Discovery of a novel megakaryopoiesis enhancer, ingenol, promoting thrombopoiesis through PI3K-Akt signaling independent of thrombopoietin. (March 2022)

• Record Type: Journal Article
• Authors: Wang, Long; Zhang, Ting; Liu, Sha; Mo, Qi; Jiang, Nan; Chen, Qi; Yang, Jing; Han, Yun-Wei; Chen, Jian-Ping; Huang, Fei-Hong; Li, Hua; Zhou, Jie; Luo, Jie-Si; Wu, Jian-Ming
• Is Part Of: Pharmacological research. Volume 177(2022)
• Publication Date: 2022-03
• Subjects: RIT Radiation-induced thrombocytopenia -- CIT Chemotherapy-induced thrombocytopenia -- US United States -- FDA Food and Drug Administration -- rHuIL-11 Recombinant human interleukin-11 -- TPO Thrombopoietin -- RAs Receptor agonists -- ITP Immune thrombocytopenia -- MKs Megakaryocytes -- BM Bone marrow -- HSCs Hematopoietic stem cells -- JAK2 Janus kinase -- STAT3 Signal transducer and activator of transcription 3 -- STAT5 Signal transducer and activator of transcription 5 -- MAPK Mitogenactivated protein kinase -- ERK Extracellular signal-regulated kinases -- PI3K Phosphoinositide 3-kinase -- Akt Protein kinase B -- TFs Transcription factors -- YRSACT Tyrosyl-tRNA synthetase -- CCL5 Chemokine ligand 5 -- IGF-1 Insulin-like growth factor-1 -- ML Machine learning -- AI Artificial intelligence -- RF Random forest -- NB Naive Bayesian -- SVM Support vector machine -- DT Decision tree -- Knn k-Nearest neighbor -- TCM Traditional Chinese medicine -- LDH Lactate dehydrogenase -- c-MPL-/- c-MPL knock-out -- RNA-seq RNA-sequencing -- MolWt Molecular weight -- MolLogP Water partition coefficient -- TPSA Topological polar surface area -- SMOTE Synthetic minority oversampling technique -- RT Room temperature -- DAPI Diamidino-2-phenylindole -- SPF Specific pathogen-free -- KM Kunming -- TBI Total body irradiation -- WT Wild-type -- BMNCs Bone marrow nuclear cells -- H&E Hematoxylin and eosin -- WBC White blood cell -- RBC Red blood cell -- FPKM Fragments Per Kilobase of transcript per Million mapped reads -- DEGs Differentially expressed genes -- EdgeR Empirical analysis of Digital Gene Expression in R -- GO Gene Ontology -- KEGG Kyoto Encyclopedia of Genes and Genomes -- PPI Protein-protein interaction -- TSS Transcription start site -- qRT-PCR Quantitative reverse transcription polymerase chain reaction -- PVDF Polyvinylidene difluoride -- HRP Horseradish peroxidase -- SDs Standard deviations -- AUC Area Under Curve -- PPF Proplatelet formation -- LncRNAs Long non-coding RNAs -- EMH Extramedullary hematopoiesis -- SCF Stem cell factor -- VEGFA Vascular endothelial growth factor A -- FLT3 Tyrosine kinase 3 ligand -- Hgh Human growth hormone -- EGR1 Early growth response gene 1 -- PMA Phorbol 12-myristate 13-acetate -- PLP platelet-like particle; Ingenol (PubChem CID: 442042); Machine learning -- Ingenol -- Megakaryocyte differentiation -- Thrombocytopenia -- Platelets; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2022.106096
• Languages: English
• ISSNs: 1043-6618

Abstract

Abstract: Thrombocytopenia, a most common complication of radiotherapy and chemotherapy, is an important cause of morbidity and mortality in cancer patients. However, there are still no approved agents for the treatment of radiation- and chemotherapy-induced thrombocytopenia (RIT and CIT, respectively). In this study, a drug screening model for predicting compounds with activity in promoting megakaryocyte (MK) differentiation and platelet production was established based on machine learning (ML), and a natural product ingenol was predicted as a potential active compound. Then, in vitro experiments showed that ingenol significantly promoted MK differentiation in K562 and HEL cells. Furthermore, a RIT mice model and c-MPL knock-out (c-MPL -/- ) mice constructed by CRISPR/Cas9 technology were used to assess the therapeutic action of ingenol on thrombocytopenia. The results showed that ingenol accelerated megakaryopoiesis and thrombopoiesis both in RIT mice and c-MPL -/- mice. Next, RNA-sequencing (RNA-seq) was carried out to analyze the gene expression profile induced by ingenol during MK differentiation. Finally, through experimental verifications, we demonstrated that the activation of PI3K/Akt signaling pathway was involved in ingenol-induced MK differentiation. Blocking PI3K/Akt signaling pathway abolished the promotion of ingenol on MK differentiation. Nevertheless, inhibition of TPO/c-MPL signaling pathway could not suppress ingenol-induced MK differentiation. In conclusion, our study builds a drug screening model to discover active compounds against thrombocytopenia, reveals the critical roles of ingenol in promoting MK differentiation and platelet production, and provides a promising avenue for the treatment of RIT. Graphical Abstract: ga1 Highlights: A drug screening model is developed to predict active compounds in hematopoiesis. Ingenol promotes megakaryocyte differentiation in K562 and HEL cells. Ingenol exhibits a great therapeutical effects on thrombocytopenia. The action of ingenol is depend on PI3K-Akt, but independ on TPO/c-MPL signaling.