Aggregation of amyloid proteins is associated with a wide range of human pathologies termed protein misfolding or deposition neurodegenerative disorders, which include Alzheimer's, Parkinson's, and Huntington's diseases. It has been shown that oligomeric species of amyloid aggregates are neurotoxic. Still, the nature of these species remains unknown. Despite this importance of oligomeric species with respect to toxicity as well as in normal physiological events, knowledge regarding the molecular mechanisms underlying proteins self-assembly is very limited. The objective of this application is to characterize each oligomer at a level that will allow us to understand the molecular mechanism of the nanoassembly process. However, the fact that oligomers are formed transiently essentially impedes their characterization. To overcome this obstacle, we propose a novel approach in which oligomers of a defined size are assembled on a polymer- based Flexible Nanoarray (FNA), which will enable the use of various methods for their characterization. Based on data obtained during the current funding period, we hypothesize that the self-assembly is driven by increased, size-dependent, intermolecular interaction and stability of the oligomers. To test this hypothesis, we will thoroughly characterize FNA-assembled oligomers by applying a set of single-molecule approaches, combined with detailed computational analyses. Guided by strong preliminary data, we will text our major hypothesis through the following three specific aims:
Aim 1) Develop a novel flexible nanoarray approach to measure interactions within oligomers;
Aim 2) Directly measure directly the lifetimes of oligomers using a novel, tethered approach;
and Aim 3) Demonstrate secondary structural analysis for individual aggregated amyloids using a Tip-Enhanced Raman Spectroscopy (TERS) approach. The rationale for the proposed aims is that understanding fundamental mechanisms of protein misfolding and aggregation has the strong potential to translate into specific approaches to control the aggregation process. These advances are expected to lead to the development of new and innovative preventative strategies and treatments for protein misfolding diseases like Alzheimer's disease. The application is innovative, because it presents a novel approach to the protein aggregation phenomenon and develops a set of new nanotechnology methods with broad biomedical applications. The proposed research is significant because the findings will lay the foundation for efficient treatments against protein misfolding diseases at the very early stages. Additionally, the availability of oligomers of select sizes assembled as FNAs opens prospects for their use as targets in the development of diagnostic tools such as immunoassays. Moreover, given that oligomers, rather than larger aggregates including fibrils, are considered neurotoxic species, the availability of oligomers with desired sizes will open realistic prospects for the development of efficient immunological preventive, diagnostic, and therapeutic strategies for Alzheimer's disease, Parkinson's disease, and other neurodegenerative disorders.

Public Health Relevance

The work proposed is relevant to public health because unraveling structural and dynamic properties of amyloid oligomers the most neurotoxic species is expected to increase the understanding of molecular mechanisms of Alzheimer's, Parkinson's and other protein misfolding diseases. The proposed research will provide knowledge on how pathologies related to protein misfolding develop as well as how to control the aggregation process. Thus, the proposed research is relevant to the part of NIH's mission that pertains to developing fundamental knowledge that will improve preventive and therapeutic treatments for diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM096039-06
Application #
9147607
Study Section
Nanotechnology Study Section (NANO)
Program Officer
Edmonds, Charles G
Project Start
2011-08-01
Project End
2019-08-31
Budget Start
2016-09-01
Budget End
2017-08-31
Support Year
6
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Nebraska Medical Center
Department
Other Basic Sciences
Type
Schools of Pharmacy
DUNS #
168559177
City
Omaha
State
NE
Country
United States
Zip Code
68198
Lv, Zhengjian; Banerjee, Siddhartha; Zagorski, Karen et al. (2018) Supported Lipid Bilayers for Atomic Force Microscopy Studies. Methods Mol Biol 1814:129-143
Lyubchenko, Yuri L (2018) Direct AFM Visualization of the Nanoscale Dynamics of Biomolecular Complexes. J Phys D Appl Phys 51:
Maity, Sibaprasad; Viazovkina, Ekaterina; Gall, Alexander et al. (2018) Polymer Nanoarray Approach for the Characterization of Biomolecular Interactions. Methods Mol Biol 1814:63-74
Pan, Yangang; Zagorski, Karen; Shlyakhtenko, Luda S et al. (2018) The Enzymatic Activity of APOBE3G Multimers. Sci Rep 8:17953
Maity, Sibaprasad; Pramanik, Apurba; Lyubchenko, Yuri L (2018) Probing Intermolecular Interactions within the Amyloid ? Trimer Using a Tethered Polymer Nanoarray. Bioconjug Chem 29:2755-2762
Sun, Zhiqiang; Hashemi, Mohtadin; Warren, Galina et al. (2018) Dynamics of the Interaction of RecG Protein with Stalled Replication Forks. Biochemistry 57:1967-1976
Zhang, Yuliang; Hashemi, Mohtadin; Lv, Zhengjian et al. (2018) High-speed atomic force microscopy reveals structural dynamics of ?-synuclein monomers and dimers. J Chem Phys 148:123322
Banerjee, Siddhartha; Hashemi, Mohtadin; Lv, Zhengjian et al. (2017) A novel pathway for amyloids self-assembly in aggregates at nanomolar concentration mediated by the interaction with surfaces. Sci Rep 7:45592
Pan, Yangang; Sun, Zhiqiang; Maiti, Atanu et al. (2017) Nanoscale Characterization of Interaction of APOBEC3G with RNA. Biochemistry 56:1473-1481
Banerjee, Siddhartha; Sun, Zhiqiang; Hayden, Eric Y et al. (2017) Nanoscale Dynamics of Amyloid ?-42 Oligomers As Revealed by High-Speed Atomic Force Microscopy. ACS Nano 11:12202-12209

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