A method for determining the risk of, or prognosis of, lung, pancreatic, and ovarian cancer, and cutaneous and uveal melanoma

Abstract

A method for predicting disease free status or overall survival in an individual diagnosed with cancer, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease free status and having an increased chance of overall survival.

Description

[0001] Title A method for determining the risk of, or prognosis of, lung, pancreatic, and ovarian cancer, and cutaneous and uveal melanoma. Field of the Invention The invention relates to a method for determining the cancer status of an individual suspecting or confirmed to have cancer, and providing a treatment to the individual if required. Specifically, the method comprises determining the status of lung, pancreatic, ovarian cancers, as well as cutaneous and uveal melanomas. Background to the Invention Ferroptosis, a form of regulated cell death triggered by iron-dependent lipid peroxidation, is a novel addition to anticancer strategies. Ferroptosis is morphologically and biochemically distinct from autophagy, apoptosis, necrosis, and necroptosis. It impedes cancer growth, bolsters chemotherapy and radiation sensitivity, and inhibits metastasis. Core ferroptosis hallmarks include: i) loss of lipid peroxide repair capacity through glutathione peroxidase 4 (GPX4), ii) bioavailability of redox-active iron, and iii) oxidation of polyunsaturated fatty acid (PUFA)-containing phospholipids and accumulation of lipid oxidation products e.g.4-HNE. Nuclear factor erythroid 2-related factor (NRF2) plays a key role in redox homeostasis. One of its major targets, heme oxygenase 1 (HO-1), is an antioxidant and detoxifying gene which can exert either a cytoprotective or detrimental action in cancer, based on the specific cellular conditions. Promoting ferroptosis can overcome cancer cells' resistance to oxidative stress, broadening therapeutic choices when combined with traditional drugs and improving clinical monitoring through the use of companion biomarkers. Lung cancer, the leading cause of cancer death globally, includes non-small-cell lung cancer (NSCLC) and small cell carcinoma (SCLC). Common treatments involve surgery, radiotherapy, chemotherapy, and targeted therapies as immunotherapy, but metastasis and drug resistance result in poor prognosis. NSCLC has a 60% diagnosis rate in advanced stages and a poor 5-year survival, indicating a need for better biomarkers and treatments. New treatments combining immunotherapy with small ferroptosis-inducing molecules show promise in overcoming resistance to existing therapies. Pancreatic cancer exhibits early and high local recurrence, resulting in <10% 5-year overall survival (OS) rates. By 2025, it's expected to be the second leading cause of cancer death in the US, surpassing colorectal cancer before 2040 and following lung cancer in mortality. Pancreatic ductal adenocarcinoma (PDAC) is usually treated with surgery and chemotherapy, but only a small percentage of patients are eligible for surgery upon diagnosis due to atypical early symptoms. Recurrence occurs in around 75% of patients after surgery, leading to lower survival rates. To improve outcomes, it is important to develop innovative and more personalized therapeutic approaches and improve postoperative monitoring. Ferroptosis-regulating strategies have garnered attention in pancreatic cancer research. Erastin has shown promise in enhancing the cytotoxicity of gemcitabine and cisplatin in pancreatic cancer cells. This is significant as FOLFIRINOX, the chemotherapy combination offering the best clinical benefits for PDAC patients and comprises folinic acid, fluorouracil (5-FU), irinotecan, and oxaliplatin, has higher toxicity (32% vs. 9%) and rapid development of chemoresistance compared to gemcitabine. Ferroptosis research has the potential to alleviate drug resistance emergence in PDAC clinical trials. Ovarian cancer (OVCA) is a highly lethal malignancy with a 5-year relative survival rate below 50%, primarily due to its frequent recurrence and limited early detection methods. The most prevalent type is epithelial ovarian cancer among OVCA's various histological forms. Current OVCA treatments involve surgery, platinum-based chemotherapies, angiogenesis inhibitors, poly ADP-ribose polymerase (PARP) inhibitors, and immunotherapies. However, standard treatments often fall short, particularly for patients with recurrent disease. Therefore, innovative OVCA treatments are urgently needed. Ferroptosis in OVCA has been explored extensively, with a focus on ovarian cancer cell lines. It was discovered that elevated Frizzled 7 expression in platinum-tolerant ovarian cancer cells. This led to increased GPX4 levels, and inhibiting GPX4 sensitised these platinum-tolerant cells to platinum, inducing ferroptosis. Bevacizumab (Avastin®), an anti-angiogenic agent targeting vascular endothelial growth factor (VEGF-A), is widely studied in OVCA. However, its efficacy diminishes after 2 years, failing to extend overall survival. Uveal melanoma (UM) is a rare eye with a global prevalence of 1-9 / 1,000,000 (www.orpha.net). UM is the most common primary intraocular malignancy in adults, arising from uveal melanocytes. Hematogenously, UM metastasizes in ~ 50% of patients, most frequently to the liver. The primary UM can be treated by surgery or radiation, but therapeutic options for metastatic UM (MUM) patients are very limited. MUM patients have a median survival time of only 6-12 months; around 8% of MUM patients survive beyond two years. Recently, Tebentafusp (Kimmtrak(R)), a bispecific fusion protein that redirects CD3+ T cells to target glycoprotein 100-positive melanoma cells, was reported to improve overall MUM survival by 6 months. Tebentafusp is approved by the United States Food and Drug Administration and by the European Medicines Agency for treatment of HLA-A*02:01-positive adults with unresectable or metastatic UM. Therefore, only a subcategory of MUM patients is eligible for Tebentafusp treatment, underlying the need for additional, more effective treatments. On August 2023, the FDA approved HEPZATO KIT (melphalan for Injection / Hepatic Delivery System) containing melphalan (HEPZATO, Delcath Systems, Inc.) as a liver- directed treatment for adult patients with uveal melanoma with unresectable hepatic metastases affecting less than 50% of the liver and no extrahepatic disease, or extrahepatic disease limited to the bone, lymph nodes, subcutaneous tissues, or lung that is amenable to resection or radiation. The main efficacy outcome measures were objective response rate (ORR) and duration of response (DoR). ORR was 36.3% (95% CI: 26.4, 47) and median DoR was 14 months (95% CI: 8.3, 17.7). Because of the risk of severe peri-procedural complications including hemorrhage, hepatocellular injury, and thromboembolic events, HEPZATO KIT is available only through a restricted program under a Risk Evaluation and Mitigation Strategy called the HEPZATO KIT REMS. Activating mutations in GNAQ or GNA11 occur in ~83% of UMs, while mutations in CYSLTR2 or PLCB4 occur in ~10% of cases. Cysteinyl leukotriene receptors 1 (CysLT1) and 2 (CysLT2), are G protein-coupled receptors, which signal to downstream effectors, such as phospholipase C-β (PLCβ), protein kinase C (PKC), ADP- ribosylation factor 6 (ARF6) and β-catenin. These cascades modulate pathways including mitogen-activated protein kinase (MAPK), PI3K / AKT, and Rho GTPase. Upstream, the receptors are activated by cysteinyl leukotrienes (CysLTs), inflammatory lipid mediators synthesized through the 5-lipoxygenase (5-LO) pathway. A role for CysLTs in cancer has recently emerged. In retrospective analyses, CysLT1 antagonists showed a dose-dependent chemo-preventative effect against 14 cancers and an overall decreased risk of cancer. of CysLT1 is observed in colorectal cancer, renal cell carcinoma, breast cancer and UM. High expression of the CysLT receptors genes, CYSLTR1 and CYSLTR2, is significantly associated with poor disease-free survival (DFS) and poor overall survival (OS) in UM patients. The CysLT1 antagonist quininib and its analogue 1,4-dihydroxy quininib significantly alter viability, long-term proliferation, secretion of inflammatory and angiogenic factors, and oxidative phosphorylation in primary and metastatic UM cell lines. CysLT1 antagonists also significantly inhibit tumour burden in zebrafish xenograft models of UM. Furthermore, in tumours from a cell line-derived mouse orthotopic xenograft model of MUM, 1,4- dihydroxy quininib significantly decreases expression levels of ATP synthase F1 β subunit (ATP5B), a protein marker of oxidative phosphorylation. Notably, high expression of ATP5F1B in primary UM is significantly associated with reduced progression-free survival and reduced OS, and patients with disomy 3 and low ATP5F1B expression have a reduced risk of metastatic disease. MUM prognosis is still very challenging, with only 1% of patients displaying metastases at the time of primary UM diagnosis. Thus, a deeper understanding of 1,4-dihydroxy quininib molecular mechanisms may also support the discovery of novel prognostication biomarkers for UM patients. It is an object of the present invention to overcome at least one of the above-mentioned problems. Summary of the Invention Expression levels of individual ferroptosis-related genes or of combinations of these genes can be clinically relevant in lung cancer, pancreatic cancer, ovarian cancer, uveal melanoma, and cutaneous melanoma. In particular, the expression levels of individual ferroptosis-related genes or of combinations of them can stratify lung cancer, pancreatic cancer, ovarian cancer, uveal melanoma, and cutaneous melanoma patients according to their risk of recurrence and / or risk of death. One of the invention’s aims was to test if CysLT receptor antagonists induce ferroptosis in lung cancer, pancreatic cancer, and ovarian cancer, and also assess whether combining these antagonists with standard treatments enhances anticancer effects of known drugs such as cisplatin, FOLRIRINOX, or bevacizumab, and whether a ferroptosis-related gene signature predicts patient survival in these cancers. Quininib and its derivative 1,4-dihydroxy-quininib display anti-angiogenic properties, which could boost the effects of bevacizumab in ovarian cancer. Furthermore, a scoring system based on 14 ferroptosis-related genes (FRGs) can predict the prognosis of ovarian cancer patients, highlighting how these signatures may represent important prognostic tool for lung, pancreatic, and ovarian cancers. The inventions is set out in the appended claims. There is provided a method for predicting disease free status or overall survival in an individual diagnosed with uveal melanoma, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or the proteins encoded by said genes, assayed from a biological sample from a subject with no cancer, the individual is predicted to have an increased probability of having disease free status or overall survival. In one aspect, the gene signature consists of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or of proteins encoded by said genes; wherein when the expression level of the gene signature consisting of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature consisting of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or the proteins encoded by said genes, assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease-free status or overall survival. In one aspect, there is a method for predicting disease free status or overall survival in an individual diagnosed with uveal melanoma, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, CTH, ACSL3, IREB2, and SLC7A11, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3 and IREB2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3 and IREB2 or the proteins encoded by said assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease-free status or overall survival. In one aspect, there is a method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) mutation status of Bap-1 in the individual or (b) expression status of Bap-1 in the individual; wherein when the individual has one or more of (a) a wildtype Bap-1 sequence or (b) normal Bap-1 expression levels, relative to a reference value for (a)-(b) from a biological sample from a subject with no metastatic uveal melanoma, and wherein when the expression levels of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value for gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma. In one aspect, the method may further comprise the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma. In one aspect, there is a method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) Bap-1 mutation status in the individual or (b) the expression status of Bap-1 in the individual; wherein when the individual has at least one of (a) the presence of mutated Bap-1 or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no metastatic uveal melanoma, and the expression levels of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from subject with no metastatic uveal melanoma, the individual is predicted to have an increased risk of developing metastatic uveal melanoma. In one aspect, the method may further comprise the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have an increased risk of developing metastatic uveal melanoma. In one aspect, there is a method of determining a 5-year survival rate or a 10-year survival rate of an individual diagnosed with an uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said gene, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) a wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate. In one aspect, the method may further comprise the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate. In one aspect, when the individual has at least one of (a) a mutated Bap-1 status or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have a decreased 5-year survival rate or 10- year survival rate. In one aspect, the method may further comprise the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate. In one aspect, there is a method of identifying an uveal melanoma patient that is suitable for treatment with a therapy for preventing recurrence or progression of the uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) mutated Bap-1 status or (b) decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from the individual following administration of a treatment, the individual is predicted not to be suitable for the treatment. In one aspect, the method may further comprise the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to not to be suitable for the treatment. In one aspect, there is a method for predicting overall survival in an individual diagnosed with lung cancer, the method comprising a step of assaying a biological sample from the individual for expression level of a gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CysLT2, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CysLT2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CysLT2, or the proteins encoded by said genes, assayed from a biological sample from the subject with no cancer, the is predicted to have a reduced chance of overall survival. In one aspect, when the individual has a status of being a smoker or a former smoker, the individual is predicted to have a reduced chance of overall survival. In one aspect, the method may further comprise the step of determining KRAS mutation status of the individual, wherein when the individual has a positive KRAS mutation status, the individual is predicted to have reduced overall survival. There is provided, a method for predicting disease free status or overall survival in an individual diagnosed with cancer, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease free status and having an increased chance of overall survival. In one aspect, there is provided method for predicting a probability of having recurrent disease or having a reduced overall survival in an individual diagnosed with cancer, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with cancer, the individual is predicted to have an increased of having recurrent disease and having a reduced chance of overall survival. In one aspect, the at least one gene, or a protein encoded by said gene, is ACSL3 and the cancer is lung cancer. In one aspect, if the at least one gene, or a protein encoded by said gene, is ACSL3 and SLC3A2, and the individual has lung cancer, and the expression of ACSL3, ATP5B, and / or SLC3A2 in the sample relative to the reference value is decreased, the individual is predicted to have a lower chance of having recurrent disease status, a higher chance of disease free status and a higher chance of overall survival. In one aspect, if the at least one gene, or a protein encoded by said gene, is ACSL3, ATP5B, and / or SLC3A2, the individual has lung cancer, and the expression of ACSL3 and SLC3A2 in the sample relative to the reference value is increased, the individual is predicted to have a recurrent disease status and a lower chance of overall survival. In one aspect, wherein when the at least one gene, or a protein encoded by said gene, is CysLT1 or CysLT2, and the individual has lung cancer, and the expression levels of CysLT1 or CysLT2 are increased relative to the reference value, the individual is predicted to have a higher chance of overall survival. In one aspect, when the at least one gene, or a protein encoded by said gene, is CysLT1 or CysLT2, and the individual has lung cancer, and the expression levels of CysLT1 or CysLT2 are decreased relative to the reference value, the individual is predicted to have a decreased or lower chance of overall survival. In one aspect, wherein when the at least one gene, or a protein encoded by said gene, is IREB2 and the cancer is ovarian cancer. In one aspect, wherein when the at least gene, or a protein encoded by said gene, is IREB2, the individual has ovarian cancer, and the expression levels of IREB2 are increased relative to the reference value, the individual is predicted to have lower risk of a recurrent disease status, and a higher chance of a disease free status. In one aspect, if the at least one gene, or a protein encoded by said gene, is IREB2, and the individual has ovarian cancer, and the expression levels of IREB2 are decreased relative to the reference the individual is predicted to have recurrent disease status, and a lower chance of disease free status. In one aspect, wherein the at least one gene, or a protein encoded by said gene, is CTH and the cancer is pancreatic cancer. In one aspect, wherein if the at least gene, or a protein encoded by said gene, is CTH, and the individual has pancreatic cancer, the expression level of CTH is decreased relative to the reference value, the individual is predicted to have a lower chance of having a recurrent disease status, and a higher chance of having a disease free status and predicted to have a greater likelihood of overall survival. In one aspect, if the at least one gene, or a protein encoded by said gene, is CTH, the individual has pancreatic cancer, and the expression level of CTH is increased relative to the reference value, the individual is predicted to have a higher chance of a recurrent disease status, and a lower chance of disease free status and a lower likelihood of overall survival. In one aspect, wherein the at least one gene, or a protein encoded by said gene, is IREB2 and the cancer is cutaneous melanoma. In one aspect, wherein if the at least gene, or a protein encoded by said gene, is IREB2, and the individual has a cutaneous melanoma, the expression levels of IREB2 are decreased relative to the reference value, the individual is predicted to have a low chance of a recurrent disease status, and a higher chance of a disease free status. In one aspect, if the at least one gene, or a protein encoded by said gene, is IREB2, the individual has a cutaneous melanoma, and the expression levels of IREB2 are increased relative to the reference value, the individual is predicted to have a recurrent disease status, and a lower chance of a disease free status. In one aspect, wherein the at least one gene, or a protein encoded by said gene, is ACSL3 and the cancer is cutaneous melanoma. In one aspect, wherein if the at least gene, or a protein encoded by said gene, is ACSL3, and the individual has a cutaneous melanoma, the expression levels of ACSL3 are increased relative to the reference value, the individual is predicted to have a greater likelihood of overall survival. In one aspect, if the at least one gene, or a protein encoded by said gene, is ACSL3, the individual has a cutaneous melanoma, and the expression levels of ACSL3 are decreased relative to the reference the individual is predicted to have a lower chance of overall survival. In one aspect, there is a method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH, in combination with determining at least one of (a) mutation status of Bap-1 in the individual or (b) expression status of Bap-1 in the individual; wherein when the individual has one or more of (a) a wildtype Bap-1 sequence or (b) normal Bap-1 expression levels, relative to a reference value for (a)-(b) from a biological sample from a subject with no metastatic uveal melanoma, and wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma. In one aspect, there is a method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status in the individual or (b) the expression status of Bap-1 in the individual; wherein when the individual has at least one of (a) the presence of mutated Bap-1 or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no metastatic uveal melanoma, expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have an increased risk of developing metastatic uveal melanoma. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have an increased risk of developing metastatic uveal melanoma. In one aspect, there is a method of determining a 5-year survival rate or a 10-year survival rate of an individual diagnosed with an uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) a wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate. In one aspect, there method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate. In one aspect, when the individual has at least one of (a) a mutated Bap-1 status or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate. In one aspect, there is a method of identifying an uveal melanoma patient that is suitable for treatment with a therapy for preventing recurrence or progression of the uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) mutated Bap-1 status or (b) decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the individual following administration of a treatment, the individual is predicted not to be suitable for the treatment. In one aspect, the method further the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to not to be suitable for the treatment. In one aspect, wherein when the individual has at least one of (a) wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the individual following administration of a treatment, the individual is predicted to be suitable for the treatment. