Targeted delivery of lipid nanoparticle-mRNA formulations modified with ligand pattern to dendritic cells based on DNA origami

Project Information

LIPIRNADO

Grant agreement ID: 101068419

DOI

10.3030/101068419

Project closed

EC signature date 9 June 2022

Start date 1 August 2023

End date 31 July 2025

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 222 727,68

Coordinated by

KAROLINSKA INSTITUTET
Sweden

Periodic Reporting for period 1 - LIPIRNADO (Targeted delivery of lipid nanoparticle-mRNA formulations modified with ligand pattern to dendritic cells based on DNA origami)

Reporting period: 2023-08-01 to 2025-07-31

Summary of the context and overall objectives of the project

Cancer immunotherapy has revolutionized the treatment of various malignancies. However, current therapeutic strategies often face limitations in precision, reproducibility, and control over immune cell activation. The LIPIRNADO project aims to address these challenges by developing DNA origami-based molecular platforms to precisely modulate T cell receptor (TCR) signaling. By mimicking natural spatial arrangements of immune ligands, this project explores how nanoscale geometry and molecular crowding affect T cell activation. The ultimate objective is to pave the way for programmable, synthetic immunotherapies with higher specificity and tunability. This fundamental research contributes to EU priorities in health, innovation, and responsible technology development.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The project designed and fabricated two types of DNA origami platforms—single-layer hexagonal and double-layer polygonal nanostructures—each equipped with a single anti-CD3 Fab fragment. These nanodevices were validated by AFM, agarose gel electrophoresis, and DNA-PAINT imaging to confirm structural integrity and binding capacity. Using Jurkat NFAT-luciferase reporter cells and hPBMCs, the project demonstrated that spatial positioning and structural rigidity of Fab-bearing origami directly influenced T cell activation strength and cytokine release. The most significant finding is that DNA origami–anchored anti-CD3 Fab, even in single copies, can trigger TCR activation in a geometry-dependent manner, providing strong evidence for kinetic segregation theory in a synthetic context.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This project represents a conceptual leap in the design of nanoscale immunomodulators. Unlike traditional antibody therapies or CAR-T platforms, these DNA origami constructs offer full spatial control of ligand display and enable programmable interactions with immune receptors. The results demonstrate a previously unachievable level of precision in modulating immune signaling, validating the potential of nanotechnology in synthetic immunology. These findings support the case for further translational research, including testing in relevant in vivo or organoid models. The work opens doors for patenting geometry-specific immunomodulators and collaboration with biotech or medtech companies for therapeutic development and commercialization.

Preparation of anti-CD3 Fab and DNA origami

Jurkat T cell activated by origami with various pillar length

Jurkat T cell activated by single- and double-layer origami-Fab conjugates

Schematic illustration of TCR kinetic segregation model by DNA origami

DNA-PAINT imaging on Jurkat T cells

Human PBMC activated by SL_Fab conjugates without pillar and with 30-nm pillar