MLK3, also known as SPRK, is a mitogen-activated protein kinase that can activate c-jun N- terminal kinase (JNK) and induce cell death in neuronal and non-neuronal cells. A broad-spectrum inhibitor of MLK family members (CEP-1347) is under clinical trials for treating neurodegenerative disorder related diseases like Parkinson's. TNF-a and ceramide have been shown to cause cell death during neurodegenerative disorders. Interestingly, ceramide has also emerged as a biochemical mediator of cell death in response to chemo- and radio- therapies. Studies from our laboratory have revealed that the cell death and differentiation inducing ligands, TNF-a and ceramide are potent agonists of MLK3 in mammalian cells. In addition, our more recent studies (both published and unpublished) indicate that (i) ceramides can directly associate, and activate MLK3, (ii) MLK3 mediates TNF-a and ceramide-induced JNK activation, (iii) TNF-a treatment of Jurkat cells triggers TRAF2 association with MLK3, (vi) overexpression of MLK3 causes cell death, and (v) Retinoic acid induced HL-60 cell differentiation is inhibited by MLKs specific inhibitor. Based on these observations, our major goals for this grant application are to elucidate the mechanisms of MLK3 activation by ceramide and TNF-a, and to define the physiological roles of ligand-activated MLK3 in cell death and differentiation pathways. These goals will be achieved following four specific aims.
In aim 1, the in vitro and in vivo mechanisms of MLK3 activation by ceramide will be elucidated.
In aim 2, the mechanism of TNF-a-induced MLK3 activation, and its synergy with ceramide-induced MLK3 activation will be examined.
In aim 3, the physiological roles of MLK3 in mediating cell death and differentiation pathways will be elucidated using both biochemical and genetic approaches.
In aim 4, the physiological role of MLK3 in neuronal apoptosis will be examined in MPTP mouse model (i.e. PD model), using recently created MLK3 KO and wild type animals. Elucidating the mechanism(s) of ceramide and TNF-ainduced MLK3 activation, and MLK3's role in modulating cell death and differentiation pathways will offer a basis for the development of targeted therapeutic interventions for pathologic neuronal loss occurring as a result of conditions, such as neurodegeneration, trauma or ischemia. Furthermore, the results from these studies will aide in the development of novel pharmacological approaches towards increasing the efficiency and specificity of chemo- or radio- sensitization for cancer therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM055835-10
Application #
7745524
Study Section
Special Emphasis Panel (ZRG1-CB-G (03))
Program Officer
Maas, Stefan
Project Start
1999-03-01
Project End
2012-11-30
Budget Start
2009-12-01
Budget End
2012-11-30
Support Year
10
Fiscal Year
2010
Total Cost
$268,374
Indirect Cost
Name
Loyola University Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
791277940
City
Maywood
State
IL
Country
United States
Zip Code
60153
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Rana, Ajay; Rana, Basabi; Mishra, Rajakishore et al. (2013) Mixed Lineage Kinase-c-Jun N-Terminal Kinase Axis: A Potential Therapeutic Target in Cancer. Genes Cancer 4:334-41
Ghosh, Anamitra; Saminathan, Hariharan; Kanthasamy, Arthi et al. (2013) The peptidyl-prolyl isomerase Pin1 up-regulation and proapoptotic function in dopaminergic neurons: relevance to the pathogenesis of Parkinson disease. J Biol Chem 288:21955-71
Leicht, Deborah T; Balan, Vitaly; Zhu, Jun et al. (2013) MEK-1 activates C-Raf through a Ras-independent mechanism. Biochim Biophys Acta 1833:976-86
Viswakarma, Navin; Jia, Yuzhi; Bai, Liang et al. (2013) The Med1 subunit of the mediator complex induces liver cell proliferation and is phosphorylated by AMP kinase. J Biol Chem 288:27898-911
Rangasamy, Velusamy; Mishra, Rajakishore; Sondarva, Gautam et al. (2012) Mixed-lineage kinase 3 phosphorylates prolyl-isomerase Pin1 to regulate its nuclear translocation and cellular function. Proc Natl Acad Sci U S A 109:8149-54
Kanthasamy, Anumantha; Jin, Huajun; Anantharam, Vellareddy et al. (2012) Emerging neurotoxic mechanisms in environmental factors-induced neurodegeneration. Neurotoxicology 33:833-7
Thylur, Ramesh P; Senthivinayagam, Subramanian; Campbell, Edward M et al. (2011) Mixed lineage kinase 3 modulates ?-catenin signaling in cancer cells. J Biol Chem 286:37470-82
Jin, Huajun; Kanthasamy, Arthi; Anantharam, Vellareddy et al. (2011) Transcriptional regulation of pro-apoptotic protein kinase Cdelta: implications for oxidative stress-induced neuronal cell death. J Biol Chem 286:19840-59
Rangasamy, Velusamy; Mishra, Rajakishore; Mehrotra, Suneet et al. (2010) Estrogen suppresses MLK3-mediated apoptosis sensitivity in ER+ breast cancer cells. Cancer Res 70:1731-40

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