Generation of gene expression profiles of neurodegenerative diseases

Coordinator:    Dr. Wilfried Nietfeld
Institution: Max-Planck-Institut für molekulare Genetik, Berlin
A major characteristic of neurodegenerative diseases (NDs) is the accumulation or aggregation of misfolded proteins that are suspected to lead to the loss of neuronal cells in various brain regions. Gene expression and subsequent misfolding and aggregation of the resulting proteins are influenced by interactions with other proteins or with chemical compounds.
In this subproject, we developed a cell-based in vivo model of protein aggregation, in which we overexpressed eight known disease-relevant proteins and their mutants: Parkinson protein 2, presenilin 1, ataxin 1, the amyloid precursor protein; superoxide dismutase, alpha-synuclein, huntingtin and TDP-43. These proteins are regarded as causative to the following neurodegenerative diseases: Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis (ALS), spinocerebellar Ataxia and Huntington’s disease. The generated cell lines were characterised and transferred to subproject 7 for use in various cell-based assays.
For gene expression studies, cell lines expression wild-type and mutant huntingtin were first investigated. Cells were incubated with 37 different chemical agents to investigate whether they modulate misfolding and aggregation of huntingtin in the cellular system. The choice of compounds was made on the basis of previous results that hinted at potential effects on huntingtin.
To increase throughput in the compound screening, a FRET assays was established. This is a highly sensitive test system based on fluorescence measurements. This assay was also used by the consortium for the validation of protein-protein interactions that were identified in high-throughput automated screenings for the disease-relevant proteins mentioned above.
In collaboration with subprojects 4 and 8, we carried out gene expression studies in blood samples from Huntington’s disease patients and healthy controls based on RNA sequencing. Also, we studied gene expression in mouse models of Alzheimer’s disease treated with chemical compounds. For individual substances, an effect on gene expression in mouse brain could be detected. These compounds were investigated in further for their therapeutic potential, including behavioural tests to determine improved memory.

Fig 1. Experimental strategy for gene expression analysis in the subproject.
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