A Biomimetic In Vitro Model of the Pro-Inflammatory Microenvironment Recapitulates Type I/II Interferon Signaling and...

Preclinical drug development suffers from a persistent translational gap, as conventional animal models fail to reproduce the human immune response, contributing to high attrition rates of drug candidates in clinical trials. To address this, we previously established an in vitro high-density culture (HDC) model of acute monocytic leukemia (AML) cells that recapitulates an aseptic pro-inflammatory microenvironment using acute monocytic leukemia (AMoL) THP-1 cells. In the present study, using RNA-seq, quantitative RT-PCR, flow cytometry, Western blotting, and multiplex cytokine analysis, we characterized how this biomimetic microenvironment shapes Type I and II interferon signaling in THP-1 cells that acquire resistance to apoptosis under HDC conditions. The model revealed predominant activation of the Type II interferon pathway, marked by elevated interferon-gamma expression, STAT1 activation, and increased production of interferon-stimulated chemokines. Protein-protein interaction network analysis identified key interferon-induced hub genes and regulatory transcription factors driving this response. Notably, the interferon-stimulated gene profile generated by this in vitro platform, including the Interferon-Related DNA Damage Resistance Signature, closely resembled gene expression patterns observed in clinical samples from patients with refractory AMoL, supporting the relevance of the HDC biomimetic system as a research model of inflammatory adaptation in leukemia cells and associated resistance. These findings support further investigation of the HDC biomimetic system as a research model of inflammatory adaptation in leukemia cells and associated resistance.