PAK4-NAMPT Dual Inhibition as a Novel Strategy for Therapy Resistant Pancreatic Neuroendocrine Tumors
Abstract
:1. Introduction
2. Results
2.1. PAK4 and NAMPT Are Overexpressed in PNET
2.2. PAK4 and NAMPT Promote PNET Survival
2.3. KPT-9274 and Analog KPT-7523 Decrease PNET Cell Survival and Growth
2.4. KPT-9274 Synergizes with Everolimus in PNET Cellular Models
2.5. Molecular Analysis of KPT-9274-Everolimus Combination
2.6. Metabolomic Analysis of PNET Cells Treated with KPT-9274
2.7. KPT-9274 Shows Single Agent Anti-Tumor Activity in PNET
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Reagents
4.2. Small Interference RNA and Transfection
4.3. MTT Assay
4.4. Colonogenic Assay
4.5. Annexin V and 7AAD Apoptosis Analysis
4.6. Immunohistochemistry Analysis
4.7. Western Blot Analysis
4.8. Quantitative Real-Time PCR
4.9. Metabolomic Analysis.
4.10. Animal Studies
4.11. Statistical Consideration
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Metabolites | Treatment Effect | p-Value |
---|---|---|
Kynurenic acid | −2.173 | 0.026 |
Fructose 1,6-bisphosphate | −1.903 | 0.016 |
Histidine | −2.356 | 0.023 |
Phenylalanine | −1.076 | 0.044 |
Arginine | −1.693 | 0.025 |
Lysine (s) | −2.729 | 0.021 |
dUMP | −1.043 | 0.025 |
1-Methylhistidine | −1.381 | 0.045 |
2-Ketohexanoic acid | 1.938 | 0.001 |
N-Alpha-acetyllysine | −1.866 | 0.047 |
ADP_new | 1.410 | 0.001 |
CMP | 1.610 | 0.023 |
DL-2-Aminooctanoic acid | −2.158 | 0.008 |
Folic acid | −2.659 | 0.007 |
Guanine | −1.380 | 0.010 |
N-Acetylornithine | −2.706 | 0.046 |
Phenyllactic acid | −1.514 | 0.047 |
Pyridoxal 5-phosphate | −2.006 | 6.05 × 10−7 |
pyridoxine | −1.682 | 0.046 |
Thymine | −1.959 | 0.027 |
Sarcosine (s) | −1.121 | 0.018 |
CDP | 2.040 | 0.003 |
GDP | 1.496 | 0.002 |
UDP | 1.875 | 0.001 |
dGMP_New 1 | 1.465 | 0.015 |
dGDP_new | 1.452 | 0.001 |
Cholesterol sulfate | −1.412 | 0.010 |
Hippurate | −1.911 | 0.027 |
p-cresyl sulfate | −1.536 | 0.049 |
Linoleyl Carnitine | 1.224 | 0.033 |
C14 Carnitine (Myristoyl-L-carnitine) | 1.307 | 0.040 |
Metabolites | Treatment Effect | p-Value |
---|---|---|
Succinyl-CoA | −1.299 | 8.87 × 10−4 |
Fructose 6-phosphate | 2.483 | 0.008 |
Arginine | −1.179 | 0.008 |
Picolinic acid | 1.633 | 0.034 |
Nicotinamide | 1.695 | 0.033 |
D-Ribose 5-phosphate | 2.741 | 6.27 × 10−6 |
Adenosine | 3.164 | 2.51 × 10−6 |
ADP_new | 1.805 | 5.26 × 10−8 |
Aminoadipic acid | 1.352 | 0.026 |
Cytosine | 1.806 | 0.007 |
Glucose 6-phosphate | 2.506 | 0.008 |
GMP_New | 2.775 | 2.08 × 10−4 |
Ornithine | 1.342 | 0.025 |
Uracil | 3.249 | 0.001 |
Uridine | 3.731 | 0.001 |
Adenosine triphosphate | −1.486 | 0.004 |
Cytidine triphosphate | −1.884 | 0.003 |
Guanosine triphosphate | −1.819 | 0.006 |
Uridine triphosphate | −1.394 | 0.008 |
dATP | −1.520 | 0.001 |
dCTP | −1.011 | 0.008 |
dGTP | −1.389 | 0.008 |
dTTP | −1.052 | 0.002 |
Argininosuccinic acid | −1.162 | 0.001 |
Sarcosine (s) | −1.266 | 0.010 |
Dimethylglycine+M191 | 1.571 | 0.001 |
Choline | 1.509 | 0.001 |
Cytidine | 2.923 | 0.001 |
CDP | 2.720 | 2.06 × 10−6 |
GDP | 1.622 | 0.004 |
UDP | 2.185 | 0.001 |
dGMP_New 1 | 3.419 | 3.40 × 10−8 |
dGDP_new | 1.811 | 2.91 × 10−6 |
D-Sedoheptulose 7-phosphate | 1.285 | 0.008 |
N-Acetyl-glucosamine 1-phosphate | 1.210 | 0.008 |
Citicoline | 3.320 | 0.005 |
Adenine | 1.726 | 0.001 |
Spermidine | 2.009 | 0.013 |
C16:0 Carnitine (Palmitoyl-L carnitine) | 3.023 | 0.013 |
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Mpilla, G.; Aboukameel, A.; Muqbil, I.; Kim, S.; Beydoun, R.; Philip, P.A.; Mohammad, R.M.; Kamgar, M.; Shidham, V.; Senapedis, W.; et al. PAK4-NAMPT Dual Inhibition as a Novel Strategy for Therapy Resistant Pancreatic Neuroendocrine Tumors. Cancers 2019, 11, 1902. https://doi.org/10.3390/cancers11121902
Mpilla G, Aboukameel A, Muqbil I, Kim S, Beydoun R, Philip PA, Mohammad RM, Kamgar M, Shidham V, Senapedis W, et al. PAK4-NAMPT Dual Inhibition as a Novel Strategy for Therapy Resistant Pancreatic Neuroendocrine Tumors. Cancers. 2019; 11(12):1902. https://doi.org/10.3390/cancers11121902
Chicago/Turabian StyleMpilla, Gabriel, Amro Aboukameel, Irfana Muqbil, Steve Kim, Rafic Beydoun, Philip A. Philip, Ramzi M. Mohammad, Mandana Kamgar, Vinod Shidham, William Senapedis, and et al. 2019. "PAK4-NAMPT Dual Inhibition as a Novel Strategy for Therapy Resistant Pancreatic Neuroendocrine Tumors" Cancers 11, no. 12: 1902. https://doi.org/10.3390/cancers11121902