Method Profile

Colorectal Cancer Immune Assembloids for IFNγ Agonist Comparison

Patient-derived colorectal organoid assembloids with stromal and immune cells used to compare recombinant IFNγ, an untargeted reference and Roche-engineered FAP-targeted IFNγ receptor agonists.

Tumor ModelsOrganoids
Cell-type maps and interferon-response analyses from the colorectal-cancer immune assembloid study.
Cell-type and interferon-response analyses in the colorectal-cancer immune assembloid study (Fig. 3). (Nina Maenhoudt, Mechthild Lütge, Melanie Obenloch et al., CC BY 4.0)

Research Applications

Comparing cellular and spatial immune responses to engineered cytokine candidates in a mixed tumor–stromal–immune model.

Model & Design

Patient-derived colorectal cancer organoids, autologous cancer-associated fibroblasts and allogeneic peripheral blood mononuclear cells in a three-dimensional matrix.

Experimental Format

Untreated, recombinant IFNγ, untargeted reference and FAP-targeted cis/trans/all-in-one conditions compared through imaging, cytokine assays, single-cell transcriptomics and T-cell migration.

Model Components

  • Component

    Tumor and stromal components

    Description

    Patient-derived colorectal tumor organoids combined with fibroblasts from the same tumors.

  • Component

    Immune and treatment components

    Description

    Allogeneic peripheral blood immune cells; recombinant IFNγ, untargeted reference and FAP-targeted receptor agonists.

Measurements & Endpoints

  • Ki67-positive tissue area
  • MHC-I and PD-L1 staining
  • CXCL10 release and inflammatory cytokines
  • CD8-positive T-cell migration
  • Cell-type-specific transcriptional states

Analysis

Contrast each treatment with untreated assembloids and distinguish recombinant cytokine from the untargeted and FAP-targeted constructs.

Published Evidence

  • Study Design

    Comparative exposure of patient-derived assembloids to recombinant and engineered IFNγ agonists; key Figure 4 comparisons at 100 nM.

    Study Sample

    Figure 4 reports three or four independent patients for histologic measures and three patient-derived lines for migration.

    Comparison

    Untreated controls, recombinant IFNγ and an untargeted receptor-binding reference.

    Finding

    Recombinant IFNγ significantly reduced Ki67 and increased CD8 migration; FAP-targeted constructs gave little PD-L1 upregulation but Ki67 and CD8 migration changes were trends that did not reach significance over untreated controls.

    Interpretation & Limits
    • Roche designed the molecules, funded and conducted the study; several authors held company stock or patents.
    • The assembloids use allogeneic immune cells and lack vascular and systemic interactions.

Sources

  1. Evaluating cytokine-based therapeutics in human 3D in vitro immune-containing assembloids. Frontiers in Cell and Developmental Biology. 2026-10-15. DOI: 10.3389/fcell.2026.1930651. ↗