Review
Lead genetic studies in Dictyostelium discoideum and translational studies in human cells demonstrate that sphingolipids are key regulators of sensitivity to cisplatin and other anticancer drugs

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Abstract

A Dictyostelium discoideum mutant with a disruption in the sphingosine-1-phosphate (S-1-P) lyase gene was obtained in an unbiased genetic analysis, using random insertional mutagenesis, for mutants with increased resistance to the widely used cancer chemotherapeutic drug cisplatin. This finding opened the way to extensive studies in both D. discoideum and human cells on the role and mechanism of action of the bioactive sphingolipids S-1-P and ceramide in regulating the response to chemotherapeutic drugs. These studies showed that the levels of activities of the sphingolipid metabolizing enzymes S-1-P lyase, sphingosine kinase and ceramide synthase, affect whether a cell dies or lives in the presence of specific drugs. The demonstration that multiple enzymes of this biochemical pathway were involved in regulating drug sensitivity provided new opportunities to test whether pharmacological intervention might increase sensitivity. Thus it is of considerable clinical significance that pharmacological inhibition of sphingosine kinase synergistically sensitizes cells to cisplatin, both in D. discoideum and human cells. Linkage to the p38 MAP kinase and protein kinase C (PKC) signaling pathways has been demonstrated. This work demonstrates the utility of D. discoideum as a lead genetic system to interrogate molecular mechanisms controlling the sensitivity of tumor cells to chemotherapeutic agents and for determining novel ways of increasing efficacy. The D. discoideum system could be easily adapted to a high throughput screen for novel chemotherapeutic agents.

Section snippets

Introduction: chemotherapy and cancer

Of the roughly 1.5 million people diagnosed with cancer in the United States this year, many will receive chemotherapy at some time during the course of their treatment [1]. Conventional chemotherapy generally consists of the systemic administration of one or more cytotoxic drugs that are known to kill the rapidly dividing cancer cells. Despite its widespread use, conventional chemotherapy suffers from significant problems including lack of specificity towards the tumors (compared to normal

Dictyostelium discoideum: its biochemistry, cell biology and genetics make it a good target for drugs and understanding drug resistance

The simple eukaryote D. discoideum has been studied extensively since its discovery in 1935 [5] as a model for multicellular morphogenesis and cytodifferentiation using a combination of genetic, biochemical and molecular methods. It has an interesting life cycle, in which cells divide mitotically and remain single until they deplete their nutrient supply, at which time they aggregate using chemotaxis into multicellular assemblies (tissues) each consisting of 105 cells. The multicellular

Selecting drug resistant mutants

The drug cisplatin (cis-diamminedichloroplatinum (II)) is widely used to treat non-Hodgkin's lymphoma, small cell and non-small cell lung, testicular, ovarian, head and neck, esophageal, and bladder cancer [26]. It is a small molecular weight molecule that primarily forms covalent adducts with adjacent purines on DNA [27]. Its anticancer effect is widely believed to be due to this chemistry, although there is a generally poor understanding of the subsequent signaling pathways that regulate its

Sphingolipid metabolism – development, cell motility, and drug sensitivity

Three of the genes identified in the mutant screen for cisplatin resistance were of particular interest. The regA cAMP phosphodiesterase had been studied. Its role in development and its regulation of protein kinase A (PKA) were well established [34] and PKA was previously linked to cisplatin resistance in CHO cells [35]. Additional work on PKA mutants in D. discoideum was done to show that cisplatin resistance was indeed PKA dependent (unpublished). Golvesin has been shown to be a Golgi

Validation of the role of S-1-P lyase and sphingosine kinase in human cells – extending the paradigm

As demonstrated above, model systems with powerful genetics are critical to identifying novel information about important biomedical problems such as the molecular basis of cisplatin resistance. Not only can un-biased genetic selections/screens be done, but further validation and mechanistic interrogation can be done rapidly and with little expense. However, these new ideas must be translated to human systems, allowing the verification of their importance to human health and physiology.

The

Ceramide synthase enzymes

Further studies have looked at the role of the other bioactive sphingolipid, ceramide, in determining cellular sensitivity to drugs. The guiding idea here was that if increased levels of S-1-P result in decreased sensitivity, then increased levels of ceramide would be expected to increase sensitivity. Moreover, this would provide an opportunity to test directly the S-1-P/ceramide rheostat model with respect to drug sensitivity. Indeed, there were reports that exogenously added soluble membrane

The future

Extensive genetic and pharmacological investigations in D. discoideum and human cells have shown that sphingolipids, and the enzymes that regulate their homeostasis, play a central role in controlling the response of a cell to specific chemotherapeutic drugs. The D. discoideum system allowed the initial blind selection of mutants, and a rapid way of making additional isogenic mutants and testing the overall hypothesis that modulating the activity of the S-1-P metabolizing enzymes could

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