4.7 Article

Whole-genome CRISPR screening identifies N-glycosylation as a genetic and therapeutic vulnerability in CALR-mutant MPNs

期刊

BLOOD
卷 140, 期 11, 页码 1291-1304

出版社

AMER SOC HEMATOLOGY
DOI: 10.1182/blood.2022015629

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资金

  1. National Heart, Lung, and Blood Institute (NHLBI)
  2. National Institutes of Health (NIH) [R01HL131835, 1K08HG010061-01A1]
  3. Gabrielle's Angel Foundation for Cancer Research
  4. German Research Foundation (DFG) [JU 3104/2-1, 3415-22]
  5. US Department of Defense [W81XWH-20-1-0904]
  6. German Research Foundation (DFG)
  7. Leukemia & Lymphoma Society [3415-22]
  8. US Department of Defense [JU 3104/2-1]
  9. NIH (National Human Genome Research Institute) [W81XWH-20-1-0904]
  10. National Center for Advancing Translational Sciences [T32GM132055]
  11. Novartis Foundation [JU 3104/2-1]
  12. Associazione Italiana Ricerca Contro Cancro (AIRC) [3415-22]
  13. [3UL1TR001085-04S1]
  14. [INC424XT1349549]
  15. [23976]

向作者/读者索取更多资源

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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