Systemic inflammation in hemorrhagic strokes—A novel neurological sign and therapeutic target? J. Cereb. Blood Flow Metab. 39, 959–988 (2019). 34. C. F. Chang et al., Erythrocyte efferocytosis modulates macrophages towards recovery after intracerebral hemorrhage. J. Clin. Invest. 128, 607–624 (2018). 35. L. Garcia-Bonilla et al., Spatio-temporal profile, phenotypic diversity, and fate of recruited monocytes into the post-ischemic brain. J. Neuroinflammation 13, 285 (2016). 36. J. Yang et al., Interleukin-4 ameliorates the functional recovery of intracerebral hemorrhage through the alternative activation of microglia/macrophage. Front. Neurosci. 10, 61 (2016). 37. I. D. Vainchtein et al., Astrocyte-derived interleukin-33 promotes microglial synapse engulfment and neural circuit development. Science 359, 1269–1273 (2018). 38. A. K. Fu et al., IL-33 ameliorates Alzheimer’s disease-like pathology and cognitive decline. Proc. Natl. Acad. Sci. U.S.A. 113, E2705–E2713 (2016). NEUROSCIENCE Downloaded at Elsevier Science London on December 23, 2020 39. Y. Gao et al., IL-33 exerts neuroprotective effect in mice intracerebral hemorrhage model through suppressing inflammation/apoptotic/autophagic pathway. Mol. Neu-robiol. 54, 3879–3892 (2017). 40. R. K. Sheean et al., Effect of thymic stimulation of CD4+ T cell expansion on disease onset and progression in mutant SOD1 mice. J. Neuroinflammation 12, 40 (2015). 41. K. Mittal et al., CD4 T cells induce A subset of MHCII-expressing microglia that at-tenuates Alzheimer pathology. iScience 16, 298–311 (2019). 42. J. T. Walsh et al., MHCII-independent CD4+ T cells protect injured CNS neurons via IL-4. J. Clin. Invest. 125, 2547 (2015). 43. E. Maier et al., Inhibition of suppressive T cell factor 1 (TCF-1) isoforms in naive CD4+ T cells is mediated by IL-4/STAT6 signaling. J. Biol. Chem. 286, 919–928 (2011). 44. H. Shen et al., Recirculating Th2 cells induce severe thymic dysfunction via IL-4/STAT6 signaling pathway. Biochem. Biophys. Res. Commun. 501, 320–327 (2018). 45. Q. Li et al., Neuroprotection of brain-permeable iron chelator VK-28 against intra-cerebral hemorrhage in mice. J. Cereb. Blood Flow Metab. 37, 3110–3123 (2017). 46. W. Ni et al., Deferoxamine reduces intracerebral hemorrhage-induced white matter damage in aged rats. Exp. Neurol. 272, 128–134 (2015). 47. X. Duan, Z. Wen, H. Shen, M. Shen, G. Chen, Intracerebral hemorrhage, oxida-tive stress, and antioxidant therapy. Oxid. Med. Cell. Longev. 2016, 1203285 (2016). 48. M. Selim et al.; i-DEF Investigators, Deferoxamine mesylate in patients with intrace-rebral haemorrhage (i-DEF): A multicentre, randomised, placebo-controlled, double-blind phase 2 trial. Lancet Neurol. 18, 428–438 (2019). 49. X. Zhao et al., Neutrophil polarization by IL-27 as a therapeutic target for intrace-rebral hemorrhage. Nat. Commun. 8, 602 (2017). 50. S. Cunha, M. H. Amaral, J. M. S. Lobo, A. C. Silva, Lipid nanoparticles for nasal/in-tranasal drug delivery. Crit. Rev. Ther. Drug Carrier Syst. 34, 257–282 (2017). 51. National Research Council, Guide for the Care and Use of Laboratory Animals (Na-tional Academies Press, Washington, DC, ed. 8, 2011). Funding Information: ACKNOWLEDGMENTS. We thank T. Kevin Hitchens and Lesley M. Foley for assistance with the MRI experiments and Patricia Strickler for administrative support. The ST2 knockout breeders were a gift from A. McKenzie (Medical Research Council as part of UK Research and Innovation). This project was supported by the University of Pittsburgh School of Medicine. J.C. is the Richard King Mellon Professor of Neurology and a recipient of a VA Senior Research Career Scientist Award. M.V.L.B. is the Sylvia and Robert S. Olnick Professor of Neuroscience.