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to be suitable for the treatment. In one aspect, there is a method for monitoring the effectiveness of treatment of uveal melanoma in an individual with uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) a wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject with uveal melanoma previously administered the treatment, the treatment is predicted to be effective and the individual is predicted to have a positive outcome. In one aspect, the method further the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have a positive outcome. In one aspect, wherein when the individual has at least one of (a) a mutated Bap-1 status or (b) decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject with uveal melanoma previously administered the treatment, the treatment is predicted to be ineffective and the individual is predicted to have a poor outcome. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a poor outcome. In one aspect, the treatment is active surveillance, a neoadjuvant therapy, an adjuvant therapy, a surgery, or a combination thereof. Preferably, the neoadjuvant therapy and adjuvant therapy is an agent selected from 1,4-dihydroxy quininib, Dacarbazine® (DTIC or imidazole carboxamide), and / or tebentafusp. In one aspect, the at least one gene selected is ATP5B. In one aspect, the at least two genes selected are selected from ATP5B and GPX4, ATP5B and SLC7A11, ATPB5 and IREB2, ATP5B and CTH, GPX4 and SLC7A11, GPX4 and IREB2, GPX4 and CTH, SLC7A11 and IREB2, SLC7A11 and CTH, or IREB2 and CTH. In one aspect, at least three genes are selected from ATP5B, GPX4, and SLC7A11; ATP5B, GPX4, and IREB2; ATP5B, GPX4, and CTH; ATP5B, SLC7A11, and IREB2; ATP5B, SLC7A11, and CTH; ATP5B, IREB2, and CTH; GPX4, SLC7A11, and IREB2; GPX4, SLC7A11, and CTH; or SLC7A11, IREB2, and CTH. In one aspect, at least four genes are selected from ATP5B, GPX4, SLC7A11, and IREB2; ATP5B, GPX4, SLC7A11, CTH; or GPX4, SLC7A11, IREB2, and CTH. In one aspect, at least five genes are selected from ATP5B, GPX4, SLC7A11, IREB2, and CTH; GPX4, SLC3A2, GCLM, NQO1, and AIFM2; CYSLTR1, CYSLTR2, ATP5B, ACSL3 and SLC3A2. In one aspect, at least eleven genes selected are GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, and CTH. In one aspect, the step of determining the Bap-1 expression status and / or the Bap-1 mutation status is preferred to the step of determining chromosome 3 status in the individual. In one aspect, there is a method for treating uveal melanoma comprising the steps of: identifying an individual with increased potential for having an uveal melanoma or a metastatic uveal melanoma by assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) the expression status of Bap-1 in the individual; wherein when the individual has at least one of (a) a mutated Bap-1 status or (b) decreased expression levels of Bap-1, and the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, and CysLT2 are elevated, and the expression levels of CTH are decreased, relative to a reference value of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject with no uveal melanoma, the individual has an increased risk of having a metastatic uveal melanoma or a recurrent uveal melanoma; and treating the individual with a therapeutically effective amount of an adjuvant therapy, a neoadjuvant therapy, or a combination thereof. In one aspect, the method further comprises the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a metastatic uveal melanoma or a recurrent uveal melanoma. In one aspect, there is a method for treating cancer comprising the steps of: identifying an individual with increased potential for having cancer by assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH are dysregulated relative to a reference value of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject with no cancer, the individual has an increased risk of having cancer; and treating the individual with a therapeutically effective amount of an adjuvant therapy, a neoadjuvant therapy, or a combination thereof. In one aspect, when the gene, or a protein encoded by said gene, is ACSL3, the cancer is lung cancer and / or cutaneous melanoma. In one aspect, when the at least one gene, or a protein encoded by said gene, is IREB2, the cancer is ovarian cancer. In one aspect, when the at least one gene, or a protein encoded by said gene, is CTH, the cancer is pancreatic cancer. In one aspect, when the at least one gene, or a protein encoded by said gene, is IREB2, the cancer is cutaneous melanoma. In one aspect, the therapy or treatment is a neoadjuvant therapy. In one aspect, the therapy is an adjuvant therapy. In one aspect, the therapy can be a combination of neoadjuvant and adjuvant therapy. In one aspect, the therapy can be an active surveillance, where the treatment plan involves closely watching an individual’s condition but not giving any treatment unless there are changes in test results that show the condition is worsening. In one aspect, the therapy can be a procedure (for example, a surgical procedure to remove a primary or a secondary tumour), or a combination of neoadjuvant therapy and surgery; or adjuvant therapy and surgery; or neoadjuvant therapy, surgery, and adjuvant therapy. In one aspect, the cancer patient may be suitable for treatment with a neoadjuvant therapy for preventing recurrence or progression of the cancer. In one aspect, the adjuvant therapy and neoadjuvant therapy is a chemotherapeutic therapy. In one aspect, with respect to uveal melanoma, the adjuvant therapy and neoadjuvant therapy is selected from 1,4-dihydroxy quininib, Dacarbazine® (DTIC or imidazole carboxamide), and / or Tebentafusp. In one aspect, with respect to lung cancer, the adjuvant therapy and neoadjuvant therapy is selected from one or more of the following: Adagrasib, Afatinib Dimaleate, Alectinib, Amivantamab-vmjw, Atezolizumab, Bevacizumab, Binimetinib, Brigatinib, Capmatinib Hydrochloride, Cemiplimab rwlc, Ceritinib, Crizotinib, Dabrafenib Mesylate, Dacomitinib, Docetaxel, Doxorubicin Hydrochloride, Durvalumab, Encorafenib, Entrectinib, Erlotinib Hydrochloride, Etoposide, Etoposide Phosphate, Everolimus, Everolimus Fam-Trastuzumab Deruxtecan-nxki, Gefitinib, Gemcitabine Hydrochloride, Ipilimumab, Lorlatinib Lurbinectedin, Methotrexate Sodium, Necitumumab, Nivolumab, Osimertinib Mesylate, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle Formulation, Pembrolizumab, Pemetrexed Disodium, Pralsetinib, Ramucirumab, Repotrectinib, Selpercatinib, Sotorasib, Tepotinib Hydrochloride, Trametinib Dimethyl Sulfoxide, Tremelimumab-actl, Topotecan Hydrochloride, and Vinorelbine Tartrate. In one aspect, a combination of carboplatin and taxol, and / or gemcitabine and cisplatin, are selected. In one aspect, in relation to pancreatic cancer, the adjuvant therapy and neoadjuvant therapy is selected from one or more of the following: Capecitabine, Erlotinib Hydrochloride, Everolimus, Fluorouracil Injection, Gemcitabine Hydrochloride, Irinotecan Hydrochloride Liposome, Mitomycin, Olaparib, Paclitaxel Albumin-stabilized Nanoparticle Formulation, and Sunitinib Malate. In one aspect, a combination of folinic acid, fluorouracil, irinotecan and oxaliplatin; gemcitabine and cisplatin; and / or gemcitabine and oxaliplatin is used. In one aspect, in relation to ovarian, tube, or primary peritoneal cancer, the adjuvant therapy and neoadjuvant therapy is selected from one or more of the following: Bevacizumab, Carboplatin, Cisplatin, Cyclophosphamide, Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, Gemcitabine Hydrochloride, Melphalan, Mirvetuximab soravtansine-gynx, Niraparib Tosylate Monohydrate, Olaparib, Paclitaxel, Rucaparib Camsylate, Thiotepa, and Topotecan Hydrochloride. In one aspect, a combination of bleomycin, etoposide phosphate, and cisplatin; carboplatin and toxol; carboplatin, etoposide phosphate, and bleomycin; cisplatin, etoposide phosphate, and bleomycin; vincristine, actinomycin D and cyclophosphamide; and / or vinblastine, ifosfamide and cisplatin is used. In one aspect, in relation to melanoma, the adjuvant therapy and neoadjuvant therapy is selected from one or more of the following: Aldesleukin, Binimetinib, Cobimetinib Fumarate, Dabrafenib Mesylate, Dacarbazine, Encorafenib, Ipilimumab, Melphalan Hydrochloride, Nivolumab, Nivolumab and Relatlimab-rmbw, Pembrolizumab, Recombinant Interferon Alfa-2b, Talimogene Laherparepvec, Tebentafusp-tebn, Trametinib Dimethyl Sulfoxide, and Vemurafenib. In one aspect, the therapy is electing not to have surgery, adjuvant therapy, and / or neoadjuvant therapy. In one aspect, the at least one gene, or protein encoded by said gene consists of GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, and CTH. In some embodiments of any of the aspects, the measurement of the expression value of a target and / or detection of the expression value or presence of a target, e.g., of an expression product (nucleic acid or polypeptide of one of the genes described herein) or a mutation can comprise a transformation. As used herein, the term “transforming” or “transformation” refers to changing an object or a substance, e.g., biological sample, nucleic acid, or protein, into another substance. The transformation can be physical, biological, or chemical. Exemplary physical transformation includes, but is not limited to, pre-treatment of a biological sample, e.g., from whole blood to blood serum by differential centrifugation. A biological / chemical transformation can involve the action of at least one enzyme and / or a chemical reagent in a reaction. For example, a DNA sample can be digested into fragments by one or more restriction enzymes, or an exogenous molecule can be attached fragmented DNA sample with a ligase. In some embodiments of any of the aspects, a DNA sample can undergo enzymatic replication, e.g., by polymerase chain reaction (PCR). Transformation, measurement, and / or detection of a target molecule, e.g., a gene, nucleotide, mRNA, or polypeptide can comprise contacting a sample obtained from a subject with a reagent (e.g., a detection reagent) which is specific for the target, e.g., a target-specific reagent. In some embodiments of any of the aspects, the target-specific reagent is detectably labelled. In some embodiments of any of the aspects, the target- specific reagent can generate a detectable signal. In some embodiments of any of the aspects, the target-specific reagent generates a detectable signal when the target molecule is present. Methods to measure gene expression products are known to a skilled artisan. Such methods to measure gene expression products, e.g., protein level, include ELISA (enzyme linked immunosorbent assay), western blot, immunoprecipitation, and immunofluorescence using detection reagents such as an antibody or protein binding agents. Alternatively, a peptide can be detected in a subject by introducing into a subject a labelled anti-peptide antibody and other types of detection agent. For example, the antibody can be labelled with a detectable marker whose presence and location in the subject is detected by standard imaging techniques. For example, antibodies for the various targets described herein are commercially available and can be used for the purposes of the invention to measure protein expression levels, e.g., anti-ATP5B (Sigma-Aldrich - HPA001520), anti-GPX4 (Abcam - ab125066), anti-SLC7A11 (Proteintech - 26864-1-AP), anti-IREB2 (Proteintech - 23829-1-AP), anti-CTH (Proteintech - 12217-1-AP), anti-SLC3A2 (Proteintech - 15193- 1-AP), anti-GCLM (Proteintech - 14241-1-AP), anti-ACSL3 (Proteintech - 20710-1-AP), anti-NQO1 (Proteintech - 11451-1-AP), anti-AIFM2 (Proteintech - 68049-1-Ig), anti- HMOX1 (Proteintech – 10701-1-AP), anti-POR (Proteintech – 29814-1-AP), anti-Bap-1 (Santa Cruz Biotechnology - sc-28383), anti-CysLT1 (Abcam – ab151484, 1:200) and anti-CysLT2 (Cayman Chemical - CAY120560, 1:500). Alternatively, since the amino acid sequences for the targets described herein are known and publicly available at the NCBI website, one of skill in the art can raise their own antibodies against these polypeptides of interest for the purpose of the methods described herein. In some embodiments of any of the immunohistochemistry (“IHC”) and immunocytochemistry (“ICC”) techniques can be used. IHC is the application of immunochemistry to tissue sections, whereas ICC is the application of immunochemistry to cells or tissue imprints after they have undergone specific cytological preparations such as, for example, liquid-based preparations. Immunochemistry is a family of techniques based on the use of an antibody, wherein the antibodies are used to specifically target molecules inside or on the surface of cells. The antibody typically contains a marker that will undergo a biochemical reaction, and thereby experience a change of colour, upon encountering the targeted molecules. In some instances, signal amplification can be integrated into the particular protocol, wherein a secondary antibody, that includes the marker stain or marker signal, follows the application of a primary specific antibody. In some embodiments of any of the aspects, the assay can be a Western blot analysis. Alternatively, proteins can be separated by two-dimensional gel electrophoresis systems. Two-dimensional gel electrophoresis is well known in the art and typically involves iso-electric focusing along a first dimension followed by SDS-PAGE electrophoresis along a second dimension. These methods also require a considerable amount of cellular material. The analysis of 2D SDS-PAGE gels can be performed by determining the intensity of protein spots on the gel or can be performed using immune detection. In other embodiments, protein samples are analysed by mass spectroscopy. Immunological tests can be used with the methods and assays described herein and include, for example, competitive and non-competitive assay systems using techniques such as Western blots, radioimmunoassay (RIA), ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoprecipitation assays, immunodiffusion assays, agglutination assays, e.g. latex agglutination, complement- fixation assays, immunoradiometric assays, fluorescent immunoassays, e.g., FIA (fluorescence-linked immunoassay), chemiluminescence immunoassays (CLIA), electrochemiluminescence immunoassay (ECLIA, counting immunoassay (CIA), lateral flow tests or immunoassay (LFIA), magnetic immunoassay (MIA), and protein A immunoassays. Methods for performing such assays are known in the art, provided an appropriate antibody reagent is available. In some embodiments of any of the aspects, the immunoassay can be a quantitative or a semi-quantitative immunoassay. An immunoassay is a biochemical test measures the concentration of a substance in a biological sample, typically a fluid sample such as blood or serum, or a sample from the eye using the interaction of an antibody or antibodies to its antigen. The assay takes advantage of the highly specific binding of an antibody with its antigen. For the methods and assays described herein, specific binding of the target polypeptides with respective proteins or protein fragments, or an isolated peptide, or a fusion protein described herein occurs in the immunoassay to form a target protein / peptide complex. The complex is then detected by a variety of methods known in the art. An immunoassay also often involves the use of a detection antibody. Enzyme-linked immunosorbent assay, also called ELISA, enzyme immunoassay or EIA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In one embodiment, an ELISA involving at least one antibody with specificity for the desired antigen (e.g., any of the targets as described herein) can also be performed. A known amount of sample and / or antigen is immobilized on a solid support (usually a polystyrene microtiter plate). Immobilization can be either non-specific (e.g., by adsorption to the surface) or specific (e.g., where another antibody immobilized on the surface is used to capture antigen or a primary antibody). After the antigen is immobilized, the detection antibody is added, forming a complex with the antigen. The detection antibody can be covalently linked to an enzyme or can itself be detected by a secondary antibody which is linked to an enzyme through bio-conjugation. Between each step the plate is typically washed with a mild detergent solution to remove any proteins or antibodies that are not specifically bound. After the final wash step the plate is developed by adding an enzymatic substrate to produce a visible signal, which indicates the quantity of antigen in the sample. Older ELISAs utilize chromogenic substrates, though newer assays employ fluorogenic substrates with much higher sensitivity. In another embodiment, a competitive ELISA is used. Purified antibodies that are directed against a target polypeptide or fragment thereof are coated on the solid phase of multi-well plate, i.e., conjugated to a solid surface. A second batch of purified antibodies that are not conjugated on any solid support is also needed. These non- conjugated purified antibodies are labelled for detection purposes, for example, labelled with horseradish peroxidase to a detectable signal. A sample (e.g., a blood sample) from a subject is mixed with a known amount of desired antigen (e.g., a known volume or concentration of a sample comprising a target polypeptide) together with the horseradish peroxidase labelled antibodies and the mixture is then added to coated wells to form competitive combination. After incubation, if the polypeptide level is high in the sample, a complex of labelled antibody reagent-antigen will form. This complex is free in solution and can be washed away. Washing the wells will remove the complex. Then the wells are incubated with TMB (3,3´,5,5´-tetramethylbenzidene) colour development substrate for localization of horseradish peroxidase-conjugated antibodies in the wells. There will be no colour change or little colour change if the target polypeptide level is high in the sample. If there is little or no target polypeptide present in the sample, a different complex in formed, the complex of solid support bound antibody reagents-target polypeptide. This complex is immobilized on the plate and is not washed away in the wash step. Subsequent incubation with TMB will produce significant colour change. Such a competitive ELSA test is specific, sensitive, reproducible, and easy to operate. There are other different forms of ELISA, which are well known to those skilled in the art. The standard techniques known in the art for ELISA are described in “Methods in Immunodiagnosis”, 2ndEdition, Rose and Bigazzi, eds. John Wiley & Sons, 1980; and Oellerich, M. 1984, J. Clin. Chem. Clin. Biochem. 22:895-904. These references are hereby incorporated by reference in their entirety. In one embodiment, the levels of a polypeptide in a sample can be detected by a lateral flow immunoassay test (LFIA), also known as the immunochromatographic assay, or strip test. LFIAs are a simple device intended to detect the presence (or absence) of antigen, e.g., a polypeptide, in a fluid sample. There are currently many LFIA tests used for medical diagnostics, either for home testing, point of care testing, or laboratory use. LFIA tests are a form of immunoassay in which the test sample flows along a solid substrate via capillary action. After the sample is applied to the test strip it encounters a coloured reagent (generally comprising antibody specific for the test target antigen) bound to microparticles which mixes with the sample and transits the substrate encountering lines or zones which have been pretreated with another antibody or antigen. Depending upon the level of target polypeptides present in the sample the coloured reagent can be captured and become bound at the test line or zone. LFIAs are essentially immunoassays adapted to operate along a single axis to suit the test strip format or a dipstick format. Strip are extremely versatile and can be easily modified by one skilled in the art for detecting an enormous range of antigens from fluid samples such as urine, blood, water, and / or homogenized tissue samples etc. Strip tests are also known as dip stick tests, the name bearing from the literal action of “dipping” the test strip into a fluid sample to be tested. LFIA strip tests are easy to use, require minimum training and can easily be included as components of point-of-care test (POCT) diagnostics to be use on site in the field. LFIA tests can be operated as either competitive or sandwich assays. Sandwich LFIAs are similar to sandwich ELISA. The sample first encounters coloured particles which are labelled with antibodies raised to the target antigen. The test line will also contain antibodies to the same target, although it may bind to a different epitope on the antigen. The test line will show as a coloured band in positive samples. In some embodiments of any of the aspects, the lateral flow immunoassay can be a double antibody sandwich assay, a competitive assay, a quantitative assay, or variations thereof. Competitive LFIAs are similar to competitive ELISA. The sample first encounters coloured particles which are labelled with the target antigen or an analogue. The test line contains antibodies to the target / its analogue. Unlabelled antigen in the sample will block the binding sites on the antibodies preventing uptake of the coloured particles. The test line will show as a coloured band in negative samples. There are several variations on lateral flow technology. It is also possible to apply multiple capture zones to create a multiplex test. The use of "dip sticks" or LFIA test strips and other solid supports have been described in the art in the context of an immunoassay for a number of antigen biomarkers. U.S. Pat. Nos. 4,943,522; 6,485,982; 6,187,598; 5,770,460; 5,622,871; 6,565,808, U. S. patent applications Ser. No. 10 / 278,676; U.S. Ser. No.09 / 579,673 and U.S. Ser. No. 10 / 717,082, which are incorporated herein by reference in their entirety, are non- limiting examples of such lateral flow test devices. Examples of patents that describe the use of “dip stick” technology to detect soluble antigens via immunochemical assays include but are not limited to US Patent Nos. 4,444,880; 4,305,924; and 4,135,884; which are incorporated by reference herein in their entireties. The apparatuses and methods of these three patents broadly describe a first component fixed to a solid surface on a “dip stick” which is exposed to a solution containing a soluble antigen that binds to the component fixed upon the “dip stick,” prior to detection of the component- antigen complex upon the stick. It is within the skill of one in the art to modify the teachings of this “dip stick” technology for the detection of polypeptides using antibody reagents as described herein. Other techniques can be used to level of a polypeptide in a sample. One such technique is the dot blot, an adaptation of Western blotting (Towbin et al., Proc. Nat. Acad. Sci.76:4350 (1979)). In a Western blot, the polypeptide or fragment thereof can be dissociated with detergents and heat and separated on an SDS-PAGE gel before being transferred to a solid support, such as a nitrocellulose or PVDF membrane. The membrane is incubated with an antibody reagent specific for the target polypeptide or a fragment thereof. The membrane is then washed to remove