Jutzi, Jonas SMarneth, Anna ECiboddo, MicheleGuerra-Moreno, AngelJiménez-Santos, María JoséKosmidou, AnastasiaDressman, James WLiang, HongyanHamel, RebeccaLozano, PatriciaRumi, ElisaDoench, John GGotlib, JasonKrishnan, AnandiElf, ShannonAl-Shahrour, FatimaMullally, Ann2024-06-122024-06-122022-09-15Blood . 2022 ;140(11):1291-1304http://hdl.handle.net/20.500.12105/19755Calreticulin (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.enghttp://creativecommons.org/licenses/by-nc-nd/4.0/CalreticulinMyeloproliferative DisordersAnimalsClustered Regularly Interspaced Short Palindromic RepeatsGlucoseGlycosylationHumansJanus Kinase 2MiceMutationReceptors, ThrombopoietinotherAttribution-NonCommercial-NoDerivatives 4.0 Internacional3576366514011129110.1182/blood.20220156291528-0020Blood9479036open access