Pulmonary arterial hypertension (PAH), a progressive fatal disease, manifests by vascular remodeling of pulmonary arteries (PA), elevated right ventricular afterload, right heart failure and death. Enhanced proliferation and impaired apoptosis of pulmonary arterial vascular smooth muscle cells (PAVSMC) are key pathophysiological components of vascular remodeling in PAH, the molecular mechanisms of which are not fully understood. This proposal focuses on two functionally distinct complexes of mammalian target of rapamycin (mTOR), mTORC1 and mTORC2, as novel regulators of PAVSMC metabolism, growth and survival in PAH. By using PAVSMC from subjects with idiopathic PAH (human PAH PAVSMC) and unused donor lungs, we show that cultured human PAH PAVSMC retain molecular and cellular abnormalities reported in PAH lungs in vivo, such as elevated proliferation, survival, and altered cellular ATP levels due to glycolytic metabolism that provides a unique tool for translational mechanistic studies. Our published study demonstrates that increased PAVSMC proliferation under chronic hypoxia requires activation of both mTORC1 and mTORC2. Our preliminary data show that mTORC1 and mTORC2 signaling pathways are up-regulated in vivo and in vitro in PAVSMC from small PA from subjects with idiopathic PAH and from rats with chronic hypoxia-induced pulmonary vascular remodeling. Our data also show that both mTORC1 and mTORC2 promote proliferation, but only mTORC2 modulates cellular energy levels and cell survival and suggest that inhibition of both mTORC1 and mTORC2 is required to suppress growth, proliferation and promote apoptosis in human PAH PAVSMC. Based on published studies and our data, we hypothesize that increased proliferation and survival of PAVSMC in PAH requires activation of both mTORC1 (promoting protein synthesis and cell growth) and mTORC2 (activating Akt, increasing cellular energy levels, down-regulating AMPK and increasing cell survival). We also propose that targeting mTOR in both mTORC1 and mTORC2 is necessary to inhibit PAVSMC growth, proliferation, promote apoptosis, and prevent or abrogate pulmonary vascular remodeling in PAH. To test this hypothesis, in Aim 1, activation of mTORC1 and mTORC2 signaling pathways will be critically tested using lung tissue samples from PAH patients and healthy donors and cultured PAVSMC from idiopathic PAH patients and unused donor lungs;
in Aim 2, we will determine whether mTORC1 and mTORC2 employ differential molecular mechanisms to modulate growth, proliferation and survival of human PAH PAVSMC. Specific roles of mTORC1 and mTORC2 in regulating cell growth, cellular ATP levels and apoptosis will be examined;
in Aim 3, we will evaluate whether suppression of both mTORC1 and mTORC2 inhibits growth, cellular energy levels, proliferation and induces apoptosis in vitro in human PAH PAVSMC and prevents or abrogates pulmonary vascular remodeling in vivo in a rat chronic hypoxia model of PH. Proposed studies will define the role of mTOR in regulating energy levels, growth, proliferation and survival of PAH PAVSMC and will explore whether mTOR could serve as a potential molecular target for treatment of human PAH.
These studies will define the specific roles of mTORC1 and mTORC2 in regulating energy levels, growth, proliferation and survival of PAVSMC in pulmonary arterial hypertension (PAH);will provide insights into the molecular mechanisms that are important in pathogenesis of PAH;and will explore whether mTOR could serve as a target molecule to inhibit PAVSMC proliferation, induce apoptosis, and prevent or abrogate pulmonary vascular remodeling in pulmonary hypertension.
|Goncharov, Dmitry A; Kudryashova, Tatiana V; Ziai, Houman et al. (2014) Mammalian target of rapamycin complex 2 (mTORC2) coordinates pulmonary artery smooth muscle cell metabolism, proliferation, and survival in pulmonary arterial hypertension. Circulation 129:864-74|
|Goncharova, Elena A (2013) mTOR and vascular remodeling in lung diseases: current challenges and therapeutic prospects. FASEB J 27:1796-807|