# S-Nitrosylation-Induced Posttranslational Modification and Aberrant Cell Signaling in Sporadic Alzheimer's Disease

> **NIH NIH R01** · SCRIPPS RESEARCH INSTITUTE, THE · 2021 · $651,737

## Abstract

SUMMARY
This collaborative R01 application between a neuroscience lab (led by Stuart Lipton at Scintllon Inst./UC San
Diego) and a chemistry lab (led by Steve Tannenbaum at MIT) will identify the redox posttranslational
modification of proteins called S-nitrosylation by developing a more effective and integrated Mass Spec-based
platform to screen for the S-nitrosoproteome and resulting alterations in protein function that contribute to the
pathogenesis of Alzheimer’s disease (AD). Our hypothesis is that entire biochemical pathways critical to
neuronal function are affected by aberrant S-nitrosylation of multiple proteins, these aberrant redox reactions
(which are located, at least in part, downstream of Aß insult) contribute to the pathogenesis of AD, and the
reactions occur in both sporadic and familial cases of the disease. Chemical and functional analysis of S-
nitrosylated proteins will be assessed by biochemical assays, and by imaging of cells and tissues, including
human AD brain and various in vitro and in vivo models of AD, ranging from transgenic mice to hiPSC-based
model systems. We will also use site-directed mutagenesis and CRISPR/Cas9 techniques to generate DNA
constructs or genes encoding proteins that that cannot be S-nitrosylated (thus forming non-nitrosylatable
proteins). Accordingly, our Specific Aims are as follows:
AIM #1. To determine the S-nitrosoproteome in human AD brain and transgenic mouse models. We will
validate our recent S-nitrosoproteome findings in the CK-p25 mouse model of AD (published in PNAS, 2016)
and determine if it generalizes to human AD brain and other transgenic mouse models of AD, e.g., hAPP-J20
and Tg2576.
AIM #2. To use hiPSC-derived cerebrocortical neurons generated from human AD patients or WT exposed to
oligomeric Aß (as a model of sporadic AD) as an in vitro model system to study the S-nitrosoproteome and
how it affects biochemical pathways. This approach will allow us to study the functional effect of SNO-proteins
in AD in a human context.
AIM #3. To screen the effects of various S-nitrosoproteins in hiPSC-based models for impact on potential
biological functions, e.g., effect on synaptic loss or neuronal cell death. This will be accomplished by
generating non-nitrosylatable constructs of proteins (e.g., substituting Ala for Cys) by replacing the underling
gene by CRISPR/Cas9 technology. For selected gene products that manifest profound effects of S-
nitrosylation on synaptic functions and neuronal cell survival in hiPSC-based models, the non-nitrosylatable
version of the gene can also be created in mice using CRISPR/Cas9 to mechanistically test its effect in vivo.

## Key facts

- **NIH application ID:** 10173585
- **Project number:** 5R01AG056259-05
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** STUART A LIPTON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $651,737
- **Award type:** 5
- **Project period:** 2017-09-30 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10173585

## Citation

> US National Institutes of Health, RePORTER application 10173585, S-Nitrosylation-Induced Posttranslational Modification and Aberrant Cell Signaling in Sporadic Alzheimer's Disease (5R01AG056259-05). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10173585. Licensed CC0.

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