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Neurotherapeutics

, Volume 16, Issue 1, pp 231–243 | Cite as

Evaluation of 3K3A-Activated Protein C to Treat Neonatal Hypoxic Ischemic Brain Injury in the Spiny Mouse

  • Stacey J. ElleryEmail author
  • Madeleine G. Goss
  • Nadine Brew
  • Hayley Dickinson
  • Nadia Hale
  • Domenic A. LaRosa
  • David W. Walker
  • Flora Y. Wong
Original Article

Abstract

Neonatal hypoxic ischemic encephalopathy (HIE) resulting from intrapartum asphyxia is a global problem that causes severe disabilities and up to 1 million deaths annually. A variant form of activated protein C, 3K3A-APC, has cytoprotective properties that attenuate brain injury in models of adult stroke. In this study, we compared the ability of 3K3A-APC and APC (wild-type (wt)) to attenuate neonatal brain injury, using the spiny mouse (Acomys cahirinus) model of intrapartum asphyxia. Pups were delivered at 38 days of gestation (term = 39 days), with an intrapartum hypoxic insult of 7.5 min (intrapartum asphyxia cohort), or immediate removal from the uterus (control cohort). After 1 h, pups received a subcutaneous injection of 3K3A-APC or wild-type APC (wtAPC) at 7 mg/kg, or vehicle (saline). At 24 h of age, pups were killed and brain tissue was collected for measurement of inflammation and cell death using RT-qPCR and histopathology. Intrapartum asphyxia increased weight loss, inflammation, and apoptosis/necrosis in the newborn brain. 3K3A-APC administration maintained body weight and ameliorated an asphyxia-induced increase of TGFβ1 messenger RNA expression in the cerebral cortex, immune cell aggregation in the corpus callosum, and cell death in the deep gray matter and hippocampus. In the cortex, 3K3A-APC appeared to exacerbate the immune response to the hypoxic ischemic insult. While wtAPC reduced cell death in the corpus callosum and hippocampus following intrapartum asphyxia, it increased markers of neuro-inflammation and cell death in control pups. These findings suggest 3K3A-APC administration may be a useful therapy to reduce cell death and neonatal brain injury associated with HIE.

Keywords

Birth Intrapartum asphyxia Brain injury Neuro-inflammation Apoptosis 

Notes

Acknowledgments

The authors would like to thank ZZ Biotech LLC (a biotechnology company with a mission to develop APC and its functional mutants for the treatment of stroke and other neurological disorders) for providing the 3K3A-APC used in this study. We also thank Ms. Mina Khalaji for her technical assistance in the laboratory, and the Hudson Institute of Medical Research’s Genomics and Histology Platforms. This work was supported by the Victorian Government’s Operational Infrastructure Support Program to the Hudson Institute of Medical Research and by the Phyllis Connor Memorial Trust. During these studies, SJE was supported by an NHMRC Early Career Fellowship; HD was supported by an NHMRC Career Development Fellowship; DWW was a senior scientist at The Ritchie Centre at the time of this study and is now a program leader in Neurodevelopment in Health and Disease (NDHD) at RMIT University, Melbourne; and FYW is supported by the NHMRC/Cerebral Palsy Alliance Career Development Fellowship.

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Supplementary material

13311_2018_661_MOESM1_ESM.pdf (498 kb)
ESM 1 (PDF 498 kb)

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Copyright information

© The American Society for Experimental NeuroTherapeutics, Inc. 2018

Authors and Affiliations

  1. 1.The Ritchie CentreHudson Institute of Medical ResearchClaytonAustralia
  2. 2.Department of Obstetrics and GynaecologyMonash UniversityClaytonAustralia
  3. 3.Women and Infants Hospital, Alpert Medical SchoolBrown UniversityProvidenceUSA
  4. 4.School of Health and Biomedical SciencesRMIT UniversityMelbourneAustralia
  5. 5.Department of PaediatricsMonash UniversityClaytonAustralia
  6. 6.Monash NewbornMonash Medical CentreClaytonAustralia

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