Our goal is to define the roles of Pten phosphorylation remodeling in hematopoietic stem cell (HSC) regulation. Pten, a tumor suppressor, has both lipid and protein phosphatase activities that inhibit PI3K/Akt and Fak-MAPK signaling respectively. It regulates many aspects of cell behavior, including proliferation, survival, adhesion and migration.1-4 The phosphatase activities of Pten are regulated by its c-terminal tail phosphorylation.5, 6 In addition, Pten many also have some phosphatase-independent functions. Many of Pten's biological functions are dependent upon protein-protein interactions which are mediated by its PDZ-motif.7 Knockout of Pten in mouse hematopoietic tissues results in abnormal activation of PI3K/Akt and Src signaling, which leads to uncontrolled HSC activation (G0 to G1 transition) and mobilization, followed by HSC decline. These mice develop myeloproliferative disorder (MPD) followed by acute myeloid/T lymphoid leukemia. Although Pten mutations are not commonly found in hematopoietic malignancies, including leukemia, p-Pten (the phosphorylated form of Pten) levels are increased in the abnormal blasts of most leukemic patients'bone marrow samples. Phosphorylation of Pten's c-terminal tail (ser380, thr382, and thr383) leads to a conformation change which may result in the blocking of its ability to bind to other partner proteins, the reduction of Pten phosphatase activity, and/or the alteration of the lifespan of the Pten protein. Our recent studies have suggested that the phosphorylation of Pten's c-terminal tail may not affect its lipid phosphatase activity but significantly compromises its protein phosphatase activity. The non-phosphorylated form of Pten (non-p-Pten) inhibits Src/Fak/p38 activity, thus repressing cell migration/invasiveness and inducing cell:cell contact inhibition of growth. p-Pten might have a dominant-negative function which induces cell-contact-related Src/Fak/p38 activation. We found that non-p-Pten is expressed in HSCs, while p- Pten levels are increased when HSCs enter the cell cycle;both of these events correspond to increased p- Src, p-Fak and p-p38 levels. Transduced over-expression of non-p-Pten preserves HSCs in a bone marrow niche-dependent manner, whereas transduced over-expression of p-Pten induces HSC/progenitors (HSC/Ps, from wild-type mice which have endogenous Pten expression) to differentiate to myeloid precursors. We propose that non-p-Pten maintains HSC quiescence and self-renewal ability through inducing cell:cell (HSCs and niche cells) contact-induced inhibition of growth by inhibiting Src/Fak/p38 signaling activities, whereas Pten's c-terminal phosphorylation alters its ability to bind to its partners and compromises its protein phosphatase activity. p-Pten promotes opposite functions to these through inducing cell:cell contact-related Src/Fak/p38 signaling. These studies will provide insights into how quiescent HSCs become activated and expand in number, and how we might be able to induce activated HSCs to revert back into quiescence in order to enhance their engraftment ability. This should greatly help our ability to expand HSCs in vitro and hence improve the outcome of clinical stem cell transplantation. It might be also help us to understand the nature of Pten c-terminal phosphorylation in leukemogenesis.

Public Health Relevance

This proposed study will provide insights into how quiescent hematopoietic stem cell be induced cell cycle entry and proliferation, and how stem cell function is regulated by PTEN c-terminal phosphorylation. The predicted outcome will greatly help our ability to expand hematopoietic stem cells in vitro and hence improve the outcome of clinical stem cell transplantation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL095896-05
Application #
8590215
Study Section
Hematopoiesis Study Section (HP)
Program Officer
Thomas, John
Project Start
2010-03-15
Project End
2014-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
5
Fiscal Year
2014
Total Cost
$333,011
Indirect Cost
$110,261
Name
Loyola University Chicago
Department
Pathology
Type
Schools of Medicine
DUNS #
791277940
City
Maywood
State
IL
Country
United States
Zip Code
60153
Shang, Na; Arteaga, Maribel; Zaidi, Ali et al. (2015) FAK is required for c-Met/?-catenin-driven hepatocarcinogenesis. Hepatology 61:214-26
Zhang, Jun; Seet, Christopher S; Sun, Clare et al. (2013) p27kip1 maintains a subset of leukemia stem cells in the quiescent state in murine MLL-leukemia. Mol Oncol 7:1069-82
Li, Zejuan; Huang, Hao; Chen, Ping et al. (2012) miR-196b directly targets both HOXA9/MEIS1 oncogenes and FAS tumour suppressor in MLL-rearranged leukaemia. Nat Commun 3:688
Toth, George; Zraly, Claudia B; Thomson, Tricia L et al. (2011) Congenital anomalies and rhabdoid tumor associated with 22q11 germline deletion and somatic inactivation of the SMARCB1 tumor suppressor. Genes Chromosomes Cancer 50:379-88
Zhang, J; Xiao, Y; Guo, Y et al. (2011) Differential requirements for c-Myc in chronic hematopoietic hyperplasia and acute hematopoietic malignancies in Pten-null mice. Leukemia 25:1857-68