unbound proteins and proteins with non-specific binding. Detectably labelled enzyme-linked secondary or detection antibodies can then be used to detect and assess the amount of polypeptide in the sample tested. A dot blot immobilizes a protein sample on a defined region of a support, which is then probed with antibody and labelled secondary antibody as in Western blotting. The intensity of the signal from the detectable label in either format corresponds to the amount of enzyme present, and therefore the amount of polypeptide. Levels can be quantified, for example by densitometry. In some embodiments of any of the aspects, the level of a target can be measured, by way of non-limiting example, by Western blot; immunoprecipitation; enzyme-linked immunosorbent assay (ELISA); radioimmunological assay (RIA); sandwich assay; fluorescence in situ hybridization (FISH); immunohistological staining; radioimmunometric assay; immunofluorescence; chromogenic assay; mass spectroscopy and / or immunoelectrophoresis assay. In certain embodiments, the gene expression products as described herein can be instead determined by determining the level of messenger RNA (mRNA) expression of the genes described herein. Such molecules can be isolated, derived, or amplified from a biological sample, such as a blood sample. Techniques for the detection of mRNA expression is known by persons skilled in the art, and can include but not limited to, PCR procedures, RT-PCR, quantitative RT-PCR Northern blot analysis, differential gene expression, RNAse protection assay, microarray-based analysis, next-generation sequencing, hybridization methods, etc. In general, the PCR procedure describes a method of gene amplification which is comprised of (i) sequence-specific hybridization of primers to specific genes or sequences within a nucleic acid sample or library, (ii) subsequent amplification involving multiple rounds of annealing, elongation, and denaturation using a thermostable DNA polymerase, and (iii) screening the PCR products for a band of the correct size. The primers used are of sufficient length and appropriate sequence to provide initiation of polymerization, i.e., each primer is specifically designed to be complementary to a strand of the genomic locus to be amplified. In an alternative embodiment, mRNA level of gene expression products described herein can be determined by reverse-transcription (RT) PCR and by quantitative RT-PCR (QRT- PCR) or real-time PCR methods. Methods of RT-PCR and QRT-PCR are well known in the art. In some embodiments of any of the aspects, the level of an mRNA can be measured by a quantitative sequencing technology, e.g., a quantitative next-generation sequence technology. Methods of sequencing a nucleic acid sequence are well known in the art. Briefly, a sample obtained from a subject can be contacted with one or more primers which specifically hybridize to a single-strand nucleic acid sequence flanking the target gene sequence and a complementary strand is synthesized. In some next-generation technologies, an adaptor (double or single-stranded) is ligated to nucleic acid molecules in the sample and synthesis proceeds from the adaptor or adaptor compatible primers. In some third-generation technologies, the sequence can be determined, e.g., by determining the location and pattern of the hybridization of probes or measuring one or more characteristics of a single molecule as it passes through a sensor (e.g., the modulation of an electrical field as a nucleic acid molecule passes through a nanopore). Exemplary methods of sequencing include, but are not limited to, Sanger sequencing, dideoxy chain termination, high-throughput sequencing, next generation sequencing, 454 sequencing, SOLiD sequencing, polony sequencing, Illumina sequencing, Ion Torrent sequencing, sequencing by hybridization, nanopore sequencing, Helioscope sequencing, single molecule real time sequencing, RNAP sequencing, and the like. Methods and protocols for performing these sequencing methods are known in the art, see, e.g., “Next Generation Genome Sequencing” Ed. Michal Janitz, Wiley-VCH; “High-Throughput Next Generation Sequencing” Eds. Kwon and Ricke, Humanna Press, 2011; and Sambrook et al., Molecular Cloning: A Laboratory Manual (4 ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012); which are incorporated by reference herein in their entireties. The nucleic acid sequences of the genes described herein have been assigned NCBI accession numbers for different species such as human, mouse and rat. For example, the human ATP5B, GPX4, SLC7A11, IREB2, CTH, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and Bap-1 nucleic acid sequences (e.g., SEQ ID NO: 1, 3, 8, 10, 16, 20, 25, 28, 61, 63, 41, 46, and 58, respectively) and mRNA is known. Accordingly, a skilled artisan can design an appropriate primer based on the known sequence for determining the nucleic acid or mRNA level of the respective gene. Nucleic acid and ribonucleic acid (RNA) molecules can be isolated from a particular biological sample using any of a number of procedures, which are well-known in the art, the particular isolation procedure chosen being appropriate for the particular biological sample. For example, freeze-thaw and alkaline lysis procedures can be useful for obtaining nucleic acid molecules from solid materials; heat and alkaline lysis procedures can be useful for obtaining nucleic acid molecules from urine; and proteinase K extraction can be used to obtain nucleic acid from blood (Roiff, A et al. PCR: Clinical Diagnostics and Research, Springer (1994)). In some embodiments of any of the aspects, one or more of the reagents (e.g., an antibody reagent and / or nucleic acid probe) described herein can comprise a detectable label and / or comprise the ability to generate a detectable signal (e.g., by catalysing reaction converting a compound to a detectable product). Detectable labels can comprise, for example, a light-absorbing dye, a fluorescent dye, or a radioactive label. Detectable labels, methods of detecting them, and methods of incorporating them into reagents (e.g., antibodies and nucleic acid probes) are well known in the art. In some embodiments of any of the aspects, detectable labels can include labels that can be detected by spectroscopic, photochemical, biochemical, immunochemical, electromagnetic, radiochemical, or chemical means, such as fluorescence, chemifluoresence, or chemiluminescence, or any other appropriate means. The detectable labels used in the methods described herein can be primary labels (where the label comprises a moiety that is directly detectable or that produces a directly detectable moiety) or secondary labels (where the detectable label binds to another moiety to produce a detectable signal, e.g., as is common in immunological labelling using secondary and tertiary antibodies). The detectable label can be linked by covalent or non-covalent means to the reagent. Alternatively, a detectable label can be linked such as by directly labelling a molecule that achieves binding to the reagent via a ligand-receptor binding pair arrangement or other such specific recognition molecules. Detectable labels can include, but are not limited to radioisotopes, bioluminescent compounds, antibodies, chemiluminescent compounds, fluorescent compounds, metal chelates, and enzymes. In other embodiments, the detection reagent is label with a fluorescent compound. When the fluorescently labelled reagent is exposed to light of the proper wavelength, its presence can then be detected due to fluorescence. In some embodiments of any of the aspects, a detectable label can be a fluorescent dye molecule, or fluorophore including, but not limited to fluorescein, phycoerythrin, phycocyanin, o-phthaldehyde, fluorescamine, Cy3TM, Cy5TM, allophycocyanine, Texas Red, peridinin chlorophyll, cyanine, tandem conjugates such as phycoerythrin-Cy5TM, green fluorescent protein, rhodamine, fluorescein isothiocyanate (FITC) and Oregon GreenTM, rhodamine and derivatives (e.g., Texas red and tetrarhodimine isothiocynate (TRITC)), biotin, phycoerythrin, AMCA, CyDyesTM, 6-carboxyfhiorescein (commonly known by the abbreviations FAM and F), 6-carboxy-2’,4’,7’,4,7-hexachlorofiuorescein (HEX), 6- carboxy-4’,5’-dichloro-2’,7’-dimethoxyfiuorescein (JOE or J), N,N,N’,N’-tetramethyl- 6carboxyrhodamine (TAMRA or T), 6-carboxy-X-rhodamine (ROX or R), 5- carboxyrhodamine-6G (R6G5 or G5), 6-carboxyrhodamine-6G (R6G6 or G6), and rhodamine 110; cyanine dyes, e.g., Cy3, Cy5 and Cy7 dyes; coumarins, e.g., umbelliferone; benzamide dyes, e.g., Hoechst 33258; phenanthridine dyes, e.g., Texas Red; ethidium dyes; acridine dyes; carbazole dyes; phenoxazine dyes; porphyrin dyes; polymethine dyes, e.g., cyanine dyes such as Cy3, Cy5, etc.; BODIPY dyes and quinoline dyes. In some embodiments of any of the aspects, a detectable label can be a radiolabel including, but not limited to3H,125I,35S,14C,32P, and33P. In some embodiments of any of the aspects, a detectable label can be an enzyme including, but not limited to horseradish peroxidase and alkaline phosphatase. An enzymatic label can produce, for example, a chemiluminescent signal, a colour signal, or a fluorescent signal. Enzymes contemplated for use to detectably label an antibody reagent include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, alpha-glycerophosphate dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-VI-phosphate dehydrogenase, glucoamylase and acetylcholinesterase. In some embodiments of any of the aspects, a detectable label is a chemiluminescent label, including, but not limited to lucigenin, luminol, luciferin, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester. In some embodiments of any of the aspects, a detectable label can be a spectral colorimetric label including, but not limited to colloidal gold or glass or plastic (e.g., polystyrene, polypropylene, and latex) beads. In some embodiments of any of the aspects, detection reagents can also be labelled with a detectable tag, such as c-Myc, HA, VSV-G, HSV, FLAG, V5, HIS, or biotin. Other detection systems can also be used, for example, a biotin-streptavidin system. In this system, the antibodies immunoreactive (i.e., specific for) with the biomarker of interest is biotinylated. Quantity of biotinylated antibody bound to the biomarker is determined using a streptavidin-peroxidase conjugate and a chromogenic substrate. Such streptavidin peroxidase detection kits are commercially available, e.g., from DAKO; Carpinteria, CA. A reagent can also be detectably labelled using fluorescence emitting metals such as152Eu, or others of the lanthanide series. These metals can be attached to the reagent using such metal chelating groups as diethylenetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA). An expression value or risk score which is less than a reference expression value or reference risk score can be an expression value which is less by at least about 10%, at least about 20%, at least about 50%, at least about 60%, at least about 80%, at least about 90%, or less relative to the reference expression value or reference risk score. In some embodiments of any of the aspects, an expression value or risk score which is less than a reference expression value or risk score can be an expression value or risk score which is statistically significantly less than the reference expression value or risk score. An expression value or risk score which is more than a reference expression value or a reference risk score can be an expression value or risk score which is greater by at least about 10%, at least about 20%, at least about 50%, at least about 60%, at least about 80%, at least about 90%, at least about 100%, at least about 200%, at least about 300%, at least about 500% or more or less than the reference expression value or reference risk score. In some embodiments of any of the aspects, an expression value or a risk score which is more or less than a reference expression value or a reference risk score can be an expression value or a risk score which is statistically significantly greater than the reference expression value or reference risk score. In some embodiments of any of the aspects, the reference can be an expression value of the target molecule in a population of subjects who do not have or are not diagnosed as having, and / or do not exhibit signs or of cancer. In some embodiments of any of the aspects, the reference can also be an expression value of the target molecule in a control sample, a pooled sample of control individuals or a numeric value or range of values based on the same. In some embodiments of any of the aspects, the reference can be the expression value of a target molecule in a sample obtained from the same subject at an earlier point in time, e.g., the methods described herein can be used to determine if a subject’s sensitivity or response to a given therapy is changing over time. In some aspects, the expression value or risk score can be used when using a combination of gene expression levels to determine the outcome or direction to take for the subject with cancer or having been diagnosed with cancer, or for the subject suspected of having cancer, or who is undergoing treatment for cancer. The “direction of effect” where genes are increased or decreased in a cancer patient are based on single gene models of the adverse pathology (AP) outcome. However, the effect of the direction of gene expression levels can change when there are multiple genes (2 or more) in a signature (also known as modified expression). A more general description that is accurate for any gene combination and implicitly incorporates the directions of effects would be using a risk score (related to modified expression). To calculate a risk score, a weighted linear combination of normalised expression values of multiple genes is used to calculate the risk score for any gene combination. The risk score estimates the probability of, for example, aggressive cancer; larger values of the risk score indicate a higher probability of aggressive cancer and vice versa. The same is true for determining the efficacy of treatment, or determining whether a treatment is suitable, or whether the outcome for the cancer patient is poor or good prognostically. For example, the term “combination of genes” or “risk score” should be understood to mean a linear combination, or weighted sum, of the normalised expression of said genes. Larger values of a risk score indicate higher risk of aggressive or recurrent cancer, a poor outcome, a lower 5-year or 10-year survival rate, treatment not working, and the like. Smaller values of a risk score indicate lower risk of aggressive or recurrent cancer, better outcome, higher rate of 5-year and / or 10-year survival rate, a treatment that is working, and the like. The risk score for a combination of N genes is calculated as follows: Risk score = W1 x G1 + W2 x G2 + … + WN x GN (Equation 1) where, W = A gene-specific real number constant that can be positive or negative; and G = A gene-specific normalised expression value In some embodiments of any of the the expression value of expression products of no more than 200 other genes is determined. In some embodiments of any of the aspects, the expression value of expression products of no more than 100 other genes is determined. In some embodiments of any of the aspects, the expression value of expression products of no more than 20 other genes is determined. In some embodiments of any of the aspects, the expression value of expression products of no more than 15 other genes is determined. In some embodiments of any of the aspects, the expression value of expression products of no more than 10 other genes is determined. In some embodiments of the foregoing aspects, the expression value of a given gene can be normalized relative to the expression value of one or more reference genes or reference proteins. In some embodiments, the reference value can be the expression value in a sample of similar cell type, sample type, sample processing, and / or obtained from a subject of similar age, sex, and other demographic parameters as the sample / subject for which the expression value of one or more of ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH (and Bap-1) is to be determined. In some embodiments, the test sample and control reference sample are of the same type, that is, obtained from the same biological source, and comprising the same composition, e.g., the same number and type of cells. The test sample can be obtained by removing a sample from a subject but can also be accomplished by using a previously isolated sample (e.g., isolated at a prior timepoint and isolated by the same or another person). In some embodiments of any of the aspects, the test sample can be an untreated test sample. As used herein, the phrase “untreated test sample” refers to a test sample that has not had any prior sample pre-treatment except for dilution and / or suspension in a solution. Exemplary methods for treating a test sample include, but are not limited to, centrifugation, filtration, sonication, homogenization, heating, freezing, and thawing, and combinations thereof. In some embodiments of any of the aspects, the test sample can be a frozen test sample, e.g., a frozen tissue. The frozen sample can be thawed before employing methods, assays and systems described herein. After thawing, a frozen sample can be centrifuged before being subjected to methods, assays and systems described herein. In some of any of the aspects, the test sample is a clarified test sample, for example, by centrifugation and collection of a supernatant comprising the clarified test sample. In some embodiments of any of the aspects, a test sample can be a pre-processed test sample, for example, supernatant or filtrate resulting from a treatment selected from the group consisting of centrifugation, filtration, thawing, purification, and any combinations thereof. In some embodiments of any of the aspects, the test sample can be treated with a chemical and / or biological reagent. Chemical and / or biological reagents can be employed to protect and / or maintain the stability of the sample, including biomolecules (e.g., nucleic acid and protein) therein, during processing. One exemplary reagent is a protease inhibitor, which is generally used to protect or maintain the stability of protein during processing. The skilled artisan is aware of methods and processes appropriate for pre-processing of biological samples required for determination of the level of an expression product as described herein. In some embodiments of any of the aspects, the methods, assays, and systems described herein can further comprise a step of obtaining or having obtained a test sample from a subject. In some embodiments of any of the aspects, the subject can be a human subject. In some embodiments of any of the aspects, the subject can be a subject in need of treatment for (e.g., having or diagnosed as having prostate cancer) or a subject at risk of or at increased risk of developing prostate cancer as described elsewhere herein. In some embodiments of any of the aspects, the sample obtained from a subject can be a biopsy sample. In some embodiments of any of the aspects, the sample obtained from a subject can be a blood or serum sample. Definitions In the specification, the term “healthy” should be understood to mean where the individual or patient has no underlying medical condition, infection, inflammatory response, condition or otherwise occurring. In the specification, the term “cancer” should be understood to mean a cancer selected from the group comprising node-negative, ER-positive breast cancer; early stage, node positive breast cancer; multiple myeloma, prostate cancer, glioblastoma, lymphoma, fibrosarcoma; myxosarcoma; liposarcoma; chondrosarcoma; osteogenic sarcoma; chordoma; angiosarcoma; lymphangiosarcoma; lymphangioendotheliosarcoma; synovioma; mesothelioma; Ewing's tumour; leiomyosarcoma; rhabdomyosarcoma; colon carcinoma; pancreatic cancer; breast cancer; ovarian cancer; fallopian tube cancer; squamous cell carcinoma; basal cell carcinoma; adenocarcinoma; sweat gland carcinoma; sebaceous gland carcinoma; papillary carcinoma; papillary adenocarcinomas; cystadenocarcinoma; medullary carcinoma; bronchogenic carcinoma; renal cell carcinoma; hepatoma; bile duct carcinoma; choriocarcinoma; seminoma; embryonal carcinoma; Wilms' tumour; cervical cancer; uterine cancer; testicular tumour; lung carcinoma; small cell lung carcinoma; non-small cell lung carcinoma; bladder carcinoma; epithelial carcinoma; glioma; astrocytoma; medulloblastoma; craniopharyngioma; ependymoma; pinealoma; hemangioblastoma; acoustic neuroma; oligodendroglioma; meningioma; melanoma; retinoblastoma; primary peritoneal cancer; and leukemias. Also included are metastases selected from the group comprising: bone metastases; lung metastases; liver metastases; bone marrow metastases; breast metastases; and brain metastases. In the specification, the term “individual” or “patient” should be understood to mean all mammals, for example, a human, primates, non-human primates, farm animals (such as pigs, horses, goats, sheep, cows (including bulls, bullocks, heifers etc.), donkey, reindeer, etc.), veterinary mammals (such as dogs, cats, rabbits, hamsters, guinea pigs, mice, rats, ferrets, etc.), and mammals kept in captivity (such as lions, tigers, elephants, zebras, giraffes, pandas, rhino, hippopotamus, etc.), and other mammals and higher mammals for which the use of the invention is practicable. The term “sample”, “biological sample”, or “test sample” as used herein denotes a sample taken or isolated from a biological organism, e.g., a blood or plasma sample from a subject. In some embodiments of any of the aspects, the present invention encompasses several examples of a biological sample. In some embodiments of any of the aspects, the biological sample is cells, or tissue, or peripheral blood, or bodily fluid. Exemplary biological samples include, but are not limited to, a biopsy, a biological sample, biofluid sample; blood; blood derivatives; serum; plasma; urine; sperm; mucus; tissue biopsy; organ biopsy; synovial fluid; bile fluid; cerebrospinal fluid; mucosal secretion; effusion; sweat; saliva; and / or tissue sample etc. The term also includes a mixture of the above-mentioned samples. The term “test sample” also includes untreated or pretreated (or pre-processed) biological samples. In some embodiments of any of the aspects, a test sample can comprise cells from a subject. In some embodiments of any of the aspects, the sample can be a urine sample or a tissue sample from a subject where the urine sample or the tissue sample may contain tumour cells. In addition to the definition summarised above, the sample can also include non-tumour tissue, a pre-cancerous tissue, conditioned media, or formalin-fixed paraffin-embedded (FFPE) tumour or non-tumour tissue. In the specification, the term “positive expression” as applied to a gene or a protein encoded by that gene should be understood to mean a level of expression of the gene or protein encoded by that gene (an expression value) that is increased above an average level of expression of the same gene or protein encoded by that same gene found in a cohort