Journal article
Whole-genome CRISPR screening identifies N-glycosylation as a genetic and therapeutic vulnerability in CALR-mutant MPNs
Blood, v 140(11), pp 1291-1304
15 Sep 2022
PMID: 35763665
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Calreticulin (CALR) mutations are frequent, disease-initiating events in myeloproliferative neoplasms (MPNs). Although the biological mechanism by which CALR mutations cause MPNs has been elucidated, there currently are no clonally selective therapies for CALR-mutant MPNs. To identify unique genetic dependencies in CALR-mutant MPNs, we performed a whole-genome clustered regularly interspaced short palindromic repeats (CRISPR) knockout depletion screen in mutant CALR-transformed hematopoietic cells. We found that genes in the N-glycosylation pathway (among others) were differentially depleted in mutant CALR-transformed cells as compared with control cells. Using a focused pharmacological in vitro screen targeting unique vulnerabilities uncovered in the CRISPR screen, we found that chemical inhibition of N-glycosylation impaired the growth of mutant CALR-transformed cells, through a reduction in MPL cell surface expression. We treated Calr-mutant knockin mice with the N-glycosylation inhibitor 2-deoxy-glucose (2-DG) and found a preferential sensitivity of Calr-mutant cells to 2-DG as compared with wild-type cells and normalization of key MPNs disease features. To validate our findings in primary human cells, we performed megakaryocyte colony-forming unit (CFU-MK) assays. We found that N-glycosylation inhibition significantly reduced CFU-MK formation in patient-derived CALR-mutant bone marrow as compared with bone marrow derived from healthy donors. In aggregate, our findings advance the development of clonally selective treatments for CALR-mutant MPNs.
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Details
- Title
- Whole-genome CRISPR screening identifies N-glycosylation as a genetic and therapeutic vulnerability in CALR-mutant MPNs
- Creators
- Jonas S Jutzi - Harvard UniversityAnna E Marneth - Harvard UniversityMichele Ciboddo - Brigham and Women's HospitalAngel Guerra-Moreno - Brigham and Women's HospitalMaría José Jiménez-Santos - Spanish National Cancer Research CentreAnastasia Kosmidou - Brigham and Women's HospitalJames W Dressman - Medical University of South CarolinaHongyan Liang - Medical University of South CarolinaRebecca Hamel - Harvard UniversityPatricia Lozano - Brigham and Women's HospitalElisa Rumi - University of PaviaJohn G Doench - Broad InstituteJason Gotlib - Stanford UniversityAnandi Krishnan - Stanford UniversityShannon Elf - Brigham and Women's HospitalFátima Al-Shahrour - Spanish National Cancer Research CentreAnn Mullally - Harvard University
- Publication Details
- Blood, v 140(11), pp 1291-1304
- Publisher
- American Society of Hematology (ASH)
- Grant note
- T32 GM132055 / NIGMS NIH HHS UL1 TR003142 / NCATS NIH HHS R01 HL131835 / NHLBI NIH HHS K08 HG010061 / NHGRI NIH HHS UL1 TR001085 / NCATS NIH HHS R25 GM113278 / NIGMS NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Medicine (Graduate)
- Web of Science ID
- WOS:000877605100011
- Scopus ID
- 2-s2.0-85138090868
- Other Identifier
- 991021932789104721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Hematology