Journal
BLOOD
Volume 140, Issue 11, Pages 1291-1304Publisher
AMER SOC HEMATOLOGY
DOI: 10.1182/blood.2022015629
Keywords
-
Categories
Funding
- National Heart, Lung, and Blood Institute (NHLBI)
- National Institutes of Health (NIH) [R01HL131835, 1K08HG010061-01A1]
- Gabrielle's Angel Foundation for Cancer Research
- German Research Foundation (DFG) [JU 3104/2-1, 3415-22]
- US Department of Defense [W81XWH-20-1-0904]
- German Research Foundation (DFG)
- Leukemia & Lymphoma Society [3415-22]
- US Department of Defense [JU 3104/2-1]
- NIH (National Human Genome Research Institute) [W81XWH-20-1-0904]
- National Center for Advancing Translational Sciences [T32GM132055]
- Novartis Foundation [JU 3104/2-1]
- Associazione Italiana Ricerca Contro Cancro (AIRC) [3415-22]
- [3UL1TR001085-04S1]
- [INC424XT1349549]
- [23976]
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In this study, the researchers identified that genes in the N-glycosylation pathway were differentially depleted in mutant CALR-transformed cells compared to control cells. Chemical inhibition of N-glycosylation impaired the growth of mutant CALR-transformed cells. The researchers also found a preferential sensitivity of Calr-mutant cells to N-glycosylation inhibition, and normalization of key MPNs disease features. CFU-MK formation in patient-derived CALR-mutant bone marrow was significantly reduced with N-glycosylation inhibition. These findings contribute to the development of clonally selective treatments for CALR-mutant MPNs.
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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