Stephanie Vrijsen, Laura Besora-Casals, Sarah van Veen, Jeffrey Zielich, Chris Van den Haute, Norin Nabil Hamouda, Christian Fischer, Bart Ghesquière, Ivailo Tournev, Patrizia Agostinis, Veerle Baekelandt, Jan Eggermont, Eric Lambie, Shaun Martin, Peter Vangheluwe
Author Information
Stephanie Vrijsen: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium. ORCID
Laura Besora-Casals: Cell and Developmental Biology, Department Biology II, Ludwig Maximilians Universität, 80539 Münich, Germany. ORCID
Sarah van Veen: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium. ORCID
Jeffrey Zielich: Cell and Developmental Biology, Department Biology II, Ludwig Maximilians Universität, 80539 Münich, Germany. ORCID
Chris Van den Haute: Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, 3000 Leuven, Belgium.
Norin Nabil Hamouda: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium.
Christian Fischer: Cell and Developmental Biology, Department Biology II, Ludwig Maximilians Universität, 80539 Münich, Germany.
Bart Ghesquière: Metabolomics Expertise Center, Vlaams Instituut voor Biotechnologie (VIB)-KU Leuven Center for Cancer Biology, Department of Oncology, KU Leuven, 3000 Leuven, Belgium. ORCID
Ivailo Tournev: Department of Neurology, Medical University-Sofia, 1431 Sofia, Bulgaria.
Patrizia Agostinis: Laboratory of Cell Death Research and Therapy, VIB-KU Leuven Center for Cancer Biology, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
Veerle Baekelandt: Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, 3000 Leuven, Belgium.
Jan Eggermont: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium. ORCID
Eric Lambie: Cell and Developmental Biology, Department Biology II, Ludwig Maximilians Universität, 80539 Münich, Germany. ORCID
Shaun Martin: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium.
Peter Vangheluwe: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, Katholieke Universiteit Leuven (KU Leuven), 3000 Leuven, Belgium; peter.vangheluwe@kuleuven.be. ORCID
Recessive loss-of-function mutations in () are associated with a spectrum of neurodegenerative disorders, including Parkinson's disease (PD). We recently revealed that the late endo-lysosomal transporter ATP13A2 pumps polyamines like spermine into the cytosol, whereas ATP13A2 dysfunction causes lysosomal polyamine accumulation and rupture. Here, we investigate how ATP13A2 provides protection against mitochondrial toxins such as Rotenone, an environmental PD risk factor. Rotenone promoted mitochondrial-generated superoxide (MitoROS), which was exacerbated by ATP13A2 deficiency in SH-SY5Y cells and patient-derived fibroblasts, disturbing mitochondrial functionality and inducing toxicity and cell death. Moreover, ATP13A2 knockdown induced an ATF4-CHOP-dependent stress response following Rotenone exposure. MitoROS and ATF4-CHOP were blocked by MitoTEMPO, a mitochondrial antioxidant, suggesting that the impact of ATP13A2 on MitoROS may relate to the antioxidant properties of spermine. Pharmacological inhibition of intracellular polyamine synthesis with α-difluoromethylornithine (DFMO) also increased MitoROS and ATF4 when ATP13A2 was deficient. The polyamine transport activity of ATP13A2 was required for lowering Rotenone/DFMO-induced MitoROS, whereas exogenous spermine quenched Rotenone-induced MitoROS via ATP13A2. Interestingly, fluorescently labeled spermine uptake in the mitochondria dropped as a consequence of ATP13A2 transport deficiency. Our cellular observations were recapitulated in vivo, in a strain deficient in the ATP13A2 ortholog These animals exhibited a basal elevated MitoROS level, mitochondrial dysfunction, and enhanced stress response regulated by , the ortholog of ATF4, causing hypersensitivity to Rotenone, which was reversible with MitoTEMPO. Together, our study reveals a conserved cell protective pathway that counters mitochondrial oxidative stress via ATP13A2-mediated lysosomal spermine export.
