Original ArticlesRecombinant methioninase in combination with doxorubicin (DOX) overcomes first-line DOX resistance in a patient-derived orthotopic xenograft nude-mouse model of undifferentiated spindle-cell sarcoma
Introduction
Undifferentiated spindle-cell sarcoma (USCS) is a recalcitrant cancer with predominant spindle-shaped cells that originates in the nerve sheath, connective tissue under the skin, in muscles, and other organs and are fibromyxoid with mesenchymal and neuroendocrine differentiation [1]. USCS of the thyroid gland [2], chest wall [3], and lung with metastasis to bone have also been described [4].
We previously established a patient-derived orthotopic xenograft (PDOX) nude mouse model of USCS with the technique of surgical orthotopic implantation (SOI) in the right biceps femoris muscle [5]. We initially evaluated the efficacy of standard first-line chemotherapy of doxorubicin (DOX), gemcitabine (GEM) combined with docetaxel (DOC), compared to pazopanib (PAZ), a multi-targeting tyrosine-kinase inhibitor, which had the greatest efficacy in the USCS PDOX model. These recent results demonstrated that the PDOX model of USCS could identify a promising novel agent with significantly greater efficacy than first-line therapy [5].
PAZ is a synthetic indazolylpyrimidine that is an inhibitor of multiple tyrosine kinases [6]. A phase III clinical trial for metastatic STS has been carried out to evaluate the efficacy of PAZ. A significant 3-month advantage in progression-free survival (PFS) was achieved by PAZ [7], which is consistent with our previously results in the PDOX model. However, it is impotant to discover more effective therapy for USCS.
Methionine dependence is due to the overuse of methionine for aberrant transmethylation reactions in cancer and is possibly the only known general metabolic defect in cancer [[8], [9], [10], [11], [12], [13], [14]].
The overuse of methionine by cancer cells for enhanced and unbalanced transmethylation may be the basis of the methionine dependence of cancer cells and is termed the “Hoffman effect”, analogous to Warburg effect of flucose overuse in cancer [12].
In order to target methionine dependence for therapy, our laboratory previously cloned l-methionine α-deamino-γ-mercaptomethane lyase [EC 4.4.1.11]) [15]. The cloned methioninase, termed recombinant methioninase, or rMETase, has been tested in mouse models of human cancer cell lines [[16], [17], [18], [19]]. rMETase effectively reduced tumor growth of a DOX-resistant Ewing's sarcoma PDOX compared to untreated control. The methionine level both of plasma and supernatants derived from sonicated tumors was lower in the rMETase group [20].
In a PDOX models of BRAF V600E mutant melanoma, first-line temozolomide (TEM) and rMETase was significantly more efficacious than either mono-therapy [21].
In the present study, we show that rMETase can overcome first-line DOX resistance in the PDOX models of USCS.
Section snippets
Mice
Athymic nu/nu nude mice (AntiCancer Inc., San Diego, CA), 4–6 weeks old, were used in this study. Animals were housed in a barrier facility on a high efficiency particulate arrestance (HEPA)-filtered rack under standard conditions of 12-h light/dark cycles. The animals were fed an autoclaved laboratory rodent diet. All animal studies were conducted in accordance with the principles and procedures outlined in the National Institute of Health Guide for the Care and Use of Animals under Assurance
Efficacy testing of first line therapy DOX on the USCS PDOX
To test the efficacy of first line therapy DOX on the USCS PDOX, we injected DOX weekly for 2 weeks. We found that the USCS PDOX was highly resistant to DOX with a tumor volume of 355 +/- 110 mm3 at 14 days compared to the control of 360 ± 85, p = .927 (Fig. 2, Fig. 3).
Efficacy of testing rMETase on the USCS PDOX
To test the efficacy rMETase on the USCS PDOX, we injected rMETase daily for 2 weeks. We found that rMETase alone inhibited the USCS PDOX growth with a tumor volume of 182 ± 57 mm, p = .0003 (Fig. 2, Fig. 3).
Efficacy testing of the combination of rMETase and DOX on the USCS PDOX
To test the efficacy
Discussion
The present study demonstrates the power of the PDOX model to identify effective therapy for recalcitrant cancer in this case USCS. A critical finding of the current study was that rMETase treatment could overcome first-line DOX resistance in the USCS PDOX model.
Toward this goal of precision, personalized oncology, our laboratory pioneered the patient-derived orthotopic xenograft (PDOX) nude mouse model with the technique of surgical orthotopic implantation (SOI), including pancreatic [[22],
Conclusions
In the present study of DOX-resistant USCS PDOX model, it was found that rMETase in combination with DOX could overcome DOX resistance. The present study further emphasizes the power of the PDOX model to distinguish therapies. The other critical observation of the present study is that rMEtase can overcome first-line-therapy DOX resistance, suggesting a future clinical strategy for this recalcitrant disease, especially since previous studies have shown minimal toxicity of rMETase and
Conflicts of interest
The authors declare that they have no competing interests.
Acknowledgements
This manuscript is dedicated to the memory of Dr. A.R. Moossa, Dr. Sun Lee and Dr. Shigeo Yagi.
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