MTMR14 protects against cerebral stroke through suppressing PTEN-regulated autophagy

Abstract

The phosphoinositide phosphatase, myotubularinrelated protein 14 (MTMR14), plays a critical role in the regulating autophagy. However, its functional contribution to neuronal autophagy is still unclear. In the present study, we attempted to explore the effects of MTMR14 on ischemic stroke progression, as well as the underlying molecular mechanisms. Oxygen-glucose deprivation/reoxygenation (OGDR)-induced primary cortical neurons and pheochromocytoma (PC12) cells, and middle cerebral artery occlusion (MCAO)-operated mice were used to establish cerebral ischemia/reperfusion (I/R) injury in vitro and in vivo, respectively. OGDR treatment markedly decreased the expression of MTMR14 expression from mRNA and protein levels in the cultured primary neurons and PC12 cells. Functional analysis showed that OGDR-reduced cell viability was further accelerated by MTMR14 knockdown. On the contrary, MTMR14 over-expression significantly rescued the cell survival in OGDR-exposed cells. Moreover, autophagic markers including LC3BII and Beclin 1 were highly up-regulated in OGDR-incubated neurons and PC12 cells, while being further exacerbated by MTMR14 deletion. However, promoting MTMR14 dramatically alleviated LC3BII and Beclin 1 expression levels stimulated by OGDR. Importantly, we found that MTMR14-regulated autophagy was through its interactions with phosphatase and tensin homolog (PTEN). MTMR14 negatively modulated PTEN protein expression levels in OGDR-exposed cells. In vivo, MCAO-operated mice exhibited significantly reduced expression of MTMR14 in the ischemic penumbra tissues. After MCAO operation, MTMR14 over-expression effectively reduced infarct volume and neurological deficits scores, along with decreased activation of LC3B in neurons. Consistently, MCAO-increased PTEN, LC3BII and Beclin 1 were repressed by MTMR14 in mice. An interaction between MTMR14 and PTEN in response to MCAO was confirmed in vivo. Together, these results indicated the neuroprotective effects of MTMR14 on modulating PTEN-dependent excessive autophagy during cerebral I/R injury. Thus, targeting MTMR14 may provide feasible therapy for ischemic stroke onset and progression.

Keywords: Autophagy; Cerebral ischemia/reperfusion (I/R); LC3B; MTMR14; PTEN.