1. Patent Search
  2. Details

METHODS FOR DETECTING OVARIAN CANCER

20210405054 · 2021-12-30

    Inventors

    Cpc classification

    International classification

    Abstract

    The present disclosure is related to the field of ovarian cancer diagnostics. It introduces novel biomarkers that can be used to detect presence of ovarian cancer and to provide a prognosis of the disease.

    Claims

    1. An in vitro screening method for assessing whether a subject is at risk to develop or is suffering from ovarian cancer comprising: (a) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group A; (b) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group B; and (c) determining that the subject is suffering from or is at an increased risk of developing ovarian cancer, if said sample contains an increased concentration of the at least one small molecule biomarker from Group A and a decreased concentration of the at least one small molecule biomarker from Group B, when compared to a control.

    2. The method of claim 1, wherein the method further comprises determining from said sample a concentration of at least one protein biomarker, wherein an increase or decrease in the concentration of said protein biomarker(s), when compared to a control, is indicative of said subject suffering from or having an increased risk of developing ovarian cancer.

    3.-4. (canceled)

    5. A method of assessing whether a subject has a decreased or poor survival prognosis for ovarian cancer comprising: (a) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group A; (b) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group B; and (c) determining that the subject has a decreased or poor survival prognosis, if said sample contains an increased concentration of the at least one small molecule biomarker from Group A and a decreased concentration of the at least one small molecule biomarker from Group B, when compared to a control.

    6. The method of claim 5, wherein the method further comprises determining from said sample a concentration of at least one protein biomarker, wherein an increase or decrease in concentration of said protein biomarker(s), when compared to a control, is indicative of said subject having a decreased or poor survival prognosis.

    7.-8. (canceled)

    9. An in vitro method of evaluating the effectiveness of ovarian cancer therapy in a subject comprising: (a) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group A; (b) assaying a sample from said subject to determine a concentration of at least one small molecule biomarker from Group B; and (c) determining that the therapy is effective, if said sample contains a decreased concentration of the at least one small molecule biomarker from Group A and an increased concentration of the at least one small molecule biomarker from Group B, when compared to a control.

    10. The method of claim 9, wherein the method further comprises determining from said sample a concentration of at least one protein biomarker, wherein an increase or decrease in concentration of said protein biomarker(s), when compared to a control, is indicative of the effectiveness of the therapy.

    11.-46. (canceled)

    47. The method of claim 1, wherein the method comprises determining the concentration of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 or more small molecule biomarkers from Group A and at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 or more small molecule biomarkers from Group B.

    48. The method of claim 1, wherein the small molecule biomarker concentration is determined by using mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy or dual polarization interferometry, an immunoassay, enzymatic assay, colorimetric assay, fluorometric assay, a rapid test and/or with a binding moiety capable of specifically binding the biomarker.

    49. (canceled)

    50. The method of claim 1, wherein the sample is a serum sample or a plasma sample.

    51. The method of claim 1, further comprising a step of spiking the sample with at least one isotope-labelled small molecule biomarker from Group A and/or at least one isotope-labelled small molecule biomarker from Group B.

    52. The method of claim 51, wherein the at least one isotope-labelled small molecule biomarker from Group A and/or at least one isotope-labelled small molecule biomarker from Group B is/are deuterium-labelled.

    53. The method of claim 1, wherein the ovarian cancer is early stage ovarian cancer.

    54. The method of claim 1, wherein the method further comprises after the determining step (c), (d) diagnosing the subject as suffering from or having an increased risk of developing ovarian cancer from the results in step (c), and (e) administering a treatment to the subject diagnosed in step (d).

    55.-57. (canceled)

    58. The method of claim 5, wherein the method further comprises after the determining step (c), (d) administering a treatment to the subject.

    59.-61. (canceled)

    62. The method of claim 9, wherein the method further comprises (d) determining that the therapy is not effective in the subject from the results in step (c), and (e) escalating the therapy of the subject.

    63. (canceled)

    64. The method of claim 54, wherein the treatment is a surgery, chemotherapy, radiation therapy, hormonal therapy, anti-angiogenic therapy, therapies targeting homologous recombination deficiency, antibody therapy, other targeted therapy utilizing ovarian cancer specific signalling pathways, or any therapeutic treatment or operation typically given to a subject having ovarian cancer; or the treatment comprises administering a pharmaceutical agent affecting lipid metabolism.

    65. (canceled)

    66. The method of claim 54, wherein the method further comprises requesting a test from a laboratory which provides the results of an assay useful for determining the concentration of the at least one small molecule biomarker from Group A and the at least one small molecule biomarker from Group B, and administering to the subject a treatment if the subject has an increased concentration of the at least one small molecule biomarker from Group A and a decreased concentration of the at least one small molecule biomarker from Group B, as compared to the control.

    67.-69. (canceled)

    70. The method of claim 2, wherein the at least one protein biomarker is CA-125 and/or HE4, and wherein an increase in the concentration of said protein biomarker(s), when compared to a control, is indicative of said subject suffering from or having an increased risk of developing ovarian cancer.

    71.-73. (canceled)

    74. The method of claim 1, wherein the at least one small molecule biomarker from Group A and/or the at least one small molecule biomarker from Group B is(are) selected from the small molecule biomarkers listed in Table 2, Table 3 and/or Table 4.

    75.-76. (canceled)

    77. The method of claim 1, wherein the subject is a premenopausal subject.

    Description

    DETAILED DESCRIPTION OF THE INVENTION

    [0174] Reference will now be made in detail to various exemplary embodiments, examples of which are discussed in the detailed description that follows. It is to be understood that the following detailed description is provided to give the reader a fuller understanding of certain embodiments, features, and details of aspects of the invention, and should not be interpreted as limiting the scope of the invention.

    Abbreviations

    [0175] Unless indicated otherwise, the abbreviations used in this description have the following meanings: ApoA1—apolipoprotein A1, AUC—area under the curve, BSTFA—N,O-bis(trimethylsilyl)trifluoroacetamide, CA-125—cancer antigen 125, carcinoma antigen 125, carbohydrate antigen 125, CE—cholesterylester, Cer—ceramide, CI—confidence interval, CRP—C-reactive protein, DAG—diacylglycerol, EI—electron ionization, FA—fatty acid, FDA—US Food and Drug Administration, FSH—follicle-stimulating hormone, Gb3 globotriasoylceramide, GC—gas chromatography, GC×GC—two dimensional gas chromatography, GIc/GalCer—glucosyl/galactosylceramide, HE4—human epididymic protein 4, HPLC—high performance liquid chromatography, HR—hazard ratio, IS—internal standard, LacCer—lactosylceramide, LC—liquid chromatography, LPC—lyosphosphatidylcholine, LPC-O—ether-linked (alkyl) lysophosphatidylcholine, LPE—lysophosphatidylethanolamine, LPL—lysophospholipid, MRM—multiple reaction monitoring, MS—mass spectrometry, NMR—nuclear magnetic resonance, PC—phosphatidylcholine, PC O—ether-linked (alkyl) phosphatidylcholine, PC P—ether-linked (alkenyl) phosphatidylcholine, PE—phosphatidylethanolamine, PE O—ether-linked (alkyl) phosphatidylethanolamine, PE P—ether-linked (alkenyl) phosphatidylethanolamine, PG—phosphatidylglycerol, PI—phosphatidylinositol, ROC—receiver operating characteristic, S1P—sphingosine-1-phosphate, SA1P—sphinganine-1-phosphate, SE—sensitivity, SIM—selected ion monitoring, sMRM—scheduled multiple reaction monitoring, SP—specificity, SM—sphingomyelin, TAG—triacylglycerol, TMS—trimethylsilyl, TOF—time-of-flight, TRF—transferrin, UHPLC—ultra-high performance liquid chromatography, WHO—World Health Organization.

