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Blood DOI:10.1182/blood-2018-05-850339

-truncating mutations confer resistance to chemotherapy and sensitivity to PPM1D inhibition in hematopoietic cells.

Publication TypeJournal Article
Year of Publication2018
AuthorsKahn, JD, Miller, PG, Silver, AJ, Sellar, RS, Bhatt, S, Gibson, C, McConkey, M, Adams, D, Mar, B, Mertins, P, Fereshetian, S, Krug, K, Zhu, H, Letai, A, Carr, SA, Doench, J, Jaiswal, S, Ebert, BL
JournalBlood
Volume132
Issue11
Pages1095-1105
Date Published2018 09 13
ISSN1528-0020
KeywordsBase Sequence, Cell Line, Tumor, CRISPR-Cas Systems, Drug Resistance, Neoplasm, Enzyme Inhibitors, Hematologic Neoplasms, Hematopoietic Stem Cells, Humans, Myeloproliferative Disorders, Neoplasm Proteins, Neoplastic Stem Cells, Protein Phosphatase 2C, Sequence Deletion
Abstract

Truncating mutations in the terminal exon of protein phosphatase Mg2/Mn2 1D () have been identified in clonal hematopoiesis and myeloid neoplasms, with a striking enrichment in patients previously exposed to chemotherapy. In this study, we demonstrate that truncating mutations confer a chemoresistance phenotype, resulting in the selective expansion of -mutant hematopoietic cells in the presence of chemotherapy in vitro and in vivo. Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein-9 nuclease mutational profiling of in the presence of chemotherapy selected for the same exon 6 mutations identified in patient samples. These exon 6 mutations encode for a truncated protein that displays elevated expression and activity due to loss of a C-terminal degradation domain. Global phosphoproteomic profiling revealed altered phosphorylation of target proteins in the presence of the mutation, highlighting multiple pathways including the DNA damage response (DDR). In the presence of chemotherapy, -mutant cells have an abrogated DDR resulting in altered cell cycle progression, decreased apoptosis, and reduced mitochondrial priming. We demonstrate that treatment with an allosteric, small molecule inhibitor of PPM1D reverts the phosphoproteomic, DDR, apoptotic, and mitochondrial priming changes observed in mutant cells. Finally, we show that the inhibitor preferentially kills -mutant cells, sensitizes the cells to chemotherapy, and reverses the chemoresistance phenotype. These results provide an explanation for the enrichment of truncating mutations in the blood of patients exposed to chemotherapy and in therapy-related myeloid neoplasms, and demonstrate that PPM1D can be a targeted in the prevention of clonal expansion of -mutant cells and the treatment of -mutant disease.

DOI10.1182/blood-2018-05-850339
Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/29954749?dopt=Abstract

Alternate JournalBlood
PubMed ID29954749
PubMed Central IDPMC6137556
Grant ListT32 HL066987 / HL / NHLBI NIH HHS / United States
P50 CA206963 / CA / NCI NIH HHS / United States
R01 HL082945 / HL / NHLBI NIH HHS / United States
/ HHMI / Howard Hughes Medical Institute / United States
R01 GM038627 / GM / NIGMS NIH HHS / United States
P01 CA108631 / CA / NCI NIH HHS / United States
T32 HL116324 / HL / NHLBI NIH HHS / United States
P01 CA066996 / CA / NCI NIH HHS / United States