Whole-genome CRISPR screening identifies genetic manipulations to reduce immune rejection of stem cell-derived islets
Human embryonic stem cells (hESCs) provide opportunities for cell replacement therapy of insulin-dependent diabetes. Therapeutic quantities of human stem cell-derived islets (SC-islets) can be produced by directed differentiation. However, preventing allo-rejection and recurring autoimmunity, withou...
Gespeichert in:
Veröffentlicht in: | Stem cell reports 2022-09, Vol.17 (9), p.1976-1990 |
---|---|
Hauptverfasser: | , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | Human embryonic stem cells (hESCs) provide opportunities for cell replacement therapy of insulin-dependent diabetes. Therapeutic quantities of human stem cell-derived islets (SC-islets) can be produced by directed differentiation. However, preventing allo-rejection and recurring autoimmunity, without the use of encapsulation or systemic immunosuppressants, remains a challenge. An attractive approach is to transplant SC-islets, genetically modified to reduce the impact of immune rejection. To determine the underlying forces that drive immunogenicity of SC-islets in inflammatory environments, we performed single-cell RNA sequencing (scRNA-seq) and whole-genome CRISPR screen of SC-islets under immune interaction with allogeneic peripheral blood mononuclear cells (PBMCs). Data analysis points to “alarmed” populations of SC-islets that upregulate genes in the interferon (IFN) pathway. The CRISPR screen
in vivo
confirms that targeting IFNγ-induced mediators has beneficial effects on SC-islet survival under immune attack. Manipulating the IFN response by depleting chemokine ligand 10 (CXCL10) in SC-islet grafts confers improved survival against allo-rejection compared with wild-type grafts in humanized mice. These results offer insights into the nature of immune destruction of SC-islets during allogeneic responses and provide targets for gene editing.
•
IFN pathway induction sets the fate of SC-islets under allogeneic immune challenge
•
“Alarm” genes drive immunogenicity of SC-islets
•
Genetically modified SC-islets were generated and evaluated for hypo-immunogenicity
•
CXCL10 depletion can reduce immune activation and SC-islet graft rejection
Using single-cell transcriptomics and whole-genome CRISPR screening, Sintov and colleagues find gene perturbation targets that can promote immune evasion of SC-islets in allogeneic transplantations. One of these targets, the early inflammatory response chemokine CXCL10, was depleted in SC-islets, which resulted in partial immune evasion
in vitro
and
in vivo
. |
---|---|
ISSN: | 2213-6711 2213-6711 |
DOI: | 10.1016/j.stemcr.2022.08.002 |