    Definitions

    [0176] In order that the present disclosure may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the present description.

    [0177] As used herein, “ovarian cancer” is a malignant tumor of the ovary. There are several types of ovarian cancers such as malignant serous tumors (low or high grade), mucinous tumors, endometrioid tumors, clear cell tumors, transitional cell tumors, epithelial-stromal tumors, adenosarcomas, carcinosarcomas, granulosa tumors, Sertoli-Leydig tumors, germ cell tumors such as teratomas and mixed germ cell tumors, unclassified tumors, metastatic tumors from nonovarian origin and also tumors with borderline malignancy.

    [0178] The terms “subject,” “host,” “patient,” and “individual” are used interchangeably herein to refer to any mammalian subject for whom diagnosis or therapy is desired, particularly humans. The subject may have previously suffered from ovarian cancer or the subject may be a healthy individual with no previous signs or symptoms of ovarian cancer. The subject may be a premenopausal or postmenopausal individual.

    [0179] As used in this description, a “small molecule biomarker” relates to the small molecule biomarkers shown in Table 2. In some embodiments, a “small molecule biomarker” relates to the small molecule biomarkers shown in Table 3 or 4.

    [0180] As used herein, a “small molecule biomarker from Group A” and a “small molecule biomarker from Group B” relate to the small molecule groups shown in Table 1.

    [0181] As used herein, a “lipid biomarker” relates to the lipid biomarkers shown in Table 4.

    [0182] As used herein, a “protein biomarker” relate to any protein used as a biomarker for ovarian cancer, such as cancer antigen 125 (CA-125), human epididymal protein-4 (HE4), prealbumin, apolipoprotein A-1 (ApoA1), beta-2-microglobulin, transferrin (TRF), follicle-stimulating hormone (FSH) and C-reactive protein (CRP).

    [0183] As used herein, a “sample” is a biological sample obtained from a subject or a group or population of subjects. The sample may be a blood sample, a serum sample, a plasma sample, a saliva sample, an urine sample or a fraction thereof. Blood serum and plasma samples are typical. The sample can be prepared with techniques well known in the art. In certain embodiments, the blood sample is a blood spot dried on a filter. Alternatively, both the sample from the subject and the control sample may also be tissue samples, e.g., ovarian tissue sample, or an ovarian cyst fluid sample.

    [0184] As used herein, a “control” may be a control sample. A control may also be a concentration determined from a sample from a single healthy individual or a subject with benign tumor or other medical condition causing similar symptoms to ovarian cancer, or the same subject before developing malignant tissue. The control may also be a sample that represents a combination of samples from a generalized population of healthy individuals. Alternatively, the control may be a control value or a set of data concerning the biomarker in a sample previously determined, calculated or extrapolated, or may have yet to be determined, calculated or extrapolated, or may also be taken from the literature.

    [0185] A control as used herein, i.e., a control value or a control sample, is typically representative of a group of subjects or a population of subjects. In this context, “representative” means that the biomarker concentration(s) reflected by said control value to which a comparison is made in the context of the present disclosure correspond(s) to the average concentration value(s) of said biomarker concentration(s) in corresponding individual samples from the subjects of said group or population. Likewise, in the case of a control sample “representative” means that the biomarker concentration(s) in said control sample to which a comparison is made in the context of the present disclosure correspond(s) to the average concentration(s) of said biomarker concentration(s) in corresponding individual samples from the subjects of said group or population. Typically, the concentrations of all biomarker concentrations in said control sample correspond to the average concentrations of said biomarker concentrations in corresponding individual samples from the subjects of said group or population. An individual with such values can be considered a “healthy individual” for the purposes of the present disclosure.

    [0186] A control sample can be particularly suitably compared to the subject's sample if it has been obtained from the same type of biological tissue or source in the same, or essentially the same, manner. For example, if the subject's sample is a serum sample or a plasma sample, a corresponding control sample will likewise be a serum sample or a plasma sample, respectively.

    [0187] It will be appreciated that a useful control value for the purposes of the present disclosure is typically one that has been, or is, obtained using any one of the suitable control samples described herein.

    [0188] As used herein, in the small molecule nomenclature X:Y, X indicates the number of total carbon atoms in the fatty acid(s) (FA) portions of the molecule, and Y the total number of double bonds in the fatty acid portion(s) of the molecule.

    [0189] The nomenclature NB indicates, for a DAG molecule, A and B types of fatty acid moieties attached to the glycerol backbone of the molecule. The fatty acid moieties A and B can be attached to any of the two bonding positions of the glycerol backbone of the molecule.

    [0190] The nomenclature (dC/A) indicates, for a molecule of Cer, Gb3, Glc/GalCer and LacCer, C the type of long-chain base with an amide-linked, and A, fatty acid moiety.

    [0191] The nomenclature A/B/C indicates A, B and C types of fatty acid moieties attached to the glycerol backbone of the molecule. The fatty acid moieties A, B and C can be attached to any of the three bonding positions of the glycerol backbone of the molecule.

    [0192] The nomenclature sn1 and sn2 indicate the sn1 and sn2 positions of the glycerol backbone, respectively, to which the fatty acid moiety is attached.

    [0193] A “treatment” and “therapy” are used interchangeably in the present description and may comprise any therapeutic treatment or operations typically given to a subject having ovarian cancer, such as, but not limited to, surgery, chemotherapy, radiation therapy, hormonal therapy, anti-angiogenic therapy, therapies targeting homologous recombination deficiency, antibody therapy or other targeted therapy utilizing ovarian cancer specific signalling pathways. The treatment may comprise treatment having a direct effect on the metabolism of the malignant tissue. Ovarian cancer therapy may comprise administering a pharmaceutical agent affecting lipid metabolism.

    [0194] The term “effectiveness of a treatment” and “effectiveness of a therapy” are taken to mean the ability of a treatment and therapy to achieve the therapeutic purpose for which it is administered.

    [0195] A “pharmaceutical”, “drug”, “medicament” and “medicine” are used interchangeably in the present description and may comprise any pharmaceutical typically given to a subject having ovarian cancer.

    [0196] As used herein, a “composition” and “kit” are used for diagnosing, predicting and detecting ovarian cancer and comprise means and elements for assaying the small molecule biomarkers described in the present disclosure.

    [0197] As used herein, a “preparation” is used in the assays determining the small molecule biomarkers described in the present disclosure for diagnosing, predicting and detecting ovarian cancer.

    [0198] For the purposes of the present disclosure, the terms “obtaining data”, “collecting data”, “obtaining information” and “collecting information” may be used interchangeably.

    [0199] The terms “the disclosure, description or invention”, “in accordance with the disclosure, description or invention”, “according to the disclosure, description or invention”, “the present disclosure, description or invention” as used herein, are intended to refer to all aspects and embodiments of the disclosure described and/or claimed herein.

    [0200] As used herein, the term “comprising” is to be construed as encompassing both “including” and “consisting of”.

