PMID- 31147807 OWN - NLM STAT- MEDLINE DCOM- 20210303 LR - 20210303 IS - 1573-0646 (Electronic) IS - 0167-6997 (Linking) VI - 38 IP - 2 DP - 2020 Apr TI - Apoptosis induction and cell cycle arrest of pladienolide B in erythroleukemia cell lines. PG - 369-377 LID - 10.1007/s10637-019-00796-2 [doi] AB - Splicing of pre-mRNA into functional mRNA, carried out by the spliceosome, represents a crucial step in eukaryotic gene expression. Mutations and other deregulation in some of the spliceosome components have been identified in multiple pathologies, including hematological malignancies. In this context, we evaluated the therapeutic potential of a splicing inhibitor, Pladienolide B (Pla-B), in two erythroleukemia cell lines. HEL and K562 cell lines were incubated with increasing doses of Pla-B in single and daily administration. Cell viability and density were evaluated using trypan blue assay. Flow cytometry was used to evaluate cell death, cell cycle, and caspase activity. NGS analysis was performed to assess the mutational status of 4 splicing-related genes (SF3B1, U2AF1, ZRSR2 and SRSF2). Expression levels of SF3B1 and unspliced DNAJB1 were evaluated by qPCR. Pla-B significantly decreased the viability and proliferation of both cell lines in time, dose, administration schedule, and cell line-dependent manner. HEL cells were more sensible to Pla-B (IC(50) = 1.5 nM) than K562 (IC(50) = 25 nM), with an IC(50) almost 17 times lower. Pla-B induced cell death, mainly by apoptosis, and cell cycle arrest in G(0)/G(1) phase. No mutations were found in any of the analyzed genes, suggesting that the observed cytotoxic effect is independent of the spliceosome mutations. Splicing modulator Pla-B showed high antitumor activity against HEL and K562 cell lines, inducing apoptosis and cell cycle arrest. These data suggest that Pla-B might represent a new therapeutic approach for erythroleukemia. FAU - Jorge, Joana AU - Jorge J AD - Laboratory of Oncobiology and Hematology, University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal. AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. FAU - Petronilho, Sara AU - Petronilho S AD - Laboratory of Oncobiology and Hematology, University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal. FAU - Alves, Raquel AU - Alves R AD - Laboratory of Oncobiology and Hematology, University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal. AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. AD - Center for Innovative Biomedicine and Biotechnology (CIBB), Coimbra, Portugal. FAU - Coucelo, Margarida AU - Coucelo M AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. AD - Center for Innovative Biomedicine and Biotechnology (CIBB), Coimbra, Portugal. AD - Clinical Hematology Department, Centro Hospitalar Universitario de Coimbra (CHUC), Coimbra, Portugal. FAU - Goncalves, Ana Cristina AU - Goncalves AC AUID- ORCID: 0000-0003-1470-4802 AD - Laboratory of Oncobiology and Hematology, University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal. acgoncalves@fmed.uc.pt. AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. acgoncalves@fmed.uc.pt. AD - Center for Innovative Biomedicine and Biotechnology (CIBB), Coimbra, Portugal. acgoncalves@fmed.uc.pt. FAU - Nascimento Costa, Jose Manuel AU - Nascimento Costa JM AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. AD - Center for Innovative Biomedicine and Biotechnology (CIBB), Coimbra, Portugal. AD - University Clinic of Oncology, Faculty of Medicine, University of Coimbra, FMUC, Coimbra, Portugal. FAU - Sarmento-Ribeiro, Ana Bela AU - Sarmento-Ribeiro AB AD - Laboratory of Oncobiology and Hematology, University Clinic of Hematology, Faculty of Medicine, University of Coimbra, FMUC, Azinhaga de Santa Comba-Celas, 3000-548, Coimbra, Portugal. AD - Coimbra Institute for Clinical and Biomedical Research (iCBR) - Group of Environment Genetics and Oncobiology (CIMAGO), FMUC, Coimbra, Portugal. AD - Center for Innovative Biomedicine and Biotechnology (CIBB), Coimbra, Portugal. AD - Clinical Hematology Department, Centro Hospitalar Universitario de Coimbra (CHUC), Coimbra, Portugal. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190531 PL - United States TA - Invest New Drugs JT - Investigational new drugs JID - 8309330 RN - 0 (Antineoplastic Agents) RN - 0 (DNAJB1 protein, human) RN - 0 (Epoxy Compounds) RN - 0 (HSP40 Heat-Shock Proteins) RN - 0 (Macrolides) RN - 0 (Phosphoproteins) RN - 0 (RNA Splicing Factors) RN - 0 (SF3B1 protein, human) RN - 0 (pladienolide B) SB - IM MH - Antineoplastic Agents/*pharmacology MH - Apoptosis/drug effects MH - Cell Cycle Checkpoints/drug effects MH - Cell Line, Tumor MH - Cell Survival/drug effects MH - Epoxy Compounds/*pharmacology MH - HSP40 Heat-Shock Proteins/genetics MH - Humans MH - Leukemia, Erythroblastic, Acute/*drug therapy/genetics MH - Macrolides/*pharmacology MH - Phosphoproteins/genetics MH - RNA Splicing Factors/genetics OTO - NOTNLM OT - Acute myeloid leukemia OT - Erythroleukemia OT - Pladienolide B OT - SF3B1 OT - Splicing inhibitor EDAT- 2019/05/31 06:00 MHDA- 2021/03/04 06:00 CRDT- 2019/06/01 06:00 PHST- 2019/04/17 00:00 [received] PHST- 2019/05/22 00:00 [accepted] PHST- 2019/05/31 06:00 [pubmed] PHST- 2021/03/04 06:00 [medline] PHST- 2019/06/01 06:00 [entrez] AID - 10.1007/s10637-019-00796-2 [pii] AID - 10.1007/s10637-019-00796-2 [doi] PST - ppublish SO - Invest New Drugs. 2020 Apr;38(2):369-377. doi: 10.1007/s10637-019-00796-2. Epub 2019 May 31.