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

Blood
Authors
Keywords
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.

Year of Publication
2018
Journal
Blood
Volume
132
Issue
11
Pages
1095-1105
Date Published
2018 09 13
ISSN
1528-0020
DOI
10.1182/blood-2018-05-850339
PubMed ID
29954749
PubMed Central ID
PMC6137556
Links
Grant list
T32 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