    [0201] As used herein “determining” in reference to a molecular biomarker or a protein biomarker as disclosed herein refers to quantitatively or relatively determining an amount of a biomarker in a sample. For quantitative determination, either the absolute or precise amount of the biomarker in a sample is determined. The relative amount or level of a biomarker in a sample, may alternatively be determined, e.g., the biomarker amount in the sample is determined to be enlarged or diminished with respect to a control as described herein.

    EXAMPLES

    [0202] The following examples are provided to illustrate various aspects of the present disclosure. They are not intended to limit the disclosure, which is defined by the accompanying claims.

    Example 1. Materials and Methods

    Description of Study Cohorts and Samples

    [0203] Analyses in two study cohorts led to the current disclosure, referred here as cohort I and II. Cohort II had more early-stage ovarian cancers than cohort I. Cohort I had 100 subjects without malignant disease (control group) and 158 ovarian cancer patients. Cohort II had 109 subjects without malignant disease (control group) and 62 ovarian cancer patients. The serum samples of both these cohorts were collected from preoperative primary ovarian cancer patients as well as from patients without ovarian cancer at the Charité Medical University (Berlin, Germany). The Ethics Committee approved the use of the samples for the study. The patient's informed consent was obtained prior to surgery or during subsequent treatment, sample collection and documentation of clinical and surgical data. The study population without ovarian cancer consisted of a group of patients with benign tumors, endometriosis, cysts, uterus myomatosus and other conditions causing similar symptoms to ovarian cancer. Blood was collected within the Tumor Bank Ovarian Cancer project (http://toc-network.de) using serum tubes containing clot activators (Vacutainer, BD, Medical-Pharmaceutical System, Franklin Lakes, N.J.). Collected blood was clotted for 30 min to 2 h at room temperature and serum was separated by centrifugation at 1200 g for 15 minutes. Serum was aliquoted and stored at −80° C.

    GC×GC-TOF Method

    [0204] For the serum samples of cohort I, 400 μl methanol and 10 μl standard mixture (valine-d8 (37.6 mg/I), heptadecanoic acid (186.5 mg/I), succinic acid-d4 (62.9 mg/I), glutamic acid-d5 (103.5 mg/I)) was added to 30 μl of the sample. The samples were vortexed for 2 min. After 30 minutes at room temperature the samples were centrifuged for 5 min at 10000 rpm. 200 μl of the supernatant was moved to a GC vial and evaporated to dryness under nitrogen. The samples were derivatized with 25 μl methoxyamine (45° C., 60 minutes) and 25 μl Nmethyltrimethylsilyltrifluoroacetamide (45° C., 60 minutes) and 50 μl of hexane with retention index compounds and injection standard (4,4′-dibromooctafluorobiphenyl) was added to samples.

    [0205] For the analysis, a Leco Pegasus 4D GC×GC-TOFMS instrument (Leco Corp., St. Joseph, Mich.) equipped with a cryogenic modulator was used. The GC part of the instrument was an Agilent 6890 gas chromatograph (Agilent Technologies, Palo Alto, Calif.), equipped with split/splitless injector. The first-dimension chromatographic column was a 10-m Rxi-5MS capillary column with an internal diameter of 0.18 mm and a stationary-phase film thickness of 0.18 μm, and the second-dimension chromatographic column was a 1.5 m BPX-50 capillary column with an internal diameter of 100 μm and a film thickness of 0.1 μm. A methyl deactivated retention gap (1.5 m×0.53 mm i.d.) was used in the front of the first column. High-purity helium was used as the carrier gas at a constant pressure mode (40 psig). A 4-s separation time was used in the second dimension. The MS spectra were measured at 45-700 atomic mass unit (amu) with 100 spectra/second. For the injection, a splitless injection (1.0 μl) at 240° C. was utilized. The temperature program was as follows: the first-dimension column oven ramp began at 50° C. with a 2 minute hold after which the temperature was programmed to 240° C. at a rate of 7° C./minute and further to 300° C. at a rate of 25° C./minute and then held at this temperature for 3 minute. The second-dimension column temperature was maintained 15° C. higher than the corresponding first-dimension column. The programming rate and hold times were the same for the two columns.

    [0206] ChromaTOF vendor software (LECO) was used for within-sample data processing, and inhouse made software Guineu (Castillo et al., 2011, Anal Chem) was used for alignment, normalization and peak matching across samples. The peaks were first filtered based on number of detected peaks in the total profile of all sample runs. The normalization for uncalibrated metabolites was performed by correction for internal standard C17:0. 27 of the metabolites were checked manually in each serum sample for correct integration and identification. Other mass spectra from the GC×GC-TOFMS analysis were searched against National Institutes of Standards and Technology 05 (NIST05) mass spectral library.

    GC-MS Method

    [0207] For the serum of cohort II, a modified method of the GC×GC-TOF method was used for sample pretreatment and instrument parameters. After samples were thawed unassisted on top of ice, 20 μl of serum was dispensed into test tube and extraction solution (600 μl) with internal standard (succinic acid-d4, 0.3 μg/ml in methanol) was added to the sample. The sample was mixed and left in the freezer for 10 minutes (−20° C.). Samples were centrifuged at 14,000 rpm for 10 minutes at 4° C., and the supernatants (300 μl) were evaporated to dryness under nitrogen. Analytes were converted into their trimethylsilyl (TMS) derivatives by adding 50 μl N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) to the sample and the mixture was heated at 50° C. for 30 minutes. After cooling samples to the room temperature, 50 μl of hexane was added and mixed with Vortex mixer for 10 seconds. The quantification was based on internal standard method.

    [0208] The instrument was Agilent Technologies GC-MS system (Agilent Technologies; Palo Alto, Calif.). The system consisted of a 7683 autosampler and a 6890N gas chromatograph coupled to a 5973N mass spectrometry. Chromatographic conditions were as follows: VF-5 ms capillary column (30 m×0.25 mm, film 0.25 μm) with a built-in guard column (EZ-Guard, 10 m) (Agilent Technologies; Palo Alto, Calif., USA) was used with pulsed splitless injection. Injection port temperature was 250° C. and injection volume was 1 μl. The oven temperature was held at 60° C. for 2 minutes, then increased to 170° C. at 10° C. per minute rate, and then increased to 300° C. at 20° C. per minute rate, and held at 300° C. for 6.5 minutes. The carrier gas was helium with constant flow of 32 cm/sec (equal to 1 ml/min). The temperatures of the MSD transfer line heater, ionization source and quadrupole were maintained at 250° C., 230° C., and 150° C., respectively. The mass spectrometer was operated in electron ionization (EI) mode with the electron energy 70 eV. The total measurement time was 26 min. A solvent delay of 8 minutes was applied. For GC/EI-MS in the selected ion monitoring (SIM) mode was used to record target and qualifier ions measurements. Dwell time was 25 milliseconds (ms) for all recorded ions.

