Gene Delivery of Adenoviral-TMBIM6 Vector Protects the Neonatal Brain after Hypoxic-Ischemic Injury

<p>Neonatal hypoxia ischemia (HI) is an injury caused to the immature brain due to reduced cerebral blood flow which is associated with life-long neurological impairments. HI causes oxidative protein folding in the endoplasmic reticulum (ER), which results in ER stress. Generation of reactive oxygen species (ROS), from cytochrome P450 members (CYP) and NADPH-P450 reductases (NPR), in combination with activation of the unfolded protein response (UPR) are two major consequences of ER stress that cause oxidative damage and cell death. Herein we identified the role of Bax Inhibitor-1 (BI-1), an evolutionary conserved protein encoded by the Transmembrane Bax inhibitor Motif Containing 6 (TMBIM6) gene, in protection from ER stress after HI injury in the neonatal rat. As BI-1 has multimodal properties that can target a wide array of pathophysiological consequences after injury, our main objective was to evaluate BI-1’s protective mechanisms by overexpressing it, using viral-mediated gene delivery of human adenoviral-TMBIM6 (Ad-TMBIM6) vector, in an in vitro and in vivo model of HI. Following the Rice-Vannuci model, ten-day old (P10) rat pups underwent rightunilateral carotid artery ligation followed by 1.5h of hypoxia. Our results showed that overexpression of BI-1 ameliorated short and long-term deficits following HI. There was </p> <p>a significant reduction in percent infarcted area which was linked with attenuation of apoptosis, reduction of ROS accumulation and inhibition of pro-inflammatory mediators. BI-1 mediated protection was observed to be via both inhibition of IRE1α signaling, a stress sensor protein part of the UPR response, and P4502E1 activity, a major contributor of ROS generation. Attenuation of apoptosis was associated with a decrease in IRE1α- XBP1 levels while decreased levels of P4502E1 coupled with upregulation of Nrf-2 and HO-1, anti-oxidant enzymes, were correlated with a reduction in ROS generation and inflammation. In conclusion we established that overexpression of BI-1 attenuated the morphological and neurological consequences post HI via inhibition of ER stress induced pathways. This new finding may help to provide a basis for BI-1 as a potential therapeutic target.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Basic Sciences

Year

2018

Author dc:creator

• Doycheva, Desislava

Contributors dc:contributor

• Zhang, John H.
• Angeles, Danilyn
• Brantely, Eileen
• Tang, Jiping
• Zhang, Lubo

• Medical Physiology
• Medicine and Health Sciences
• Gene Therapy; Hypoxia-Ischemia; Brain; Fetal Hypoxia; Infant; Newborn; Fetal Brain
• Neonatal Hypoxia-Ischemia; Cerebral Blood Flow; Neurological Impairment

Statement dc:rights

• This title appears here courtesy of the author, who has granted Loma Linda University a limited, non-exclusive right to make this publication available to the public. The author retains all other copyrights.

Language dc:language

English

Repository record dc:identifier

https://scholarsrepository.llu.edu/etd/509

OAI identifier oai:identifier

oai:scholarsrepository.llu.edu:etd-1520