of matched control individuals with cancer (the “control group”). The cohort of matched individuals may consist of individuals who did not experience a recurrence of a cancer following surgery to remove the cancer. In relation to controls, the usual practise for one skilled in the art would be to use a ‘standard’ control, for example, for Immunohistochemistry (IHC), a cell line or cell lines where the expression level of the biomarker is known, or for qPCR (quantitative Polymerase Chain Reaction), a similar standard control or a pool of a number of samples is known. In the specification, the term “dysregulated expression” or “modified expression” should be understood to mean a normalised expression value that is increased over or decreased below a normalised expression value of the same gene or combination of genes found in a cohort of matched individuals with cancer that did not recur following surgery to remove the cancer or following non-surgical treatment, or found in a cohort of matched individuals who are cancer free or who have never been diagnosed with cancer. Put another way, the terms should be understood to mean that a risk score calculated from the expression of a gene or combination of genes is larger or smaller than the risk score calculated from the expression of the same gene or combination of genes in individuals with cancer that did not recur following surgery to remove the cancer or following non-surgical treatment, or found in a cohort of matched individuals who are cancer free or who have never been diagnosed with cancer. The terms “normal expression” as applied to a gene or protein should be understood to mean a level of expression of the gene (or protein encoded by that gene) that is equivalent to a level of expression of the same gene or protein encoded by that same gene found in a cohort of matched control individuals with cancer, or matched controls without cancer. The cohort of matched may consist of individuals who did not experience a recurrence of a cancer following surgery to remove the cancer. The method used to set thresholds is different for the microarray analysis, qRT-PCR analysis, and protein expression. For microarrays, the threshold is relative (samples were split into three equal groups, so the threshold is dataset dependent), and for the qPCR and protein expression it is set at specific points. For RNA (microarrays), expression levels of ‘low’, ‘moderate’ and ‘high’ refer to expression values that fall within the lower, middle or upper third of the expression range; or alternatively, ‘low’ and ‘high’ expression can refer to expression values that fall within the lower or upper half of the expression range. For qRT-PCR and protein expression levels, specific thresholds have been set, but in general, the term “dysregulated” refers to tumours with expression values falling above or below set values in the range of expression. For the terms “moderate” and “normal”, the terms refer to tumours with expression values falling within set values in the range of expression. For example, the gene expression values (or proteins) of the at least one of the listed genes are first normalised against the gene expression values (or proteins) of the reference genes in the same biological sample, and the Ct value of each of the listed genes is obtained. The mean expression level of all reference genes is calculated, and the mean RefCt value is obtained. The normalised expression level (ΔCTgene1) of each of the listed genes is calculated by subtracting the expression level of the gene of interest (Ctgene1) from the mean RefCt value (mean RefCt - Ctgene1= ΔCTgene1). The ΔCT for at least one of the listed genes is then obtained and the combined normalised expression value calculated. Typically, the normalised qRT-PCR thresholds for ‘modified’ expression are 0.7 and 1.99. Typically, the normalised protein thresholds (using IHC) are 1% and 50% of positive cells. That is, a modified score here refers to a tumour with >1% and <50% tumour cells positive for the normalised gene expression values of the at least one of the listed genes (or proteins). These values may be adjusted based on any new data, but the same theory applies for the terms “normal”, “moderate” and “dysregulated” with respect to expression levels of the at least one of the listed genes (or proteins). In the specification, the term “adjuvant therapy” should be understood to mean any treatment given after primary treatment to increase the chances of long-term survival. In the specification, the term “neoadjuvant therapy” should be understood to mean treatment given before primary treatment to increase the chances of long-term survival. Primary treatment is generally surgery. therapy and neoadjuvant therapy are generally selected from chemotherapy, hormonal therapy, targeted therapy, radiation therapy, immunotherapy or a combination thereof. In the specification, the term “active surveillance” should be understood to mean monitoring the uveal melanoma closely. Usually this includes a doctor visit with imaging or liver functions tests. Active surveillance may be best suited if the subject has a low cancer score (usually 2 or lower), which indicates a less aggressive, slower growing form of cancer. Detection of expression generally involves measuring the RNA expression of a selected panel of prognostic genes in formalin-fixed paraffin-embedded (FFPE) tissue specimens from uveal melanoma biopsies using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assays. Other means or measuring include immunohistological staining of a tumour biopsy tissue or a control biopsy tissue using suitable means such as immunohistochemical staining. Many other means of detecting the biomarkers of the invention will be apparent to those skilled in the art. For example, RNA sequencing, quantitative polymerase chain reaction (qPCR), reverse transcriptase PCR (RT-PCR), quantitative real time RT-PCR (qRT-PCR), ELISA, Western Blot, protein determination on polyacrylamide gels, and the like. In the specification, the term “recurrence” should be understood to mean the recurrence of the uveal melanoma cancer, which is being sampled in the patient, in which the uveal melanoma cancer has returned to the sampled area after treatment. The term should also be understood to mean recurrence of a primary cancer whose site is different to that of the cancer initially sampled, that is, the cancer has returned to a non-sampled area after treatment, such as non-locoregional recurrences. In this specification, the term “poor outcome” should be understood to mean that the chances of disease-free survival are low. In the specification, the term “indolent” should be understood to mean a cancer that is typically slow-growing (lazy) and which would not become symptomatic in an individual's lifetime and would not contribute to death. In the specification, the term “survival should be understood to mean the period of time during which a patient diagnosed with uveal melanoma, will likely survive. The survival rate is expressed as a 5-year survival rate, a 10-year survival rate, a 15-year survival rate, a 20-year survival rate, a 25-year survival rate, a 30-year survival rate, a 35-year survival rate, a 40-year survival rate, a 45-year survival rate, or a 50-year survival rate. Ideally, the survival rate is expressed as a 5-year survival rate or a 10- year survival rate. In this specification, the term “treatment” should generally encompass its accepted meaning which includes prohibiting, preventing, restraining, and slowing, stopping or reversing progression or severity of a metastatic, recurrent or existing cancer phenotype. In this specification, the term “at least one” should be understood to mean and encompass that at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, or all genes can be selected from the group consisting of ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH. Preferably, the at least one gene is one, two or all of ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH. Bap-1 expression levels and gene status is also determined in addition to the listed genes. In this specification, the term “at least two” should be understood to mean and encompass that at least two, at least three, at least four, or all genes can be selected from the group consisting of ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH. Preferably, the at least two genes are ATP5B and GPX4, ATP5B and SLC7A11, ATPB5 and IREB2, ATP5B and CTH, GPX4 and SLC7A11, GPX4 and IREB2, GPX4 and CTH, SLC7A11 and IREB2, SLC7A11 and CTH, IREB2 and CTH, or a combination of those pairs. Preferably the at least three genes are ATP5B, GPX4, and SLC7A11; ATP5B, GPX4, and IREB2; ATP5B, GPX4, and CTH; ATP5B, SLC7A11, and IREB2; ATP5B, SLC7A11, and CTH; ATP5B, IREB2, and CTH; GPX4, SLC7A11, and IREB2; GPX4, SLC7A11, and CTH; SLC7A11, IREB2, and CTH, or a combination of those triplets. Preferably, the at least four genes selected are ATP5B, GPX4, SLC7A11, and IREB2; ATP5B, GPX4, SLC7A11, CTH; GPX4, SLC7A11, IREB2, and CTH; ATP5B, SLC7A11, GPX4, and CysLT1. the at least 5 genes, or proteins encoded by said genes, are GPX4, SLC3A2, GCLM, NQO1, and AIFM2; CYSLTR1, CYSLTR2, ATP5B, ACSL3 and SLC3A2; or ATP5B, GPX4, SLC7A11, IREB2, and CTH. Bap-1 expression levels and gene status is also determined in addition to the above combinations or replacing one of the markers in the combinations. In the specification, the term “variant” should be understood to mean nucleic acids encoding a protein having amino acid sequences that are substantially identical to the construct protein encoded by the nucleic acid defined in the sequences disclosed herein. Thus, for example, the term should be taken to include proteins or polypeptides that are altered in respect of one or more amino acid residues. Preferably such alterations involve the insertion, addition, deletion and / or substitution of 5 or fewer amino acids, more preferably of 4 or fewer, even more preferably of 3 or fewer, most preferably of 1 or 2 amino acids only. Insertion, addition, and substitution with natural and modified amino acids is envisaged. The variant may have conservative amino acid changes, wherein the amino acid being introduced is similar structurally, chemically, or functionally to that being substituted. Typically, the construct protein which have been altered by substitution or deletion of catalytically-important residues will be excluded from the term “variant”. Generally, the variant will have at least 70% amino acid sequence homology, preferably at least 80% sequence homology, more preferably at least 90% sequence homology, and ideally at least 95%, 96%, 97%, 98% or 99% sequence homology with the construct protein encoded by, for example, SEQ ID NO: 1. In this context, sequence homology comprises both sequence identity and similarity, i.e. a polypeptide sequence that shares 70% amino acid homology with the construct protein encoded by, for example, SEQ ID NO: 2 is one in which any 70% of aligned residues are either identical to, or conservative substitutions of, the corresponding residues in the construct protein encoded by, for example, SEQ ID NO: 2. The function of the peptide or protein translated from the variant nucleic acid sequence does not lose any function (biological activity) when compared to the original or wild type sequence. The terms “polypeptide,” “peptide” and “protein” refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acids. As used herein, “variant may comprise conservatively substituted sequences, meaning that one or more amino acid residues is replaced by different residues, and that the conservatively substituted polypeptide retains a desired biological activity, that is essentially equivalent to that of the native polypeptide. Examples of conservative substitutions include substitution of amino acids that do not alter the secondary and / or tertiary structure of the polypeptide. Other examples involve substitution of amino acids that have not been evolutionarily conserved. One or more polypeptide sequences from non-human species can be aligned with, for example, human using methods well known to one of ordinary skill in the art to determine which residues are conserved and which tolerate more variability. Advantageously, in some embodiments, these conserved amino acids are not altered when generating conservatively substituted sequences. Any given amino acid may be replaced by a residue having similar physiochemical characteristics, e.g., substituting one aliphatic residue for another (such as Ile, Val, Leu, or Ala for one another), or substitution of one polar residue for another (such as between Lys and Arg; Glu and Asp; or Gln and Asn). Other such conservative substitutions, e.g., substitutions of entire regions having similar hydrophobicity characteristics, are well known. Amino acids may be grouped according to similarities in the properties of their side chains (in A. L. Lehninger, in Biochemistry, second ed., pp.73-75, Worth Publishers, New York (1975)): (1) non-polar: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) uncharged polar: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic: Asp (D), Glu (E); (4) basic: Lys (K), Arg (R), His (H). Alternatively, naturally occurring residues may be divided into groups based on common side-chain properties: (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr, Phe. Non-conservative substitutions will entail exchanging a member of one of these classes for another class. Conservative substitutions may include, for example: Ala into Gly or into Ser; Arg into Lys; Asn into Gln or into His; Asp into Glu; Cys into Ser; Gln into Asn; Glu into Asp; Gly into Ala or into Pro; His into Asn or into Gln; Ile into Leu or into Val; Leu into Ile or into Val; Lys into Arg, into Gln or into Glu; Met into Leu, into Tyr or into Ile; Phe into Met, into Leu or into Tyr; Ser into Thr; Thr into Ser; Trp into Tyr; Tyr into Trp; and / or Phe into Val, into Ile or into Leu. Any cysteine residue not involved in maintaining the proper conformation of the polypeptide also be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, cysteine bond(s) can be added to the polypeptide to improve its stability or facilitate oligomerization. As used herein, the term “nucleic acid” or “nucleic acid sequence” refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analog thereof. The nucleic acid can be either single- stranded or double-stranded. A single-stranded nucleic acid can be one strand nucleic acid of a denatured double- stranded DNA. Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the template nucleic acid is DNA. In another aspect, the template is RNA. Suitable nucleic acid molecules are DNA, including genomic DNA, ribosomal DNA and cDNA. Other suitable nucleic acid molecules are RNA, including mRNA, rRNA and tRNA. The nucleic acid molecule can be naturally occurring, as in genomic DNA, or it may be synthetic, i.e., prepared based up human action, or may be a combination of the two. The nucleic acid molecule can also have certain modification such as 2'-deoxy, 2'-deoxy-2'-fluoro, 2'-O- methyl, 2'-O-methoxyethyl (2'-O-MOE), 2'-O-aminopropyl (2'-O-AP), 2'-O- dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), 2'-O- dimethylaminoethyloxyethyl (2'-O-DMAEOE), or 2'-O--N-methylacetamido (2'-O-NMA), cholesterol addition, and phosphorothioate backbone as described in US Patent Application 20070213292; and certain ribonucleoside that are is linked between the 2’- oxygen and the 4’-carbon atoms with a methylene unit as described in US Pat No. 6,268,490, wherein both patent and patent application are incorporated hereby reference in their entirety. As used here, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. As used here, the term "pharmaceutically-acceptable carrier" means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject agent from one or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminium hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and / or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23) other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, colouring agents, release agents, coating agents, sweetening agents, flavouring agents, perfuming agents, preservative, and antioxidants can also be present in the formulation. The amount of agent which can be combined with a carrier material to produce a single dosage form will generally be that amount of the agent which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.1% to 99% of agent, preferably from about 5% to about 70%, most preferably from 10% to about 30%. Formulations suitable for parenteral administration conveniently include sterile aqueous preparation of the active agent which is preferably isotonic with the blood of the recipient. Thus, such formulations may conveniently contain distilled water, 5% dextrose in distilled water or saline. Useful formulations also include concentrated solutions or solids containing the agent which upon dilution with an appropriate solvent give a solution suitable for parental administration above. For enteral administration, an agent can be incorporated into an inert carrier in discrete units such as capsules, cachets, tablets, or lozenges, each containing a predetermined amount of the active agent; as a granules; or a suspension or solution in an aqueous liquid or non-aqueous liquid, e.g., a syrup, an elixir, an emulsion or a draught. Suitable carriers may be starches or sugars and include lubricants, flavourings, binders, and other materials of the same nature. A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active agent in a free-flowing form, e.g., a powder or granules, optionally mixed with accessory ingredients, e.g., binders, lubricants, inert diluents, surface active or dispersing agents. Moulded tablets may be made by moulding in a suitable machine, a mixture of the powdered active agent with any suitable carrier. A syrup or suspension may be made by adding the active agent to a concentrated, aqueous solution of a sugar, e.g., sucrose, to which may also be added any accessory ingredients. Such accessory ingredients may include flavouring, an agent to retard crystallization of the sugar or an agent to increase the solubility of any other ingredient, e.g., as a polyhydric alcohol, for example, glycerol or sorbitol. Formulations for rectal administration may be presented as a suppository with a conventional carrier, e.g., cocoa butter or Witepsol S55 (trademark of Dynamite Nobel Chemical, Germany), for a suppository base. Formulations for oral administration may be presented with an enhancer. Orally- acceptable absorption enhancers include surfactants such as sodium lauryl sulfate, palmitoyl carnitine, Laureth-9, phosphatidylcholine, cyclodextrin and derivatives thereof; bile salts such as sodium deoxycholate, sodium taurocholate, sodium glycocholate, and sodium fusidate; chelating agents including EDTA, citric acid and salicylates; and fatty acids (e.g., oleic acid, lauric acid, acylcarnitines, mono- and diglycerides). Other oral absorption enhancers include benzalkonium chloride, benzethonium chloride, CHAPS (3-(3-cholamidopropyl)-dimethylammonio-1-propanesulfonate), Big-CHAPS (N, N- bis(3-D-gluconamidopropyl)-cholamide), chlorobutanol, octoxynol-9, benzyl alcohol, phenols, cresols, and alkyl alcohols. In one embodiment, the oral absorption enhancer may be sodium lauryl sulfate. As used herein, the term “administer(s)” or “administering” refers to the placement of a composition into a subject by a method or route which results in at least partial localization of the composition at a site such that desired effect is produced. A agent or composition described herein can be administered by any appropriate route known in the art including, but not limited to, oral or parenteral routes, including intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, nasal, rectal, and topical (including buccal and sublingual) administration. Exemplary modes of administration include, but are not limited to, injection, infusion, instillation, inhalation, or ingestion. “Injection” includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal, intracerebro spinal, and intrasternal injection and infusion. The dosage may be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment. Generally, the compositions are administered so that the agent is given at a dose from 1 µg / kg to 150 mg / kg, 1 µg / kg to 100 mg / kg, 1 µg / kg to 50 mg / kg, 1 µg / kg to 20 mg / kg, 1 µg / kg to 10 mg / kg, 1µg / kg to 1mg / kg, 100 µg / kg to 100 mg / kg, 100 µg / kg to 50 mg / kg, 100 µg / kg to 20 mg / kg, 100 µg / kg to 10 mg / kg, 100µg / kg to 1mg / kg, 1 mg / kg to 100 mg / kg, 1 mg / kg to 50 mg / kg, 1 mg / kg to 20 mg / kg, 1 mg / kg to 10 mg / kg, 10 mg / kg to 100 mg / kg, 10 mg / kg to 50 mg / kg, or 10 mg / kg to 20 mg / kg. It is to be understood that ranges given here include all intermediate ranges, for example, the range 1 mg / kg to 10 mg / kg includes 1mg / kg to 2 mg / kg, 1mg / kg to 3 mg / kg, 1mg / kg to 4 mg / kg, 1mg / kg to 5 mg / kg, 1mg / kg to 6 mg / kg, 1mg / kg to 7 mg / kg, 1mg / kg to 8 mg / kg, 1mg / kg to 9 mg / kg, 2mg / kg to 10mg / kg, 3mg / kg to 10mg / kg, 4mg / kg to 10mg / kg, 5mg / kg to 10mg / kg, 6mg / kg to 10mg / kg, 7mg / kg to 10mg / kg,8mg / kg to 10mg / kg, 9mg / kg to 10mg / kg etc. It is to be further understood that the ranges intermediate to the given above are also within the scope for use in methods and pharmaceutical compositions described herein, for example, in the range 1mg / kg to 10 mg / kg, dose ranges such as 2mg / kg to 8 mg / kg, 3mg / kg to 7 mg / kg, 4mg / kg to 6mg / kg etc. With respect to duration and frequency of treatment, it is typical for skilled clinicians to monitor subjects to determine when the treatment is providing therapeutic benefit, and to determine whether to increase or decrease dosage, increase or decrease administration frequency, discontinue treatment, resume treatment or make other alteration to treatment regimen. The schedule can vary from once a week to daily depending on several clinical factors, such as the subject's sensitivity. The desired dose can be administered at one time or divided into subdoses, e.g., 2-4 subdoses and administered over a period of time, e.g., at appropriate intervals through the day or other appropriate schedule. Such sub-doses can be administered as unit dosage forms. In some embodiments, administration is chronic, e.g., one or more doses daily over a period of weeks or months. Examples of dosing schedules are administration daily, twice daily, three times daily or four or more times daily over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months or more. The pharmaceutical compositions can be administered during infancy (between 0 to about 1 year of life), childhood (the period of life between infancy and puberty) and during puberty (between about 8 years of life to 18 years of life). The pharmaceutical compositions can also be administered to treat adults (greater than about 18 years of life). A dose administered at least once, may be provided as a bolus, a continuous administration or sustained release. Multiple administration over a period of weeks or months may be preferable. It may also be preferable to administer the dose at least once / week and even more frequent