    LC-MS/MS Lipidomics Analysis

    [0209] Lipidomic analyses of cohorts I and II were performed using two platforms, a global screening method and a phosphosphingolipid platform. Lipids for the screening method were extracted using a modified Folch extraction (Folch et al., 1957, J. Biol Chem) and protein precipitation in methanol was used for the extraction of phosphosphingolipids. Prior to extraction, samples were thawed at +4° C., and Hamilton MICROLAB STAR system (Hamilton Robotics, Switzerland) was used for the extraction. For the screening method, samples (10 μl) were aliquoted into a 96-well plate, and internal standard mixture (20 μL) containing a known amount of synthetic internal standards was added followed by chloroform (450 μl). Organic phase separation was facilitated by adding 20 mM acetic acid and centrifuging the plate for 5 minutes at 500×g. The lower organic phase (360 μl) was transferred into a new 96-well plate. The remaining water-containing phase was washed with additional chloroform (360 μl) followed by centrifugation and removal of the remaining organic phase. The two organic phases were pooled and evaporated under N.sub.2 until dryness. The lipid extracts were then re-dissolved in chloroform:methanol (1:2, v/v). For the analysis of phosphosphingolipids, samples (25 μl) were aliquoted into a 96-well plate, and ice-cold methanol containing 0.1% butylated hydroxytoluene (500 μL) was added to each sample, followed by internal standard mixture (25 μL) containing a known amount of synthetic standards. Samples were mixed and incubated for 10 minutes. After centrifugation, supernatant (450 μl) was transferred into a new 96-well plate, evaporated under N.sub.2 until dryness and re-dissolved in methanol (200 μL).

    [0210] Lipidomics screening and phosphosphingolipid platforms were both analyzed on a hybrid triple quadrupole/linear ion trap mass spectrometer (QTRAP 5500, AB Sciex, Concors, Canada) equipped with an ultra-high performance liquid chromatography (UHPLC) (Nexera-X2, Shimadzu). Chromatographic separation of the lipidomics screening platform was performed on Acquity BEH C18, 2.1×50 mm id. 1.7 μm column (Waters, Mass., USA). Mobile phases consisted of (A) 10 mM ammonium acetate in LC-MS grade water with 0.1% formic acid, and (B) 10 mM ammonium acetate in acetonitrile:2-propanol (3:4, VN) with 0.1% formic acid (FA). Chromatographic separation of phosphosphingolipid platform was performed on AQUASIL C18, 2.1×50 mm, 5 μm (Thermo Fisher, Massachusetts, USA), column set at 60° C. Mobile phases consisted of (A) 10 mM ammonium acetate in LC-MS grade water with 0.1% formic acid, and (B) 10 mM ammonium acetate in methanol:2-Propanol (1:2) with 0.1% formic acid.

    [0211] For the MS analysis, a targeted approach in positive ion mode was used for both platforms. Data was collected using scheduled multiple reaction monitoring (sMRM™) algorithm for the lipidomics screening platform (Weir et al., 2013, J Lipid Res) and multiple reaction monitoring (MRM) for phosphosphingolipids. Mass spectrometer parameters were optimized based on lipid class. Lipidomics data were processed using Analyst and MultiQuant 3.0 software (QTRAP 5500, AB Sciex, Concors, Canada), area or height ratios of analyte and its corresponding internal standard (IS) peak were normalized with IS amount and sample volume.

    Statistical Analyses

    [0212] All statistical analyses were performed using R, version x64 3.3.2. For the two-group comparisons unpaired t-tests were performed after log 2 transformation of the data. In addition, mean relative differences were calculated between the patients with malignant tumors and control group. Association of small molecule biomarkers to overall survival was analysed by cox proportional hazards regression test. AUC values were calculated using R package ROCR. Sensitivity and specificity values were calculated with a cut-off value where the sum of sensitivity and specificity was at maximum. Predictive models combining small molecule biomarker ratios and CA-125 were binary logistic regression models.

    Example 2. Results

    Increased and Decreased Small Molecule Biomarkers

    [0213] Table 2 shows statistically significantly (p<0.05) increased or decreased small molecule biomarkers in ovarian cancer patients as compared to control group. The results were derived from cohort I, except for acetoacetic acid, where its concentration was determined by more accurate method in cohort II.

