In 2016, the U.S. Food and Drug Administration warned that repeated or lengthy use of general anesthetics in children below the age of three might affect their brain development. This warning raises serious concerns regarding the safety of pediatric anesthesia. There are two main barriers in the research field of anesthetic-induced developmental neurotoxicity (AIDN): 1) So far, most of the evidence for AIDN was obtained from animal studies. The results from human studies remain inconclusive. 2) The mechanisms are largely unknown. The goal of this proposed study is to address both of these barriers. First, we established a new in vitro system of three-dimensional (3D) human mini brains using induced pluripotent stem cells for modeling human brain development. Human mini brains are more similar to developing human brains, both structurally and functionally, than the widely used 2D neurons. Thus, application of human mini brains in AIDN research field helps bridge the gap between the animal and human studies. Our preliminary data provided the first evidence showing that clinically relevant doses of either propofol or sevoflurane, two commonly used pediatric anesthetics, induced cell death in human mini brains. Second, we recently used an unbiased approach to screen the expression of 24,881 long non-coding RNAs (lncRNAs) and 35,923 messenger RNAs in neonatal mouse hippocampi. We discovered that the expression levels of the lncRNA AK156531 gene, and its nearby protein-coding gene Neuronal Per Arnt Sim domain protein 4 (NPAS4), were dramatically decreased following propofol exposure. One of the known functions of lncRNAs is to regulate their nearby gene expression. We found that knockdown of AK156531 decreased NPAS4 levels in both human mini brains and mouse brains, strongly suggesting that AK156531 might regulate NPAS4 expression. NPAS4 is involved in excitatory/inhibitory (E/I) balance, learning and memory, and neuroprotection. We also found that neonatal propofol exposure caused multiple adverse effects in mice (E/I imbalance, neuronal death, and impaired memory function). These exciting findings, combined with the reported function of NPAS4, suggest that the abnormally expressed AK156531 might directly contribute to AIDN. Thus, we propose to utilize AK156531 gain- and loss-of-function approaches to examine the role and mechanism of AK156531 in AIDN in mice, and to facilitate the translation of these findings to humans by using human mini brains. We hypothesize that downregulation of AK156531 contributes to E/I imbalance, neuronal death and cognitive dysfunction via NPAS4 signaling. For the first time in this field, human mini brains will be combined with AK156531 knockdown and overexpression mouse models to investigate the novel mechanisms of lncRNA involvement. This proposal is expected to provide new mechanistic insights into the neurodevelopmental consequences of pediatric anesthetic exposure. This will further aid in the development of more rational neuroprotective strategies related to pediatric anesthetic use, and movement towards a better assurance of safety for pediatric anesthesia use.

Public Health Relevance

The aim of this study is to dissect the novel molecular mechanisms of the long non-coding RNA AK156531 in anesthetic-induced developmental neurotoxicity (AIDN). Thus, we propose to utilize AK156531 gain- and loss- of-function approaches to examine the role and mechanism of AK156531 in AIDN in mice and to facilitate the translation of these findings to humans by using stem cell-derived developing 3D human mini brains. This proposal is expected to provide new mechanistic insights into the neurodevelopmental consequences of pediatric anesthetic exposure, further aiding in the development of more rational neuroprotective strategies related to pediatric anesthetic use, and moving towards a better assurance of safety for anesthetic use in young children.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM112696-07
Application #
9980423
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Justinova, Zuzana
Project Start
2014-05-15
Project End
2023-05-31
Budget Start
2020-06-01
Budget End
2021-05-31
Support Year
7
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Medical College of Wisconsin
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
937639060
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Logan, Sarah; Jiang, Congshan; Yan, Yasheng et al. (2018) Propofol Alters Long Non-Coding RNA Profiles in the Neonatal Mouse Hippocampus: Implication of Novel Mechanisms in Anesthetic-Induced Developmental Neurotoxicity. Cell Physiol Biochem 49:2496-2510
Yan, Yasheng; Qiao, Shigang; Kikuchi, Chika et al. (2017) Propofol Induces Apoptosis of Neurons but Not Astrocytes, Oligodendrocytes, or Neural Stem Cells in the Neonatal Mouse Hippocampus. Brain Sci 7:
Liu, Yanan; Yan, Yasheng; Inagaki, Yasuyoshi et al. (2017) Insufficient Astrocyte-Derived Brain-Derived Neurotrophic Factor Contributes to Propofol-Induced Neuron Death Through Akt/Glycogen Synthase Kinase 3?/Mitochondrial Fission Pathway. Anesth Analg 125:241-254
Sedlic, Filip; Muravyeva, Maria Y; Sepac, Ana et al. (2017) Targeted Modification of Mitochondrial ROS Production Converts High Glucose-Induced Cytotoxicity to Cytoprotection: Effects on Anesthetic Preconditioning. J Cell Physiol 232:216-24
Bosnjak, Zeljko J; Logan, Sarah; Liu, Yanan et al. (2016) Recent Insights Into Molecular Mechanisms of Propofol-Induced Developmental Neurotoxicity: Implications for the Protective Strategies. Anesth Analg 123:1286-1296
Canfield, Scott G; Zaja, Ivan; Godshaw, Brian et al. (2016) High Glucose Attenuates Anesthetic Cardioprotection in Stem-Cell-Derived Cardiomyocytes: The Role of Reactive Oxygen Species and Mitochondrial Fission. Anesth Analg 122:1269-79
Olson, Jessica M; Yan, Yasheng; Bai, Xiaowen et al. (2015) Up-regulation of microRNA-21 mediates isoflurane-induced protection of cardiomyocytes. Anesthesiology 122:795-805
Twaroski, Danielle; Bosnjak, Zeljko J; Bai, Xiaowen (2015) MicroRNAs: New Players in Anesthetic-Induced Developmental Neurotoxicity. Pharm Anal Acta 6:357
Kikuchi, Chika; Bienengraeber, Martin; Canfield, Scott et al. (2015) Comparison of Cardiomyocyte Differentiation Potential Between Type 1 Diabetic Donor- and Nondiabetic Donor-Derived Induced Pluripotent Stem Cells. Cell Transplant 24:2491-504
Twaroski, Danielle M; Yan, Yasheng; Zaja, Ivan et al. (2015) Altered Mitochondrial Dynamics Contributes to Propofol-induced Cell Death in Human Stem Cell-derived Neurons. Anesthesiology 123:1067-83

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