Targeting of mTORC1/2 by the mTOR kinase inhibitor PP242 induces apoptosis in AML cells under conditions mimicking the bone marrow microenvironment

• Published in: Blood 2012-09, Vol.120 (13), p.2679-2689
• Main Authors: Zeng, Zhihong, Shi, Yue Xi, Tsao, Twee, Qiu, YiHua, Kornblau, Steven M., Baggerly, Keith A., Liu, Wenbin, Jessen, Katti, Liu, Yi, Kantarjian, Hagop, Rommel, Christian, Fruman, David A., Andreeff, Michael, Konopleva, Marina
• Format: Article
• Language: English
• Subjects: Animals; Antibiotics, Antineoplastic - therapeutic use; Antineoplastic Combined Chemotherapy Protocols; Apoptosis - drug effects; Blotting, Western; Bone Marrow - metabolism; Bone Marrow - pathology; Cell Proliferation; Coculture Techniques; Flow Cytometry; Humans; Indoles - therapeutic use; Leukemia, Experimental - mortality; Leukemia, Experimental - pathology; Leukemia, Experimental - prevention & control; Leukemia, Myeloid, Acute - mortality; Leukemia, Myeloid, Acute - pathology; Leukemia, Myeloid, Acute - prevention & control; Mechanistic Target of Rapamycin Complex 1; Mechanistic Target of Rapamycin Complex 2; Mesenchymal Stem Cells - metabolism; Mesenchymal Stem Cells - pathology; Mice; Mice, SCID; Multiprotein Complexes - antagonists & inhibitors; Multiprotein Complexes - metabolism; Myeloid Neoplasia; Phosphorylation - drug effects; Protein Array Analysis; Protein Kinase Inhibitors - pharmacology; Purines - therapeutic use; Real-Time Polymerase Chain Reaction; Receptors, CXCR4 - genetics; Receptors, CXCR4 - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; Signal Transduction - drug effects; Sirolimus - therapeutic use; TOR Serine-Threonine Kinases - antagonists & inhibitors; TOR Serine-Threonine Kinases - metabolism
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2011-11-393934

The interactions between the bone marrow (BM) microenvironment and acute myeloid leukemia (AML) is known to promote survival of AML cells. In this study, we used reverse phase-protein array (RPPA) technology to measure changes in multiple proteins induced by stroma in leukemic cells. We then investigated the potential of an mTOR kinase inhibitor, PP242, to disrupt leukemia/stroma interactions, and examined the effects of PP242 in vivo using a mouse model. Using RPPA, we confirmed that multiple survival signaling pathways, including the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR), were up-regulated in primary AML cells cocultured with stroma. PP242 effectively induced apoptosis in primary samples cultured with or without stroma. Mechanistically, PP242 attenuated the activities of mTORC1 and mTORC2, sequentially inhibited phosphorylated AKT, S6K, and 4EBP1, and concurrently suppressed chemokine receptor CXCR4 expression in primary leukemic cells and in stromal cells cultured alone or cocultured with leukemic cells. In the in vivo leukemia mouse model, PP242 inhibited mTOR signaling in leukemic cells and demonstrated a greater antileukemia effect than rapamycin. Our findings indicate that disrupting mTOR/AKT signaling with a selective mTOR kinase inhibitor can effectively target leukemic cells within the BM microenvironment.