The United States is experiencing a demographic sea change owing to the rapidly growing elderly and 'Baby Boomer'populations. Our astonishing biomedical advances in the last half-century have greatly increased our life expectancy. But as a consequence of living longer, our population now faces an uptick in the incidence of neurodegenerative diseases. These truly disastrous disorders include Alzheimer's, Huntington's, Parkinson's, amyotrophic lateral sclerosis (ALS) and the frontal temporal dementias. Interestingly, though disparate in their pathophysiology, many of these diseases share a common theme manifest in the accumulation of insoluble protein aggregates in the brain and nervous system. Deciphering the mechanisms causing these proteins to misfold and aggregate and identifying the genes and cellular pathways affected by misfolded human disease proteins will aid the development of new therapeutic approaches, which we are in dire need of. The protein TDP-43 was recently identified as the major disease protein in pathological inclusions in both ALS and frontal temporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). Moreover, mutations in the TDP-43 gene have now been identified in sporadic and familial ALS patients. Pathology and genetics both converge on TDP-43 being central to the pathogenesis of these diseases. The study of tau, alpha-synuclein, and amyloid beta in neurodegenerative disorders has revolutionized our understanding of their respective disease mechanisms. A similar intense effort to understand TDP-43 biology and its role in pathology will be beneficial to the development of therapies to treat ALS and FTLD-U. Nothing is yet known about how TDP-43 causes disease (loss- or gain-of-function mechanism). We have generated in vitro and in vivo TDP-43 proteinopathy models to explore TDP-43 and our preliminary data demonstrate: 1) a critical role for the RNA recognition motif and carboxy-terminal region of TDP-43 in mediating aggregation and cellular toxicity, 2) increased aggregation and toxicity caused by a disease-linked TDP-43 mutation, and 3) a pilot genetic screen identified multiple RNA binding proteins as potent toxicity modifiers. We hypothesize that TDP-43 causes disease by a toxic gain-of-function mechanism involving RNA binding. Thus, with the goal to define TDP-43 disease mechanisms from multiple angles we propose three Specific Aims: 1) Directly testing the requirements for RNA binding by TDP-43 on aggregation and toxicity and testing the effect of ALS-linked TDP-43 mutations in vitro and in vivo;2) Performing two genome wide screens for genetic modifiers of TDP-43 aggregation and toxicity, and;3) Collaborating with a team of experts to develop animal models (zebrafish, Drosophila, C. elegans) to explore TDP-43 disease mechanisms and validating modifier genes from our genetic screens.

Public Health Relevance

As our population continues to age, neurodegenerative disease will increase in prevalence and thus pose a daunting challenge to public health worldwide. The protein TDP-43 has recently been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontal temporal lobar degeneration with ubiquitin- positive inclusions (FTLD-U), but very little is known about the mechanisms by which TDP-43 causes neurodegeneration. Our project harnesses multiple in vitro and in vivo models and techniques to gain mechanistic insight into how TDP-43 contributes to human disease, which will suggest novel avenues for therapeutic intervention.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Research Project (R01)
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Cellular and Molecular Biology of Neurodegeneration Study Section (CMND)
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Gubitz, Amelie
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Stanford University
Schools of Medicine
United States
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Couthouis, Julien; Raphael, Alya R; Siskind, Carly et al. (2014) Exome sequencing identifies a DNAJB6 mutation in a family with dominantly-inherited limb-girdle muscular dystrophy. Neuromuscul Disord 24:431-5
Figley, Matthew D; Bieri, Gregor; Kolaitis, Regina-Maria et al. (2014) Profilin 1 associates with stress granules and ALS-linked mutations alter stress granule dynamics. J Neurosci 34:8083-97
Figley, Matthew D; Thomas, Anna; Gitler, Aaron D (2014) Evaluating noncoding nucleotide repeat expansions in amyotrophic lateral sclerosis. Neurobiol Aging 35:936.e1-4
Jovi?i?, Ana; Gitler, Aaron D (2014) TDP-43 in ALS: stay on target…almost there. Neuron 81:463-5
Raphael, Alya R; Couthouis, Julien; Sakamuri, Sarada et al. (2014) Congenital muscular dystrophy and generalized epilepsy caused by GMPPB mutations. Brain Res 1575:66-71
Couthouis, Julien; Raphael, Alya R; Daneshjou, Roxana et al. (2014) Targeted exon capture and sequencing in sporadic amyotrophic lateral sclerosis. PLoS Genet 10:e1004704
Kim, Hyung-Jun; Raphael, Alya R; LaDow, Eva S et al. (2014) Therapeutic modulation of eIF2? phosphorylation rescues TDP-43 toxicity in amyotrophic lateral sclerosis disease models. Nat Genet 46:152-60
Aguzzi, Adriano; Gitler, Aaron D (2013) A template for new drugs against Alzheimer's disease. Cell 154:1182-4
Li, Yun R; King, Oliver D; Shorter, James et al. (2013) Stress granules as crucibles of ALS pathogenesis. J Cell Biol 201:361-72
Chesi, Alessandra; Staahl, Brett T; Jovicic, Ana et al. (2013) Exome sequencing to identify de novo mutations in sporadic ALS trios. Nat Neurosci 16:851-5

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