# Development of a screening strategy to identify A-beta resistance genes

> **NIH NIH R03** · UNIVERSITY OF COLORADO DENVER · 2020 · $155,500

## Abstract

PROJECT SUMMARY
Genetic factors play an important role in the pathogenesis and progression of Alzheimer's disease (AD). One
hallmark of AD is the development of amyloid ß (Aß) neuritic plagues and fibrils, which is believed to play a
central role in neuronal cell loss. Most people of advanced age develop amyloid plagues and fibrils in their
brains, but only a subset of the aging population develops clinical signs of AD. This finding suggests the
existence of gene regulatory pathways controlling disease development and potentially disease severity.
Protective mechanisms against AD appear to exist, but have not been characterized at the genetic or
molecular level. Herein, we propose to screen for genes/pathways in mammalian cells that confer protection
against AD by using resistance to Aß cytotoxicity as a surrogate. Our laboratory has generated a library of
mouse mutant embryonic stem (ES) cells, comprising 42,000 independent mutants mutagenized by piggyBac
(PB) transposon-mediated gene entrapment. We will utilize Aß42 oligomers for the selection of resistance
mutants in a forward genetic screen via a high-throughput ES cell platform. This novel approach will allow for
an unbiased interrogation of the genome, potentially revealing novel functional pathways protective against AD,
which may provide new insights into therapeutic targets for AD treatment. Our long-term plan is to screen the
entire library (42,000 mutants) to isolate Aß resistance mutants and identify the underlying genes and
pathways. In this application, we will establish the necessary techniques required to select Aß resistance and
to identify resistance genes; a pilot screening will be conducted as a proof of principle. Aß affects neurons, so
we will differentiate our mutant ES cells into neurons, followed by selection for Aß resistance in 96-well plates.
Mutants showing a significant increase in survival in the presence of Aß will be isolated for extensive
characterizations, including the identification of the mutated gene(s), the nature of the mutational events (null,
partial loss of function, or gain of function), and the confirmation of the causal relationship between the
disrupted gene and the Aß resistance phenotype. Once those genes are identified, their functions can also be
confirmed in human neurons by engineering the mutations using CRISPR. This study pursues an innovative
approach encompassing the use of stem cells, a transposon, and forward genetics to identify genes/pathways
protective against AD. Furthermore, the ES cells carrying AD-protective mutations may be used in subsequent
experiments to generate the corresponding mice for in vivo studies.

## Key facts

- **NIH application ID:** 9902296
- **Project number:** 5R03AG063095-02
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** Wallace Siu Hung Chick
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $155,500
- **Award type:** 5
- **Project period:** 2019-04-01 → 2021-09-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9902296

## Citation

> US National Institutes of Health, RePORTER application 9902296, Development of a screening strategy to identify A-beta resistance genes (5R03AG063095-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9902296. Licensed CC0.

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