Amphetamines promote mitochondrial dysfunction and DNA damage in pulmonary hypertension
Pin-I Chen, Aiqin Cao, Kazuya Miyagawa, Nancy F. Tojais, Jan K. Hennigs, Caiyun G. Li, Nathaly M. Sweeney, Audrey S. Inglis, Lingli Wang, Dan Li, Matthew Ye, Brian J. Feldman, Marlene Rabinovitch
Pin-I Chen, Aiqin Cao, Kazuya Miyagawa, Nancy F. Tojais, Jan K. Hennigs, Caiyun G. Li, Nathaly M. Sweeney, Audrey S. Inglis, Lingli Wang, Dan Li, Matthew Ye, Brian J. Feldman, Marlene Rabinovitch
View: Text | PDF
Research Article Cell biology Vascular biology

Amphetamines promote mitochondrial dysfunction and DNA damage in pulmonary hypertension

Abstract

Amphetamine (AMPH) or methamphetamine (METH) abuse can cause oxidative damage and is a risk factor for diseases including pulmonary arterial hypertension (PAH). Pulmonary artery endothelial cells (PAECs) from AMPH-associated-PAH patients show DNA damage as judged by γH2AX foci and DNA comet tails. We therefore hypothesized that AMPH induces DNA damage and vascular pathology by interfering with normal adaptation to an environmental perturbation causing oxidative stress. Consistent with this, we found that AMPH alone does not cause DNA damage in normoxic PAECs, but greatly amplifies DNA damage in hypoxic PAECs. The mechanism involves AMPH activation of protein phosphatase 2A, which potentiates inhibition of Akt. This increases sirtuin 1, causing deacetylation and degradation of HIF1α, thereby impairing its transcriptional activity, resulting in a reduction in pyruvate dehydrogenase kinase 1 and impaired cytochrome c oxidase 4 isoform switch. Mitochondrial oxidative phosphorylation is inappropriately enhanced and, as a result of impaired electron transport and mitochondrial ROS increase, caspase-3 is activated and DNA damage is induced. In mice given binge doses of METH followed by hypoxia, HIF1α is suppressed and pulmonary artery DNA damage foci are associated with worse pulmonary vascular remodeling. Thus, chronic AMPH/METH can induce DNA damage associated with vascular disease by subverting the adaptive responses to oxidative stress.

Authors

Pin-I Chen, Aiqin Cao, Kazuya Miyagawa, Nancy F. Tojais, Jan K. Hennigs, Caiyun G. Li, Nathaly M. Sweeney, Audrey S. Inglis, Lingli Wang, Dan Li, Matthew Ye, Brian J. Feldman, Marlene Rabinovitch

×

Figure 3

Amphetamine exaggerates hypoxia-induced DNA damage in a p-Akt– and caspase-3–dependent manner.

Options: View larger image (or click on image) Download as PowerPoint
Amphetamine exaggerates hypoxia-induced DNA damage in a p-Akt– and caspa...
(A) Pulmonary artery endothelial cells (PAECs) were treated daily for 3 days with the indicated 0.5 mM amphetamine (AMPH), and then cultured with vehicle (Veh) or AMPH under 0.5% O2 hypoxia (Hx) or normoxia (Nx) for 48 hours, immunoblotted for p-Akt, cleaved caspase-3 (cCasp3), and β-actin (loading control). (B) PAECs were transfected with vector (Vec) or HA-tagged myristoylated Akt1 (myr-Akt) construct. Twenty-four hours after transfection, cells were treated with AMPH and Hx/Nx as in A. Cell lysates were immunoblotted for γH2AX, cCasp3, β-actin (for normalization), and HA, to verify the expression of HA-myr-Akt. (C) PAECs were treated as in A together with the indicated concentrations of zVAD-FMK, and lysates were immunoblotted for γH2AX, cCasp3, β-actin (loading control), and p-Akt. Dot plots represent mean ± SEM, n = 3–4. (A–C) *P < 0.05, **P < 0.005, ****P < 0.0001 vs. Nx+Veh; #P < 0.05, ##P < 0.005, ####P < 0.0001 vs. Hx+Veh or Vec+Hx+Veh; &P < 0.05, &&P < 0.005, &&&&P < 0.0001 vs. Hx+AMPH or Vec+Hx+AMPH; by 2-way ANOVA, Bonferroni’s post-test.