    TABLE-US-00002 TABLE 2 Increased (group A) (a) and decreased (group B) (b) small molecule biomarkers in ovarian cancer patients. Abbreviations are described in the detailed description of the present disclosure. CHANGE (%) P-VALUE CLASS Table 2a. Group A molecules Cysteine 27.3 4.9E−02 Amino acids I Glutamic acid 25.7 1.6E−03 Amino acids I Glutamine 10.6 2.2E−02 Amino acids I Glycine 56.1 4.1E−04 Amino acids I 2,3-Dihydroxybutyric acid 29.3 5.8E−04 Butyric acids 2,4-Dihydroxybutyric acid 29.8 3.2E−04 Butyric acids 2-Aminobutyric acid 99.7 2.1E−04 Butyric acids 2-Hydroxybutyric acid 48.8 1.5E−09 Butyric acids 3,4-Dihydroxybutyric acid 90.7 1.8E−25 Butyric acids 3-Hydroxybutyric acid 196.6 4.7E−20 Butyric acids Cer(d16:1/18:0) 24.6 1.3E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/16:0) 27.0 3.2E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/18:0) 49.7 5.7E−05 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/20:0) 22.0 1.8E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/24:1) 16.4 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/16:0) 26.7 1.6E−05 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/18:0) 71.4 7.2E−12 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/20:0) 39.7 9.9E−08 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/22:0) 11.9 2.1E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/24:1) 30.5 1.1E−06 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/16:0) 15.2 1.2E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/18:0) 26.2 5.2E−04 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/24:1) 13.5 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/22:0) 21.6 2.8E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/24:1) 43.9 2.1E−07 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Eicosenoic acid (C20:1) 11.6 2.1E−02 Fatty acids Lauric acid (C12:0) 14.9 4.1E−02 Fatty acids Linolenic acid (C18:3) 37.3 2.3E−05 Fatty acids Myristic acid (C14:0) 20.8 5.7E−03 Fatty acids Oleic acid (C18:1) 16.7 6.0E−04 Fatty acids Palmitelaidic acid (C16:1) 27.6 2.9E−04 Fatty acids Palmitic acid (C16:0) 8.0 1.4E−02 Fatty acids LPC 16:0_sn1 40.7 6.3E−03 LPL I LPC 20:1_sn2 20.6 3.8E−02 LPL I LPC 20:5_sn1 18.6 2.6E−02 LPL I LPC 22:6_sn1 20.5 3.7E−02 LPL I LPC O-18:1 35.9 1.9E−02 LPL I LPC P-18:1 29.2 4.4E−02 LPL I LPE 18:0_sn1 35.5 1.4E−03 LPL I LPE 18:0_sn2 33.5 2.6E−03 LPL I 3-Hydroxyisovaleric acid 63.8 2.3E−04 Other I 4-Hydroxyphenyllactic acid 12.6 1.5E−02 Other I Acetoacetic acid 271.2 1.6E−08 Other I Adipic acid 35.9 7.6E−07 Other I Alpha-ketoglutaric acid 52.1 4.8E−02 Other I Ethanolamine 49.2 3.1E−07 Other I Glycerol 64.2 9.7E−14 Other I PE O-36:1 62.4 1.9E−05 PE O I PE O-36:2 93.3 6.5E−08 PE O I PE O-36:3 66.0 9.3E−05 PE O I Arabinose 52.5 1.3E−05 Sugars Erythritol 34.3 3.4E−05 Sugars Glucopyranose 17.0 2.6E−02 Sugars Maltose 814.4 5.0E−23 Sugars Mannonic acid 103.7 6.0E−14 Sugars Melibiose 392.4 6.6E−08 Sugars Myo-Inositol 32.4 4.2E−07 Sugars Turanose 434.5 2.4E−03 Sugars Xylitol 30.1 5.0E−09 Sugars TAG(16:0/18:1/18:1) 9.2 3.7E−02 TAG I TAG(18:0/18:2/18:2) 18.1 1.2E−02 TAG I TAG(18:1/18:1/18:1) 8.2 3.9E−02 TAG I TAG(18:1/18:1/20:4) 28.4 6.7E−06 TAG I TAG(18:1/18:1/22:6) 54.9 5.4E−08 TAG I TAG(18:2/18:2/18:2) 118.1 3.3E−07 TAG I Table 2b. Group B molecules Alanine −44.7 3.0E−19 Amino acids II Methionine −25.9 5.7E−17 Amino acids II Phenylalanine −13.2 1.2E−03 Amino acids II Proline −21.9 3.0E−05 Amino acids II Serine −13.1 1.5E−03 Amino acids II Threonine −36.3 1.4E−15 Amino acids II Tryptophan −66.8 2.1E−21 Amino acids II Tyrosine −24.9 1.5E−03 Amino acids II Valine −8.2 1.2E−02 Amino acids II CE 14:0 −22.3 4.1E−06 CE CE 14:1 −22.0 4.7E−05 CE CE 15:0 −16.8 2.0E−04 CE CE 16:0 −12.5 1.4E−04 CE CE 16:2 −21.1 2.0E−07 CE CE 17:0 −16.5 1.2E−04 CE CE 17:1 −10.1 2.1E−02 CE CE 18:0 −26.7 4.9E−10 CE CE 18:1 −12.7 1.9E−04 CE CE 18:2 −21.9 7.2E−10 CE CE 18:3 −23.2 3.8E−10 CE CE 19:1 −0.9 4.6E−02 CE CE 20:2 −23.3 2.6E−07 CE CE 20:3 −21.3 5.8E−06 CE CE 20:4 −7.4 5.0E−02 CE CE 22:3 −15.8 3.9E−02 CE CE 22:4 −14.9 1.6E−03 CE CE 22:5 −35.3 2.0E−08 CE CE 24:6 −22.7 1.2E−04 CE Cer(d16:1/23:0) −24.9 5.7E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/24:0) −29.3 6.1E−08 