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Series GSE225540 Query DataSets for GSE225540
Status Public on Jul 04, 2023
Title Gene Editing of Hematopoietic Stem Cells Restores T Cell Response in a Familial Hemophagocytic Lymphohistiocytosis Model [Amplicon]
Organism Mus musculus
Experiment type Other
Summary Hemophagocytic lymphohistiocytosis (HLH) is a hyperinflammatory disorder characterized by a life-threatening cytokine storm and immunopathology. Familial HLH type 3 (FHL3) accounts for 30% of all inborn HLH cases worldwide. It is caused by mutations in the UNC13D gene, which result in impaired processing of cytotoxic vesicles and hence compromised T and NK cell-mediated killing. Current treatment protocols, including allogeneic hematopoietic stem cell (HSC) transplantation, still show 30-40% mortality. As a first step to meet this medical need, we developed and tested a curative genome editing strategy in the FHL3 Jinx mouse model. Jinx mice harbor a cryptic splice donor site (cSD) in Unc13d intron 26 and develop the clinical symptoms of human FHL3 upon infection with lymphocytic choriomeningitis virus (LCMV). Here, we employed CRISPR-Cas technology to delete the disease-underlying mutation in HSCs, and transplanted the Unc13d-edited stem cells into busulfan-conditioned Jinx recipient mice. Genotyping, phenotyping and CAST-Seq based off-target analyses of cells isolated from transplanted mice revealed efficient gene editing (>95%), polyclonality of the T cell receptor repertoire, and neither signs of off-target effects nor leukemogenesis. Unc13d transcription levels of edited and wildtype cells were comparable. LCMV challenge of the transplanted mice resulted in acute HLH in Jinx mice transplanted with mock-edited HSCs, while Jinx mice grafted with Unc13d-edited cells showed rapid virus clearance and protection from HLH. In sum, our study demonstrates that transplantation of CRISPR-Cas edited HSCs supports the development of a functional polyclonal T cell response in the absence of genotoxicity-associated clonal outgrowth or leukemogenesis.
Overall design Mouse bone marrow HSPCs were treated with two RNP complexes (1:3 molar ratio of Cas9:sgRNA). Genotyping was performed on various tissues pre and post transplantation.
Contributor(s) Dettmer-Monaco V, Weißert K, Monaco G, Rhiel M, Aichele P, Cathomen T
Citation(s) 37595758
Submission date Feb 17, 2023
Last update date Oct 09, 2023
Contact name Gianni Monaco
Organization name Freiburg Medical Center
Street address Breisacher Straße
City Freiburg
ZIP/Postal code 79106
Country Germany
Platforms (1)
GPL16417 Illumina MiSeq (Mus musculus)
Samples (19)
GSM7050012 M31 Amplicon-NGS on treated cells (post transplantation)
GSM7050013 M32 Amplicon-NGS on treated cells (post transplantation)
GSM7050014 M33 Amplicon-NGS on treated cells (post transplantation)
This SubSeries is part of SuperSeries:
GSE225541 Gene Editing of Hematopoietic Stem Cells Restores T Cell Response in a Familial Hemophagocytic Lymphohistiocytosis Model
BioProject PRJNA936144

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Supplementary file Size Download File type/resource
GSE225540_RAW.tar 11.1 Mb (http)(custom) TAR (of XLSX)
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Raw data are available in SRA
Processed data provided as supplementary file

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