# TNBC Ligand-displaying Exosomes Using RNA Nanotechnology for Targeted Cytosol Delivery of RNAi without Endosome Entrapment

> **NIH NIH R01** · OHIO STATE UNIVERSITY · 2022 · $324,643

## Abstract

SUMMARY
 Triple negative breast cancer (TNBC) is a heterogeneous, complex, and aggressive breast cancer subtype.
TNBC patients respond poorly to chemotherapy, leading to high mortality rates and a worsening prognosis.
RNAi therapeutics, including siRNA and miRNA, have shown tremendous potential for TNBC cancer therapy.
Developing a safe targeted delivery system with endosomal avoidance of the payload is crucial in terms of
realizing the full potential of RNAi therapeutics and could revolutionize clinical treatment of TNBC.
 Our group has demonstrated that delivery of anti-miR21 to TNBC can efficiently inhibit TNBC cell proliferation
(Shu D, et al. ACS Nano. 2015, 27:9731; Yin H, et al. Shu D. Mol Therapy. 2019, 27:1252). We have also
recently reported the use of RNA nanoparticle orientation for decorating exosome surfaces with targeting ligands
to deliver siRNA loaded exosomes for TNBC treatment. We proved that exosomes can be utilized as
nanocarriers to deliver siRNA to TNBC cancer cells very efficiently to inhibit cancer growth (Pi F, Binzel D et al.
Nat. Nanotechnol. 2018;13:82). Preliminary data from the investigation with KB cell models in vitro has shown
the mechanism behind the high efficiency of cancer inhibition to be the cytosolic delivery of siRNA via exosomes
without endosomal trapping (Zhen Z, et al. J Control Release. 2019,311:43). In this study, we will investigate
the targeting and delivery mechanisms of RNAi to TNBC cells by exosomes displayed with TNBC specific
ligands for targeted delivery. It’s our goal to select a lead TNBC therapeutic candidate to move towards potential
clinical translation. We will combine the targeting and drug delivery capabilities of RNA nanotechnology and
exosomes for targeted delivery of RNAi to cell cytosols without endosomal entrapment.
 We will construct and evaluate the multi-functional exosomal RNA nanoparticle complexes harboring
targeting ligands including RNA aptamers (EGFRapt, or alternatively, CD133apt, CD44apt, and nAHRsapt) or
chemical ligand (Methotrexate) and tumor suppressing RNAi therapeutics (siRNA, suppressor miRNAs and anti-
oncogenic miRNA). We will further investigate the pathways of internalization and intracellular trafficking in
addition to investigating whether the payloads in the exosome are delivered to cytosol via fusion or to the
endosome via endocytosis. In depth studies of the release and cellular processing of RNAi cargo loaded into
exosomes will be completed. We will investigate the PK/PD parameters, delivery mechanisms, antitumor
efficacy, and safety of the therapeutic RNA nanoparticles in order to select a lead candidate for preclinical
translation. The preclinical studies will give a clear understanding of the viability of exosome-RNA nanoparticle
complexes for TNBC therapy and provide data to move towards an Investigational New Drug (IND) application
that will facilitate advancement to clinical trials.

## Key facts

- **NIH application ID:** 10404055
- **Project number:** 5R01CA257961-02
- **Recipient organization:** OHIO STATE UNIVERSITY
- **Principal Investigator:** Daniel W Binzel
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $324,643
- **Award type:** 5
- **Project period:** 2021-05-11 → 2026-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10404055

## Citation

> US National Institutes of Health, RePORTER application 10404055, TNBC Ligand-displaying Exosomes Using RNA Nanotechnology for Targeted Cytosol Delivery of RNAi without Endosome Entrapment (5R01CA257961-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10404055. Licensed CC0.

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