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/26:0) −17.1 4.0E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/23:0) −17.4 3.8E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/24:0) −14.8 2.5E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/14:0) −10.3 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/24:0) −11.9 8.6E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/23:0) −18.5 5.6E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/24:0) −22.4 1.1E−05 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/26:0) −14.2 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 DAG(14:0/18:1) −39.1 6.9E−04 DAG DAG(14:0/18:2) −38.3 1.6E−03 DAG DAG(18:1/20:3) −31.3 1.1E−02 DAG DAG(16:0/16:1) −27.6 1.3E−02 DAG DAG(18:0/20:4) −22.0 3.9E−02 DAG Gb3(d18:1/16:0) −14.8 3.0E−04 Gb3 Gb3(d18:1/18:0) −13.4 2.2E−02 Gb3 Gb3(d18:1/22:0) −19.5 4.5E−04 Gb3 Gb3(d18:1/23:0) −16.2 5.6E−03 Gb3 Gb3(d18:1/24:0) −23.1 8.6E−05 Gb3 Glc/GalCer(d16:1/20:0) −16.8 9.7E−04 Glc/GalCer Glc/GalCer(d16:1/22:0) −23.4 1.5E−05 Glc/GalCer Glc/GalCer(d16:1/23:0) −19.3 3.8E−04 Glc/GalCer Glc/GalCer(d16:1/24:0) −23.1 5.0E−06 Glc/GalCer Glc/GalCer(d18:1/16:0) −13.3 1.1E−02 Glc/GalCer Glc/GalCer(d18:1/18:0) −16.1 7.5E−04 Glc/GalCer Glc/GalCer(d18:1/20:0) −19.7 9.2E−05 Glc/GalCer Glc/GalCer(d18:1/22:0) −25.9 2.6E−07 Glc/GalCer Glc/GalCer(d18:1/23:0) −23.8 7.8E−07 Glc/GalCer Glc/GalCer(d18:1/24:0) −24.2 4.7E−07 Glc/GalCer Glc/GalCer(d18:1/26:0) −21.0 9.4E−05 Glc/GalCer Glc/GalCer(d18:2/20:0) −23.3 1.7E−04 Glc/GalCer Glc/GalCer(d18:2/22:0) −17.9 5.0E−04 Glc/GalCer Glc/GalCer(d18:2/23:0) −21.6 5.2E−05 Glc/GalCer Glc/GalCer(d18:2/24:0) −23.9 4.8E−08 Glc/GalCer LacCer(d16:1/16:0) −21.1 5.3E−05 LacCer LacCer(d18:1/16:0) −23.5 2.0E−08 LacCer LacCer(d18:1/22:0) −27.7 7.6E−10 LacCer LacCer(d18:1/23:0) −26.0 1.7E−07 LacCer LacCer(d18:1/24:0) −23.2 1.5E−08 LacCer LacCer(d18:1/24:1) −21.0 2.8E−06 LacCer LacCer(d18:2/16:0) −25.2 5.5E−08 LacCer LacCer(d18:2/22:0) −18.2 3.9E−02 LacCer LacCer(d18:2/24:0) −19.7 4.0E−03 LacCer LacCer(d18:2/24:1) −10.7 2.6E−02 LacCer LPC 14:0_sn1 −19.1 1.6E−03 LPL II LPC 14:0_sn2 −30.3 2.3E−05 LPL II LPC 18:2_sn1 −29.7 1.9E−07 LPL II LPC 18:2_sn2 −34.7 2.3E−12 LPL II LPC 20:0_sn1 −16.6 3.4E−03 LPL II LPC 20:0_sn2 −14.6 1.1E−02 LPL II LPC 20:2_sn1 −5.7 3.7E−02 LPL II LPC 20:2_sn2 −16.4 3.2E−05 LPL II LPC 20:3_sn1 −16.1 2.3E−03 LPL II LPC 20:3_sn2 −24.4 1.4E−06 LPL II LPC 22:0_sn1 −28.7 6.3E−10 LPL II LPC 22:4_sn1 −11.1 1.7E−02 LPL II LPC 22:4_sn2 −19.6 4.9E−05 LPL II LPC 24:0_sn1 −28.7 1.5E−12 LPL II LPC 24:0_sn2 −28.9 2.3E−13 LPL II LPC 0-20:0 −15.5 7.1E−04 LPL II LPC 0-22:0 −25.3 4.8E−11 LPL II LPC 0-22:1 −17.5 1.9E−03 LPL II LPC 0-24:0 −26.6 2.9E−10 LPL II LPC 0-24:1 −17.8 8.2E−05 LPL II LPC 0-24:2 −33.0 1.2E−08 LPL II LPE 18:2_sn1 −40.1 1.5E−11 LPL II LPE 18:2_sn2 −35.4 4.6E−10 LPL II LPE 20:4_sn1 −16.4 2.0E−03 LPL II 2-oxo-3-methylpentanoic acid −26.9 7.0E−09 Other II 2-Oxoisovaleric acid −18.5 2.3E−02 Other II Cholesterol −31.3 1.8E−06 Other II Glyceric acid −28.1 6.5E−04 Other II Glycerol-3-phosphate −21.4 8.2E−03 Other II lndole-3-acetic acid −27.2 3.0E−03 Other II Ketoleucine −45.2 7.0E−19 Other II Lactic acid −29.0 3.2E−11 Other II Malic acid −28.5 2.1E−06 Other II Pyruvic acid −32.0 3.2E−02 Other II PC 28:0 −59.0 1.5E−15 PC PC 30:0 −43.4 4.0E−15 PC PC 30:1 −52.1 2.4E−13 PC PC 30:2 −61.4 2.1E−14 PC PC 31:0 −26.0 1.6E−07 PC PC 31:1 −36.9 1.9E−08 PC PC 32:0 −15.5 4.3E−05 PC PC 32:1 −29.0 1.9E−06 PC PC 32:2 −52.5 4.0E−19 PC PC 32:3 −58.5 4.2E−17 PC PC 33:0 −17.1 2.0E−03 PC PC 33:1 −25.2 5.2E−06 PC PC 33:2 −35.6 6.9E−13 PC PC 33:3 −42.0 4.2E−11 PC PC 34:0 −11.9 3.6E−03 PC PC 34:1 −10.2 1.9E−02 PC PC 34:2 −28.6 4.5E−10 PC PC 34:3 −37.9 9.7E−14 PC PC 34:4 −52.1 5.6E−16 PC PC 34:5 −41.3 1.4E−05 PC PC 35:0 −21.3 5.5E−04 PC PC 35:1 −15.5 4.0E−03 PC PC 35:2 −26.3 2.6E−06 PC PC 35:3 −39.8 3.1E−15 PC PC 35:4 −25.8 1.5E−07 PC PC 36:0 −21.7 1.3E−07 PC PC 36:2 −35.1 2.0E−13 PC PC 36:3 −36.0 2.2E−14 PC PC 36:5 −32.1 1.4E−07 PC PC 36:6 −46.4 2.0E−09 PC PC 36:7 −27.4 2.1E−05 PC PC 37:1 −20.5 4.6E−06 PC PC 37:2 −36.3 6.0E−13 PC PC 37:3 −30.4 7.4E−09 PC PC 37:6 −20.3 8.9E−03 PC PC 38:0 −29.1 5.0E−11 PC PC 38:1 −9.8 2.8E−02 PC PC 38:2 −27.9 3.7E−08 PC PC 38:3 −30.6 1.5E−09 PC PC 38:4 −24.1 6.4E−06 PC PC 38:5 −15.7 6.6E−05 PC PC 38:6 −35.0 2.6E−10 PC PC 38:7 −30.9 7.8E−05 PC PC 39:0 −18.7 9.9E−07 PC PC 39:4 −19.5 5.6E−06 PC PC 39:5 −12.0 3.5E−02 PC PC 40:1 −22.9 3.6E−08 PC PC 40:3 −15.8 1.6E−04 PC PC 40:4 −29.9 9.0E−12 PC PC 40:5 −14.8 1.4E−03 PC PC 40:8 −33.4 6.8E−10 PC PC O-32:0 −24.5 2.7E−08 PC O/P PC O-32:1 −31.3 7.1E−10 PC O/P PC O-34:0 −20.9 1.1E−06 PC O/P PC O-34:1 −22.2 1.2E−08 PC O/P PC O-34:2 −39.7 6.7E−17 PC O/P PC O-36:1 −27.6 1.5E−07 PC O/P PC O-36:2 −36.3 4.2E−14 PC O/P PC O-36:3 −38.7 4.8E−16 PC O/P PC O-36:4 −23.1 2.4E−09 PC O/P PC O-38:0 −21.3 8.0E−09 PC O/P PC O-38:1 −27.9 5.8E−12 PC O/P PC O-38:2 −28.3 4.8E−09 PC O/P PC O-38:3 −31.7 2.9E−11 PC O/P PC O-38:4 −31.5 7.2E−12 PC O/P PC O-38:5 −16.0 3.4E−05 PC O/P PC O-38:6 −13.5 7.2E−03 PC O/P PC O-40:1 −23.6 4.1E−06 PC O/P PC O-40:3 −37.5 2.5E−10 PC O/P PC O-40:4 −18.1 8.5E−04 PC O/P PC O-40:5 −20.3 1.3E−04 PC O/P PC P-36:2 −26.8 6.2E−08 PC O/P PC P-32:0 −20.2 8.1E−07 PC O/P PC P-32:1 −21.4 1.6E−05 PC O/P PC P-34:1 −13.9 3.6E−04 PC O/P PC P-34:2 −33.3 2.7E−13 PC O/P PC P-36:3 −17.8 2.4E−06 PC O/P PC P-36:4 −20.0 8.1E−07 PC O/P PC P-38:1 −16.5 4.7E−04 PC O/P PC P-38:2 −17.0 2.7E−03 PC O/P PC P-38:4 −21.4 2.1E−05 PC O/P PC P-38:5 −11.3 1.5E−03 PC O/P PC P-38:6 −22.9 5.9E−04 PC O/P PC P-40:1 −15.9 5.1E−04 PC O/P PC P-40:2 −21.6 1.3E−05 PC O/P PC P-40:3 −23.1 3.4E−07 PC O/P PC P-40:4 −21.8 2.8E−07 PC O/P PC P-40:5 −17.3 2.4E−05 PC O/P PC P-40:6 −6.2 3.7E−02 PC O/P PE 32:1 −39.1 2.3E−04 PE PE 34:2 −39.2 1.0E−06 PE PE 34:3 −49.7 8.1E−07 PE PE 36:2 −37.9 1.5E−06 PE PE 36:3 −53.8 2.0E−07 PE PE 36:4 −30.7 2.9E−04 PE PE 36:5 −26.5 2.3E−02 PE PE 38:1 −37.4 7.9E−05 PE PE 38:2 −29.4 2.0E−02 PE PE 38:3 −40.4 6.8E−07 PE PE 38:4 −47.6 7.3E−07 PE PE 38:5 −33.5 5.1E−06 PE PE 40:4 −34.8 1.2E−03 PE PE 40:5 −21.8 8.3E−04 PE PE 40:7 −42.4 1.3E−03 PE PE O-34:1 −12.4 4.7E−02 PE 0 II PE O-36:4 −49.1 4.9E−11 PE 0 II PE O-38:4 −48.9 1.5E−12 PE 0 II PE O-38:5 −38.5 1.3E−09 PE 0 II PE O-38:6 −16.2 4.4E−02 PE 0 II PE P-34:2 −20.6 1.4E−03 PE P PE P-36:2 −41.7 7.0E−10 PE P PE P-36:4 −31.6 1.0E−06 PE P PE P-38:4 −36.4 7.4E−10 PE P PE P-38:6 −31.2 1.4E−07 PE P PE P-40:6 −20.6 1.6E−03 PE P PG 34:1 −22.9 1.1E−02 PG PG 36:2 −25.6 3.5E−04 PG PI 32:0 −49.6 3.0E−04 PI PI 34:0 −45.7 5.7E−04 PI PI 34:1 −31.3 4.7E−04 PI PI 34:2 −31.3 6.4E−06 PI PI 36:1 −40.1 3.8E−07 PI PI 36:2 −27.9 5.2E−06 PI PI 36:3 −51.5 9.9E−10 PI PI 36:4 −24.3 2.7E−03 PI PI 38:2 −40.5 3.5E−07 PI PI 38:3 −33.8 1.6E−09 PI PI 38:5 −24.2 4.3E−05 PI PI 40:4 −19.9 1.1E−03 PI S1P d16:1 −20.0 1.4E−08 S1P/SA1P S1P d18:1 −19.3 1.3E−07 S1P/SA1P S1P d18:2 −29.3 1.2E−13 S1P/SA1P SA1P d18:0 −24.8 5.3E−13 S1P/SA1P SM 30:2 −35.8 2.2E−11 SM SM 31:0 −12.2 4.2E−02 SM SM 31:1 −24.7 1.2E−07 SM SM 31:2 −12.3 7.5E−03 SM SM 32:0 −24.2 8.6E−08 SM SM 32:1 −17.3 7.4E−06 SM SM 32:2 −28.0 3.3E−11 SM SM 33:1 −11.6 4.7E−03 SM SM 33:2 −9.2 4.4E−02 SM SM 34:0 −15.1 5.0E−04 SM SM 34:1 −13.2 4.6E−04 SM SM 34:2 −15.8 2.3E−05 SM SM 35:0 −25.3 3.2E−06 SM SM 35:1 −10.3 9.7E−03 SM SM 35:2 −9.3 9.1E−03 SM SM 36:3 −14.6 2.7E−03 SM SM 37:0 −10.1 1.6E−02 SM SM 37:1 −14.8 1.9E−04 SM SM 37:2 −34.8 1.4E−13 SM SM 38:0 −22.5 2.0E−05 SM SM 38:1 −20.4 1.0E−06 SM SM 38:2 −19.1 2.8E−06 SM SM 38:3 −20.2 8.5E−05 SM SM 39:0 −18.0 7.1E−03 SM SM 39:1 −27.5 1.3E−10 SM SM 39:2 −21.6 1.5E−03 SM SM 40:0 −27.0 5.9E−07 SM SM 40:1 −23.7 4.2E−08 SM SM 40:2 −27.3 4.5E−12 SM SM 40:3 −19.8 2.6E−06 SM SM 41:0 −32.5 1.3E−09 SM SM 41:1 −25.2 8.0E−12 SM SM 41:2 −17.0 7.1E−05 SM SM 42:0 −30.9 3.8E−10 SM SM 42:1 −31.1 2.6E−13 SM SM 42:3 −15.8 3.9E−05 SM SM 44:1 −23.7 1.1E−08 SM SM 44:3 −16.6 1.3E−04 SM TAG(14:0/16:0/18:1) −37.3 1.0E−04 TAG II TAG(14:0/16:0/18:2) −36.5 2.3E−04 TAG II TAG(14:0/16:1/18:1) −44.3 1.5E−04 TAG II TAG(14:0/16:1/18:2) −50.9 2.4E−06 TAG II TAG(14:0/17:0/18:1) −22.6 4.5E−02 TAG II TAG(14:0/18:0/18:1) −44.4 1.7E−04 TAG II TAG(14:0/18:2/18:2) −47.7 3.3E−06 TAG II TAG(14:1/16:0/18:1) −35.4 4.8E−03 TAG II TAG(14:1/16:1/18:0) −28.7 1.6E−02 TAG II TAG(14:1/18:0/18:2) −25.3 1.4E−02 TAG II TAG(14:1/18:1/18:1) −33.0 1.2E−04 TAG II TAG(16:1/16:1/16:1) −43.7 5.2E−05 TAG II TAG(16:1/16:1/18:0) −40.5 6.6E−05 TAG II TAG(16:1/18:1/18:2) −13.9 3.8E−02 TAG II