administrations (e.g., daily). Subsequent doses may be administered as indicated. As used herein, the term "treatment" or "treating" refers to an intervention (e.g., the administration of an agent to a subject) which cures, ameliorates or lessens the symptoms of a disease or removes (or lessens the impact of) its cause(s). In this case, the term is used synonymously with the term “therapy”. Additionally, the terms “treatment” or “treating” refers to an intervention (e.g., the administration of an agent to a subject) which prevents or delays the onset or progression of a disease or reduces (or eradicates) its incidence within a treated population. In this case, the term treatment is used synonymously with the term “prophylaxis”. The term “consist essentially of” should be understood to mean all fourteen, genes, or eleven genes, or ten genes, or nine genes, or eights genes, or seven genes, or six genes, or five genes, or four genes, or three genes, or two genes selected from ATP5B, GPX4, SLC7A11, IREB2, CTH, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, and CysLT2. Bap-1 can also be included with one or more of the fourteen genes selected from ATP5B, GPX4, SLC7A11, IREB2, CTH, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, and CysLT2. In the specification, the term therapy” should be understood to mean treatment given before primary treatment to increase the chances of long-term survival. Primary treatment is generally surgery. Neoadjuvant therapies are generally selected from chemotherapy, hormonal therapy, targeted therapy, radiation therapy, immunotherapy or a combination thereof. In the specification, the term “adjuvant therapy” should be understood to mean any treatment given after primary treatment to increase the chances of long-term survival. Primary treatment is generally surgery. Adjuvant therapies are generally selected from chemotherapy, hormonal therapy, targeted therapy, radiation therapy, immunotherapy, or a combination thereof. Active surveillance can also be considered an adjuvant therapy. In the specification, the term “test platform” in relation to determining the levels of the biomarkers should be understood to mean determining biomarker concentrations by immunoassay-based methods such as lateral flow immunoassays (LFAs), enzyme- linked immunosorbent assay (ELISA) and immunoturbidimetric assays. In the specification, the term “KRAS status” should be understood to mean determining whether a patient with lung cancer has KRAS-positive or KRAS-negative lung cancer. KRAS-positive lung cancer, refers to any lung cancer that tests positive for a KRAS mutation biomarker. KRAS mutation biomarkers are present in approximately 15-30% of patients with non-small cell lung cancer (NSCLC). KRAS gene mutations tend to occur in people who currently or have formerly smoked, but KRAS mutations have been found in people with diverse backgrounds. The prognosis for advanced KRAS- mutant lung cancers has been poor for decades, with a median survival of 1.2 years. In the specification, the term “smoking status” should be understood to mean whether or not an individual is a former smoker, current smoker, or was never a smoker. The smoking status of the individual can influence the type of lung cancer they have. Brief Description of the Drawings The invention will be more clearly understood from the following description of an embodiment thereof, given by way of example only, with reference to the accompanying drawings, in which:- Figure 1 illustrates the analysis of the correlation between expression of individual ferroptosis-related genes (FRGs) and (A) overall survival or (B) disease free survival in lung cancer patients in the TCGA database. Genes significantly associated with survival are highlighted in red (with a border). Figure 2 illustrates the bioinformatic analysis of the correlation between expression of individual ferroptosis-related genes (FRGs) and overall survival in ovarian cancer patients in the TCGA database. Genes significantly associated with survival are highlighted in red (with a border). Figure 3 illustrates the bioinformatic analysis of the correlation between expression of individual ferroptosis-related genes (FRGs) and (A) overall survival or (B) disease free survival in pancreatic cancer patients in the TCGA database. Genes significantly associated with survival are highlighted in red (with a border). Figure 4 illustrates the bioinformatic analysis of the correlation between expression of individual ferroptosis-related genes (FRGs) and (A) overall survival or (B) disease free survival in cutaneous melanoma patients in the TCGA database. Genes significantly associated with survival are highlighted in red (with a border). Figure 5 illustrates the analysis of proteome profiling uncovered ferroptosis as a biological process affected by 20 µM 1,4 dihydroxy quininib in OMM2.5 cells. (A) Heatmap chart depicting all the differentially expressed proteins between 0,5% DMSO or 20 µM 1,4 dihydroxy quininib (Q7) treated OMM2.5 cells after 4, 8 or 24 hours of treatment. The heat maps show n = 4 for each time point with the respective colour scale located below each figure. The blue and red refer to down-regulated and up- regulated proteins, respectively. The 4 hours heat map (left panel) represents 66 differentially expressed proteins where 41 proteins are up-regulated and 25 proteins are downregulated. The 8 hours heat map (middle panel) represents 164 differentially expressed proteins where 72 proteins are up-regulated and 92 proteins are downregulated. The 24 hours heat map (right panel) represents 95 differentially expressed proteins where 49 proteins are up-regulated and 46 proteins are downregulated. (B) Venn diagram analyses showing the unique and shared downregulated proteins between 0,5% DMSO or 20 µM Q7 treated OMM2.5 cells at 4, 8 and 24 hours (upper panel), the unique and shared upregulated proteins between 0,5% DMSO or 20 µM Q7 treated OMM2.5 cells at 4, 8 and 24 hours (middle panel), and the total amount of unique and shared differentially expressed proteins between 0,5% DMSO or 20 µM Q7 treated OMM2.5 cells at 4, 8 and 24 hours (lower panel). (C) Volcano plots depicting the differentially expressed proteins between 0.5% DMSO and 20 µM Q7 treated OMM2.5 cells at 4 hours (left panel), 8 hours (middle panel) and 24 hours (right panel). The proteins in red represent the proteins with a Student’s T-Test Difference ≥ 0.5. The proteins highlighted in blue represent those with a Student’s T-Test Difference ≤ - 0.5. The most consistently upregulated protein after 4, 8 or 24 hours of treatment is heme oxygenase 1 (HO-1). (D-H) Gene ontology GO classification of proteomic data for differentially expressed proteins between 0.5% DMSO or 20 µM Q7-treated OMM2.5 cells after 8 hours (D, E) and 24 hours (F, G) of treatment. (D) KEGG pathway analysis of significantly upregulated proteins in 20 µM Q7-treated vs 0.5% DMSO-treated OMM2.5 cells after 8 hours of treatment, showing the most enriched categories in biological process. (E) Protein-protein interaction network showing the significantly upregulated proteins in 20 µM Q7-treated vs 0.5% DMSO-treated OMM2.5 cells associated to ferroptosis, after 8 hours of treatment. (F) KEGG pathway analysis of significantly upregulated proteins in 20 µM Q7-treated vs 0.5% DMSO-treated OMM2.5 cells after 24 hours of treatment, showing the most enriched categories in biological process. (G) Protein-protein interaction network showing the significantly upregulated proteins in 20 µM Q7-treated vs 0.5% DMSO- treated OMM2.5 cells associated to ferroptosis, after 24 hours of treatment. The panel also shows upregulated (red nodes) and downregulated (blue node) proteins associated to fluid shear stress and atherosclerosis process. HO-1 upregulation is common to both pathways. (H) DAVID pathway analysis displaying significantly upregulated proteins at 8 hours (red stars) and 24 hours (red triangles) in 20 µM Q7- treated vs 0.5% DMSO-treated OMM2.5 cells in ferroptosis process. Image obtained from KEGG (102). Q7 = 1,4-dihydroxy quininib; HMOX1 = HO-1; h = hours. Figure 6 illustrates that the most significantly altered proteins identified by proteomic profiling following 1,4 dihydroxy quininib treatment in OMM2.5 cells and immunoblot validation (A-D). (A) Immunoblot analysis of HO-1 and IREB2 in protein extracts from OMM2.5 cells treated with 0.5% DMSO or 20 µM Q7 for 4, 8 or 24 h (B) Immunoblot analysis of GDF-15 in protein extracts from OMM2.5 cells treated with 0.5% DMSO or 20 µM Q7 OMM2.5 cells for 4, 8 or 24 hours. (C) Densitometric quantification of HO-1 and IREB2 normalized to beta actin, as determined by n = 4 independent western blot experiments as in (A) (*, p < 0.05; ****, p < 0.0001). (D) Densitometric quantification of GDF-15 normalized to beta actin, as determined by at least three independent western blot experiments as in (C) (*, p < 0.05). The histograms report mean ± SEM. Q7 = 1,4-dihydroxy quininib; h = hours. Figure 7 illustrates that treatment with 20 µM 1,4 dihydroxy quininib affects ROS, biliverdin, GSH, LOOH levels and NRF2, GPX4, GCLM expression in OMM2.5 cells. (A) Evaluation of the effects of 20 µM Q7 on ROS levels in OMM2.5 cells after 8 (n = 4 independent experiments) and (n = 3 independent experiments) hours of treatment (*, p < 0.05). (B) Western blot analysis of NRF2 expression in 0.5% DMSO or 20 µM Q7 treated OMM2.5 cells after 4 (n = 6 independent experiments), 8 (n = 6 independent experiments) and 24 (n = 5 independent experiments) hours of treatment (*, p < 0.05; ** p < 0.01). (C) Densitometric quantification of NRF2 vs beta-actin as determined by at least three independent western blot experiments as in (B). The results are expressed as means ± SEM. (D) Measurement of intracellular biliverdin in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4, 8 and 24 hours of treatment (** p < 0.01). N = 3 independent experiments. (E) Western blot showing GPX4 expression in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4 (n = 3 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 7 independent experiments) hours. (F) Densitometric quantification of GPX4 vs b-actin as determined by at least three independent western blot experiments as in (E) (*, p < 0.05; *** p < 0.001). (G) Measurement of non-protein thiol group (RSH) concentrations after 4 (n = 4 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 3 independent experiments) hours of treatment (**, p < 0.01; ***, p < 0.001). (H) Evaluation of LOOH levels after 4 (n = 4 independent experiments), 8 (n = 4 independent experiments) and 24 (n = 3 independent experiments) hours of treatment (**, p < 0.01). The results are expressed as means ± SEM. (I) Western blot showing GCLM expression in 0.5% DMSO- or 20 µM Q7- treated OMM2.5 cells after 4 (n = 6 independent experiments), 8 (n = 5 independent experiments), and 24 (n = 4 independent experiments) hours. (J) Densitometric quantification of GCLM normalized to beta-actin as determined by at least three independent western blot experiments as in (I) (** p < 0.01). The histograms report mean ± SEM. Q7 = 1,4-dihydroxy quininib; h = hours. Figure 8 illustrates that Quininib drugs and CysLT receptor antagonists exert overlapping and distinct effects on ferroptosis markers, while erastin reduces OMM2.5 cells metabolic activity. (A) Western blot showing GPX4 and GCLM expression in 0.5% DMSO- or 20 µM Q7-, 50 µM montelukast-, 20 µM quininib-, 50 µM HAMI 3379- treated OMM2.5 cells after 8 hours of treatment. (B) Densitometric quantification of GCLM (left) and GPX4 (right) vs b-actin as determined by three independent western blot experiments as in (A). (C) Western blot showing GPX4 and GCLM expression in 0.5% DMSO- or 20 µM Q7-, 50 µM montelukast-, 20 µM quininib-, 50 µM HAMI 3379- treated OMM2.5 cells after 24 hours of treatment. (D) Densitometric quantification of GCLM (left) and GPX4 (right) vs b-actin as determined by three independent western blot experiments as in (C) (*, p < 0.05; ** p < 0.01). (E) A significant dose-dependent decrease of OMM2.5 cell metabolic was observed following 96 hours of erastin treatment in comparison to 0.5% DMSO control. N = 3-4 independent experiments (F) IC50 value of erastin in OMM2.5 cell metabolic activity assays. Metabolic activity of cells was determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. All the data are expressed as mean ± SEM (**, p < 0.01; **** p < 0.0001). Q1 = quininib; Q7 = 1,4-dihydroxy quininib; h = hours. Figure 9 illustrates that 1,4-dihydroxy quininib significantly increases HO-1 and 4-HNE expression in MUM OPDX explants. (A) Schematic of the explant culture protocol to evaluate 1,4 dihydroxy quininib effects. Rodent xenograft models of MUM have been generated by transplanting fresh patient’s tumour samples into the liver of mouse models. MUM tumours were removed from mouse liver and then dissected into 3 fragments, arbitrarily named left (L), middle (M), right (R). Each fragment was divided into 2 pieces and grown in complete media with 20 μM 1,4-dihydroxy quininib (Q7) or vehicle (0.5% DMSO) for 72 hours. Tumour tissue was snap-frozen for protein isolation and western blot experiments. (B) Table showing the 2 MUM patients’ clinical characteristics. Tumours from these patients were used to generate OPDX mouse models, named UVM 4 and UVM 7. (C) Western blot analysis of HO-1 and GPX4 expression in 0.5% DMSO or 20 µM Q7 treated MUM tumours cultured ex vivo for 72 hours (*, p < 0.05). (D) Densitometric quantification of HO-1 and GPX4 vs beta-actin as determined by at least three independent western blot experiments as in (C) (*, p < 0.05). Data represent the mean ± SEM. N = 3 MUM OPDX models for each type of tumor, i.e., tumors from 3 UVM4 and 3 UVM7 mouse models. For each tumor fragments L and R were used to perform Western blot. Triangles represent data from UVM 4 and circles represent data from UVM 7. (E) Western blot analysis of 4-HNE expression in 0.5% DMSO or 20 µM Q7 treated MUM tumours cultured ex vivo for 72 hours (*, p < 0.05). (F) Densitometric quantification of 4-HNE vs beta-actin as determined by at least three independent western blot experiments as in (E) (*, p < 0.05). Data represent the mean ± SEM. N = 3 MUM OPDX models for each type of tumour, i.e., tumours from 3 UVM4 and 3 UVM7 mouse models. For each tumour fragments L and R were used to perform Western blot. Triangles represent data from UVM 4 and circles represent data from UVM 7. Q7 = 1,4-dihydroxy quininib. Figure 10 illustrates that high expression levels of key genes involved in ferroptosis modulation are associated with a worse prognosis in the TCGA-UVM cohort. (A) Graphs showing the statistically significant relationship between expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and disomy 3 or monosomy 3 in the TCGA-UVM cohort. (B) Graphs showing the statistically significant between expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and the normal BAP1 (BAP1_mut NO) or mutated BAP1 (BAP1_mut YES) TCGA-UVM cohort. (C) Kaplan–Meier survival curves showing the statistically significant relationship between the expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and overall survival in TCGA-UVM patients presenting chromosome 3 disomy (chr3_status: disomy) or chromosome 3 monosomy (chr3_status: monosomy) UM patients. (D) Kaplan–Meier survival curves showing the statistically significant relationship between the expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and overall survival probability in normal BAP1(BAP1_mut: NO) or in mutated BAP1 (BAP1_mut: YES) TCGA-UVM samples. (E) Kaplan–Meier survival curves showing the statistically significant relationship between the expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and DFS in TCGA-UVM patients presenting chromosome 3 disomy (chr3_status: disomy) or chromosome 3 monosomy (chr3_status: monosomy). (F) Kaplan–Meier survival curves showing the statistically significant relationship between the expression levels of GPX4, SLC7A11, SLC3A2, HMOX1, GCLM, CTH, NQO1 ACSL3, IREB2, POR, AIFM2 and DFS in normal BAP1(BAP1_mut: NO) or in mutated BAP1 (BAP1_mut: YES) TCGA- UVM samples. Figure 11 illustrates that high expression of a combination of genes inhibiting ferroptosis (iFERR) correlates with reduced overall and disease-free survival in the GSE84976 and TCGA-UVM cohorts. (A) Graphs showing the statistically significant relationship between expression levels of a novel ferroptosis signature (iFERR: GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3, IREB2, NQO1, AIFM2) and disomy 3 or monosomy 3 in the GSE22138, GSE27831, GSE84976 and TCGA-UVM cohorts. (B) Kaplan-Meier curves revealing the statistically significant relationship between iFERR and OS in the GSE84976 and TCGA-UVM cohorts. (C) Kaplan-Meier graphs showing the statistically significant relationship between iFERR and DFS in the GSE22138, GSE27831, GSE84976 and TCGA-UVM cohorts. Figure 12 illustrates a schematic model summarising alterations in the expression of the indicated ferroptosis hallmarks in MUM samples after 1,4-dihydroxy quininib treatment. (A) Figure showing the specific factors modulated by 1,4-dihydroxy quininib in a time-dependent fashion. Created with BioRender.com. Subscription: Institution (University College Dublin) (B). Table summarising the significant differences in ferroptosis hallmarks after 1,4-dihydroxy quininib. Non-significant differences are represented with “=”, significant with “↓” and significant upregulation with “↑”. Q7 = quininib; Pi, phosphorylated. Figure 13 illustrates that high expression of a combination of 5 genes inhibiting ferroptosis (iFERR5g: GPX4, SLC3A2, GCLM, NQO1, AIFM2) correlates with reduced overall and disease-free survival in the GSE27831, GSE84976 and TCGA-UVM cohorts. (A) Graphs showing the statistically significant relationship between expression levels of a novel ferroptosis signature (iFERR5g: GPX4, SLC3A2, GCLM, NQO1, AIFM2) and disomy 3 or monosomy 3 in the GSE22138, GSE84976 and TCGA-UVM cohorts. (B) Kaplan-Meier curves revealing the statistically significant relationship between iFERR5g and OS in the GSE84976 and TCGA-UVM cohorts. (C) Kaplan-Meier graphs showing the statistically significant relationship between iFERR5g and DFS in the GSE27831, GSE84976 and TCGA-UVM cohorts. (D) ROC curves showing the accuracy of iFERR5g at predicting overall survival, alone or in combination with chromosome 3 status. Figure 14 illustrates that high expression of a combination of 7 genes inhibiting ferroptosis (iFERR7g: GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3, IREB2) correlates with reduced disease-free survival in the GSE22138 cohort. Kaplan-Meier graphs show the statistically significant relationship between iFERR7g and DFS in the GSE22138 cohort. Figure 15 illustrates that expression of IFERR4 gene signature is associated with patient overall survival, tumour stage, KRAS status and smoking status in lung cancer. Expression levels of genes in the IFERR4 signature (ACSL3, ATP5F1B, SLC3A2, and CYSLTR2) were analysed in the TCGA-LUAD lung cancer dataset (n=566) using R version 4.4. (A) Kaplan-Meier curve showing association of the IFERR4 signature score with overall survival (p = 0.00037). (B) The combined signature score was significantly higher in tumour tissue compared to matched normal tissue (paired Welch’s t-test, p < 0.0001). (C) The combined signature score was significantly higher in T2 tumours (1-way ANOVA, p = 0.0028) and T3 tumours (p = 0.0030) compared to T1 tumours. (D) The combined signature score was significantly higher in patients with KRAS mutations compared to patients with wild-type KRAS (Welch’s t-test, p = 0.0088). (E) Kaplan-Meier curves showing the association between IFERR4 gene expression and patient overall survival in patients with wild-type and mutant KRAS. The association with survival was stronger in patients with KRAS mutations. (F) There was no significant difference in the IFERR signature score between smokers and non-smokers (Welch’s t-test, p = 0.27). (G) Kaplan-Meier curves showing the association between gene expression and patient overall survival in smokers and non-smokers. The association with survival was stronger in smokers. Detailed Description of the Drawings Materials and Methods TCGA data analysis Clinical and mutational data of primary lung, pancreatic, ovarian, and cutaneous tumours included in The Cancer Genome Atlas were collected from GDC data portal using the “TCGAbiolinks” package in R. Specifically, data were collected from the TCGA-LUAD lung cancer dataset (n=566), the TCGA-PAAD pancreatic cancer dataset (n=184), the TCGA-OV ovarian cancer dataset (n=585), and the TCGA-SKCM cutaneous melanoma dataset (n=442). Annotated mutational data was downloaded from the cBioPortal (www.cbioportal.org). RNA-seq data were downloaded in fragments per kilobase per million (FPKM), then converted to log2 scale. Samples were divided into high and low expression groups for each gene using the median expression level as the cut-off. Survival analysis was performed using Overall Survival (OS) or Disease Free Survival (DFS) as endpoints. Kaplan–Meier curves were generated using the “survival” and “survminer” packages in R, and the log-rank test was used to compare the groups. Differential expression analysis was conducted using the “rstatix” package in R. Paired t-tests were employed for comparisons of matched samples, while Welch’s unpaired t-tests were used to analyse differences between two patient subgroups. In cases where extreme outliers were identified, the Wilcoxon test was applied. For comparisons involving more than two subgroups, one-way ANOVA was performed, followed by Tukey’s HSD post-hoc test for pairwise comparisons. Clinical and mutational data of primary uveal tumours was collected from TCGA-UVM dataset included in The Cancer Genome Atlas (n=80). Annotated mutational data was downloaded from the cBioPortal. RNA-seq was downloaded in fragments per kilobase per million (FPKM), then converted to log2 scale. Bulk RNA-seq data together with clinical annotation from the studies by Laurent et al., Gangemi et al., van Essen et al. (99-101) were obtained from the GEO database (dataset identifiers: GSE22138, GSE27831 and GSE84976, respectively). Expression between chromosome 3 monosomic or disomic patients was statistically compared using the U-MannWhitney test. Survival analysis was performed using Overall Survival (OS) or Disease Free Survival (DFS) from annotation. Kaplan–Meier curves were plotted to represent the result and log-rank test was computed. Cell culture UM cell line OMM2.5 derived from a metastatic UM tumor and Mel285 derived from primary UM tumor were kindly provided by Dr. Martine Jager (Leiden, The Netherlands) (89). Cell cultures were maintained at 37°C / 5% CO2 in RPMI 1640 Medium (Gibco, Gaithersburg, MD, USA) supplemented with 10% FBS and 2% Penicillin / Streptomycin. OMM2.5 and Mel285 cells were authenticated by short-tandem repeat (STR) profiling performed by American Type Culture Collection (ATCC) and tested for absence of mycoplasma contamination. MUM OPDX mouse models Human tumors were aseptically isolated and placed at room temperature in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS plus 50 U / ml penicillin and 50 mg / ml streptomycin. Tumours were orthotopically implanted into the liver of two six-week-old male athymic nude mice (strain Hsd:Athymic Nude-Foxn1nu) purchased from Envigo. Briefly, mice were anesthetized with a continuous flow of 1% to 3% isoflurane and oxygen mixture (2 L / min). After performing a median laparotomy, the tumour fragment was anchored with a Prolene 7-0 suture into a small pocket created in the anterior hepatic lobe, and the abdominal incision will be closed with surgical staples. After implantation, mice were inspected twice a week, and at euthanasia, OPDX were harvested, cut into small fragments and serially transplanted into new animals for tumour perpetuation and / or experimental procedures. Animals were housed in a sterile environment, cages and water were autoclaved and bedding and food was γ-ray sterilized. Explant culture of MUM liver tumours from OPDX mouse model Immediately following dissection, MUM tissue was placed into complete culture medium at room temperature. Upon arrival at UCD, the tissue was washed three times in sterile PBS wash buffer (PBS and 2% Penicillin / Streptomycin). Using a sterile scalpel and forceps, the tumour was cut into 3 pieces, arbitrarily renamed as left (L), middle (M) or right (R). (Figure 9A).2 fragments of each piece were incubated in 20 μM 1,4- dihydroxy quininib or 0.5% DMSO, made up to 1 ml in complete culture medium in a 12-well plate. Explants were incubated for 72 h at 37°C / 5% CO2. Plates were wrapped in parafilm to prevent evaporation of medium during the incubation period. After 72 hours, the explant tissue was snap-frozen in liquid nitrogen and stored at –80°C. Drug preparation for use in cell and culture Quininib, 1,4-dihydroxy quininib, montelukast (Sigma-Aldrich, St. Louis, MO, USA #SML0101), HAMI 3379 (Cayman Chemical, Ann Arbor, MI, USA #10580) and Erastin (Sigma-Aldrich; St. Louis, MO, USA #E7781) were dissolved in 100% DMSO and stored as (10–50 mM) stock solutions. Working solutions (100 μM) were prepared fresh prior to each experiment in complete cell culture medium as described above. Drugs were made to final test concentrations by adding the required volume of the working solution to cells in complete media. 