PEN photoporation for the genetic engineering of therapeutic mesenchymal stromal cells and T cells

Informazioni relative al progetto

Penphomet

ID dell’accordo di sovvenzione: 101158879

DOI

10.3030/101158879

Data della firma CE 26 Febbraio 2024

Data di avvio 1 Marzo 2024

Data di completamento 28 Febbraio 2027

Finanziato da

• The European Innovation Council (EIC)

Costo totale

€ 2 497 711,25

Contributo UE

€ 2 497 711,25

Coordinato da

TRINCE
Belgium

Periodic Reporting for period 1 - Penphomet (PEN photoporation for the genetic engineering of therapeutic mesenchymal stromal cells and T cells)

Periodo di rendicontazione: 2024-03-01 al 2025-02-28

Sintesi del contesto e degli obiettivi generali del progetto

Adoptive cell therapy has emerged as a promising strategy to treat cancer. It relies on patient-derived cells, such as T cells and mesenchymal stromal cells (MSCs), which are genetically engineered to become better equipped to fight cancer cells. While ex vivo genetic modification of T cells and MSCs has traditionally been performed with viral vectors, they come with concerns about safety, sustainable production and high development costs. Electroporation is a non-viral alternative transfection technology, but can lead to significant gene expression changes, phenotypic alterations, and decreased therapeutic potency. Recently, photoporation with electrospun photothermal nanofibers (PEN photoporation) was demonstrated to provide a safer alternative with minimal impact on the cell’s functionality and phenotype. The technology makes use of photothermal nanofibers which, upon stimulation with laser light, can transiently permeabilize cells to allow gene-modifying effector molecules to enter the cells. Having been thoroughly demonstrated and validated in a research setting (TRL4), this project aims to bring the PEN photoporation technology to TLR6 by developing hard- and software for automated high-throughput transfections of T cells (>1B cells/h) and MSCs (>10M cells/h). The technology will be extensively tested and validated in the cGMP compliant laboratories of the project partners for the genetic engineering of T cells and MSCs. At the same time, steps will be taken to prepare for commercialization and market deployment. By the end of the project a fully automated and validated high-throughput prototype system will be available for installation at centralized cell production facilities or ready for integration in point-of-care cell manufacturing equipment. This project aligns with the Micro-Nano-Bio challenge as it combines nanotechnology with microfluidics to enhance genetically engineered cell therapy products.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

During the first project year, we have successfully developed a cell culture plate containing photothermal nanofibers for the photoporation of cells. Experiments demonstrated, however, that photoporation with biodegradable polymeric particles was more efficient than with photothermal nanofibers. Therefore, it was decided to continue with those sensitizers instead, for which we could demonstrate efficient transfection of T cells and MSCs with relevant biological molecules, such as mRNA.

In-house built photoporation equipment was installed at the research facilities of the project partners, with which they could demonstrate that results on the transfection of T cells and MSCs are reproducible.

Hard- and software is being developed for high-throughput photoporation of cells.

It was demonstrated that photoporation of T cells and MSCs happens equally efficiently in microfluidic chambers as in traditional well plates.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

The possibility of transfecting T cells and MSCs in microfluidic devices opens up the possibility of developing closed and automated cGMP compliant photoporation equipment, ready for integration in cell therapy manufacturing facilities. Further work will focus on upscaling and demonstrating efficient transfection of these cells with relevant biomolecules other than mRNA, such as pDNA and gene editing enzymes.