Juvenile myelomonocytic leukemia (JMML) is a fatal childhood myeloproliferative neoplasm in which myeloid cells are overproduced and hematopoietic progenitors are hypersensitive to granulocyte-macrophage colony stimulating factor (GM-CSF). The only curative treatment is allogeneic bone marrow transplant, but even this rigorous therapy yields only a 50% relapse-free 5-year survival. A majority of patients have hyperactive Ras signaling, with the most common mutation being somatic gain-of-function (GOF) mutations in PTPN11, which encodes the protein tyrosine phosphatase Shp2. Previous work in the Chan lab has shown that the PI3K catalytic subunit p110? is needed for both Akt and Erk hyperactivation, and promotes GOF Shp2-induced GM- CSF hypersensitivity and hyperproliferation, contributing to the progression of JMML. Based on the significant role of p110? in promoting GOF Shp2-induced leukemia, we investigated potential tyrosine kinases that can cooperate with p110? to promote Akt and Erk activation and lead to hyperproliferation of myeloid cells. Bruton's Tyrosine Kinase (BTK) has been identified as a critical molecule in lymphoid malignancies and the BTK inhibitor, ibrutinib, was recently approved for use in patients with B cell leukemias. As BTK has been well- studied in the context of B cell receptor signaling, it is known that BTK is activated downstream of p110? and that active BTK phosphorylates B cell adaptor for PI3K (BCAP), allowing phospho-BCAP to bind to the regulatory p85? subunit and promote activation of PI3K. Based on what is known in B cells, we hypothesize that BTK and BCAP are critical players in the GOF Shp2-mediated hyperactivation of p110? signaling in JMML. To address this hypothesis, we propose three aims: (1) we will use murine cells expressing the GOF Shp2 mutation, E76K, with genetic knockout of BTK to perform proliferation and biochemical analysis in vitro, as well as examine the progression of myeloproliferative disease in p110? inhibitor-treated mice in vivo; (2) we will use cells from mice lacking expression of p85? along with p85? site-specific mutant constructs to explore the physical interaction between BCAP and p85?; and (3) we will test the efficacy of BTK (ACP-196) and p110? (ACP-319) inhibitors provided by AcertaPharma on Shp2E76K-expressing mice in vivo, as well as on primary JMML patient cells in vitro. These studies are timely and significant, as the AcertaPharma inhibitors are currently being tested in combination for safety and efficacy in chronic lymphocytic leukemia patients, and therefore have the potential to be a novel treatment in patients with JMML.

Public Health Relevance

Juvenile Myelomonocytic Leukemia is a deadly blood cancer in children. Our laboratory has developed a series of experiments to help us understand how changes in a protein called Shp2 along with another protein called Bruton's Tyrosine Kinase can cause normal blood cells to become cancerous in the hope of developing improved treatments for this disease.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
Project #
5F30CA210518-03
Application #
9525929
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Damico, Mark W
Project Start
2016-08-01
Project End
2020-07-31
Budget Start
2018-08-01
Budget End
2019-07-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Genetics
Type
Schools of Medicine
DUNS #
603007902
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Deng, Lisa; Richine, Briana M; Virts, Elizabeth L et al. (2018) Rapid development of myeloproliferative neoplasm in mice with Ptpn11 D61Y mutation and haploinsufficient for Dnmt3a. Oncotarget 9:6055-6061
Deng, Lisa; Virts, Elizabeth L; Kapur, Reuben et al. (2017) Pharmacologic inhibition of PI3K p110? in mutant Shp2E76K-expressing mice. Oncotarget 8:84776-84781
Tarnawsky, Stefan P; Yoshimoto, Momoko; Deng, Lisa et al. (2017) Yolk sac erythromyeloid progenitors expressing gain of function PTPN11 have functional features of JMML but are not sufficient to cause disease in mice. Dev Dyn 246:1001-1014
Li, Xing Jun; Deng, Lisa; Brandt, Stephanie L et al. (2016) Role of p85? in neutrophil extra- and intracellular reactive oxygen species generation. Oncotarget 7:23096-105