# The role of CSPalpha in Adult onset neuronal Lipofuscinosis pathogenesis

> **NIH NIH R21** · BETH ISRAEL DEACONESS MEDICAL CENTER · 2022 · $490,512

## Abstract

Autosomal dominant adult-onset neuronal ceroid lipofuscinosis (ANCL) is a rapidly progressing fatal  neurodegenerative dementia with no treatment currently available. Our group and others simultaneously  reported that mutations in the DNAJC5 gene are the most common cause of autosomal dominant ANCL. We  also showed that DNAJC5 mutations reduce lysosomal function and autophagic flux while causing the  accumulation of autofluorescent storage material (AFSM) and high molecular weight aggregates (HMWA) in  patient-derived fibroblasts. However, the mechanisms leading to neurodegeneration in ANCL are not known.  Furthermore, there are no studies done in relevant patient-derived brain cellular models. Unprecedented  advances in high-throughput and hypothesis-free “omics” technologies can generate highly detailed molecular  atlas for ANCL. We hypothesize that CSPα plays a role in the endo-lysosomal pathway and that CSPα mutations  lead to neuronal and microglial dysfunction and neurodegeneration. To test this hypothesis, we plan to perform  bulk-RNA Seq, single nucleus RNAseq and targeted proteomics in four brain regions with and without DNAJC5 mutations on differentially expressed genes and genes co-expressed with DNAJC5 mutants in a cell-specific  manner. Our preliminary data demonstrate that DNAJC5 transcript levels are higher in microglial cells than  neurons. We will perform bulk-RNAseq and proteomics in human iPSC-derived neurons and microglia with  DNAJC5 mutations and an isogenic control generated by CRISPR/Cas9 to define the impact of DNAJC5 on cell  autonomous-specific pathways. We will apply snRNA-seq to multiple brain areas affected differently by ANCL  pathology, thus tracing pseudotemporal trajectories of pathology progression, defining DEG in major brain cell  types and cell-type-specific transcriptional states for the first time in brains of carriers of DNAJC5 mutations.  (Aim 1). To determine the cell-autonomous effect of DNAJC5 mutations on ANCL pathology in human iPSC-derived neurons and microglia (Aim 2), we will use genetic (CRISPR/Cas9) and pharmacologic approaches in  iPSC-derived neurons and microglia to determine how DNAJC5 mutants impact the phagocytic capacity, cell  viability, lysosomal function, and the accumulation of pathogenic HMWA and AFSM. We will use proteomics in  human iPSC-derived neurons and microglia to test the effect of DNAJC5 mutations on the secretome and the  levels of prone-to-aggregate and proinflammatory proteins. We will harmonize RNA-seq and proteomic data  across the brain regions and iPSC-derived cells to determine DNAJC5-associated neurodegenerative pathways.

## Key facts

- **NIH application ID:** 10592180
- **Project number:** 1R21NS127211-01A1
- **Recipient organization:** BETH ISRAEL DEACONESS MEDICAL CENTER
- **Principal Investigator:** Bruno A. Benitez
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $490,512
- **Award type:** 1
- **Project period:** 2022-09-20 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10592180

## Citation

> US National Institutes of Health, RePORTER application 10592180, The role of CSPalpha in Adult onset neuronal Lipofuscinosis pathogenesis (1R21NS127211-01A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10592180. Licensed CC0.

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