# Mass spectrometry and multiplexed immunofluorescence imaging of metabolic and proteomic contributors to selective neuronal vulnerability in Alzheimer's disease

> **NIH NIH R01** · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · 2024 · $71,342

## Abstract

We aim to uncover metabolic and protein signaling pathways contributing to the regional vulnerability of
neocortical pyramidal neurons in Alzheimer’s disease and to identify novel targets for detection and
intervention. We will compare the prefrontal cortex, a neocortical brain area afflicted by neuropathology
early in Alzheimer’s disease, with the primary visual cortex, a brain area that is relatively spared. In the
funded grant, we are using mass spectrometric imaging in combination with segmentation analyses, to
identify spatial changes of small molecules and proteins in postmortem brain sections prepared from
these neocortical areas from donors at various clinical and pathological stages of Alzheimer’s disease
compared to controls. In the proposed supplement, we will apply multiplexed immunofluorescence
imaging on sections from the same specimens, to obtain information on cellular and microenvironment
changes, specifically astrocytic changes, in and around vulnerable neocortical neurons during disease
progression, correlated with the clinical severity, degree and location of neuropathological changes, and
the risk genotype. We will characterize cortical layer-specific changes in astrocytic coverage of blood
vessels, determine how astrocytic cell state and morphology changes in Alzheimer’s disease, and
ascertain how astrocyte heterogeneity correlates spatially to neuronal vulnerability. We will optimize cell
co-culture systems and transcriptional analysis for validation studies. Further, in the parent grant, we will
register the data from both imaging techniques to identify metabolic pathways and protein signaling
changes at the regional, laminar and cellular level and to locate covariation in molecular and cellular
phenotypes contributing to Alzheimer’s disease vulnerability. In addition to generating a comprehensive
dataset of the cellular and molecular changes at various stages of Alzheimer’s disease, these studies will
involve the development, validation, and dissemination of novel tools for analysis of large datasets
generated using two powerful imaging tools, one that detects hundreds of analytes with the possibility of
detecting previously unknown contributors to disease and the other that provides higher resolution with
a select set of known markers. In the long term, the data generated from these studies could provide the
basis for testing novel disease-modifying treatments by cell-type specific targeting of identified metabolic
pathways using experimental models, such as brain organoids to replicate cortical lamination with human
neurons or humanized mouse chimeras to model interactions between neurons and non-neuronal cell
types.

## Key facts

- **NIH application ID:** 11017225
- **Project number:** 3R01AG078755-03S1
- **Recipient organization:** ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
- **Principal Investigator:** PARAG Kumar MALLICK
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $71,342
- **Award type:** 3
- **Project period:** 2022-09-15 → 2024-12-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11017225

## Citation

> US National Institutes of Health, RePORTER application 11017225, Mass spectrometry and multiplexed immunofluorescence imaging of metabolic and proteomic contributors to selective neuronal vulnerability in Alzheimer's disease (3R01AG078755-03S1). Retrieved via AI Analytics 2026-09-03 from https://api.ai-analytics.org/grant/nih/11017225. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*