A modular platform for targeted RNAi therapeutics

Abstract

Previous studies have identified relevant genes and signalling pathways that are hampered in human disorders as potential candidates for therapeutics. Developing nucleic acid-based tools to manipulate gene expression, such as short interfering RNAs (siRNAs), opens up opportunities for personalized medicine. Yet, although major progress has been made in developing siRNA targeted delivery carriers, mainly by utilizing monoclonal antibodies (mAbs) for targeting, their clinical translation has not occurred. This is in part because of the massive development and production requirements and the high batch-to-batch variability of current technologies, which rely on chemical conjugation. Here we present a self-assembled modular platform that enables the construction of a theoretically unlimited repertoire of siRNA targeted carriers. The self-assembly of the platform is based on a membrane-anchored lipoprotein that is incorporated into siRNA-loaded lipid nanoparticles that interact with the antibody crystallizable fragment (Fc) domain. We show that a simple switch of eight different mAbs redirects the specific uptake of siRNAs by diverse leukocyte subsets in vivo. The therapeutic potential of the platform is demonstrated in an inflammatory bowel disease model by targeting colon macrophages to reduce inflammatory symptoms, and in a Mantle Cell Lymphoma xenograft model by targeting cancer cells to induce cell death and improve survival. This modular delivery platform represents a milestone in the development of precision medicine.

A Combinatorial Library of Lipid Nanoparticles for RNA Delivery to Leukocytes

“…All the lipids were synthesized using standard organic synthesis procedures (see Supporting Information) and characterized by NMR and mass spectroscopy (see Supporting Information). LNPs were composed of ionizable lipid, cholesterol, 1,2‐distearoyl‐sn‐glycero‐3‐phosphocholine (DSPC), and PEGylated lipids in the molar ratios of 50:38.5:10:1.5, respectively, and assembled using a microfluidic mixing device, Nanoassemblar, as previously reported . The produced LNPs were small and uniformly distributed as evidenced by hydrodynamic diameter and polydispersity index (PDI).…”

“…PLK1 is overexpressed in a variety of tumor types and has a significant role in the maintenance of mitotic integrity. The silencing of PLK1 will arrest the cell cycle leading ultimately to cell death . The U266 cells were treated with different LNP‐siPLK1 at a dose of 0.06 µM and the cell viability was measured by the XTT assay.…”

Integrating Artificial Intelligence and Nanotechnology for Precision Cancer Medicine

“…A frequently asked question when discussing precision medicine treatment's is: can we personalize a nanomedicine for every patient? Although present techniques enable versatile conjugation of any desired antibody to nanoparticles, as well as variability in the choice of cargo (e.g., a specific siRNA sequence that matches the chosen gene for inhibition), many further obstacles remain in order to make this approach realizable . Besides the complicated clinical approval process for personalized nanomedicine, limitations of current fabrication techniques and the high costs of nanomedicine development must be addressed.…”

Tapping the RNA world for therapeutics

“…3). Some of these strategies, which have not yet been tested in the clinic, show exquisite targeting specificity and provide the possibility of knocking down genes in narrowly defined subsets of immune cells (i.e., only activated lymphocytes or only regulatory T cells) 88,93 or only in cancer cells 94 . Targeted gene knockdown is also likely to reduce bystander cell toxicity and require lower doses.…”