Description ALS is a fatal progressive paralysis affecting 34,000 Americans and killing 8,000 per year. The discovery in 1993 of dominant mutations to Cu,Zn-superoxide dismutase (Cu,Zn-SOD) linked to 2-7% of ALS cases led to hopes that new treatments for ALS based on antioxidants might be forthcoming. Whereas there is broad agreement that ALS mutant SODs have impaired stability, the specific toxic gain-of-function associated with these mutations is still hotly debated. The two leading explanations are the aggregation hypothesis that aggregation of mutant SODs is a direct cause of disease and the zinc-deficient hypothesis that disease is due to increased redox activity of SOD that has lost zinc but retains copper in its active site. We propose, based on extensive preliminary evidence, that the two hypotheses are intimately interconnected with aggregation being paradoxically protective by removing zinc-deficient SOD. The zinc-deficient hypothesis also raises the intriguing possibility that SOD could contribute to sporadic ALS. We have developed innovative mass spectrometric methods to quantitively the metal content of SOD across the spinal cord, which allow us to critically evaluate the zinc-deficient and aggregation hypotheses.
Aim 1 will characterize how the dimer interface and the intramolecular disulfide of SOD are affected by ALS-associated mutations to affect zinc and copper binding as well as propensity for aggregation of SOD.
This aim will explore the physical basis underlying metal loss and aggregation to understand how mutant SODs are more prone to losing zinc and why mutant SODs are dominant in inheritance.
Aim 2 will assess how the concentrations of zinc-deficient SOD are modulated by CCS (the copper chaperone for SOD) and wild-type Cu,Zn SOD in transgenic animals both of which accelerate disease in vivo -- and how to most effectively pharmacologically decrease zinc-deficient SOD in vivo.
Aim 3 will map the anatomical distribution of the different metal states of SOD in disease-affected versus unaffected regions in human sporadic ALS patients, testing whether zinc-deficient SOD may be the common connection between sporadic and familial SOD. New preliminary data demonstrate a vast improvement in sensitivity with the FTICR that makes Aim 3 feasible. Completion of the proposed experiments will critically test how SOD mutations lead to the development of ALS and whether loss of zinc occurs from wild-type SOD in sporadic ALS.
Only a small percentage of patients with Lou Gehrig's disease (also known as ALS) carry mutations to a common antioxidant defense enzyme called SOD. Completion of the proposed experiments will help explain how SOD can be involved in the vast majority of sporadic ALS patients who do not have SOD mutations, which will point to new ways to treat the disease. We specifically are testing the hypothesis that this enzyme becomes toxic when it loses one zinc atom and have developed new mass spectrometric methods that directly measure the loss of zinc directly from the spinal cord.
|Williams, Jared R; Trias, Emiliano; Beilby, Pamela R et al. (2016) Copper delivery to the CNS by CuATSM effectively treats motor neuron disease in SOD(G93A) mice co-expressing the Copper-Chaperone-for-SOD. Neurobiol Dis 89:1-9|
|Trias, Emiliano; Ibarburu, Sofía; Barreto-Núñez, Romina et al. (2016) Post-paralysis tyrosine kinase inhibition with masitinib abrogates neuroinflammation and slows disease progression in inherited amyotrophic lateral sclerosis. J Neuroinflammation 13:177|
|Roemeling, Margo D; Williams, Jared; Beckman, Joseph S et al. (2015) Imidazole catalyzes chlorination by unreactive primary chloramines. Free Radic Biol Med 82:167-78|
|Viera, Liliana; Radmilovich, Milka; Vargas, Marcelo R et al. (2013) Temporal patterns of tyrosine nitration in embryo heart development. Free Radic Biol Med 55:101-8|
|Gandelman, Mandi; Levy, Mark; Cassina, Patricia et al. (2013) P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway. J Neurochem 126:382-8|
|Rhoads, Timothy W; Williams, Jared R; Lopez, Nathan I et al. (2013) Using theoretical protein isotopic distributions to parse small-mass-difference post-translational modifications via mass spectrometry. J Am Soc Mass Spectrom 24:115-24|
|Franco, Maria Clara; Ye, Yaozu; Refakis, Christian A et al. (2013) Nitration of Hsp90 induces cell death. Proc Natl Acad Sci U S A 110:E1102-11|
|Drechsel, Derek A; Estévez, Alvaro G; Barbeito, Luis et al. (2012) Nitric oxide-mediated oxidative damage and the progressive demise of motor neurons in ALS. Neurotox Res 22:251-64|
|Trumbull, Kari A; McAllister, Donna; Gandelman, Mandi M et al. (2012) Diapocynin and apocynin administration fails to significantly extend survival in G93A SOD1 ALS mice. Neurobiol Dis 45:137-44|
|Rhoads, Timothy W; Lopez, Nathan I; Zollinger, Daniel R et al. (2011) Measuring copper and zinc superoxide dismutase from spinal cord tissue using electrospray mass spectrometry. Anal Biochem 415:52-8|
Showing the most recent 10 out of 18 publications