Mixed Lineage Kinase 3 (MLK3) is a stress-activated MAP Kinase Kinase Kinase member, whose biological function is still elusive. While dissecting the signaling pathway mediated via MLK3, we observed a strong interaction between mammalian homolog of yeast Ste20 member, p21 activated kinase (Pak1) and MLK3. Initially, we speculated that Pak1 might regulate MLK3 kinase activities similar to its yeast counterparts. However, the observed results were counterintuitive to our speculation, and instead MLK3 directly phosphorylated and activated Pak1, which subsequently lead to NF-?B activation. These results were quite intriguing because Pak1 activation has been implicated in mammary gland hyperplasia, and recently it has been identified as a major oncogene in breast cancer. Our preliminary investigation revealed that Pak1 activity was directly regulated in a MLK3-dependent manner in breast cancer cell lines. We also observed that in primary human breast tumors, the MLK3 activities directly correlated with Pak1 and NF-?B activations, exclusively in ER and PR negative breast tumors and Pak1 was overexpressed in ER- PR- breast tumors. If these results are true then the available MLKs inhibitor, CEP-1347/CEP-11004 should induce cell death in ER- breast cancer cells? Indeed, treatment of ER-, but not ER+ breast cancer cell lines with CEP-1347 caused cell death. Based on our observations, we hypothesize that in ER- breast cancer cells, MLK3 activates Pak1 and its downstream NF-?B, leading to ER- tumorigenesis. Therefore targeting MLK3 or other MLKs could abrogate ER- breast tumors. To achieve our goals, we will determine that: (1) activation of Pak1 by MLK3 induces ER- breast cell tumorigenesis, (2) disruption of MLK3-Pak1 signaling cascade attenuates ER- breast cell tumorigenesis;and (3) the mechanism of MLK3, Pak1 and NF-?B activation in ER- breast cell tumorigenesis. It is expected that by defining the role of MLK3 in Pak1 and NF-?B activation and tumorigenesis, we might control/treat ER- breast cancers by using available inhibitors of this pathway. Interestingly, the specific inhibitor of MLK3 family, CEP-1347 has been used in clinical trials for treating neurodegenerative diseases. It has also been suggested that MLKs inhibitors might serve as a treatment for specific type of cancer, underscoring our unexpected novel results related to MLK3 role in ER- breast cancer. Taken together, the present study will lead to the identification of new prognostic factors and specific targeted therapies for difficult to treat ER- breast cancer.

Public Health Relevance

Gonadal hormone estrogen and its cognate receptor, ER plays a pivotal role in the pathogenesis, of ER positive breast (ER+) cancer. The agents that target ER-mediated pathways are in clinical practice to treat ER+ breast cancer, however, for ER- breast cancer, lack of definitive target leads to high mortality. Our work has unraveled a new signaling pathway (i.e. MLK3-Pak1-NF-kB axis) which is constitutive active in ER- breast cancer only. Our preliminary results show that inhibition of this axis by using MLK3/MLKs inhibitor, promotes cell death, specifically in ER- breast cancer cells. Thus it is expected that understanding this novel signaling axis could lead to future therapeutics for ER- breast cancer treatment.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA176846-01
Application #
8517343
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Yassin, Rihab R,
Project Start
2013-04-01
Project End
2018-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
1
Fiscal Year
2013
Total Cost
$311,744
Indirect Cost
$85,110
Name
Loyola University Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
791277940
City
Maywood
State
IL
Country
United States
Zip Code
60153
Ramakrishnan, Gopalakrishnan; Davaakhuu, Gantulga; Kaplun, Ludmila et al. (2014) Sirt2 deacetylase is a novel AKT binding partner critical for AKT activation by insulin. J Biol Chem 289:6054-66