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Minnelide Overcomes Oxaliplatin Resistance by Downregulating the DNA Repair Pathway in Pancreatic Cancer

  • 2015 SSAT Plenary Presentation
  • Published:
Journal of Gastrointestinal Surgery Aims and scope

Abstract

Introduction

Oxaliplatin is part of pancreatic cancer therapy in the FOLFIRINOX or GEMOX/XELOX regimen. DNA damage repair is one of the factors responsible for oxaliplatin resistance that eventually develops in this cancer. Triptolide/Minnelide has been shown to be effective against pancreatic cancer in preclinical trials. In this study, we evaluated the efficacy of combination of triptolide and oxaliplatin against pancreatic cancer.

Methods

Highly aggressive pancreatic cancer cells (MIA PaCa-2 and PANC-1) were treated with oxaliplatin (0–10 μM), low-dose triptolide (50 nM), or a combination of both for 24–48 h. Cell viability, apoptosis, and DNA damage were evaluated by appropriate methods. Nucleotide excision repair pathway components were quantitated using qPCR and Western blot. Combination of low doses of Minnelide and oxaliplatin was tested in an orthotopic murine model of pancreatic cancer.

Results

Proliferation of pancreatic cancer cells was markedly inhibited by combination treatment. Triptolide potentiated apoptotic cell death induced by oxaliplatin and sensitized cancer cells towards oxaliplatin-induced DNA damage by suppressing the oxaliplatin-induced DNA damage repair pathway. Combination of low doses of Minnelide and oxaliplatin inhibited tumor progression by inducing significant apoptotic cell death in these tumors.

Conclusions

Combination of low doses of Minnelide and oxaliplatin has immense potential to emerge as a novel therapeutic strategy against pancreatic cancer.

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Acknowledgments

This study was funded by NIH grants R01-CA170946 and CA124723 (to AKS); NIH grant R01-CA184274 (to SB); UAB/UMN SPORE in Pancreatic Cancer-P50 CA101955 (to SM); Katherine and Robert Goodale foundation support (to AKS); and Minneamrita Therapeutics LLC (to AKS).

Author information

Authors and Affiliations

  1. Division of Basic and Translational Research, Department of Surgery, University of Minnesota, 420 Delaware Street SE, MMC195, Minneapolis, MN, 55455, USA

    Shrey Modi, Devika Kir, Bhuwan Giri, Kaustav Majumder, Nivedita Arora, Vikas Dudeja, Sulagna Banerjee & Ashok K Saluja

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  1. Shrey Modi
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  2. Devika Kir
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Corresponding author

Correspondence to Ashok K Saluja.

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Conflict of Interest

The University of Minnesota has a patent for Minnelide, which has been licensed to Minneamrita Therapeutics, LLC. AKS is the co-founder and the Chief Scientific Officer of this company. SB is a consultant with Minneamrita Therapeutics LLC. This relationship has been reviewed and managed by the University of Minnesota in accordance with its conflict of interest policies.

Additional information

Primary Discussant

Jeffrey B. Matthews, M.D. (Chicago, IL): Despite advances in surgical therapy for pancreatic cancer, we have made precious little impact on overall survival in this disease due in part to slow progress in identifying effective chemotherapeutic agents, or combinations of agents, against this difficult tumor. You have shown that Minnelide, a synthetic prodrug of triptolide, increased cell death and reduced drug resistance to oxaliplatin. These results are interesting and suggest a novel approach to combat resistance to platinum-based chemotherapy. Nevertheless, our enthusiasm is always tempered by past experience. The road to successful cancer drug development is unfortunately littered with compounds that were effective in cell lines and murine models only to fail in the human disease. We eagerly await the results of your Phase 1 clinical trial.

I have two questions for you. First, because triptolide is derived from a Chinese herb, I am wondering

1) Whether the concentrations used in your experiments and trials are similar to what would be achieved in its use as a traditional Chinese medicine? Are there any specific toxicities and side effects?

2) Regarding mechanism of action. In previous publications from your lab, you suggested that heat shock protein 70 was involved in the pro-apoptotic action of Minnelide, and other work suggests involvement of reactive oxidant species and NfKB signaling. In today’s work, you focus on the nucleotide excision repair pathway. Can you connect the dots for us? Is there a single molecular target for Minnelide that might explain both its own apoptotic action and its effect on platinum resistance?

Closing Discussant

Dr. Modi

1. The extracts of Tripterygium wilfordii hook F (TWHF) contain around 80 active components with triptolide being one of them. Therefore, the direct comparison between the concentrations of triptolide achieved by administration of various extracts of Chinese herbal medicine and Minnelide is not feasible.

In the Phase I clinical trial, 27 patients have been enrolled so far, with 24 evaluable for toxicity. The therapy has been generally well tolerated with the only common toxicity being hematologic, but one patient experienced reversible cerebellar dysfunction at the highest dose. Hematologic toxicity has been notable for rapid onset and rapid recovery from neutropenia—often within 2–3 days of dose interruption. We have used these reference doses obtained from the trial in our experiments.

2. The exact molecular target of triptolide in a cancer cell is still not completely known. The cellular pathways are an intricate network of interconnected signaling. We have previously shown that triptolide inhibits Sp1 activity leading to decreased NF-κB signaling and thereby decreased HSP 70 levels. AP1 has also been shown to regulate pro-survival pathways like NF-κB and vice versa. Promoter regions of AP1 are predicted to have Sp1 and NF-κB binding sites as well (www.genecards.org). So, it is possible that triptolide affects these upstream events that culminate into AP1 inhibition leading to triggering of apoptotic pathways and overcoming platinum resistance.

The proposed mechanism of action of how triptolide inhibits AP1.

figure a

Electronic supplementary material

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Supplementary Figure 1

mRNA expression of genes not inhibited by TPL treatment. MIA PaCa-2 cells were treated with TPL 100 nM for 24 h and data is represented as fold change over untreated cells. (PDF 1671 kb)

Supplementary Figure 2

Equi-effective doses of paclitaxel (PXL) and rapamycin (Rapa) do not affect components of NER pathway demonstrating the pathway’s specificity as a resistance mechanism to DNA damaging agents like oxaliplatin (Ox). MIA PaCa-2 (a) and PANC-1 (b) cells were treated with ED25 doses of Ox (5 μM), PXL (9.1 nM) and Rapa (248.1 nM) for 24 h and mRNA levels of ERCC1, XPA, XPB, XPC, XPD, XPF and XPG were analyzed. Data is represented as fold change over untreated cells. (PDF 2932 kb)

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Modi, S., Kir, D., Giri, B. et al. Minnelide Overcomes Oxaliplatin Resistance by Downregulating the DNA Repair Pathway in Pancreatic Cancer. J Gastrointest Surg 20, 13–24 (2016). https://doi.org/10.1007/s11605-015-3000-3

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  • DOI: https://doi.org/10.1007/s11605-015-3000-3

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