0.5% DMSO was used as a control for all other drug treatment experiments. Proteomics sample preparation OMM2.5 cells were seeded at a density of 1 x 106cells in triplicate wells and drug treated for 4, 8 or 24 hours with 0.5% DMSO or 20 µM 1,4-dihydroxy quininib. Four independent experiments were performed. Proteins were isolated using PreOmics iST 8X for protein / proteomics preparation kit (PreOmics GmbH; Martinsried, Germany) according to manufacturer’s protocol. The samples were analyzed by the UCD Conway Institute Mass Spectrometry Resource (MSR) on a Thermo Fisher Scientific Inc. Q Exactive mass spectrometer connected to a Dionex Ultimate 3000 (RSLCnano) chromatography system. Peptides were separated on C18 home-made column (C18-AQ Dr. Maisch Reprosil-Pur 100 x 0.075 mm x 3 μm) over 120 min at a flow rate of 250 nL / min with a linear gradient of increasing ACN from 1% to 27%. The mass spectrometer was operated in data dependent mode; a high resolution (70,000) MS scan (300-1600 m / z) was performed to select the fourteen most intense ions and fragmented using high energy C-trap dissociation for MS / MS analysis. Raw data from the Q Exactive was processed using MaxQuant (version 2.0.3.0) incorporating the Andromeda search engine, as described in (90). Briefly, to identify peptides and proteins, MS / MS spectra were matched against Uniprot homo sapiens database (2021_03) containing 78,120 entries. All searches were performed using the default setting of MaxQuant, with trypsin as specified enzyme allowing two missed cleavages and a false discovery rate of 1% on the peptide and protein level. The database searches were performed with carbamidomethyl (C) as fixed modification and acetylation (protein N terminus) and oxidation (M) as variable modifications. For the generation of label free quantitative (LFQ) ion intensities for protein profiles, signals of corresponding peptides in different HPLC MS / MS runs were matched by MaxQuant in a maximum time window of 1 min. Perseus software was used to process the data and create heatmaps. After screening a large number of proteins simultaneously an important statistical concept is multiple hypothesis testing, in which many statistical tests are conducted at the same time and detection of a protein with a relatively small p-value may be a false discovery. To address this problem, Storey and Tibshirani propose a new metric, the q-value (91). The q-value is intended to be analogous to the p-value but takes into account multiple testing corrections. The p- value produces false discoveries but the q-values fail to detect some true discoveries, as revealed by immunoblots (Figure 6A-D). In light of this, in the present work, the results are discussed without the multiple testing corrections (i.e. the p-values) but include the results with the multiple testing corrections (i.e. the q-values). ClueGo (v2.5.8) and Cluepedia (v1.5.8) plugins in Cytoscape (v3.8.2) with the Homo sapiens (9606) marker set was utilized for GO:Biological processes pathway analysis of enriched proteins. Functional Enrichment analysis tool (FunRich v3.1.3) was used to create Venn diagrams using associated gene names identified. The mass spectrometry proteomics data is deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD046822 and 10.6019 / PXD046822. Explant total protein determination Total protein was extracted from each piece of tumour explant tissue. Each individual explant was placed in a tube with 400 μl of ice-cold T-PER lysis reagent (Thermo Fisher Scientific, Rockford, IL, USA) supplemented with 10 μl / ml protease inhibitor and a 3 mm stainless steel bead. Tubes were placed in a TissueLyser II (Qiagen) for 2.5 min to homogenize the tissue. The tissue lysate was centrifuged at 14,000 rpm for 30 min at 4°C. The supernatant was used immediately for protein determination or stored at −80°C. The BCA (Thermo Fisher Scientific, Rockford, IL, USA) kit was used to quantify the total protein extracted from explant tissue in μg / ml as per the manufacturer’s instructions. Western blot OMM2.5 cells were seeded at 1 × 106cells per well of a 6-well plate and left to adhere for 24 hours. Cells were treated with 0.5% DMSO or 20 μM 1,4-dihydroxy quininib for 4, 8, or 24 hours. Mel285 cells were seeded at 1 × 106cells per well of a 6-well plate, left to adhere for 24 hours and then treated with 0.5% DMSO or 20 μM 1,4-dihydroxy quininib for 24 hours. Total protein was extracted from cells as described (18). Cells protein concentrations were measured BCA protein assay kit (ThermoFisher Scientific; Waltham, MA, United States) in accordance with manufacturer’s instructions, and 10 μg of protein was loaded per lane (N = 3, at least). Explants protein concentrations were determined as described above and 8 μg of protein was loaded per lane. 3 tumours (= 3 animals) for each OPDX model were used for these experiments, i.e. 3 tumours for UVM4 and 3 tumours for UVM7. For each tumour, fragments “L” and “R” of were used for western blot experiments. Blots were probed for HMOX1 (10701-1-AP, Proteintech, IREB2 (23829-1-AP, Proteintech), GDF15 (27455- 1-AP, Proteintech), NRF2 (16396-1-AP, Proteintech), GPX4 (67763-1-Ig Proteintech), GCLM (14241-1-AP, Proteintech), 4-HNE (JaICA, MHN-100P) and β-actin (A5441, Sigma-Aldrich). Anti-rabbit IgG, HRP-linked Antibody (1:3000; #7074s, Cell Signaling Technology) and anti-mouse IgG, HRP-linked Antibody (1:3000; #7076s, Cell Signaling Technology) were used as secondary antibodies. Signal was detected with enhanced chemiluminescence substrate (Pierce™ ECL Western Blotting Substrate; ThermoFisher Scientific). For 4-HNE, the 48 kDa band was considered for the quantification, based on previous evidence showing that a 48 kDa protein exhibits a strong reactivity with the antibody in hepatocytes treated with 10 µM HNE (93). Measurement of ROS Levels Levels of reactive oxygen species (ROS) were determined using the fluorescent probe 2,7-dichlorofluorescein diacetate (DCFH-DA, Sigma-Aldrich, St. Louis, MO, USA #D6883). Cells were seeded at a density of 2 x 104into 96-well plates and allowed to adhere overnight. They were then treated for 8 or 24 hours with 0.5% DMSO or 20 µM 1,4-dihydroxy quininib in RPMI 1640 Medium (Gibco, Gaithersburg, MD, USA) supplemented with 10% FBS and 2% Penicillin / Streptomycin. Media was removed and cell were washed with a 0.1% Triton solution in complete media to enhance cellular probe permeation.100 µL of a DCFH-DA working solution (200 µM) prepared in RPMI 1640 Medium (Gibco, Gaithersburg, MD, USA) supplemented 2% Penicillin / Streptomycin but no FBS was added to each well and incubated at at 37 °C / 5% CO2 for 30 min int he dark. After incubation, the solution was removed and wells were washed with in 1X PBS.200 µl of radioimmunoprecipitation assay (RIPA, Sigma- Aldrich, St. Louis, MO, USA, #R0278) buffer were added to each well. The plate was incubated on ice for 5 min, then cell lysate was collected into 1.5 mL tubes. Tubes underwent centrifugation at 21,130 × g for 10 min at 4 °C.100 μL of supernatant were transferred to a 96 well plate and measure the fluorescence intensity using a fluorescence a microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. The protocol was used on wells without cells, which represented the negative control.10 μL of the supernatant were used to perform the Bicinchoninic Acid (BCA) Protein Assay (#23227, ThermoFisher Scientific name of kit). Six replicate wells were used for each group. The mean fluorescence intensity value of wells without cells was subtracted to each fluorescence intensity value obtained for the other conditions. The final results are expressed as fluorescence intensity (AU) / proteins (mg / mL). Experiments were conducted in triplicate (24 hour- treatment) or quadruplicate (8 hour-treatment). The results are expressed as percentage of the control. Biliverdin measurement OMM2.5 cells were seeded at 1 × 106cells per well of a 6-well plate and left to adhere for 24 hours. Cells were treated for 4, 8, or 24 hours with 0.5% DMSO or 20 μM 1,4- dihydroxy quininib. At the end of the treatment cells were detached with trypsin, resuspended in medium, centrifuged at 800 g for 5 minutes, washed once in PBS, and centrifuged again. The pellet was frozen at -80°C. Each cell pellet was resuspended in 0.65 mL of lysis buffer consisting of 1% Triton-X100, 0.4 g / L bovine serum albumin in PBS, pH 8.5. The lysed pellets were divided into 3 aliquots for the determination of autofluorescence (blank), bilirubin, and biliverdin. For fluorescence calibration, bilirubin standard solutions (2, 5, 10, 20 nM) were prepared in lysis buffer. Aliquots from the autofluorescence wells were used for protein determination. Quantification of intracellular bilirubin was performed using a high-throughput fluorometric method based on the high-affinity selective binding of bilirubin to the recombinant protein HELP -UnaG (HUG) (94). The assay was adapted for the measurement of biliverdin by adding biliverdin reductase (BVR) to the assay buffer as described by Tramer and colleagues (94-96). Experiments were conducted in triplicate. Determination of Thiol Groups The concentration of non-protein thiol groups (RSH), reflecting about 90% of the GSH cellular content, was measured in total cell lysates. OMM2.5 cells were seeded at a density of 1 x 106cells per well of a 6-well plate and left adhere for 24 hours. Cells were then treated for 4, 8 or 24 hours with 0.5% DMSO or 20 µM 1,4-dihydroxy quininib in RPMI 1640 Medium (Gibco, Gaithersburg, MD, USA) supplemented with 1% FBS and 2% Penicillin / Streptomycin. Media was removed and cells were washed in 1X PBS.1 ml of trypsin was added to each well to promote cells detachment.1 ml of fresh media was then added to each well to neutralize the trypsin activity. Cells underwent centrifugation at 1200 rpm for 4 supernatant was removed, the cell pellet was washed in 1 ml of PBS media and centrifugated at 1200 rpm for 4 minutes. The last step was performed twice. Cell pellet was snap frozen in liquid nitrogen and stored at -80° C. RSH levels were evaluated by a spectrophotometric assay based on the reaction of thiol groups with 2,2-dithio-bis-nitrobenzoic acid (DTNB). A DTNB solution and the samples were mixed and incubated at room temperature for 20 min in the dark until the noticeable appearance of a yellow colour. After incubation, samples were centrifuged at 3000 rpm for 10 min. The supernatant was collected and set in a black 96-well plate for measurement of the absorbance in a microplate reader (Biotek Synergy-HT, Winooski, VT, USA) at λ = 412 nm. Experiments were conducted in quadruplicate, with 3 technical replicates per condition. The results are expressed as percentage of the control. Measurement of Lipid Peroxidation Levels of LOOH were evaluated through the oxidation of Fe2+to Fe3+in the presence of xylenol orange. OMM2.5 cells were seeded at a density of 1 x 106cells per well of a 6- well plate and treated for 4, 8 or 24 hours with 0.5% DMSO or 20 µM 1,4-dihydroxy quininib, as above (see the method Determination of Thiol Groups). The assay mixture contained 200 μg of the sample (total cell lysate), 100 μM xylenol orange, 250 μM ammonium ferrous sulphate, 90% ethanol, 4 mM butylated hydroxytoluene and 25 mM H2SO4. Samples were incubated at room temperature for 30 min, and the absorbance was finally measured at λ = 560 nm using a microplate reader (Biotek Synergy-HT, Winooski, VT, USA). Experiments were conducted in quadruplicate. The results are expressed as a percentage of the control. MTT Assay Cell metabolism, an indirect measure of viability, was determined using MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (#M5655), Sigma-Aldrich; St. Louis, MO, USA) assay (97). Briefly, 5 × 103cells / well were seeded into 96-well plates and allowed to adhere for 24 hours. Cells were treated in triplicates with either 0.5% DMSO (vehicle control), 15% H2O2 (positive control) or 1, 5, 10, 20, 50 µM erastin (#E7781, Sigma-Aldrich; St. Louis, MO, USA), prepared in complete media for 96 hours. Once drug solution was removed, MTT dye and serum-free media were added in a 1:10 ratio, to each well and incubated in the dark for 2 hours at 37°C. Then, 100% DMSO (1:1 ratio) was added to each well and absorbance was measured at 570 nm using a SpectraMax® M2 microplate reader (Molecular Devices Corporation, Sunnyvale, CA, USA). Experiments conducted in triplicate. The results are expressed as percentage of the control. Statistics Statistical analyses were performed using GraphPad Prism v7.00 (GraphPad Software, San Diego, CA, USA). The normal distribution was assessed using the Shapiro-Wilk test. For non-normally distributed sample data, comparisons between two groups were analysed using Mann-Whitney U test and comparisons between three or more groups were analysed using Kruskal-Wallis test. In the case of normally distributed data, two- group comparisons employed a two-tailed t-test, while comparisons involving three or more groups utilized a one-way ANOVA. The Brown-Forsythe test accompanied the one-way ANOVA to evaluate the homogeneity of variance among different groups. The obtained p-values for all tests indicated homogeneity of variance across groups. Sample sizes were chosen based on previously published experiments. Error bars depict the standard error of the mean (SEM), indicating the variability among samples. Central values are presented as the mean. The investigators were not blinded to allocation during experiments and outcome assessment. Significance was set at p- value less than 0.05. Sequences SEQ ID NO: 1 (human ATP5B nucleic acid) – NCBI Reference Sequence: NC_000012.12:c56645984-56638175 SEQ ID NO: 2 (human ATP5B amino acid) – NCBI Reference Sequence: NP_001677.2 SEQ ID NO: 3 (human GPX4 nucleic acid) - NCBI Reference Sequence: NG_050621.1:5069-7854 SEQ ID NO: 4 (human GPX4 amino acid isoform B) – NCBI Reference Sequence: NP_001034936.1 SEQ ID NO: 5 (human GPX4 amino acid isoform C) – NCBI Reference Sequence: >NP_001034937.1 SEQ ID NO: 6 (human GPX4 amino acid isoform D) – NCBI Reference Sequence: NP_001354761.1 SEQ ID NO: 7 (human GPX4 amino acid isoform A) – NCBI Reference Sequence: NP_002076.2 SEQ ID NO: 8 (human SLC7A11 nucleic acid) - NCBI Reference Sequence: NC_000004.12:c138312671-138164097 - NP_001295182.1 glutamate--cysteine ligase regulatory subunit [Homo sapiens] SEQ ID NO: 27 (human GCLM amino isoform 1) - NCBI Reference Sequence: NP_002052.1 glutamate--cysteine ligase regulatory subunit [Homo sapiens] SEQ ID NO: 28 (human ACSL3 nucleic acid) – NCBI Reference Sequence: NC_000002.12:222861036-222944639 SEQ ID NO: 29 (human ACSL3 amino acid) – NCBI Reference Sequence: NP_001341087.1 SEQ ID NO: 30 (human ACSL3 amino acid) – NCBI Reference Sequence: NP_001341088.1 SEQ ID NO: 31 (human ACSL3 amino acid) – NCBI Reference Sequence: NP_004448.2 SEQ ID NO: 32 (human ACSL3 amino acid) – NCBI Reference Sequence: NP_976251.1 SEQ ID NO: 33 (human NQO1 nucleic acid) NCBI Reference Sequence: NC_000016.10:c69726560-69709401 SEQ ID NO: 34 (human NQO1 amino acid isoform a) NCBI Reference Sequence: NP_000894.1 SEQ ID NO: 35 (human NQO1 amino acid isoform b) NCBI Reference Sequence: NP_001020604.1 SEQ ID NO: 36 (human NQO1 amino acid isoform c) NCBI Reference Sequence: NP_001020605.1 SEQ ID NO: 37 (human NQO1 amino acid isoform d) NCBI Reference Sequence: NP_001273066.1 SEQ ID NO: 38 (human AIFM2 nucleic acid) NCBI Reference Sequence: NC_000010.11:c70132825-70112271 SEQ ID NO: 39 (human AIFM2 amino acid) – NCBI Reference Sequence: NP_001185625.1 SEQ ID NO: 40 (human AIFM2 amino acid) – NCBI Reference Sequence: NP_116186.1 SEQ ID NO: 41 (human CysLT1 nucleic acid) - NCBI Reference Sequence: NC_000023.11:c78327611-78271468 SEQ ID NO: 42 (human CysLT1 amino acid) - NCBI Reference Sequence: NP_001269115.1 SEQ ID NO: 43 (human CysLT1 amino acid) - NCBI Reference Sequence: NP_001269116.1 cysteinyl leukotriene receptor 1 [Homo sapiens] SEQ ID NO: 44 (human CysLT1 amino acid) - NCBI Reference Sequence: NP_001269117.1 – NP_002124.1 dbSNP ID: rs121913530 Results Dysregulated expression of genes modulating ferroptosis correlates with reduced OS and DFS in patients with primary lung, pancreatic, ovarian cancers and cutaneous melanoma. To interrogate the significance of ferroptotic markers in primary lung, pancreatic, ovarian and cutaneous melanoma, their gene expression was analysed in The Cancer Genome Atlas (TCGA). The markers selected were ACSL3, AIFM2, ATP5F1B, CTH, CYSLTR1, CYSLTR2, GCLM, GPX4, IREB2, NQO1, SLC3A2, SLC7A11. All the selected genes were expressed in the TCGA samples, confirming their potential disease relevance. The analysis revealed that ACSL3 (p = 0.0014), ATP5F1B (p = 0.045), CYSLTR1 (p = 0.0025), CYSLTR2 (p = 0.0033), SLC3A2 (p = 0.017) expression significantly correlates with OS in lung cancer and that ACSL3 (p = 0.0029), expression significantly correlates with DFS in lung cancer (Figure 1). IREB2 (p = 0.019), expression significantly correlates with DFS in ovarian cancer (Figure 2). CTH (p = 0.029), expression significantly correlates with OS (p = 0.029), and DFS (p = 0.014) in pancreatic cancer (Figure 3). IREB2 (p = 0.0066) expression significantly correlates with DFS in cutaneous melanoma (Figure 4) and ACSL3 (p = 0.03) expression significantly correlates with OS in cutaneous melanoma (Figure 4). 1,4-dihydroxy quininib significantly proteomic markers of ferroptosis in OMM2.5 cells. To understand the molecular changes induced by 1,4-dihydroxy quininib in OMM2.5 cells, proteome-profiling was performed on whole cell extracts at 4, 8 and 24 hours post-treatment (hpt. A total of 4381 proteins were detected across all samples. Applying stringency cut-offs of p value < 0.05 and fold-change > + 1.2, at 4 hpt 66 differentially expressed proteins (DEPs) were identified, with 41 proteins significantly upregulated and 25 proteins significantly downregulated. At 8 hpt, 164 DEPs were detected: 72 significantly upregulated and 92 significantly downregulated. 95 DEPs were uncovered at 24 hpt, with 49 significantly upregulated and 46 significantly downregulated proteins (Figure 5A-C). Additional analysis revealed 157 proteins showing a significant time-dependent expression upon 1,4-dihydroxy quininib treatment. The most consistently upregulated protein after 4, 8 or 24 hours treatment was heme oxygenase 1 (HO-1), with changes of 3.3, 8.2 or 6-fold respectively (Figure 5C, 6A,B; Tables A-C). The HO-1 enzyme catabolizes cellular heme to biliverdin, carbon monoxide, and free iron. HO-1 has dual roles in cancer cells, since in several malignant human neoplastic diseases it promotes cell growth, proliferation and invasion, but in other cancers it displays anti-tumoral effects. KEGG pathway analysis revealed that at 8 and 24 hpt, upregulated DEPs were associated with specific cellular processes including: protein processing in endoplasmic reticulum, ferroptosis, biosynthesis of cofactors, RNA degradation, one carbon pool by folate at 8 hpt (Figure 5D,E) and proteasome, metabolic pathways, biosynthesis of amino acids, biosynthesis of cofactors, Parkinson’s disease, ferroptosis, chemical carcinogenesis - receptor activation, amino sugar and nucleotide sugar metabolism, fluid shear stress and atherosclerosis, fatty acid metabolism, Alzheimer’s disease, gastric acid secretion, and progesterone-mediated oocyte maturation at 24 hpt (Figure 5F,G). Interestingly, the analysis highlighted that ferroptosis upregulation was present at both 8 and 24 hpt with 1,4-dihydroxy quininib (Figure 5D-H). Protein-protein interaction network analysis elucidated which significantly DEPs were associated with ferroptosis, including HO-1 (Figure 5E,G) which mediates protective or detrimental effects via ferroptosis induction. At 8 hpt, additional DEPs linked to ferroptosis were solute carrier family 3 member 2 (SCL3A2) and glutamate-cysteine ligase regulatory subunit (GCLM) (Figure 5E). HO-1 regulation was also associated to fluid shear stress and atherosclerosis processes at 24 hpt (Figure 5G). GCLM was differentially expressed after 1,4-dihydroxy quininib also at 24 hpt, together with the ferroptosis marker acyl-CoA synthetase long chain family member 3 (ACSL3) (Figure 5G). To corroborate the proteomics data, some of the 10 most DEPs after 1,4-dihydroxy quininib treatment were selected and performed immunoblotting (Tables A-C below). Table A shows the 10 most up- and down-regulated proteins in OMM2.5 cells after 4 h of treatment with 20 µM Q7. Table B shows the 10 most up- and down-regulated proteins in OMM2.5 cells after 8 h of treatment with 20 µM Q7. Table C shows the 10 most up- down- regulated proteins in OMM2.5 cells after 24 h of treatment with 20 µM Q7. Protein names coloured in red and blue in Tables A, B and C are Q7-upregulated and downregulated proteins, respectively, known to be associated to the ferroptosis pathway. Significant HO-1 upregulation at 8 and 24 hpt p = 0.0286 and p = 0.0463, respectively), iron responsive element binding protein 2 (IREB2) downregulation at 8 hpt (p = 0.0412) and growth / differentiation factor-15 (GDF15) upregulation at 8 hpt (p = 0.0416) were observed (Figure 6A-D).

