# Development of cell free messenger RNA-based non-invasive diagnostic biomarker for Alzheimer's Disease

> **NIH NIH R44** · SUPERFLUID DX, INC. · 2024 · $843,061

## Abstract

Alzheimer’s disease (AD) is the most common cause of dementia, affecting over 40 million people and is
projected to triple by 2050. Although post-mortem examination is the gold standard for establishing AD
pathology, assessment of amyloid β (Aβ) and tau levels in cerebrospinal fluid (CSF) and/or positron-emission
tomography (PET) are currently used widely as surrogates. However, imaging modalities are costly and CSF
procedure is invasive. Therefore, there is a need for highly accessible and cost-effective non-invasive diagnostic
tests for AD and patients with mild cognitive impairment (MCI) who will eventually develop AD. Over the last two
decades, circulating nucleic acids have emerged as a viable source of liquid biopsy in multiple diseases. Recent
whole-transcriptome characterization studies using post-mortem human brain tissue samples have
demonstrated that the transcriptional profile of AD patients differ significantly from healthy individuals. Therefore,
circulating messenger RNA (mRNA) appear to be a promising non-invasive diagnostic biomarker source for AD.
Although quantification of cell-free mRNAs (cf-mRNAs) was once considered challenging due to their low
abundance in the circulation, we and others have developed a robust next generation sequencing (NGS) based
platform for accurate quantification of cf-mRNA. Subsequently, we have conducted preliminary cf-mRNA
profiling studies in multiple diseases including neurodegenerative diseases, liver diseases and cancers and
demonstrated its utility as a platform to develop non-invasive biomarkers for diagnosis and monitoring of
diseases. In particular, we have recently profiled plasma cf-mRNA transcriptomes of 126 AD subjects and
compared to those of 115 age matched controls. We discovered that a substantial number of genes were
dysregulated in plasma of AD subjects and some of these genes correlated with severity of cognitive impairment
in AD subjects. These AD associated dysregulated cf-mRNA transcripts were able to robustly identify AD
subjects and demonstrated the potential of cf-mRNA as a source for non-invasive biomarker development. In
this proposal, we will first optimize and establish clinical-grade protocols to minimize potential variabilities for the
downstream cf-mRNA Seq profiling. We will then profile the cf-mRNA transcriptome of well-characterized AD
and MCI subject samples as well as other neurological diseases and build diagnostic and prognostic classifiers
for AD and MCI using comprehensive bioinformatic approaches. We will subsequently validate the performance
of these classifiers in intended-use clinical validation cohorts.

## Key facts

- **NIH application ID:** 10916443
- **Project number:** 5R44AG078038-02
- **Recipient organization:** SUPERFLUID DX, INC.
- **Principal Investigator:** Shusuke Toden
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $843,061
- **Award type:** 5
- **Project period:** 2023-09-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10916443

## Citation

> US National Institutes of Health, RePORTER application 10916443, Development of cell free messenger RNA-based non-invasive diagnostic biomarker for Alzheimer's Disease (5R44AG078038-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10916443. Licensed CC0.

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