# Molecular dissection and imaging of extracellular vesicles to define their origin and targets

> **NIH NIH UH3** · MASSACHUSETTS GENERAL HOSPITAL · 2021 · $1,100,083

## Abstract

Extracellular vesicles (EVs) and the RNAs contained within them (EV-RNAs) are secreted into biofluids by every
cell type. EV-RNAs have emerged as potential prognostic or predictive biomarkers of a wide range of diseases,
providing a targetable, accurate real-time representation of the disease state. However, advancement of EV-
RNAs in the clinic as biomarkers of disease has been impeded by challenges in their isolation and
characterization. Most notably, there is a lack of tools and techniques to i) isolate and precisely characterize
tissue-specific EV-RNA populations; ii) define heterogeneity in surface markers and RNA content in tissue-
specific EV populations; and iii) determine changes in EV-RNAs associated with disease state. In this multi-PI
proposal, we will use a collaborative and innovative approach to advance technology that would allow isolation
and granular characterization of EV populations from hematopoietic cells, brain, and heart.
 Our objective in the UG3 phase is to identify cell/tissue specific markers for isolation of EVs using
computational analysis, transcriptomics, and EV-tracking in genetic mouse models; this approach would allow
for fluorescence/antibody-based identification of EVs in a cell-specific manner. Information will also be obtained
on individual EVs with a novel quantitative single molecule localization microscopy (qSMLM) approach. qSMLM,
a sensitive fluorescence-based imaging method, will be used to quantify the number of affinity isolated EVs, their
size, and key RNA content using molecular beacons. The identified tissue-specific EV-markers and EV-RNAs
will be used in the UH3 phase for validation in a variety of human models. Our objective in the UH3 phase is to
determine cellular/tissue contribution to EV-RNAs from Tissue-Chip effluents; and assess dynamic changes in
EVs from human plasma from subjects with acute disease (coronary ischemia or cerebrovascular accident) or
physiological processes (exercise). We will validate key EV-RNAs (using nano-flow cytometry and molecular
beacons) and use qSMLM with molecular beacons to provide a quantitative profile of EVs at a single vesicle
resolution. Notably, proposed experiments will help determine contribution of different tissues to the plasma
biofluid RNA landscape at baseline, and in response to physiological or disease stressors.
Together, the tools and techniques developed in the UG3 phase, and validated in the UH3 phase would serve
as a road-map for the discovery and development of EV markers and EV-RNAs specific to other tissues.
Ultimately, the use of tissue-specific EV-RNAs to probe disease state would provide a dynamic window into
disease progression or regression with higher sensitivity and fidelity compared to currently available
technologies.

## Key facts

- **NIH application ID:** 10350010
- **Project number:** 4UH3TR002878-03
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** Saumya Das
- **Activity code:** UH3 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $1,100,083
- **Award type:** 4N
- **Project period:** 2019-09-16 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10350010

## Citation

> US National Institutes of Health, RePORTER application 10350010, Molecular dissection and imaging of extracellular vesicles to define their origin and targets (4UH3TR002878-03). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10350010. Licensed CC0.

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