An intestinal organoid–based platform that recreates susceptibility to T-cell–mediated tissue injury

Abstract

A goal in precision medicine is to use patient-derived material to predict disease course and intervention outcomes. Here, we use mechanistic observations in a preclinical animal model to design an ex vivo platform that recreates genetic susceptibility to T-cell–mediated damage. Intestinal graft-versus-host disease (GVHD) is a life-threatening complication of allogeneic hematopoietic cell transplantation. We found that intestinal GVHD in mice deficient in Atg16L1, an autophagy gene that is polymorphic in humans, is reversed by inhibiting necroptosis. We further show that cocultured allogeneic T cells kill Atg16L1-mutant intestinal organoids from mice, which was associated with an aberrant epithelial interferon signature. Using this information, we demonstrate that pharmacologically inhibiting necroptosis or interferon signaling protects human organoids derived from individuals harboring a common ATG16L1 variant from allogeneic T-cell attack. Our study provides a roadmap for applying findings in animal models to individualized therapy that targets affected tissues.

Mouse models usher in precision medicine

“…1 RIPK3 deficiency in the ATG16L1-deficient recipient or administration of RIPK1 inhibitor GSK547 reduced GVHD severity and increased intestinal Paneth cell and epithelial cell survival. 1 Ex vivo intestinal organoid culture with alloreactive donor T cells induced necroptosis of intestinal organoid cells from the ATG16L1-deficient mice via interferon-g (IFN-g) and/or tumor necrosis factor-a (TNF-a). 1 These observations indicate that inflammatory cytokines from alloreactive T cells can trigger necroptosis in recipients with a defect in protective genes such as ATG16L1.…”

“…1 Ex vivo intestinal organoid culture with alloreactive donor T cells induced necroptosis of intestinal organoid cells from the ATG16L1-deficient mice via interferon-g (IFN-g) and/or tumor necrosis factor-a (TNF-a). 1 These observations indicate that inflammatory cytokines from alloreactive T cells can trigger necroptosis in recipients with a defect in protective genes such as ATG16L1. These studies also emphasize the involvement of host tissue in regulating GVHD, opening a new area of research to evaluate other protective genes in the parenchymal cells of GVHD target tissues.…”

“…As depicted in the diagram (see figure), mouse recipients with ATG16L1 deficiency in intestinal epithelium had augmented gut-GVHD with enhanced intestinal epithelial necroptosis after allo-HCT, which could be recapitulated in an ex vivo organoid culture system. 1 The death of the cultured organoid mediated by necroptosis was associated with an IFN signature, as measured with RNA-Seq analysis. 1 Blockade of the IFN signaling pathway by the JAK2 inhibitor Ruxolitinib or blockade of necroptosis pathway by RIPK1 inhibitor GSK547 markedly reduced necroptosis and increased survival of cultured intestinal organoids derived from ATG16L1deficient mice.…”

“…1 The death of the cultured organoid mediated by necroptosis was associated with an IFN signature, as measured with RNA-Seq analysis. 1 Blockade of the IFN signaling pathway by the JAK2 inhibitor Ruxolitinib or blockade of necroptosis pathway by RIPK1 inhibitor GSK547 markedly reduced necroptosis and increased survival of cultured intestinal organoids derived from ATG16L1deficient mice. 1 Matsuzawa-Ishimoto et al were also able to set up intestinal organoid cultures from fresh as well as frozen human intestinal tissues.…”

“…1 Blockade of the IFN signaling pathway by the JAK2 inhibitor Ruxolitinib or blockade of necroptosis pathway by RIPK1 inhibitor GSK547 markedly reduced necroptosis and increased survival of cultured intestinal organoids derived from ATG16L1deficient mice. 1 Matsuzawa-Ishimoto et al were also able to set up intestinal organoid cultures from fresh as well as frozen human intestinal tissues. Coculture with donor T cells or TNF-a induced death of the cultured human organoids.…”