[0002] Table A: The 10 most up- and down- proteins in OMM2.5 cells after 4 h of treatment with 20 µM Q7. UNIPROT Protein name -log p- T-test Fold value Difference Change Upregulated P09601 Heme oxygenase 1 2.95 1.72 3.30 Q7Z422 SUZ domain-containing protein 1 1.52 1.19 2.27 Inhibitor of nuclear factor kappa-B Q70UQ0 kinase-interacting protein 1.43 0.78 1.72 Q96MU7 YTH domain-containing protein 1 1.34 0.72 1.65 Q99988 Growth / differentiation factor 15 1.93 0.69 1.61 Probable E3 ubiquitin-protein ligase Q5GLZ8 HERC4 1.86 0.58 1.49 Gamma-tubulin complex component Q9BSJ2 2 1.54 0.44 1.36 Ribosome biogenesis regulatory Q15050 protein homolog 2.53 0.43 1.34 A0A087WX97 Bcl-2-like protein 13 1.82 0.38 1.30 Nuclear ubiquitous casein and Q9H1E3 cyclin-dependent kinase substrate 1 1.39 0.37 1.29 Downregulated Q14139 Ubiquitin conjugation factor E4 A 1.52 -1.39 -2.61 A0A087WYS3 Protein kinase C-binding protein 1 1.40 -0.83 -1.77 Inositol polyphosphate 5- Q01968 phosphatase OCRL 1.92 -0.7 -1.63 Chitinase domain-containing protein Q9BWS9 1 1.34 -0.56 -1.47 Q96C23 Galactose mutarotase 1.63 -0.4 -1.32 Dolichyl-phosphate beta- A0A7P0T882 glucosyltransferase 3.07 -0.37 -1.29 Haloacid dehalogenase-like hydrolase domain-containing protein Q9H0R4 2 1.48 -0.36 -1.28 Q9HD67 Unconventional myosin-X 1.83 -0.35 -1.28 Prolactin regulatory element-binding Q9HCU5 protein 1.43 -0.35 -1.28 Q9H3K6 BolA-like protein 2 1.40 -0.34 -1.27 Table B: The 10 most up- and down- proteins in OMM2.5 cells after 8 h of treatment with 20 µM Q7. UNIPROT Protein name -log p- T-test Fold value Difference Change Upregulated P09601 Heme oxygenase 1 3.93 3.04 8.22 Ragulator complex protein Q9UHA4 LAMTOR3 1.68 1.52 2.87 Q99988 Growth / differentiation factor 15 2.15 1.31 2.48 P13645 Keratin, type I cytoskeletal 10 1.37 1.27 2.41 Eukaryotic translation initiation O60739 factor 1b 1.38 1.03 2.05 Oxidative stress-induced growth Q9UJX0 inhibitor 1 1.67 0.92 1.89 Q9NQA3 WAS protein family homolog 6 1.45 0.8 1.74 Signal peptide peptidase-like 2A H0YNA7 (Fragment) 2.74 0.8 1.74 P48723 Heat shock 70 kDa protein 13 2.42 0.79 1.73 Signal transducer and activator of A0A494C0T4 transcription 1.97 0.62 1.54 Downregulated Transmembrane 9 superfamily A0A0C4DFM1 member 1.47 -3.75 -13.44 Q71UM5 40S ribosomal protein S27-like 1.71 -2.98 -7.89 Iron-responsive element-binding P48200 protein 2 1.73 -0.51 -1.43 B8ZZQ6 Prothymosin alpha 2.36 -0.48 -1.4 Q9BW19 Kinesin-like protein KIFC1 3.02 -0.44 -1.36 Membrane-associated transporter Q9UMX9 protein 1.7 -0.41 -1.33 DnaJ homolog subfamily C Q8WXX5 member 9 2.05 -0.38 -1.3 Q9NQW6 Anillin 1.54 -0.38 -1.3 Chromosome-associated kinesin O95239 KIF4A 2 -0.37 -1.29 Q86WB0 Nuclear-interacting partner of ALK 1.56 -0.36 -1.28 Table C: The 10 most up- down- regulated proteins in OMM2.5 cells after 24 h of treatment with 20 µM Q7. UNIPROT Protein name -log p- T-test Fold value Difference Change Upregulated P09601 Heme oxygenase 1 3.01 2.58 5.97 Q5QPM7 Proteasome inhibitor PI31 subunit 1.55 1.58 3 P32929 Cystathionine gamma-lyase 2.26 1.47 2.76 ELKS / Rab6-interacting / CAST family G8JLD3 member 1 1.6 1.24 2.37 Serine / threonine-protein Q96HS1 phosphatase PGAM5, mitochondrial 2.11 1.17 2.25 Glutamate--cysteine ligase P48507 regulatory subunit 2.57 0.91 1.88 Q9UJF2 Ras GTPase-activating protein nGAP 2.02 0.89 1.86 Hydroperoxy icosatetraenoate A4F3V8 dehydratase (Fragment) 1.49 0.89 1.85 E9PGT3 Ribosomal protein S6 kinase 1.41 0.8 1.75 P15559 NAD(P)H dehydrogenase [quinone]1 2.97 0.79 1.73 Downregulated m-AAA protease-interacting Q8WWC4 protein 1, mitochondrial 1.46 -1.73 -3.32 P20338 Ras-related protein Rab-4A 1.51 -1.33 -2.52 Myosin phosphatase Rho- Q6WCQ1 interacting protein 1.57 -1.11 -2.15 A0A4W8VX11 Pericentriolar material 1 protein 1.49 -0.55 -1.46Sterol 26-hydroxylase, Q02318 mitochondrial 1.8 -0.55 -1.46 Cleft lip and palate O96005 transmembrane protein 1 2.03 -0.54 -1.46 Q99661 Kinesin-like protein KIF2C 2.05 -0.53 -1.44 Q8WXH0 Nesprin-2 1.93 -0.51 -1.42 SRA stem-loop-interacting RNA- Q9GZT3 binding protein, mitochondrial 2.38 -0.47 -1.39 A0A087X0G7 NF-kappa-B essential modulator 1.65 -0.46 -1.37 5 1,4-dihydroxy quininib modulates the 1 and the GSH-GPX4-LOOH axes in OMM2.5 cells in a time dependent manner. Since HO-1 expression is regulated by NRF2, a transcription factor involved in cellular detoxification processes, it was investigated whether HO-1 upregulation correlated to increased reactive oxygen species (ROS) and NRF2 levels. ROS levels significantly increased after 8 hours of 1,4-dihydroxy quininib treatment (p = 0.0419), but did not change following the 24 hours one (p = 0.1624) (Figure 7A). Interestingly, increased ROS amount after 8 hours was associated with a significant (p = 0.0096) increase in phosphorylated NRF2. This form of NRF2 preferentially localizes in the nucleus, and binds within regulatory regions of target genes (e.g., HO-1, glutamate-cysteine ligase catalytic subunit (GCLC), GCLM, multidrug-resistant proteins (MRPs), and sequestosome-1 protein (p62)). Phosphorylated NRF2 was also significantly increased after 4 hours of treatment (p = 0.0337), while no significant change was observed with the 24 hour-treatment (Figure 7B,C). To evaluate whether increased HO-1 expression correlated to increased HO-1 enzymatic activity, biliverdin levels were measured (Figure 7D). Intriguingly, we detected a significant increase in biliverdin levels following 4 hours of treatment (p = 0.0083), while no differences were observed with the 8 or 24-hour treatment (Figure 7D). It was then investigated whether 1,4-dihydroxy quininib modulates additional ferroptosis hallmarks in OMM2.5 cells. Western blot analysis revealed significantly (p = 0.0444) increased GPX4 expression after 4 hours of 1,4-dihydroxy quininib treatment, while the 24-hour treatment significantly decreased GPX4 levels (p = 0.0006) (Figure 7E,F). No statistically significant differences were observed after 8 hours of treatment (Figure 7E,F).1,4-dihydroxy quininib did not affect glutathione (GSH) content after 4 hours of treatment, but led to a significant increase in GSH levels when administered to OMM2.5 cells for 8 or 24 hours (p = 0.0023; p = 0.0008, respectively) (Figure 7G). Concurrently, lipid hydroperoxide (LOOH) levels were significantly upregulated following 4 (p = 0.0043) and 8 (p = 0.0013) hours of 1,4-dihydroxy quininib treatment, but not after 24 hours (Figure 7H). Proteomic analysis uncovered an upregulation of GCLM (Figure 5E,G), a regulatory subunit of the glutamate cysteine ligase (GCL), the rate-limiting enzyme in GSH synthesis. Increased GCLM expression enhances capacity for GSH synthesis. In agreement, Western blot analysis showed a significant increase in GCLM levels after 24 hours of 1,4-dihydroxy quininib (p = 0.022) and a slight, but not significant (p = 0.0607), increase after 8 hour-treatment. No significant changes were observed at 4 hours (Figure 7I,J). Therefore, the increase in GSH content after 8 and 24 hours may be related to an upregulation in GCLM expression, as a consequence of the prolonged treatments. Together these results suggest that 1,4-dihydroxy quininib activates the NRF2 / HO-1 axis in OMM2.5 cells after 4 and 8 hours of treatment, with a significant increase in the expression of phosphorylated NRF2 and HO-1 at 4 and 8 hours and in the amount of intracellular biliverdin following a short treatment (4 hours). In addition, 1,4-dihydroxy quininib alters redox homeostasis, leading to lipid peroxidation and a concomitant upregulation of GPX4 expression after 4 hours of treatment. Interestingly, lipid peroxidation was not present following the 24 hour-treatment, but a significant decrease in GPX4 expression and an accumulation of GSH were detected, highlighting how 1,4-dihydroxy quininib modulates specific ferroptosis hallmarks depending on the duration of treatment. Notably, a significant increase in HO-1 expression (p = 0.0167) and a significant decrease (p = 0.0102) in GPX4 expression were observed also in Mel285 cells, a primary UM cell line, supporting the ability of 1,4-dihydroxy quininib to modulate ferroptosis also in primary UM cells. CysLT1antagonists and CysLT2antagonist have overlapping and distinct effects on specific ferroptosis markers, while erastin reduces OMM2.5 cells metabolic activity. To investigate whether other CysLT receptor antagonists modulate ferroptosis in OMM2.5 cells, CysLT1 antagonists quininib and montelukast and CysLT2 antagonist HAMI 3379 were tested, and analysed GPX4 and GCLM expression levels (Figure 8A- D). 50 µM montelukast, 20 µM quininib and 50 µM HAMI 3379 did not significantly affect GCLM or GPX4 expression after 8 hours, similarly to 1,4-dihydroxy quininib (Figure 8A,B). Intriguingly, while 20 µM 1,4-dihydroxy quininib, 50 µM montelukast, 20 µM quininib and 50 µM HAMI 3379 all significantly reduced GPX4 expression (p = 0.0066, p = 0.0059, p = 0.00390, p = 0.0395, respectively) at 24 hours, only 20 µM 1,4- dihydroxy quininib significantly increased GCLM expression (p = 0.0496) (Figure 8A- D). These results suggest that 1,4-dihydroxy quininib modulates the GSH / GPX4 axis in OMM2.5 cells differently from other CysLT1 or CysLT2 antagonists, highlighting a unique mechanism of action in these cells. It was next evaluated whether erastin, established ferroptosis inducer, exerts anti- cancer effects on OMM2.5 cells (Figure 8E,F). Significant dose-dependent reductions of cells metabolic activity were observed following 96 hours of 10 μM (p = 0.0035), 20 μM (p = 0.0036) or 50 μM (p <0.0001) erastin treatment in comparison to vehicle control (Figure 8E), with a calculated IC50 of 5.243 μM (Figure 8F). 1,4-dihydroxy quininib significantly increases HO-1 and 4-HNE levels in tumours from orthotopic patient-derived xenograft MUM mouse models. To translate these in vitro findings into more clinically relevant UM tissues, the effects of 1,4-dihydroxy quininib on MUM OPDX (Figure 9) obtained from a HLA-A*02:01- positive female (UVM4) and a HLA-A*02:01-negative male (UVM7) patients (Figure 9A,B) were analysed. MUM OPDX tumour samples grown in the mice liver were dissected into explant pieces, arbitrarily renamed as left (L), middle (M) or right (R) (Figure 9A). Western blot data showed a significant upregulation of HO-1 expression (p = 0.0151) (Figure 9C,D), no significant difference in GPX4 expression (p = 0.4033) (Figure 9C,D) and a significant increase in 4-HNE levels (p = 0.0371) (Figure 9E,F) after 1,4-dihydroxy quininib treatment of MUM tissues from both UVM4 and UVM7. 4- HNE is a reactive aldehyde derived from the oxidative cleavage of lipid peroxides. Therefore, the data suggest that 1,4-dihydroxy quininib induces ferroptosis pathways in patient-derived MUM explants. High expression of genes inhibiting ferroptosis correlates with reduced OS and DFS in primary UM patients. To further interrogate the significance of ferroptotic markers modulated by 1,4- dihydroxy quininib in primary UM, their gene expression was analysed in 80 primary UMs from The Cancer Genome Atlas (TCGA). The markers selected were SLC3A2, HMOX1, GCLM, cystathionine gamma-lyase (CTH), NAD(P)H Quinone Dehydrogenase 1 (NQO1), ACSL3, IREB2, cytochrome P450 oxidoreductases (POR), and apoptosis inducing factor mitochondria associated 2 (AIFM2). These markers were chosen due to their up- or down-regulation after the 1,4-dihydroxy quininib treatment, as revealed by the OMM2.5 proteome-profiling (Figure 5, Figure 6), and to their link with the ferroptosis process. In addition, GPX4 and solute carrier family 7 member 11 (SLC7A11) transcripts were analysed. All the selected genes were expressed in the TCGA UM samples, confirming their potential disease relevance. As chromosome 3 status is an existing predictor of UM DFS and OS, the data was stratified based on whether patients presented with monosomy or disomy 3 and high or low expression of each selected marker gene. 3 is observed together with alterations in BRCA1 Associated Protein 1 (BAP1) in >80% of patients, therefore the data based on the presence / absence of BAP1 alterations (indicated as mutated / normal BAP1, respectively) were also stratified. The analysis revealed that GPX4 (p = 0.00476), SLC7A11 (p = 0.0169), HMOX1 (p = 0.00686), NQO1 (p = 3.82e-08), POR (p = 9.17e- 06) and AIFM2 (p = 6.81e-10) are significantly increased in primary UM samples from monosomy 3 patients compared to those with disomy 3. Conversely, disomy 3 primary UM patient samples present with significantly increased CTH (p = 1.02e-05), compared with those with monosomy 3 (Figure 10A). Significantly higher levels of GPX4 (p = 0.0275), NQO1 (p = 0.00363), POR (p = 0.0358) and AIFM2 (p = 0.011), are observed in primary UM patient samples presenting with mutated BAP1, compared to those with normal BAP1 (Figure 10B). A statistically significant difference in OS probability was detected between primary UM patient samples with monosomy 3 and high SLC7A11, compared to those with monosomy 3 and low SLC7A11 (p = 0.0083) (Figure 10C). In particular, high levels of SLC7A11 correlate with a decreased survival probability. In the primary UM cases with normal BAP1, it was observed that high levels of SLC7A11 (p = 0.00014), HMOX1 (p = 0.001), NQO1 (p < 0.0001), POR (p < 0.0001), AIFM2 (p < 0.0001) and low levels of CTH (p = 0.011) correlate with a significant decrease in survival probability (Figure 10D). We detected a significant difference in DFS between patients presenting chromosome 3 disomy and low AIFM2 (p = 0.0031) and those with chromosome 3 disomy and high AIFM2. In addition, patients with monosomy 3 and high SLC7A11 (p = 0.0017) or high CTH (p = 0.01) expression show a significantly decreased DFS when compared to patients with chromosome 3 monosomy and low SLC7A11 or low CTH expression (Figure 10E). In primary UM patient samples with normal BAP1, high levels of SLC7A11 (p = 0.013), ACSL3 (p = 0.0014), NQO1 (p = 0.00013), POR (p < 0.0001) and AIFM2 (p < 0.0001) correlate with a significant decrease in DFS. A significant difference in DFS was also observed between UM patients with mutated BAP1 and low SLC3A2 expression versus those with mutated BAP1 and high SLC3A2 expression (p = 0.042) (Figure 10F). The inventors hypothesised that combinations of these ferroptosis markers may produce more significant UM prognostication value. Therefore, they created a “ferroptosis signature”, the iFERR, and investigated its relevance in the TCGA plus other UM databases. These included GSE22138, with transcriptomic data of 63 primary UM enucleations untreated patients, (https: / / www.ncbi.nlm.nih.gov / geo / query / acc.cgi?acc=GSE22138); GSE27831, with ; (https: / / www.ncbi.nlm.nih.gov / geo / query / acc.cgi?acc=GSE84976). The iFERR SLC7A11, SLC3A2, was stratified based on whether patients presented with monosomy or disomy 3 and high or low expression of the signature. The results showed a significantly higher iFERR signature in monosomy 3 patients in both the GSE84976 (p = 0.00491) and TCGA (p = 0.0188) databases, suggesting that in those UM cohorts a high expression of genes inhibiting ferroptosis correlates with the chromosome alteration highly associated with disease relapse (Figure 11A). The correlation between the expression level of iFERR and OS in GSE84976 and TCGA databases were then analysed and a high iFERR signature was found to correlate with a significant decrease in OS (p = 0.0026, p = 0.03, respectively), once again indicating that increased levels of genes blocking ferroptosis correlate with a worse clinical outcome (Figure 11B). Finally, the correlation between iFERR and DFS were analysed in the 4 databases. Intriguingly, high iFERR also correlates with a significant decrease in DFS in GSE84976 (p = 0.0026) and TCGA (p = 0.0019) databases (Figure 11C). Collectively, these results suggest that monosomy 3 patients may present mechanisms involved in preventing ferroptosis and that high levels of genes involved in ferroptosis inhibition are associated with worse clinical outcome in UM patients. Expression of the iFERR5g gene signature correlates with reduced overall and disease-free survival in UM. To evaluate the potential of ferroptosis-related genes as biomarkers in UM, a novel ferroptosis signature, named iFERR5g, was developed, consisting of GPX4, SLC3A2, GCLM, NQO1, and AIFM2. The expression level of the iFERR5g signature was significantly associated with chromosome 3 status in the GSE22138, GSE84976 and TCGA-UVM datasets (Figure 13A). Additionally, iFERR5g expression was significantly associated with patient overall survival in the GSE84976 and TCGA-UVM datasets (Figure 13B) and with patient disease-free survival in the GSE27831, GSE84976 and TCGA-UVM datasets (Figure 13C). ROC curve analysis determined that the iFERR5g signature alone had an AUC of 0.91 predicting overall survival. Furthermore, combining the iFERR5g signature with chromosome 3 status increased the AUC to 0.94 (Figure 13D). Expression of the iFERR7g gene signature correlates with reduced disease-free survival in UM. To further assess the potential of ferroptosis-related genes as biomarkers in UM, a novel ferroptosis signature, named iFERR7g, was developed, consisting of GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3, and IREB2. The expression level of the iFERR5g signature was significantly associated with patient disease-free survival in the GSE22138 dataset (Figure 14). Expression of the IFERR4 gene signature is associated with patient overall survival, tumour stage, KRAS status and smoking status in lung cancer. To evaluate the potential of ferroptosis-related genes as biomarkers in lung cancer, genes were tested in combination for their association with overall survival in the TCGA-LUAD dataset. A signature, named IFERR4, was developed using 4 genes whose combined score showed the strongest association with overall survival: ACSL3, ATP5F1B, SLC3A2, and CYSLTR2 (Figure 15A). Expression of the IFERR4 signature was analysed in primary lung tumour tissue and matched normal tissue from 58 patients. The IFERR4 combined score was significantly higher in primary tumour tissue compared to matched normal tissue (Figure 15B). Next, expression of the IFERR4 signature was analysed in tissue from tumours of different stages: T1 (n = 169), T2 (n = 278), T3 (n = 47) and T4 (n = 19). The IFERR4 combined score was significantly higher in T2 and T3 tumours compared to T1 tumours (Figure 15C). Expression of the IFERR4 signature was also analysed in samples from patients with KRAS mutations (n = 155) versus wild-type KRAS (n = 356). The IFERR4 score was higher in patients with KRAS mutations (Figure 15D) and showed a stronger association with overall survival in patients with KRAS mutations compared to those with wild-type KRAS (Figure 15E). Finally, expression of the IFERR4 gene signature was analysed in samples from smokers (n = 118) and non-smokers (n = 101). While no significant difference in the combined score was observed between smokers and non-smokers (Figure 15F), the signature showed a stronger association with overall survival in smokers compared to non-smokers (Figure 15G). Discussion Lung, pancreatic, ovarian cancers and cutaneous melanoma are challenging diseases, which every year cause a significant number of diagnoses and deaths. Ferroptosis can be a reliable prognostic marker for these cancer types, by examining specific ferroptosis-related gene signatures and their correlation with patient survival. Here, we identified several ferroptosis-associated genes which correlate with lung, pancreatic, ovarian or cutaneous melanoma patient survival or disease recurrence, by analysing the TCGA database. These findings can advance the knowledge and expedite the development of more effective treatments and diagnostic tools for these devastating cancers. In UM, metastases are associated with poor prognosis, ineffective treatments and death within 12 months. There is an unmet need for better biomarkers for MUM prognosis plus more effective and affordable treatments. Here, ferroptosis was identified as a mechanism of the anti-MUM effects mediated by 1,4-dihydroxy quininib and central to iFERR, a novel biomarker signature for prognostication of MUM. Previously, small molecule drugs preferentially targeting CysLT1 over CysLT2 exerted anti-UM phenotypes. Quininib, a CysLT1 antagonist, is an anti-inflammatory and anti- angiogenic molecule with anti-cancer activity in human ex vivo colorectal cancer patient tumour explants and colorectal cancer xenograft models. Quininib and its analogue 1,4- dihydroxy quininib significantly inhibit survival, long-term proliferation and oxidative phosphorylation in primary and MUM cell lines. Furthermore, 1,4-dihydroxy quininib alters the secretion of inflammatory factors in primary UM samples and decreases the expression of an oxidative phosphorylation marker in a cell line-derived mouse orthotopic xenograft model of MUM. However, the fundamental molecular mechanism of action of quininib drugs in these suite of models was poorly understood. Ferroptosis is a new, druggable target for MUM. Modulation of ferroptosis can inhibit cancer cell growth, improve the sensitivity of chemotherapy and radiotherapy and expand treatment options for different cancer types. Thus the evidence reported here linking, for the first time, 1,4-dihydroxy quininib with ferroptosis modulation in UM is original and significant, since it provides a potential opportunity to fundamentally change the clinical management of this rare and aggressive cancer type. OMM2.5 cells treated with 1,4- for 8 and 24 hours showed increased levels of key ferroptosis and anti-oxidant modulators e.g. HO-1, GCLM, oxidative stress induced growth inhibitor 1 (OSGIN1) and thioredoxin (TXN) (Figure 5, 6). These are regulated by NRF2 and oxidative stress. In agreement, 1,4-dihydroxy quininib significantly increased the levels of ROS and phosphorylated NRF2 after the 8 hour- treatment (Figure 7B, C). HO-1, one of the main targets of NRF2, is the first rate- limiting enzyme in the conversion of heme into ferrous ions, carbon monoxide, and biliverdin. HO-1 can act either in a cytoprotective or cytotoxic manner depending on specific cellular conditions. In particular, HO-1 overexpression is associated with malignant cancer cell growth, proliferation and invasion, but its induction can also reduce tumour cell proliferation. Indeed, increased HO-1 activity can promote the accumulation of the intracellular labile iron pool (LIP) essential for lipid peroxidation and ferroptosis. LIP is a cofactor for enzymes producing ROS, such as nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOXs), POR, nitric oxide synthases and lipoxygenases. Notably, it was observed by the inventors that POR expression was significantly increased after the 8-hour 1,4-dihydroxy quininib treatment. POR, by transferring electrons from NAD(P)H to oxygen, generates hydrogen peroxide, which reacts with iron generating reactive hydroxyl radicals for peroxidation of PUFA chains in membrane phospholipids, thereby disrupting membrane integrity during ferroptosis. 1,4-dihydroxy quininib increased HO-1 enzymatic activity and downstream ferroptosis in OMM2.5 cells based on biliverdin content and LOOH levels (Figure 7D,H). Intriguingly, intracellular biliverdin significantly increased after the 4-hour treatment and higher LOOH levels were observed after the 4- and 8-hour treatment. In parallel, at 8 hpt IREB2 levels decreased (Table B and Figure 6D,E). IREB2 is degraded when intracellular iron is excessive, thus, indirectly confirming accumulation of cytosolic Fe2+in OMM2.5 cells. Collectively, these data indicate that the 4- and 8-hour 1,4-dihydroxy quininib treatment induces ferroptosis in OMM2.5 cells through the NRF2 / HO-1 axis. GPX4 is a fundamental anti-oxidant system, which blocks lipid peroxidation. 1,4- dihydroxy quininib significantly increased GPX4 levels after 4 hours, but not after 8 hours (Figure 7E,F). This may reflect the cells trying to escape ferroptosis by increasing lipid repair capacity via activation of the xc- system. This is further corroborated by increased GSH content and GCLM expression (Figure 5E;7G,I). Indeed, GSH is a cofactor and synthetic for GPX4 and derives from cystine, which is imported intracellularly through the xc- system. At 24 hours, phosphorylated NRF2, biliverdin content and LOOH did not change after 1,4-dihydroxy quininib exposure (Figure 7B,C,D,H). However, the inventors observed a significant decrease in GPX4 expression, therefore, the core defence mechanism against ferroptosis is affected by the longest treatment duration (Figure 7E,F). Intriguingly, we detected a significantly increase in GSH content and GCLM expression (Figure 7G), suggesting again that OMM2.5 cells are adopting an enhanced antioxidant capacity. CysLT1 antagonists quininib and montelukast and CysLT2 antagonist HAMI 3379 did reduce GPX4 but did not affect GCLM expression at 24 hours (Figure 8C,D), suggesting that the 1,4-dihydroxy quininib mechanism of action in OMM2.5 cells differs from those of the other CysLT1 / 2antagonists. Human SH-SY5Y neuroblastoma cells activate an adaptive response of the GSH system to prolonged iron loads through increased expression of GCLC and the GCLM subunits of GCL (56). Therefore, our results suggest that 24-hour 1,4-dihydroxy quininib treatment may promote ferroptosis by specifically inhibiting GPX4, similarly to observations with RSL3, which inhibits GPX4 directly. Altogether, our evidence suggests a dynamic effect of 1,4-dihydroxy quininib on ferroptosis in OMM2.5 cells, potentially due to transient activity or upregulation of compensatory resistance pathways (Figure 12). Importantly, the inventors demonstrated that 1,4-dihydroxy quininib increases HO-1 and 4-HNE levels in tumours explants from MUM OPDX mouse models. These are models in which human MUM is orthotopically implanted in the liver, thus they recapitulate the genetic heterogeneity of the original MUM patients’ tumours. 4-HNE is a toxic product deriving from the oxidative cleavage of lipid peroxides, therefore it increases during ferroptosis. The evidence obtained with MUM tumour explants confirms the significant role of 1,4- dihydroxy quininib in modulating ferroptosis in more clinically relevant models of MUM. Indeed, tumour biopsies better recapitulate the physiology of the tumour microenvironment, allowing cell-to-cell contact and cell-to-matrix synthesis as well as the development of the oxygen, nutrient, and hormone levels typically found in patient’s tumours. At the same time, the upregulation of HO-1 and lipid peroxidation, observed both in OMM2.5 cells and in MUM tumour biopsies, validate OMM2.5 cells as a good model where to investigate MUM biology and carry out drug screening. The quininib series of drugs antagonise CysLT1 receptors. In cell-based assays, the salt form of 1,4 dihydroxy quininib exerted its greatest antagonistic potency at CysLT1, with moderate antagonism of CysLT2, and inhibited VEGFR2 and VEGF3 in reconstituted kinase assays. Considering CysLT1 as the prototypical target, a possible explanation for 1,4 dihydroxy quininib inducing ferroptosis is via modulation of a STAT3 / GSK-3β / β-catenin / GPX4 pathway. In mouse embryonic stem cells, CysLT1signalling promoted STAT3 and GSK-3β phosphorylation but inhibited β-catenin phosphorylation. CysLT signalling ultimately led to the activation of β-catenin in intestinal epithelial cells and in colon cancer cells. Intriguingly, beta-catenin conferred ferroptosis resistance in gastric cancer by binding to the promoter region of GPX4 and inducing its expression. This highlighted how aberrant Wnt / β-catenin activation led to ferroptosis resistance, suggesting a therapeutic option to enhance chemo-sensitivity for advanced gastric cancer patients. In gastric cancer, STAT3 directly bound to the promoters of negative ferroptosis regulators (GPX4, SLC7A11, and FTH1) and modulated their expression. Genetic ablation of STAT3 activity triggered ferroptosis through lipid peroxidation and Fe2+accumulation in gastric cancer cells. Thus, inhibition of the STAT3-ferroptosis inhibitory axis reduced tumour growth and alleviated chemoresistance. Further proof of ferroptosis induction by impairing STAT3 / NRF2 / GPX4 signalling is observed in osteosarcoma cells, where STAT3 inhibitors reactivated ferroptosis and increased cisplatin sensitivity. Overall, CysLT1antagonism may lead to a downregulation in STAT3 and GSK-3β phosphorylation, reducing β-catenin activation and decreasing GPX4 expression (e.g. after a 24 hours of 1,4-dihydroxy quininib treatment) or promoting Fe2+accumulation and lipid peroxidation (e.g. after the 4- and 8- hour 1,4-dihydroxy quininib treatment). The timely activation of the different components in the STAT3 / GSK-3β / β- Catenin / GPX4 pathway may explain the observed dynamic effect of 1,4-dihydroxy quininib, and further evidence will be fundamental to understand the specific antioxidant programs used by MUM cells. This will be key to uncover potential resistance mechanisms and thus extend drug efficacy. Importantly, ferroptosis induction-based treatment can also be combined with immune checkpoint blockade options. Upregulation of CysLT1, which enhances Wnt / β-catenin signalling, led to endogenous and IFNγ-induced PD-L1 expression in colorectal cancer cells. Inhibiting CysLT1 reduced both Wnt / β-catenin and PD-L1 expression, thus elucidating the importance of targeting CysLT1 to reach beneficial outcomes of immune checkpoint blockade in colorectal cancer patients. Immunotherapy in combination with ferroptosis induction represents a promising option, since the two treatment modalities mutually potentiate each other, triggering synergistic anticancer effects. Here, several ferroptosis associated-genes which correlate with UM patient survival are reported, by analysing the TCGA database and stratifying the data based on chromosome 3 and BAP1 status. Furthermore, a novel ferroptosis signature (iFERR) including negative regulators of ferroptosis (GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3, IREB2, NQO1 and AIFM2) was identified by the inventors, which can predict OS and DFS. In particular, a novel gene signature (iFERR5) comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2 significantly correlates with worse outcome in patients with uveal melanoma. Furthermore, a bespoke gene signature (iFERR9) comprising GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3, IREB2, NQO1 and AIFM2 correlates with worse outcome in patients with uveal melanoma. In lung cancer, a novel gene signature (iFERR4) comprising ACSL3, ATP5F1B, SLC3A2 and CYSLTR2, significantly correlates with worse outcomes. In particular, an increased iFERR signature, thus increased levels of genes involved in ferroptosis inhibition, correlates with a worse clinical outcome. Importantly, the reliability of this signature was validated in independent cohorts. Thus, the iFERR signature displays potential as a biomarker for determining lung, pancreatic, ovarian or cutaneous melanoma cancer patient survival or disease recurrence, as well as UM prognostication and treatment. Overall, this study elucidated for the first time the role of 1,4-dihydroxy quininib in modulating ferroptosis hallmarks in MUM samples and revealed a novel ferroptosis gene signature to predict UM patient outcomes. This evidence justifies additional translational studies in clinically relevant in vivo and ex vivo UM models, aimed to improve the clinical management and treatment of this challenging cancer type. In the specification the terms "comprise, comprises, comprised and comprising" or any variation thereof and the terms “include, includes, included and including" or any variation thereof are considered to be totally interchangeable and they should all be afforded the widest possible interpretation and vice versa. The invention is not limited to the embodiments hereinbefore described but may be varied in both construction and detail.

