Mitochondrial ion and energy metabolism in LETM1 pathologies

Project Background:

Mitochondria are organelles with essential functions in cell biology, and their dysfunction contribute to various metabolic and neurodegenerative diseases. Here we focus on LETM1, a seizure candidate gene implicated in Wolf-Hirschhorn Syndrome. Recent findings have unveiled bi-allelic mutations in LETM1, manifesting severe neuromuscular and metabolic symptoms with dire prognoses among newborns and infants. LETM1 protein locates at the inner membrane of mitochondria and functions as K+/H+ antiporter. However, LETM1’s diverse impacts on mitochondrial morphology, cation regulation and energy metabolism pose significant challenges on identifying the primary driver of pathogenicity among its pleiotropic effects. The identification of pathogenic mutations within LETM1's core domain presents an unprecedented opportunity to differentiate its primary role from secondary effects and unravel the molecular mechanisms underpinning disease pathology. Through a comprehensive investigation of the LETM1 core domain, the project aims to shed light on the key pathways affected by LETM1 in normal physiology and disease and provide valuable insights into the development of targeted therapies.

Objectives and Methodology

Generation of fibroblasts with LETM1KD and fibroblasts with clinical relevant mutations in the LETM1 core domain and comprehensive phenotypic analysis; Phenotypic rescue analyses of LETM1KD by the LETM1 core domain; in vitro study of K+ or Ca2+ transport functions of recombinant LETM1 core domain.

The methodological focus will be on CRISPR-mediated knockin, cloning, qPCR, western blotting, K+ and Ca2+ transport assay, enzymatic assays for metabolic functions in OXPHOS and PDH activity and NAD+/H, ROS measurements, IF and EM.

Qualification:

We invite enthusiastic applicants with a Master study in a relevant field such as molecular biology, biomedical science or biotechnology, theoretical and practical knowledge in cellular biology, experience in biochemical methods and high commitment.

Application:

Please send your motivation letter and CV by 30.04.2024 to karin.nowikovsky(at)vetmeduniwien.ac.at

Starting date as soon as possible

Salary according to FWF guidelines