Mitochondrial diseases manifesting as encephalopathies occur at a rate of 1 in 5000 live births and are often fatal by ~5 years old. Mitochondrial diseases are respiratory chain disorders in which the mitochondria are no longer operating efficiently to produce ATP, usually due to a problem with one or more components of the electron transport chain (ETC). Fortunately, genetic sequencing has identified a large number of the mutations in mitochondrial or nuclear DNA which cause these encephalomyopathies. However, in most cases there is still no clear metabolic link between the genetic defect and the neuropathology, and very few effective treatments. The innovative studies described in this proposal are expected to reveal a novel metabolic link between reduced ETC activity and neural pathology. Previously, we have detected a new post-translational modification of proteins, S-(2-succino)cysteine (2SC), which is formed by reaction of the Krebs cycle intermediate fumarate with reactive cysteine residues in protein. Both fumarate and succination of proteins are increased in adipocytes in diabetes, disturbing protein function and turnover. The increase in fumarate develops as a result of excess fuel supply, accumulation of NADH, and feedback inhibition of the Krebs cycle. In a novel, lateral extension of these observations we propose that a similar inhibition of the ETC, e.g. in Complex I deficiency during Leigh Syndrome, would result in increased NADH, fumarate and succination in mitochondrial disease. In Preliminary Studies, we demonstrate that increased succination of proteins is detectable on several proteins in the brainstem of a mouse model of Leigh syndrome (Ndufs4 knockout (KO) mouse) in association with neurodegeneration. We hypothesize that mitochondrial stress results in the accumulation of fumarate and that succination alters protein structure or function contributing to disease pathology. We will confirm this in Specific Aim 1. We have identified several succinated targets already and we plan to mechanistically address how succination of these leads to further reductions in mitochondrial function in Specific Aim 2.
In Specific Aim 3 we will use a molecular strategy to distinguish the bioenergetic defect from protein succination and investigate therapeutic strategies designed to reduce fumarate and succination leading to improvements in mitochondrial function and the disease phenotype. Overall, these foundational studies will demonstrate that succination is a mechanistic link between mitochondrial stress and neuropathology, with important implications for the elucidation of novel therapeutic avenues for the treatment of mitochondrial diseases.

Public Health Relevance

Mitochondrial diseases e.g. Leigh Syndrome affect 1 in 5000 live births. This work will directly assess how a protein modification derived from a mitochondrial metabolite contributes to the development of neuropathology during Leigh Syndrome. By demonstrating the significance of this pathway in the progression of disease pathology, we will be able to design novel targeted therapeutic strategies for the effective treatment of mitochondrial diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS092938-03
Application #
9268095
Study Section
Therapeutic Approaches to Genetic Diseases Study Section (TAG)
Program Officer
Lavaute, Timothy M
Project Start
2015-08-15
Project End
2019-05-31
Budget Start
2017-06-01
Budget End
2018-05-31
Support Year
3
Fiscal Year
2017
Total Cost
$215,669
Indirect Cost
$54,119
Name
University of South Carolina at Columbia
Department
Pharmacology
Type
Schools of Medicine
DUNS #
041387846
City
Columbia
State
SC
Country
United States
Zip Code
29208
Ruecker, Nadine; Jansen, Robert; Trujillo, Carolina et al. (2017) Fumarase Deficiency Causes Protein and Metabolite Succination and Intoxicates Mycobacterium tuberculosis. Cell Chem Biol 24:306-315
Manuel, Allison M; Walla, Michael D; Faccenda, Adam et al. (2017) Succination of Protein Disulfide Isomerase Links Mitochondrial Stress and Endoplasmic Reticulum Stress in the Adipocyte During Diabetes. Antioxid Redox Signal 27:1281-1296
Piroli, Gerardo G; Manuel, Allison M; Clapper, Anna C et al. (2016) Succination is Increased on Select Proteins in the Brainstem of the NADH dehydrogenase (ubiquinone) Fe-S protein 4 (Ndufs4) Knockout Mouse, a Model of Leigh Syndrome. Mol Cell Proteomics 15:445-61
Yang, Hao; Wu, Jiang W; Wang, Shu P et al. (2016) Adipose-Specific Deficiency of Fumarate Hydratase in Mice Protects Against Obesity, Hepatic Steatosis, and Insulin Resistance. Diabetes 65:3396-3409