Apolipoprotein-E (ApoE) is known to maintain the structure of lipoprotein particles and mediate lipid transport. Among three ApoE isoforms (i.e., ApoE2, ApoE3, and ApoE4) present in human, ApoE4 was identified as a major genetic risk factor for late onset Alzheimer?s disease (AD), whereas ApoE2 is thought to be protective. ApoE3 represents the most prevalent form and is thereby considered the standard for the general population. The three variants of ApoE differ only in their Cys/Arg composition at 112 and 158. Namely, ApoE2 contains Cys112 and Cys158; ApoE3 includes Cys112 and Arg158; and ApoE4 has Arg112 and Arg158. Although how ApoE2 protects against AD onset remains incompletely understood, the presence of two Cys residues in ApoE2 seems to engender a distinct structure (e.g., disulfide-linked dimers) and function, possibly explaining the protective effects that ApoE2 exerts towards AD. Notably, recent studies demonstrated that ApoE acts as a ligand for TREM2, another AD risk factor expressed in microglia, thus facilitating uptake and degradation of aggregated A?. In the current application, we propose that the redox-mediated posttranslational modification, S-nitrosylation, contributes to the protective effect of ApoE by forming SNO-ApoE2. Protein S-nitrosylation is a modification involving the covalent addition of a nitric oxide (NO)-related species (e.g., nitrosonium cation [NO+]) to a thiolate anion of cysteine. A critical mechanism for generation of SNO-proteins in vivo entails transfer of an NO-related species (i.e., transnitrosylation) from one protein to another. Our group and others have extensively demonstrated that S-nitrosylation produced by physiological levels of NO mediates neuroprotective effects, while, NO in excess elicits neurotoxicity in models of neurodegeneration via aberrant S-nitrosylation. Notably, our Preliminary Studies and published reports show that, unlike ApoE4, ApoE2 efficiently undergoes S-nitrosylation, leading to conformational changes. The change in SNO-ApoE conformation affects binding to interacting partners such as TREM2 and other receptors. Accordingly, we found that SNO-ApoE formation influences Ab uptake and degradation in hiPSC-derived microglia. Moreover, our Preliminary Studies show that SNO-ApoE2 can transnitrosylate TREM2 (producing SNO-TREM2), implying that ApoE2 may affect TREM2-dependent neuroprotective signaling pathways via this transnitrosylation reaction. In the current R01 application, we propose to investigate the mechanism whereby S-nitrosylation affects the biochemical properties of ApoE2 to regulate TREM2-mediated Ab uptake and degradation (Specific Aim 1). Finally, we will characterize the neuroprotective effects of SNO-ApoE2-TREM2 transnitrosylation on synaptic integrity and neuronal survival in AD transgenic mouse models as well as in hiPSC AD models (Specific Aims 2 and 3).

Public Health Relevance

While apolipoprotein E4 (ApoE4) represents the most significant genetic risk factor for developing late-onset Alzheimer?s disease (AD), ApoE2 has been suggested as a longevity-associated variant that provides neuroprotection against neurocognitive decline as seen in AD. Our preliminary data found evidence that ApoE2 undergoes a redox-based post-translational modification, known as S-nitrosylation, that can affect its biochemical properties, suggesting that this type of regulation is critical for the neuroprotective function of ApoE2. Here, using comprehensive biochemical and cellular approaches as well as our innovative Mass Spectrometry technique, we will look for new S-nitrosylation-related and ApoE2/TREM2-dependent molecular mechanisms that contribute to maintenance of synaptic function and neuronal survival, possibly identifying biomarkers and therapeutic targets for AD and other age-related cognitive disorders. !

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG061845-03
Application #
9949615
Study Section
Special Emphasis Panel (ZAG1)
Program Officer
Petanceska, Suzana
Project Start
2018-09-30
Project End
2023-05-31
Budget Start
2020-06-01
Budget End
2021-05-31
Support Year
3
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037