PY - 2020/12/22
Y1 - 2020/12/22
N2 - Intracerebral hemorrhage (ICH) is a devastating form of stroke affecting millions of people worldwide. Parenchymal hematoma triggers a series of reactions leading to primary and secondary brain injuries and permanent neurological deficits. Microglia and macrophages carry out hematoma clearance, thereby facilitating functional recovery after ICH. Here, we elucidate a pivotal role for the interleukin (IL)-4)/signal transducer and activator of transcription 6 (STAT6) axis in promoting long-term recovery in both bloodand collagenase-injection mouse models of ICH, through modulation of microglia/macrophage functions. In both ICH models, STAT6 was activated in microglia/macrophages (i.e., enhanced expression of phospho-STAT6 in Iba1+cells). Intranasal delivery of IL- 4 nanoparticles after ICH hastened STAT6 activation and facilitated hematoma resolution. IL-4 treatment improved long-term functional recovery in young and aged male and young female mice. In contrast, STAT6 knockout (KO) mice exhibited worse outcomes than WT mice in both ICH models and were less responsive to IL-4 treatment. The construction of bone marrow chimera mice demonstrated that STAT6 KO in either the CNS or periphery exacerbated ICH outcomes. STAT6 KO impaired the capacity of phagocytes to engulf red blood cells in the ICH brain and in primary cultures. Transcriptional analyses identified lower level of IL- 1 receptor-like 1 (ST2) expression in microglia/macrophages of STAT6 KO mice after ICH. ST2 KO diminished the beneficial effects of IL-4 after ICH. Collectively, these data confirm the importance of IL-4/STAT6/ST2 signaling in hematoma resolution and functional recovery after ICH. Intranasal IL-4 treatment warrants further investigation as a clinically feasible therapy for ICH.
AB - Intracerebral hemorrhage (ICH) is a devastating form of stroke affecting millions of people worldwide. Parenchymal hematoma triggers a series of reactions leading to primary and secondary brain injuries and permanent neurological deficits. Microglia and macrophages carry out hematoma clearance, thereby facilitating functional recovery after ICH. Here, we elucidate a pivotal role for the interleukin (IL)-4)/signal transducer and activator of transcription 6 (STAT6) axis in promoting long-term recovery in both bloodand collagenase-injection mouse models of ICH, through modulation of microglia/macrophage functions. In both ICH models, STAT6 was activated in microglia/macrophages (i.e., enhanced expression of phospho-STAT6 in Iba1+cells). Intranasal delivery of IL- 4 nanoparticles after ICH hastened STAT6 activation and facilitated hematoma resolution. IL-4 treatment improved long-term functional recovery in young and aged male and young female mice. In contrast, STAT6 knockout (KO) mice exhibited worse outcomes than WT mice in both ICH models and were less responsive to IL-4 treatment. The construction of bone marrow chimera mice demonstrated that STAT6 KO in either the CNS or periphery exacerbated ICH outcomes. STAT6 KO impaired the capacity of phagocytes to engulf red blood cells in the ICH brain and in primary cultures. Transcriptional analyses identified lower level of IL- 1 receptor-like 1 (ST2) expression in microglia/macrophages of STAT6 KO mice after ICH. ST2 KO diminished the beneficial effects of IL-4 after ICH. Collectively, these data confirm the importance of IL-4/STAT6/ST2 signaling in hematoma resolution and functional recovery after ICH. Intranasal IL-4 treatment warrants further investigation as a clinically feasible therapy for ICH.
KW - Bone marrow chimera
KW - Intracerebral hemorrhage
KW - Macrophages
KW - Microglia
KW - Phagocytosis
UR - http://www.scopus.com/inward/record.url?scp=85098187763&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85098187763&partnerID=8YFLogxK
U2 - 10.1073/pnas.2018497117
DO - 10.1073/pnas.2018497117
M3 - Article
C2 - 33293423
AN - SCOPUS:85098187763
VL - 117
SP - 32679
EP - 32690
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 51
ER -