Multi-omics Analysis of Serum Samples Demonstrates Reprogramming of Organ Functions Via Systemic Calcium Mobilization and Platelet Activation in Metastatic Melanoma

Abstract

Pathophysiologies of cancer-associated syndromes such as cachexia are poorly understood and no routine biomarkers have been established, yet. Using shotgun proteomics, known marker molecules including PMEL, CRP, SAA, and CSPG4 were found deregulated in patients with metastatic melanoma. Targeted analysis of 58 selected proteins with multiple reaction monitoring was applied for independent data verification. In three patients, two of which suffered from cachexia, a tissue damage signature was determined, consisting of nine proteins, PLTP, CD14, TIMP1, S10A8, S10A9, GP1BA, PTPRJ, CD44, and C4A, as well as increased levels of glycine and asparagine, and decreased levels of polyunsaturated phosphatidylcholine concentrations, as determined by targeted metabolomics. Remarkably, these molecules are known to be involved in key processes of cancer cachexia. Based on these results, we propose a model how metastatic melanoma may lead to reprogramming of organ functions via formation of platelet activating factors from long-chain polyunsaturated phosphatidylcholines under oxidative conditions and via systemic induction of intracellular calcium mobilization. Calcium mobilization in platelets was demonstrated to alter levels of several of these marker molecules. Additionally, platelets from melanoma patients proved to be in a rather exhausted state, and platelet-derived eicosanoids implicated in tumor growth were found massively increased in blood from three melanoma patients. Platelets were thus identified Serum is the most important diagnostic sample type because of its minimal invasive access, a relatively high stability and its comprehensive representation of the physiological state of an individual. The latest technological development of mass spectrometric instruments, such as high resolution orbitrap instruments, improved substantially the quality and reliability of serum proteomics data (1). Shotgun proteomics analysis using the orbitrap technology is mainly applied for protein screening purposes (2). This method allows performing untargeted analyses, applicable for hypothesis-generating clinical applications. Targeted proteomics using triple-quadrupole mass spectrometric instruments, represents a complementary approach which is applied for protein quantification, hypotheses verification and validation (3). Enormous efforts have been invested in the last decades focusing on serum biomarker discovery (4). However, mass spectrometrybased proteomics analyses hardly managed to cope with biological variation. As a consequence, almost no clinically validated biomarker has emerged from these investigations yet, and a lot of questions about pathophysiological mechanisms remain unanswered.Using mass spectrometry-based proteomics, we have previously investigated melanoma and neighboring stroma cells, focusing on the identification of biomarkers associated with intrinsic and extrinsic drug resistance (5). In the present article, we followed the question how metastatic melanoma may reprogram distant organ functions, and how these...

The Challenge of Classifying Metastatic Cell Properties by Molecular Profiling Exemplified with Cutaneous Melanoma Cells and Their Cerebral Metastasis from Patient Derived Mouse Xenografts

“…First, clinically relevant melanoma marker molecules PMEL and MLANA (21) were detected in all cells and found significantly down-regulated in the YDFR-CB variant. All detected melanoma markers varied substantially between the cells of the different donors, whereas CSPG4 (21,22) was found uniformly increased in all four CB variants lacking significance in all of them. Proteins potentially regulating immune responses of the host such as class II histocompatibility antigens HLA-DRB1 and CD74 (23), and the innate immune response regulator ANXA1 were found specifically altered between the variants and between the donors with no consistent expression pattern.…”

Integrated omics: tools, advances and future approaches

“…An integrated omics approach in cancer may provide information for improved diagnosis of carcinoma subtype pathogenesis. Recently, Muqaku et al (2017), using both label-free and targeted proteomics, lipidomics and metabolomics efforts followed by data integration in human serum samples from patients with metastatic melanoma, proposed a model on reprogramming of organ functions induced by metastatic melanoma through formation of platelet activating factors from long-chain polyunsaturated phosphatidylcholines under oxidative conditions. -Buschart et al (2016) used a multi-omics approach integrating metagenomic, metatranscriptomic and metaproteomic data from the gastrointestinal microbiome to identify intra-and inter-individual variation in subjects with type 1 diabetes mellitus (T1DM).…”

The “Janus Face” of Platelets in Cancer

“…Detrimental effects of increased thrombin generation seem to rely on its ability in promoting, in concert with its targets (among them, protease-activated receptor-1 and -4), cancer adhesion to platelets or endothelium via up-regulation of pro-inflammatory cytokines, adhesion molecules, angiogenic factors, and matrix-degrading proteases, thereby dramatically increasing tumor growth, angiogenesis, invasion, and metastasis [98]. Proofs of the "tumor cell-induced platelet aggregation" also come from quantitative analysis of circulating neutrophil elastase (which proteolytically activates integrin αIIbβ3) and serglycin (a pro-apoptotic and half-life cytokine regulating protein) in melanoma patients; while levels of the former were found up-regulated in cancer subjects, content of the latter was low [99], thus indicating the role of activated platelets in promoting cancer progression.…”

“…Eicosanoids are crucially involved in several pathophysiological conditions [139], and the intersection between changes in certain platelet-derived eicosanoids and cancer appears particularly intriguing. For example, a multi-omics analysis of serum from metastatic melanoma patients revealed a rise in the concentration of 12-hydroxyeicosatraenoic acid (HETE) and 15-HETE eicosanoids, which are respectively produced by platelet 12-LOX and COX-1 [99]. These two platelet-derived eicosanoids have also been shown to exert pro-malignant effects in several cancer types, by activating mitogen and angiogenic pathways [139][140][141][142].…”