# Application of 4D proteomics and super-resolution microscopy in extracellular vesicle and particle-borne biomarker discovery for early pancreatic cancer detection

> **NIH NIH R01** · WEILL MEDICAL COLL OF CORNELL UNIV · 2024 · $453,986

## Abstract

Project Summary/Abstract
Pancreatic adenocarcinoma (PDAC) ranks among the most lethal cancers due to a late diagnosis and
ineffective treatments. Extracellular vesicles and particles (EVPs) are secreted by most cells, including tumor
cells, and package selective molecules, including proteins, lipids, nucleic acids, and metabolites. EVPs are
actively released into the circulation and mounting evidence suggests that circulating EVPs can serve as
biomarkers for early cancer detection. Mass spectrometry (MS) has been extensively utilized for biomarker
discovery in liquid biopsies, including EVP protein characterization. However, the scope and depth of the
information obtained is limited by (i) the sensitivity and resolution of the analytic technologies and (ii) the
proteomic complexity resulting from highly abundant serum-derived contaminants. The objective of this study
is to apply an optimized reproducible EVP isolation method in conjunction with asymmetric-flow field-flow
fractionation (AF4) technology to isolate EVP subsets with significantly improved purity and to employ three
novel analytic technologies, including extremely sensitive timsTOF 4D proteomic MS, super-resolution
dSTORM imaging analysis of single EVPs, and photocatalytic proximity labeling-proteomics (µMap) technology,
to discover and validate novel, circulating EVP protein biomarkers for early detection of PDAC. In Aim 1, by
employing the label-free timsTOF MS and using samples (blood plasma and tumor tissues) collected from
patients with newly diagnosed PDAC, we will identify novel circulating EVP protein biomarkers that correlate
with early stage disease. Top-ranked candidates will be further validated by robust absolute quantitation assays
employing targeted parallel reaction monitoring (PRM) MS. In Aim 2, we will determine the percent
representation and structural location of specific EVP biomarker proteins identified in Aim 1 at the single EVP
level by utilizing the super-resolution dSTORM imaging analysis. The performance of single EVP analysis will
be compared to the bulk analysis of individual protein targets via western blotting and/or ELISA analysis. We
will further explore the potential to apply this analytic tool directly to plasma samples without prior EVP isolation.
Lastly, in Aim 3, we will define protein-protein interactions (PPIs) of individual biomarkers by employing our
recently developed photocatalytic proximity labeling-proteomics (µMap) technology. Three potential biomarker
candidates identified in our previous study and novel candidates discovered in this study Aim 1 and Aim 2 will
be subjected to the PPI analysis. These interactions will be further validated by super-resolution imaging
analysis at the single-EVP, single-molecule level. We will establish if the presence or absence of these
interactions provides a more robust approach for early cancer detection. We predict that combined application
of these technologies will greatly facilitate novel biomarker ...

## Key facts

- **NIH application ID:** 10917237
- **Project number:** 5R01CA218513-07
- **Recipient organization:** WEILL MEDICAL COLL OF CORNELL UNIV
- **Principal Investigator:** Jacob Geri
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $453,986
- **Award type:** 5
- **Project period:** 2017-07-15 → 2028-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10917237

## Citation

> US National Institutes of Health, RePORTER application 10917237, Application of 4D proteomics and super-resolution microscopy in extracellular vesicle and particle-borne biomarker discovery for early pancreatic cancer detection (5R01CA218513-07). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/10917237. Licensed CC0.

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