Thomas Arendt Lab

Functional characterization of long non-coding RNAs in Alzheimer´s disease



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he project will establish the specific pathogenetic role of long ncRNAs (lncRNAs) in Alzheimer´s di­sease (AD). To this end, we will identify and characterize functionally relevant target interactions of lncRNAs, previously identified in a genome-wide approach. In our preliminary work, applying a combination of tiling array and custom array platform (“humBrainChip”) to a cohort of 40 AD patients and controls we established the genome-wide pattern of lncRNA expression differences between AD and control brain. Enrichement analyses and adjustment with datasets on cell-cycle dependent expression of lncRNAs revelead 20 lncRNAs related to chromatin-association and cell cycle regulation which supports our concept on the critical involvement of neuronal differentiation control in AD.

The experimental pipeline is structured into five aims: (i) Functionally relevant DNA and/or RNA target-sequences of lncRNAs will be identified, adapting the technique of “Chromatin Isolation by RNA Purification-ChIRP”. (ii) DNA loci, regulated by lncRNA will be identified combining DNA-sequencing with local methylation and expression analyses and the adjustment to previously obtained datasets on AD-specific transcriptom changes (“humBrainChip”). (iii) RNA-binding proteins (RBPs) will be identified by a proteomic. (iv) Multicomponent complexes, involving lncRNA, DNA and potentially RBPs will further be characterized by specific binding assays (e.g. EMSA). (v) lncRNA-target interactions will functionally be characterized by esiRNA technology with respect to their role in cellular programmes with an established pathophysiological role in AD.

The following questions will be answered: (1) Are lncRNA specifically involved in the regulation of pathophysiological programmes with an established role in AD or is the AD-associated change of lncRNAs epiphenomenal? (2) What are the molecular targets of lncRNAs relevant to this function? (3) What is the mode of action of AD-related lncRNAs and how relates this to cell death?

Key Technologies:
- Human brain bank
- Morphometry and Stereology (NeurolucidaTM, NeuroExplorerTM, Stereo InvestigatorTM)
- Multi-dye fluorescence microscopy, confocal microscopy, two-photon microscopy
- time-lap-microscopy for cell- and tissue cultures
- In situ hybridization FISH/CISH
- Laser scanning cytometry
- Laser microdissection
- quantitative RT-PCR
- FPLC
- Electromobility shift assays (EMSA)
- ChIRP

Last Publications

Ueberham U, Rohn S, Ueberham E, Wodischeck S, Hilbrich I, Holzer M, Brückner MK, Gruschka H, Arendt T. Pin1 promotes degradation of Smad proteins and their interaction with phosphorylated tau in Alzheimer's disease. Neuropathol Appl Neurobiol. 2014 Dec;40(7):815-32

Arendt T, Bullmann T. Neuronal plasticity in hibernation and the proposed role of the microtubule-associated protein tau as a "master switch" regulating synaptic gain in neuronal networks. Am J Physiol Regul Integr Comp Physiol. 2013 Sep;305(5):R478-89.

Ueberham U, Hilbrich I, Ueberham E, Rohn S, Glöckner P, Dietrich K, Brückner MK, Arendt T. Transcriptional control of cell cycle-dependent kinase 4 by Smad proteins-implications for Alzheimer's disease. Neurobiol Aging. 2012 Dec;33(12):2827-
40.

Fischer HG, Morawski M, Brückner MK, Mittag A, Tarnok A, Arendt T. Changes in neuronal DNA content variation in the human brain during aging. Aging Cell. 2012 Aug;11(4):628-33.

Arendt T, Brückner MK, Mosch B, Lösche A. Selective cell death of hyperploid neurons in Alzheimer's disease. Am J Pathol. 2010, 177(1):15-20.

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