# Mechanisms and Functional Consequences of Selective miRNA transfer via extracellular vesicles

> **NIH NIH P01** · VANDERBILT UNIVERSITY · 2021 · $354,769

## Abstract

exRNA in Colorectal Carcinoma: Biogenesis and Function
Project 2. Mechanisms and Functional Consequences of Selective miRNA transfer via
extracellular vesicles
James G. Patton - Investigator
Summary
Increasing evidence supports the hypothesis that extracellular vesicles (EVs) constitute a novel
form of cell-cell communication through the transfer of protein, RNA and lipid cargo. Project 2
will focus on selective EV miRNA transport and uptake by recipient cells. We previously
showed that EVs from KRAS mutant cells are enriched in miR-100 and miR-125b and we have
now shown in 3D culture that EVs enriched in miR-100 and miR-125b can confer cetuximab
resistance in recipient cells. However, it remains unknown how miR-100 and miR-125b (as well
as other miRNAs) are selectively targeted for secretion while other miRNAs are retained in cells.
Here, we will focus on determining whether transfer of miR-100 and miR-125b can alter the
tumor microenvironment using a novel in vivo xenograft model in zebrafish. We will also use an
adaptation of CRISPR-Display to test the hypothesis that specific RNA sequences and/or base
modifications regulate selective miRNA export. The same cell culture assay will be used to
identify RNA binding proteins (in concert with Project 1) that recognize sequence motifs or
modified bases to drive secretion of specific miRNAs which will then be extended to in vivo
effects using the zebrafish xenograft model. Lastly, our current hypothesis is that transfer of
miR-100 and miR-125b results in the activation of Wnt signaling but the full range of mRNA
targets for these miRNAs remains unknown. Thus, we will use RIP-USE to combine
immunoprecipitation of Ago2 associated miRNAs with differential expression analysis using
Unbiased Sequence Enrichment (RIP-USE) to identify all targets of miR-100 and miR-125b.
Normal cell-cell interactions and stem cell niches in the colonic crypt appear to result from the
secretion of EVs that set up opposing gradients of Wnt and EGFR signaling, our analyses will
identify potential therapeutic target genes whose expression is altered when proper cell-cell
communication is altered during colorectal cancer.

## Key facts

- **NIH application ID:** 10087490
- **Project number:** 5P01CA229123-02
- **Recipient organization:** VANDERBILT UNIVERSITY
- **Principal Investigator:** James G. Patton
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $354,769
- **Award type:** 5
- **Project period:** 2020-01-22 → 2024-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10087490

## Citation

> US National Institutes of Health, RePORTER application 10087490, Mechanisms and Functional Consequences of Selective miRNA transfer via extracellular vesicles (5P01CA229123-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10087490. Licensed CC0.

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