Ruprecht-Karls-Universität Heidelberg
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Hecker_Wagner0118 - Scientist (f/m) / PhD position
Project no:
Hecker_Wagner0118

Project leader:

Project supervisor:
Hecker, Markus
Application deadline:
30. Sep 2018
Start of PhD project:
1. Jan 2019

Project description:

Title:
The role of lipid hydroperoxide-induced ferroptosis in diabetic nephropathy
Summary:
Recent results from our group suggest that under conditions of a diabetic metabolic state pro-inflammatory cytokines released from dysfunctional glomerular endothelial cells cause downregulation of glutathione peroxidase-4 (GPx-4) on the protein level. GPx-4, one of the most important ROS-scavenging selenoproteins, is a unique antioxidant enzyme that can directly reduce phospholipid hydroperoxides in the adjacent podocytes. Since no other glutathione peroxidise, such asGPx-1 (1), can effectively detoxify these reactive lipids, the resulting lipid hydroperoxide stress may trigger ferroptosis (2) in podocytes, ultimately leading to podocyte loss, a hallmark of diabetic nephropathy. In fact, in patients with type 1 and in particular type 2 diabetes, GPx-4 abundance is decreased in both podocytes and tubular epithelial cells (3). The main objective of the proposed project is to induce diabetes in podocyte-specific conditional GPx-4 knockout mice by streptozotocin treatment and to study the consequences of a drastically reduced antioxidative capacity for podocyte function and survival. Furthermore it is planned to employ a specifically designed adeno-associated virus 2 (AAV2) delivery approach to the kidney cortex to re- or overexpress GPx-4 in these mice to establish GPx-4 as a novel therapeutic target in combatting diabetic nephropathy.
References:
1. Sultan CS, Saackel A, Stank A, Fleming T, Fedorova M, Hoffmann R, Wade RC, Hecker M, Wagner AH. Impact of carbonylation on glutathione peroxidase-1 activity in human hyperglycemic endothelial cells. Redox Biol. 2018 Jun;16:113-122.

2. Friedmann Angeli JP et al. Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice.
Nat Cell Biol. 2014 Dec;16(12):1180-91.

3. Wiedenmann T, Dietrich N, Fleming T, Altamura S, Deelman LE, Henning RH, Muckenthaler MU, Nawroth PP, Hammes HP, Wagner AH, Hecker M. Modulation of glutathione peroxidase activity by age-dependent carbonylation in glomeruli of diabetic mice. J Diabetes Complications. 2018 Feb;32(2):130-138.
Methods that will be used:
• Cell culture techniques (cell lines, isolation and culture of primary podocytes, co-culture models for evaluating cell-cell interactions)
• Animal models
• Protein biochemistry methods (Western blot, detection of posttranslational protein modifications, )
• high resolution mass spectrometry imaging of lipid hydroperoxides in kidney tissue, ESI-mass spectrometry of phospholipids
• Molecular biology techniques (incl. genotyping, RT-PCR for expression profiling)
• Laser capture microdissection (LCM)
• Immunohistochemistry
• Microscopy (immunofluorescence)

Cooperation partners:
Thomas Fleming (Internal Medicine I, Heidelberg), Hans-Hermann Gröne (DKFZ Heidelberg), Oliver Müller (Internal Medicine III, Kiel), Bernhard Spengler (Anorganic and Analytical Chemistry, Gießen), Nadine Volk (NCT, Heidelberg)
Personal qualifications:
• Qualified master or diploma in biology, biochemistry, pharmacy or related fields
• Advanced knowledge of biochemical, cell and molecular biological techniques as well as animal experimentation (FELASA B) or high motivation to acquire the necessary skills and certificates
• Team-minded and flexible
• Ability to rapidly familiarize him/herself with new research areas in theory and praxis
Keywords:
Diabetes, nephropathy, mouse models, ferroptosis, podocytes, reversed genetics, gene transfer, AAV