# A novel microfluidic platform to study exosome biology in PAH.

> **NIH NIH R21** · STANFORD UNIVERSITY · 2022 · $196,750

## Abstract

The endothelium is the cellular monolayer that covers the inner lining of the entire circulatory system. Endothelial
dysfunction is a feature of pulmonary arterial hypertension (PAH), a life-threatening disease associated with
abnormally high pulmonary pressures and chronic right heart failure. Due to the limitations of available static cell
culture and animal models, our understanding of the mechanisms that orchestrate the initiation and perseverance
of endothelial dysfunction in PAH remains incomplete. Given that endothelial dysfunction is a common finding in
PAH, an understanding of the mechanism behind maladaptive endothelial responses could help accelerate the
discovery of novel therapies for PAH. Presently, it is believed that endothelial derived exosomes contribute to
PAH by carrying signals that trigger maladaptive endothelial responses in the setting of injury. Exosomes are
cell-derived small (~30-150 nm) extracellular vesicles that carry proteins, metabolites and nucleic acids involved
in a variety of physiological and pathological processes. While it is known that exosomes carry molecular and
genetic factors associated with angiogenesis, inflammation and vasoreactivity, a comprehensive assessment of
exosome cargo of healthy and dysfunctional PMVECs has been hindered by current low-yield exosome isolation
techniques. These techniques cannot perform real-time dynamic exosome isolation from pulmonary
microvascular endothelial cells (PMVECs) exposed to PAH-associated stressors. To address this unmet need,
we have designed the MFES (Multifunctional Exosome Sorter) that can dissect the whole exosome population
into subpopulations based on size and surface markers. MFES is the first lab-on-a-chip platform that integrates:
1) a vessel-on-a-chip module for real-time characterization of PMVEC functional responses across a wide range
of physiological and pathological parameters, 2) a module for high-yield exosome size-based isolation, 3) a
surface marker based exosome sorting using magnetic beads, and 4) multi-omics phenotyping of exosomes of
PMVECs. Here, we are proposing a technology that can enable broadly to investigate the two main defining
characteristics of exosomal subtypes, i.e., size and surface markers, both separately independently, and in
combination sequentially. We will characterize changes in exosome cargo in healthy and PAH PMVECs exposed
to shear stress-related conditions in the MFES. We will isolate subpopulations of exosomes based on size and
surface markers and characterize them for their cargo (Aim 1). Then, we will determine whether exosomes
derived from stressed PMVECs can induce pathological changes in healthy PMVECs cultured in a microfluidic
culture chip (Aim 2). This technological innovation enables to study endothelial exosome biology in a setting that
represents the flow dynamics associated with PAH. Further, the use of cutting-edge -omics technologies,
bioinformatic analysis integrated with machine learning algorith...

## Key facts

- **NIH application ID:** 10378161
- **Project number:** 5R21HL156761-02
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** VINICIO A DE JESUS PEREZ
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $196,750
- **Award type:** 5
- **Project period:** 2021-03-25 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10378161

## Citation

> US National Institutes of Health, RePORTER application 10378161, A novel microfluidic platform to study exosome biology in PAH. (5R21HL156761-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10378161. Licensed CC0.

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