The goal of this project is to determine the mechanistic differences in MEK/ERK signaling that mediates growth arrest versus cell proliferation in response to aberrant Ras and/or Raf signals. Aberrant activation of the Ras/Raf/MEK/ERK pathway is a central feature in many cancers. However, paradoxically, sustained activation of the pathway induces cell cycle arrest and senescence in normal cells and certain cancer types. It has been proposed that growth arrest acts as a defense mechanism against Ras/Raf-mediated carcinogenesis and that overcoming this """"""""growth arrest barrier"""""""" is a necessary step in tumor progression. Our understanding of this key event in carcinogenesis, controlled at the level of MEK/ERK, is currently quite limited. We have begun to address the underlying mechanisms of MEK/ERK signaling using normal and Ras/Raf-responsive tumor cells as models. The first intriguing finding is that ERK can mediate growth arrest by utilizing not only its """"""""canonical"""""""" kinase activity but also, as yet unidentified, non-catalytic functions. We recently reported that oncogenic Raf signal-induced growth arrest is abrogated by ERK1/2 depletion but introduction of catalytically disabled mutant ERK into ERK1/2- depleted cells can selectively restore the growth arrest phenomenon. Based upon a hypothesis that MEK/ERK would interact with specific proteins to mediate growth arrest signaling, we conducted tandem affinity purification and identified mortalin as a potential negative regulator of MEK/ERK-growth arrest signaling. Our preliminary studies show that mortalin binds to inactive, but not active, MEK and that mortalin depletion increases basal as well as Raf-induced MEK/ERK activity. In addition, mortalin depletion promotes growth inhibitory signaling whereas mortalin overexpression exerts the opposite effects. These preliminary studies suggest that MEK/ERK utilizes a unique signaling mechanism to mediate growth arrest, for which mortalin has a negative-regulatory role possibly via its physical interaction with the pathway and ERK modulates a mechanism that requires its non-catalytic function. To test these hypotheses, we propose to (i) determine the role of mortalin in the regulation of Raf/MEK/ERK-mediated growth arrest signaling by gain or loss of function studies in normal and K-Rasor B-Raf mutated tumor cells and by comparing mortalin expression levels with altered MEK/ERK activity in tumor tissue specimens;(ii) determine whether mortalin regulates the Raf/MEK/ERK pathway by differentially sequestering MEK1 or MEK2;and (iii) determine molecular mechanisms underlying the non-catalytic ERK functions. This study will enhance our knowledge of the specific mechanisms of MEK/ERK signaling that interrupts Ras/Raf-driven carcinogenesis.

Public Health Relevance

Activation of the MEK/ERK pathway induces terminal growth arrest in certain types of cancer lines derived from small cell lung carcinoma, medullary thyroid carcinoma, prostate carcinoma, pheochromocytoma, glioma, and gastrointestinal carcinoid. Exploration of the possibility that these cancer types can be effectively suppressed by modulating the pathway via mortalin into the direction of growth arrest may allow development of new therapeutic strategies to control them.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA138441-04
Application #
8460092
Study Section
Tumor Cell Biology Study Section (TCB)
Program Officer
Watson, Joanna M
Project Start
2010-07-01
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
4
Fiscal Year
2013
Total Cost
$263,187
Indirect Cost
$90,038
Name
Medical College of Wisconsin
Department
Biochemistry
Type
Schools of Medicine
DUNS #
937639060
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Wu, Pui-Kei; Hong, Seung-Keun; Park, Jong-In (2017) Steady-State Levels of Phosphorylated Mitogen-Activated Protein Kinase Kinase 1/2 Determined by Mortalin/HSPA9 and Protein Phosphatase 1 Alpha in KRAS and BRAF Tumor Cells. Mol Cell Biol 37:
Hong, Seung-Keun; Starenki, Dmytro; Wu, Pui-Kei et al. (2017) Suppression of B-Raf(V600E) melanoma cell survival by targeting mitochondria using triphenyl-phosphonium-conjugated nitroxide or ubiquinone. Cancer Biol Ther 18:106-114
Starenki, Dmytro; Hong, Seung-Keun; Wu, Pui-Kei et al. (2017) Vandetanib and cabozantinib potentiate mitochondria-targeted agents to suppress medullary thyroid carcinoma cells. Cancer Biol Ther 18:473-483
Klionsky, Daniel J (see original citation for additional authors) (2016) Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12:1-222
Starenki, D; Hong, S-K; Lloyd, R V et al. (2015) Mortalin (GRP75/HSPA9) upregulation promotes survival and proliferation of medullary thyroid carcinoma cells. Oncogene 34:4624-34
Starenki, Dmytro; Park, Jong-In (2015) Pediatric Medullary Thyroid Carcinoma. J Pediatr Oncol 3:29-37
Wu, Pui-Kei; Hong, Seung-Keun; Yoon, Seung-Hee et al. (2015) Active ERK2 is sufficient to mediate growth arrest and differentiation signaling. FEBS J 282:1017-30
Wu, Pui-Kei; Park, Jong-In (2015) MEK1/2 Inhibitors: Molecular Activity and Resistance Mechanisms. Semin Oncol 42:849-62
Starenki, Dmytro; Park, Jong In (2015) Selective Mitochondrial Uptake of MKT-077 Can Suppress Medullary Thyroid Carcinoma Cell Survival In Vitro and In Vivo. Endocrinol Metab (Seoul) 30:593-603
Karkhanis, Mansi; Park, Jong-In (2015) Sp1 regulates Raf/MEK/ERK-induced p21(CIP1) transcription in TP53-mutated cancer cells. Cell Signal 27:479-86

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