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| 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.
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| 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.
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| 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 |
| E-mail(s) |
mongianni1@gmail.com
|
| Organization name |
Freiburg Medical Center
|
| Street address |
Breisacher Straße
|
| City |
Freiburg |
| ZIP/Postal code |
79106 |
| Country |
Germany |
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| Platforms (1) |
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| Samples (19)
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| 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) |
| GSM7050015 |
M34 Amplicon-NGS on treated cells (post transplantation) |
| GSM7050016 |
a Amplicon-NGS on treated cells (post transplantation) |
| GSM7050017 |
b Amplicon-NGS on treated cells (post transplantation) |
| GSM7050018 |
c Amplicon-NGS on treated cells (post transplantation) |
| GSM7050019 |
d Amplicon-NGS on treated cells (post transplantation) |
| GSM7050020 |
e Amplicon-NGS on treated cells (post transplantation) |
| GSM7050021 |
f Amplicon-NGS on treated cells (post transplantation) |
| GSM7050022 |
4L Amplicon-NGS on treated cells (post transplantation) |
| GSM7050023 |
5L Amplicon-NGS on treated cells (post transplantation) |
| GSM7050024 |
6L Amplicon-NGS on treated cells (post transplantation) |
| GSM7050025 |
4R Amplicon-NGS on treated cells (post transplantation) |
| GSM7050026 |
5R Amplicon-NGS on treated cells (post transplantation) |
| GSM7050027 |
6R Amplicon-NGS on treated cells (post transplantation) |
| GSM7050028 |
X Amplicon-NGS on treated cells (without transplantation) |
| GSM7050029 |
Y Amplicon-NGS on treated cells (without transplantation) |
| GSM7050030 |
Z Amplicon-NGS on treated cells (without transplantation) |
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| This SubSeries is part of SuperSeries: |
| GSE225541 |
Gene Editing of Hematopoietic Stem Cells Restores T Cell Response in a Familial Hemophagocytic Lymphohistiocytosis Model |
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| Relations |
| BioProject |
PRJNA936144 |
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