Article Cite This: Mol. Pharmaceutics XXXX, XXX, XXX−XXX
Difluoromethylornithine Combined with a Polyamine Transport Inhibitor Is Effective against Gemcitabine Resistant Pancreatic Cancer Sarah B. Gitto,†,∥ Veethika Pandey,†,∥,§ Jeremiah L. Oyer,† Alicja J. Copik,† Frederick C. Hogan,† Otto Phanstiel, IV,‡ and Deborah A. Altomare*,† †
Burnett School of Biomedical Sciences, University of Central Florida, 6900 Lake Nona Blvd., Orlando, Florida 32827, United States Department of Medical Education, University of Central Florida, 12722 Research Parkway, Orlando, Florida 32826, United States
‡
ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) is highly chemo-resistant and has an extremely poor patient prognosis, with a survival rate at five years of 0.05).
increased survival compared to the untreated group or the combination treatment. In comparison, DFMO + PTI treated mice survival was significantly increased (Log-rank Mantel-Cox test, p < 0.0009). The mean survival time of mice in the combination treatment group (34 days post-treatment) was nearly doubled compared to the untreated group (18.5 days post-treatment). These experiments indicated that the Trimer44NMe PTI shows improved efficacy, when combined with DFMO, in prolonging survival in this PDAC model.
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DISCUSSION DFMO is currently being tested in several Phase I and II clinical trials, alone and in combination with various other therapeutics, for treatment of neuroblastoma and other neoplasms. However, cancer cells treated with DFMO may overcome polyamine depletion by the uptake of polyamines from extracellular sources. Hence, the experiments here tested the Trimer44NMe PTI, which previously was shown to exhibit low toxicity against normal cells, decrease sensitivity to exogenous addition of amine oxidases and potently inhibit the uptake of spermidine (1 μM) in DFMO-treated L3.6pl human pancreatic cancer cells.7 Trimer44NMe PTI has minimal toxicity when tested alone, especially when used on cell lines that were previously shown to have high sensitivity to DFMO.7 Tumor size or weight alone was not a good indicator of response to the combination DFMO and Trimer44NMe. Immunohistochemical markers and survival studies show the benefit of using a combination treatment with DFMO and Trimer44NMe.
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CONCLUSIONS In summary, a combination therapy of DFMO + Trimer44NMe PTI was shown to nearly double the survival of tumor-bearing mice orthotopically injected with Pan02 cells. F
DOI: 10.1021/acs.molpharmaceut.7b00718 Mol. Pharmaceutics XXXX, XXX, XXX−XXX
Article
Molecular Pharmaceutics
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These experiments demonstrated that targeting both polyamine biosynthesis and import has therapeutic value in the treatment of Gemcitabine-resistant PDAC as evidenced by decreased expression of proliferation markers like Ki67 and reduced cMyc expression compared to untreated mice.
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AUTHOR INFORMATION
Corresponding Author
*Phone: (407) 266-7040. E-mail:
[email protected]. ORCID
Otto Phanstiel IV: 0000-0001-7101-1311 Deborah A. Altomare: 0000-0002-7014-0397 Present Address §
Mayo Clinic, Griffin Cancer Research Building, 4500 San Pablo Road, Jacksonville, Florida 32224, United States. Author Contributions ∥
These authors contributed equally.
Notes
The authors declare no competing financial interest.
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ACKNOWLEDGMENTS The investigators thank Dr. Laurence von Kalm (UCF) for project discussions, Dr. Isaiah Fidler (University of Texas, M. D. Anderson Cancer Center) and Dr. Cheryl Baker (BioCurity Holdings, Inc., Orlando, FL) for the gift of L3.6pl cells, and Meera Rathod for technical help with immunohistochemistry to assess DFMO in the L3.6pl tumor model. UCF Burnett School of Biomedical Sciences shared core equipment resources for animal care, histology, flow cytometry, and imaging used for this study. This work was supported by a Department of Defense Congressionally Directed Medical Research Program (CDMRP) Peer Review Cancer Research Program (PRCRP) Discovery Award CA110724 to O.P. and D.A.A.
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ABBREVIATIONS PDAC, pancreatic ductal adenocarcinoma; PTI, polyamine transport inhibitor; DFMO, alpha-difluoromethylornithine; PTS, polyamine transport system; ODC, ornithine decarboxylase; PBS, phosphate buffered saline; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfenyl)-2H tetrazolium, inner salt
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DOI: 10.1021/acs.molpharmaceut.7b00718 Mol. Pharmaceutics XXXX, XXX, XXX−XXX
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DOI: 10.1021/acs.molpharmaceut.7b00718 Mol. Pharmaceutics XXXX, XXX, XXX−XXX