Claims

Claims 1. A method for predicting disease free status or overall survival in an individual diagnosed with uveal melanoma, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or the proteins encoded by said genes, assayed from a biological sample from a subject with no cancer, the individual is predicted to have an increased probability of having disease free status or overall survival.

2. The method of Claim 1, wherein the gene signature consists of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or of proteins encoded by said genes; wherein when the expression level of the gene signature consisting of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature consisting of GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2, or the proteins encoded by said genes, assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease-free status or overall survival.

3. A method for predicting disease free status or overall survival in an individual diagnosed with uveal melanoma, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, CTH, ACSL3, IREB2, and SLC7A11, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3 and IREB2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC7A11, SLC3A2, GCLM, CTH, ACSL3 and IREB2 or the proteins encoded by said genes, assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease- free status or overall survival.

4. A method of predicting of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) mutation status of Bap-1 in the individual or (b) expression status of Bap-1 in the individual; wherein when the individual has one or more of (a) a wildtype Bap-1 sequence or (b) normal Bap-1 expression levels, relative to a reference value for (a)-(b) from a biological sample from a subject with no metastatic uveal melanoma, and wherein when the expression levels of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value for gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma.

5. The method of Claim 4, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma.

6. A method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) Bap-1 mutation status in the individual or (b) the expression status of Bap-1 in the individual; wherein when the individual has at least one of (a) the presence of mutated Bap-1 or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no metastatic uveal melanoma, and the expression levels of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have an increased risk of developing metastatic uveal melanoma.

7. The method of Claim 6, further the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have an increased risk of developing metastatic uveal melanoma.

8. A method of determining a 5-year survival rate or a 10-year survival rate of an individual diagnosed with an uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said gene, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) a wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have an increased 5-year survival rate or 10- year survival rate.

9. The method of Claim 8, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate.

10. The method of Claim 8, wherein when the individual has at least one of (a) a mutated Bap-1 status or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from a subject who survived for 5 or 10 yearsfollowing having an uveal melanoma the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate.

11. The method of Claim 10, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate.

12. A method of identifying an uveal melanoma patient that is suitable for treatment with a therapy for preventing recurrence or progression of the uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of a gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) mutated Bap-1 status or (b) decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, is dysregulated relative to a reference value of the gene signature comprising GPX4, SLC3A2, GCLM, NQO1 and AIFM2, or a protein encoded by said genes, assayed from a biological sample from the individual following administration of a treatment, the individual is predicted not to be suitable for the treatment.

13. The method of Claim 12, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to not to be suitable for the treatment.

14. A method for predicting overall survival in an individual diagnosed with lung cancer, the method comprising a step of assaying a biological sample from the individual for expression level of a gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CYSLTR2, or of proteins encoded by said genes; wherein when the expression level of the gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CYSLTR2, or of the proteins encoded by said genes, is dysregulated relative to a reference value for the gene signature comprising ACSL3, ATP5F1B, SLC3A2 and CYSLTR2, or the proteins encoded by said genes, assayed from a biological samplefrom the subject with no cancer, the is predicted to have a reduced chance of overall survival.

15. The method of Claim 14, wherein when the individual has a status of being a smoker or a former smoker, the individual is predicted to have a reduced chance of overall survival.

16. The method of Claim 14 or Claim 15, further comprising the step of determining KRAS mutation status of the individual, wherein when the individual has a positive KRAS mutation status, the individual is predicted to have reduced overall survival.

17. A method for predicting disease free status or overall survival in an individual diagnosed with cancer, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no cancer, the individual is predicted to have an increased probability of having disease free status and having an increased chance of overall survival.

18. A method for predicting a probability of having recurrent disease or having a reduced overall survival in an individual diagnosed with cancer, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH; wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1,AIFM2, HMOX1, POR, CysLT1, CTH assayed from a biological sample from the subject with cancer, the individual is predicted to have an increased probability of having recurrent disease and having a reduced chance of overall survival.

19. The method of Claim 17 or Claim 18, wherein the at least one gene, or a protein encoded by said gene, is ACSL3 and the cancer is lung cancer.

20. The method of Claim 17, Claim 18 or Claim 19, wherein if the at least one gene, or a protein encoded by said gene, is ACSL3 and SLC3A2, the individual has lung cancer, and the expression of ACSL3, ATP5B, and / or SLC3A2 in the sample relative to the reference value is decreased, the individual is predicted to have a lower chance of having recurrent disease status, a higher chance of disease free status and a higher chance of overall survival.

21. The method of one of Claims 17 to 19, wherein if the at least one gene, or a protein encoded by said gene, is ACSL3, ATP5B, and / or SLC3A2, the individual has lung cancer, and the expression of ACSL3 and SLC3A2 in the sample relative to the reference value is increased, the individual is predicted to have a recurrent disease status and a lower chance of overall survival.

22. The method of Claim 17 or Claim 18, wherein when the at least one gene, or a protein encoded by said gene, is CysLT1or CysLT2, and the individual has lung cancer, and the expression levels of CysLT1 or CysLT2 are increased relative to the reference value, the individual is predicted to have a higher chance of overall survival.

23. The method of Claim 17 or Claim 18, wherein when the at least one gene, or a protein encoded by said gene, is CysLT1 or CysLT2, and the individual has lung cancer, and the expression levels of CysLT1 or CysLT2 are decreased relative to the reference value, the individual is predicted to have a decreased or lower chance of overall survival.

24. The method of Claim 17 or Claim 18, wherein if the at least one gene, or a protein encoded by said gene, is IREB2, and the individual has ovarian cancer, and the expression levels of IREB2 are increased relative to the reference value, the individual is predicted to have lower risk of a recurrent disease status, and a higher chance of a disease free status.

25. The method of Claim 17 or 18, wherein if the at least one gene, or a protein encoded by said gene, is IREB2, and the individual has ovarian cancer, and the expression levels of IREB2 are decreased relative to the reference value, the individual is predicted to have recurrent disease status, and a lower chance of disease free status.

26. The method of Claim 17 or Claim 18, wherein if the at least one gene, or a protein encoded by said gene, is CTH, the individual has pancreatic cancer, and the expression level of CTH is decreased relative to the reference value, the individual is predicted to have a lower chance of having a recurrent disease status, and a higher chance of having a disease free status and predicted to have a greater likelihood of overall survival.

27. The method of Claim 17 or Claim 18, wherein if the at least one gene, or a protein encoded by said gene, is CTH, the individual has pancreatic cancer, and the expression level of CTH is increased relative to the reference value, the individual is predicted to have a higher chance of a recurrent disease status, and a lower chance of disease free status and a lower likelihood of overall survival.

28. The method of Claim 17 or Claim 18, wherein if the at least one gene, or a protein encoded by said gene, is IREB2, the individual has a cutaneous melanoma, and the expression levels of IREB2 are decreased relative to the reference value, the individual is predicted to have a low chance of a recurrent disease status, and a higher chance of a disease free status; and wherein if the expression levels of IREB2 are increased relative to the reference value, the individual is predicted to have a recurrent disease status, and a lower chance of a disease free status.

29. The method of Claim 17 or Claim 18, wherein if the at least one gene, or a protein encoded by said gene, is ACSL3, the individual has a cutaneous melanoma, and the expression levels of ACSL3 are increased relative to the reference value, the individual is predicted to have a greater likelihood of overall survival; and wherein if the expression levels of ACSL3 are decreased relative to the reference value, the individual is predicted to have a lower chance of overall survival.

30. A method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene,selected from ATP5B, GPX4, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2 and CTH, in combination with determining at least one of (a) mutation status of Bap-1 in the individual or (b) expression status of Bap-1 in the individual; wherein when the individual has one or more of (a) a wildtype Bap-1 sequence or (b) normal Bap-1 expression levels, relative to a reference value for (a)-(b) from a biological sample from a subject with no metastatic uveal melanoma, and wherein when the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the subject with no metastatic uveal melanoma, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma.

31. The method of Claim 30, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have a reduced risk of developing metastatic uveal melanoma.

32. A method of predicting occurrence of metastatic uveal melanoma in an individual, the method comprising a step of assaying a biological sample from the individual for expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status in the individual or (b) the expression status of Bap-1 in the individual; wherein when the individual has at least one of (a) the presence of mutated Bap-1 or (b) has decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no metastatic uveal melanoma, and the expression levels of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value for at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1,CysLT2, and CTH assayed from a sample from the subject with no metastatic uveal melanoma, the individual is predicted to have an increased risk of developing metastatic uveal melanoma.

33. The method of Claim 32, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have an increased risk of developing metastatic uveal melanoma.

34. A method of determining a 5-year survival rate or a 10-year survival rate of an individual diagnosed with an uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) a wild-type Bap-1 status or (b) normal Bap-1 expression levels relative to a reference value from a biological sample from a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate.

35. The method of Claim 34, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a disomy status for chromosome 3, the individual is predicted to have an increased 5-year survival rate or 10-year survival rate.

36. The method of Claim 34, wherein when the individual has at least one of (a) a mutated Bap-1 status or (b) has decreased Bap-1 expression levels relative to areference value from a biological a subject who survived for 5 or 10 years following an uveal melanoma diagnosis, and the expression level of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from a subject who survived for 5 or 10 years following having an uveal melanoma diagnosis, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate.

37. The method of Claim 36, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomy status for chromosome 3, the individual is predicted to have a decreased 5-year survival rate or 10-year survival rate.

38. A method of identifying an uveal melanoma patient that is suitable for treatment with a therapy for preventing recurrence or progression of the uveal melanoma, the method comprising: assaying a biological sample from the individual for expression of at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2and CTH, in combination with determining at least one of (a) Bap-1 mutation status or (b) Bap-1 expression status of the individual; wherein when the individual has at least one of (a) mutated Bap-1 status or (b) decreased Bap-1 expression levels relative to a reference value from a biological sample from a subject with no uveal melanoma, and the expression levels of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH is dysregulated relative to a reference value of the at least one gene, or a protein encoded by said gene, selected from ATP5B, GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, CysLT1, CysLT2, and CTH assayed from a biological sample from the individual following administration of a treatment, the individual is predicted not to be suitable for the treatment.

39. The method of Claim 38, further comprising the step of determining chromosome 3 status of the individual, wherein when the individual has a monosomystatus for chromosome 3, the is predicted to not to be suitable for the treatment.

40. The method according to any one of Claims 17 to 39, wherein the at least one gene, or a protein encoded by said gene, selected is ATP5B.

41. The method according to any one of Claims 17 to 40, wherein at least two genes, or proteins encoded by said genes, are selected, and are selected from ATP5B and GPX4, ATP5B and SLC7A11, ATPB5 and IREB2, ATP5B and CTH, GPX4 and SLC7A11, GPX4 and IREB2, GPX4 and CTH, SLC7A11 and IREB2, SLC7A11 and CTH, or IREB2 and CTH.

42. The method according to any one of Claims 17 to 41, wherein at least three genes, or proteins encoded by said genes, are selected, and are selected from ATP5B, GPX4, and SLC7A11; ATP5B, GPX4, and IREB2; ATP5B, GPX4, and CTH; ATP5B, SLC7A11, and IREB2; ATP5B, SLC7A11, and CTH; ATP5B, IREB2, and CTH; GPX4, SLC7A11, and IREB2; GPX4, SLC7A11, and CTH; or SLC7A11, IREB2, and CTH.

43. The method according to any one of Claims 17 to 42, wherein at least four genes, or proteins encoded by said genes, are selected and are selected from ATP5B, GPX4, SLC7A11, and IREB2; ATP5B, GPX4, SLC7A11, CTH; or GPX4, SLC7A11, IREB2, and CTH.

44. The method according to any one of Claims 17 to 43, wherein at least five genes, or proteins encoded by said genes, are selected and are selected from ATP5B, GPX4, SLC7A11, IREB2, and CTH; GPX4, SLC3A2, GCLM, NQO1, and AIFM2; CysLT1, CysLT2, ATP5B, ACSL3 and SLC3A2; and GPX4, SLC3A2, GCLM, NQO1 and AIFM2.

45. The method according to any one of Claims 17 to 44, wherein at least seven genes, or proteins encoded by said genes, are selected, and are GPX4, SLC3A2, GCLM, CTH, ACSL3, IREB2, and SLC7A11.

46. The method according to any one of Claims 17 to 45, wherein at least nine genes, or proteins encoded by said genes, are selected, and are GPX4, SLC3A2, GCLM, NQO1, AIFM2, SLC7A11, CTH, ACSL3, and IREB2.

47. The method according to any of Claims 17 to 46, wherein at least eleven genes, or proteins encoded by said genes, are selected and are GPX4, SLC7A11, IREB2, SLC3A2, GCLM, ACSL3, NQO1, AIFM2, HMOX1, POR, and CTH.

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