# Method for Detection and Quantification of CLN3 Protein

> **NIH NIH R03** · UNIVERSITY OF KENTUCKY · 2021 · $153,000

## Abstract

Project Summary/Abstract
 CLN3 disease (Juvenile Neuronal Ceroid Lipofuscinosis or juvenile Batten disease; OMIM #204200) is a
fatal neurodegenerative disease. This autosomal recessive disease is caused by mutations in a single gene,
Ceroid Lipofuscinosis Neuronal 3 (CLN3). The majority of patients are homozygous for a 1 kb deletion mutation
in CLN3. Despite two decades of extensive research on CLN3 disease after discovery of the CLN3 gene, there
is still no cure; and even the function of CLN3 protein, and etiology and pathogenesis of the disease are still ill-
defined. Research on CLN3 disease and development of therapeutics are greatly hindered by lack of a working
antibody for detecting either wild type (WT) or disease-causing mutant CLN3 proteins. For example, it is unknown
if WT and mutant CLN3 protein levels and subcellular distributions correlate with disease severity; and the
effectiveness of CLN3 gene therapy cannot be fully evaluated without monitoring CLN3 protein levels in patients.
Therefore, there is a greatly unmet need for detection and quantification of WT and mutant CLN3 proteins. Here
we propose to develop a mass spectrometry-based method to reliably detect WT and mutant CLN3 proteins and
quantify their absolute levels. Our preliminary studies provide compelling proof-of-concept evidence that we have
built the framework of such a method that is capable of detection and quantification of endogenous WT CLN3
protein in human and mouse cell lines as well as mouse brain. In Aim 1, we will further develop this method to
detect and quantify WT (1A) and mutant (1B) CLN3 proteins in vitro, with applications on human and mouse cell
lines, human primary fibroblasts with and without CLN3 disease, and neurons differentiated from both isogenic
control and CLN3 mutant human induced pluripotent stem (iPS) cells. In Aim 2, we will expand this method to
detect and quantify WT and mutant CLN3 protein in vivo, with applications on human and mouse tissues (2A),
as well as 1 kb deletion mutant mice (Cln3∆exon7/8) that undergo AAV9-mediated CLN3 gene therapy (2B).
Successful completion of this project will provide an essential tool that removes the bottleneck of CLN3 disease
research and therapeutic development.

## Key facts

- **NIH application ID:** 10303283
- **Project number:** 1R03NS120081-01A1
- **Recipient organization:** UNIVERSITY OF KENTUCKY
- **Principal Investigator:** Qingjun Wang
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $153,000
- **Award type:** 1
- **Project period:** 2021-07-01 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10303283

## Citation

> US National Institutes of Health, RePORTER application 10303283, Method for Detection and Quantification of CLN3 Protein (1R03NS120081-01A1). Retrieved via AI Analytics 2026-06-16 from https://api.ai-analytics.org/grant/nih/10303283. Licensed CC0.

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