Impaired Proteolysis of Noncanonical RAS Proteins Drives Clonal Hematopoietic Transformation
- Description
This study demonstrated that loss of LZTR1, or leukemia-associated mutants in the LZTR1 substrate and RAS GTPase RIT1 that escape degradation, drives hematopoietic stem cell expansion and leukemia in vivo. For the study, genome-scale CRISPR/Cas9 screen performed in different variants of the TF-1 cell line genetically-modified to express mutant forms of RIT1 (M90I or F82C) or depleted in LZTR1 by CRISPR/Cas9, then infected with the Brunello single guide RNA (sgRNA) library and cells collected following 7 days of selection (day 0) or an additional 14 days in culture (day 14) to assess drop-out of sgRNAs between days 0 and day 14 by next-generation sequencing. In addition, single cell RNA sequencing was used to assess changes in gene expression within murine bone marrow associated with Rit1-M90I mutation or Lztr1 knock-out compared to a wild-type control.
Access
- Restrictions
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Free to All
- Instructions
- CRISPR screen and single cell RNA sequencing data have been deposited in the Gene Expression Omnibus (GEO).
- Grant Support
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Takeda Science Foundation/Takeda Science FoundationLeukemia & Lymphoma Society/Leukemia & Lymphoma SocietyW81XWH-20-1-0391/Congressionally Directed Medical Research ProgramsMSD Life Science Foundation/MSD Life Science FoundationKanae Foundation for the Promotion of Medical Science/Kanae Foundation for the Promotion of Medical ScienceJP20H00537/JSPS KAKENHIInoue Foundation for Science/Inoue Foundation for ScienceGeoffrey Beene Cancer Research Center/Memorial Sloan Kettering Cancer CenterEdward P. Evans MDS Foundation/Edward P. Evans MDS FoundationLady Tata Memorial Trust/Lady Tata Memorial TrustAmerican Society of Hematology/American Society of HematologyASH Research Training Award for Fellows/American Society of Hematology