As new modalities enter clinical trials, the demand for human-relevant in vitro models is increasing. Traditional pre-clinical animal models often fail to accurately predict drug efficacy and toxicity, leading to high late-stage drug attrition rates. 

New modalities, such as oligonucleotide therapies, are highly specific to humans and require a more human-centric development approach. Non-human species are frequently unsuitable for testing, and the most relevant options, like non-human primates (NHPs), are costly and ethically challenging. Therefore, human-relevant new approach methodologies (NAMs), such as organ-on-a-chip (OOC) technologies, are essential for improving the predictive power of pre-clinical testing. OOC platforms, also known as microphysiological systems (MPS), are designed to replicate the functional biomarkers of cells and tissues in a more physiologically relevant manner by culturing primary human cells on perfused 3D scaffolds (Rubiano et al., 2021). MPS have the potential to offer a more physiological approach to evaluating and designing oligonucleotide therapies before first-in-human trials.

Aim

We introduce the PhysioMimix Liver MPS to study the delivery of oligonucleotides into the liver and their uptake by PHHs. The Liver MPS produces functional and stable PHH microtissues, enabling the study of gene knockdown and dosing strategies over 14 days. To address the growing need for appropriate in vitro testing of such therapeutics, we target PHHs with fluorescently labelled siRNA or ASOs, with and without GalNAc conjugation, in our Oligonucleotide delivery assay. Our results show effective uptake by PHHs and subsequent knockdown of the target genes of interest.

Key messages

• Oligonucleotide therapies are highly specific to humans and require a more human-centric development approach.
• MPS replicates the structural and functional biomarkers of cells and tissues in a more physiologically relevant manner.
• Our study demonstrates the effectiveness of a Liver MPS and Oligonucleotide delivery assay in examining the uptake and gene knockdown, of oligonucleotides targeting the liver.
• MPS insights facilitate the development of new modality drugs where animal models are less suited, enabling better informed first-in-human studies.