Systemic Targeting of Therapeutic RNA to Tumors via a Cell-Penetrating and RNA-Binding Antibody
Date of Award
Spring 2023
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Genetics
First Advisor
Glazer, Peter
Abstract
With the success of RNA-based vaccines, there is intense interest in the advancement of RNAs as rationally designed therapeutic agents. In oncology, a major focus is the use of RNAs to stimulate pattern recognition receptors (PRRs) to leverage innate immune responses. However, delivery of RNAs in a tumor-specific manner to avoid systemic toxicity has been a challenge. Here we report that a tumor-targeting, cell-penetrating, and RNA-binding antibody, cD31N, can form non-covalent antibody/RNA complexes (ARCs) that mediate highly specific and functional delivery of RNAs into tumors following intravenous injection. Using 3p-hpRNA, an agonist of the RNA-sensing PRR, retinoic acid-inducible gene-I (RIG-I), we observed anti-tumor efficacy of systemically administered cD31N/3p-hpRNA complexes in multiple tumor models in mice including subcutaneous melanomas (B16.F10 OVA) and orthotopic medulloblastoma (DAOY) and pancreatic cancers (KPC). Synergy between cD31N/3p-hpRNA ARCs and anti-PD-1 therapy was further observed in breast (EMT6) and colon (MC38) cancer models. Immunologic correlates consistent with RIG-I pathway activation were detected in multiple assays, and specific delivery of 3p-hpRNA was confirmed by RT-PCR, demonstrating a 1000-fold difference in RNA uptake into tumor cells versus non-malignant cells within the tumor microenvironment. These studies establish that cD31N-based ARCs effectively deliver immune stimulating RNA payloads to tumors, providing a novel platform to advance rationally designed RNAs for cancer therapy. In follow-up work, we have demonstrated that cD31N can deliver alternative RIG-I payloads and messenger RNA (mRNA). Using covalent conjugation, we have also demonstrated the ability of cD31N to deliver phosphorodiamidate morpholino oligomers (PMOs), which are not amenable to complexation. In vitro and in vivo, cD31N-PMOs demonstrated potential to induce skipping of exon 23 in a mouse model of skeletal muscle disease.
Recommended Citation
Quijano, Elias, "Systemic Targeting of Therapeutic RNA to Tumors via a Cell-Penetrating and RNA-Binding Antibody" (2023). Yale Graduate School of Arts and Sciences Dissertations. 871.
https://elischolar.library.yale.edu/gsas_dissertations/871
