Angiotensin II Regulates Mitochondrial mTOR Pathway Activity Dependent on Acyl-CoA Synthetase 4 in Adrenocortical Cells

Abstract

Two well-known protein complexes in mammalian cells, mTOR type 1 and type 2 (mTORC1/2) are involved in several cellular processes such as protein synthesis, cell proliferation and commonly dysregulated in cancer. An acyl-CoA synthetase type 4 (ACSL4) is one of the most recently mTORC1/2 regulators described, in breast cancer cells. The expression of ACSL4 is hormone-regulated in adrenocortical cells and required for steroid biosynthesis. mTORC1/2 have been reported to be crucial in the proliferation of human adrenocortical tumor cells H295R and interestingly reported at several subcellular locations, which has brought cell biology to the vanguard of the mTOR signaling field. In the present work, we study the regulation of mTORC1/2 activation by angiotensin II (Ang II) –the trophic hormone for adrenocortical cells–, the subcellular localization of mTORC1/2 signaling proteins and the role of ACSL4 in the regulation of this pathway, in H295R cells. Ang II promotes activation by phosphorylation of mTORC1/2 pathway proteins in a time-dependent manner. Mitochondrial pools of ribosomal protein S6, Akt in threonine 308 and serine 473 and Rictor are phosphorylated and activated. Glycogen synthase kinase type 3 (GSK3) is phosphorylated and inactivated in mitochondria, favoring mTORC1 activation. Epidermal growth factor, a classic mTORC1/2 activator, promoted unique activation kinetics of mTORC1/2 pathway, except for Akt phosphorylation. Here, we demonstrate that ACSL4 is necessary for mTORC1/2 effectors phosphorylation and H295R proliferation, triggered by Ang II. Ang II promotes activation of mitochondrial mTORC1/2 signaling proteins, through ACSL4, with a direct impact on adrenocortical cellular proliferation.

Acyl-CoA synthetase 4 modulates mitochondrial function in breast cancer cells

“…As mentioned above, our laboratory showed that overexpression of ACSL4 in MCF-7 cells results in mTORC1/2 and AKT activation, which was confirmed in MDA-MB-231 shACSL4 cells compared to their controls [ 10 ]. Moreover, activation of mTORC1/2 by angiotensin II is mediated by ACSL4 in adrenocortical human H295R cells [ 72 ]. In particular, mTORC1 is known to control cellular metabolism mainly by regulating the translation and transcription of metabolic genes, including peroxisome proliferator-activated receptor γ coactivator-1 α (PGC-1α), an upstream regulator of NRF-1/2 [ 73 ].…”

The ACSL4 Network Regulates Cell Death and Autophagy in Diseases

“…ACSL4 can also regulate intracellular autophagic signaling pathways, such as the AMPK and mTOR pathways, and thus promote or inhibit autophagy. For example, angiotensin II (Ang II) promotes the activation of the mitochondrial mTOR1/2 signaling protein and promotes the proliferation of human H295R adrenocortical cells in an ACSL4-dependent manner [ 108 ]. Similarly, ACSL4 promotes ferroptosis in SH-SY5Y cells through the AMPK/mTOR pathway, although the detailed mechanism remains unclear [ 109 ].…”