    [0214] Table 3 shows statistically significantly (p<0.05) increased or decreased small molecule biomarkers in ovarian cancer patients as compared to control group according to some embodiments of all aspects of the present disclosure. The results were derived from cohort I, except for acetoacetic acid, where its concentration was determined by more accurate method in cohort II.

    TABLE-US-00003 TABLE 3 Increased (group A) (a) and decreased (group B) (b) small molecule biomarkers in ovarian cancer patients according to some embodiments of the present disclosure. Abbreviations are described in the detailed description of the present disclosure. CHANGE (%) P-VALUE CLASS Table 3a. Group A molecules Cysteine 27.3 4.9E−02 Amino acids I Glutamic acid 25.7 1.6E−03 Amino acids I Glutamine 10.6 2.2E−02 Amino acids I 2-Aminobutyric acid 99.7 2.1E−04 Butyric acids Cer(d16:1/18:0) 24.6 1.3E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/16:0) 27.0 3.2E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/18:0) 49.7 5.7E−05 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/20:0) 22.0 1.8E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/24:1) 16.4 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/16:0) 26.7 1.6E−05 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/18:0) 71.4 7.2E−12 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/20:0) 39.7 9.9E−08 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/22:0) 11.9 2.1E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/24:1) 30.5 1.1E−06 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/16:0) 15.2 1.2E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/18:0) 26.2 5.2E−04 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/24:1) 13.5 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/22:0) 21.6 2.8E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/24:1) 43.9 2.1E−07 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Eicosenoic acid (C20:1) 11.6 2.1E−02 Fatty acids Lauric acid (C12:0) 14.9 4.1E−02 Fatty acids Linolenic acid (C18:3) 37.3 2.3E−05 Fatty acids Myristic acid (C14:0) 20.8 5.7E−03 Fatty acids Oleic acid (C18:1) 16.7 6.0E−04 Fatty acids Palmitelaidic acid (C16:1) 27.6 2.9E−04 Fatty acids LPC 20:1_sn2 20.6 3.8E−02 LPL I LPC 20:5_sn1 18.6 2.6E−02 LPL I LPC 22:6_sn1 20.5 3.7E−02 LPL I LPC O-18:1 35.9 1.9E−02 LPL I LPC P-18:1 29.2 4.4E−02 LPL I Alpha-ketoglutaric acid 52.1 4.8E−02 Other I Ethanolamine 49.2 3.1E−07 Other I Glycerol 64.2 9.7E−14 Other I PE O-36:1 62.4 1.9E−05 PE O I PE O-36:2 93.3 6.5E−08 PE O I PE O-36:3 66.0 9.3E−05 PE O I Arabinose 52.5 1.3E−05 Sugars Erythritol 34.3 3.4E−05 Sugars Glucopyranose 17.0 2.6E−02 Sugars Maltose 814.4 5.0E−23 Sugars Mannonic acid 103.7 6.0E−14 Sugars Melibiose 392.4 6.6E−08 Sugars Myo-Inositol 32.4 4.2E−07 Sugars Turanose 434.5 2.4E−03 Sugars Xylitol 30.1 5.0E−09 Sugars TAG(16:0/18:1/18:1) 9.2 3.7E−02 TAG I TAG(18:0/18:2/18:2) 18.1 1.2E−02 TAG I TAG(18:1/18:1/18:1) 8.2 3.9E−02 TAG I TAG(18:1/18:1/20:4) 28.4 6.7E−06 TAG I TAG(18:1/18:1/22:6) 54.9 5.4E−08 TAG I TAG(18:2/18:2/18:2) 118.1 3.3E−07 TAG I Table 3b. Group B molecules Methionine −25.9 5.7E−17 Amino acids II Phenylalanine −13.2 1.2E−03 Amino acids II Serine −13.1 1.5E−03 Amino acids II Threonine −36.3 1.4E−15 Amino acids II Tryptophan −66.8 2.1E−21 Amino acids II Tyrosine −24.9 1.5E−03 Amino acids II Valine −8.2 1.2E−02 Amino acids II CE 14:0 −22.3 4.1E−06 CE CE 14:1 −22.0 4.7E−05 CE CE 15:0 −16.8 2.0E−04 CE CE 16:0 −12.5 1.4E−04 CE CE 16:2 −21.1 2.0E−07 CE CE 17:0 −16.5 1.2E−04 CE CE 17:1 −10.1 2.1E−02 CE CE 18:0 −26.7 4.9E−10 CE CE 18:1 −12.7 1.9E−04 CE CE 18:2 −21.9 7.2E−10 CE CE 19:1 −0.9 4.6E−02 CE CE 20:2 −23.3 2.6E−07 CE CE 20:3 −21.3 5.8E−06 CE CE 20:4 −7.4 5.0E−02 CE CE 22:3 −15.8 3.9E−02 CE CE 22:4 −14.9 1.6E−03 CE CE 22:5 −35.3 2.0E−08 CE CE 24:6 −22.7 1.2E−04 CE Cer(d16:1/23:0) −24.9 5.7E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/24:0) −29.3 6.1E−08 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/26:0) −17.1 4.0E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/23:0) −17.4 3.8E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/24:0) −14.8 2.5E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/14:0) −10.3 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/24:0) −11.9 8.6E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/23:0) −18.5 5.6E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/24:0) −22.4 1.1E−05 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/26:0) −14.2 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 DAG(14:0/18:1) −39.1 6.9E−04 DAG DAG(14:0/18:2) −38.3 1.6E−03 DAG DAG(18:1/20:3) −31.3 1.1E−02 DAG DAG(16:0/16:1) −27.6 1.3E−02 DAG DAG(18:0/20:4) −22.0 3.9E−02 DAG Gb3(d18:1/16:0) −14.8 3.0E−04 Gb3 Gb3(d18:1/18:0) −13.4 2.2E−02 Gb3 Gb3(d18:1/22:0) −19.5 4.5E−04 Gb3 Gb3(d18:1/23:0) −16.2 5.6E−03 Gb3 Gb3(d18:1/24:0) −23.1 8.6E−05 Gb3 Glc/GalCer(d16:1/20:0) −16.8 9.7E−04 Glc/GalCer Glc/GalCer(d16:1/22:0) −23.4 1.5E−05 Glc/GalCer Glc/GalCer(d16:1/23:0) −19.3 3.8E−04 Glc/GalCer Glc/GalCer(d16:1/24:0) −23.1 5.0E−06 Glc/GalCer Glc/GalCer(d18:1/16:0) −13.3 1.1E−02 Glc/GalCer Glc/GalCer(d18:1/18:0) −16.1 7.5E−04 Glc/GalCer Glc/GalCer(d18:1/20:0) −19.7 9.2E−05 Glc/GalCer Glc/GalCer(d18:1/22:0) −25.9 2.6E−07 Glc/GalCer Glc/GalCer(d18:1/23:0) −23.8 7.8E−07 Glc/GalCer Glc/GalCer(d18:1/24:0) −24.2 4.7E−07 Glc/GalCer Glc/GalCer(d18:1/26:0) −21.0 9.4E−05 Glc/GalCer Glc/GalCer(d18:2/20:0) −23.3 1.7E−04 Glc/GalCer Glc/GalCer(d18:2/22:0) −17.9 5.0E−04 Glc/GalCer Glc/GalCer(d18:2/23:0) −21.6 5.2E−05 Glc/GalCer Glc/GalCer(d18:2/24:0) −23.9 4.8E−08 Glc/GalCer LacCer(d16:1/16:0) −21.1 5.3E−05 LacCer LacCer(d18:1/16:0) −23.5 2.0E−08 LacCer LacCer(d18:1/22:0) −27.7 7.6E−10 LacCer LacCer(d18:1/23:0) −26.0 1.7E−07 LacCer LacCer(d18:1/24:0) −23.2 1.5E−08 LacCer LacCer(d18:1/24:1) −21.0 2.8E−06 LacCer LacCer(d18:2/16:0) −25.2 5.5E−08 LacCer LacCer(d18:2/22:0) −18.2 3.9E−02 LacCer LacCer(d18:2/24:0) −19.7 4.0E−03 LacCer LacCer(d18:2/24:1) −10.7 2.6E−02 LacCer LPC 20:0_sn1 −16.6 3.4E−03 LPL II LPC 20:0_sn2 −14.6 1.1E−02 LPL II LPC 20:2_sn1 −5.7 3.7E−02 LPL II LPC 20:2_sn2 −16.4 3.2E−05 LPL II LPC 20:3_sn1 −16.1 2.3E−03 LPL II LPC 20:3_sn2 −24.4 1.4E−06 LPL II LPC 22:0_sn1 −28.7 6.3E−10 LPL II LPC 22:4_sn1 −11.1 1.7E−02 LPL II LPC 22:4_sn2 −19.6 4.9E−05 LPL II LPC 24:0_sn1 −28.7 1.5E−12 LPL II LPC 24:0_sn2 −28.9 2.3E−13 LPL II LPC O-20:0 −15.5 7.1E−04 LPL II LPC O-22:0 −25.3 4.8E−11 LPL II LPC O-22:1 −17.5 1.9E−03 LPL II LPC O-24:0 −26.6 2.9E−10 LPL II LPC O-24:1 −17.8 8.2E−05 LPL II LPC O-24:2 −33.0 1.2E−08 LPL II 2-oxo-3-methylpentanoic acid −26.9 7.0E−09 Other II 2-Oxoisovaleric acid −18.5 2.3E−02 Other II Cholesterol −31.3 1.8E−06 Other II Glyceric acid −28.1 6.5E−04 Other II Glycerol-3-phosphate −21.4 8.2E−03 Other II Indole-3-acetic acid −27.2 3.0E−03 Other II Ketoleucine −45.2 7.0E−19 Other II Lactic acid −29.0 3.2E−11 Other II Malic acid −28.5 2.1E−06 Other II Pyruvic acid −32.0 3.2E−02 Other II PC 28:0 −59.0 1.5E−15 PC PC 30:0 −43.4 4.0E−15 PC PC 30:1 −52.1 2.4E−13 PC PC 30:2 −61.4 2.1E−14 PC PC 31:0 −26.0 1.6E−07 PC PC 31:1 −36.9 1.9E−08 PC PC 32:0 −15.5 4.3E−05 PC PC 32:1 −29.0 1.9E−06 PC PC 32:2 −52.5 4.0E−19 PC PC 32:3 −58.5 4.2E−17 PC PC 33:0 −17.1 2.0E−03 PC PC 33:1 −25.2 5.2E−06 PC PC 33:2 −35.6 6.9E−13 PC PC 33:3 −42.0 4.2E−11 PC PC 34:0 −11.9 3.6E−03 PC PC 34:3 −37.9 9.7E−14 PC PC 34:4 −52.1 5.6E−16 PC PC 34:5 −41.3 1.4E−05 PC PC 35:0 −21.3 5.5E−04 PC PC 35:1 −15.5 4.0E−03 PC PC 35:2 −26.3 2.6E−06 PC PC 35:3 −39.8 3.1E−15 PC PC 35:4 −25.8 1.5E−07 PC PC 36:0 −21.7 1.3E−07 PC PC 36:2 −35.1 2.0E−13 PC PC 36:3 −36.0 2.2E−14 PC PC 36:5 −32.1 1.4E−07 PC PC 36:6 −46.4 2.0E−09 PC PC 36:7 −27.4 2.1E−05 PC PC 37:1 −20.5 4.6E−06 PC PC 37:2 −36.3 6.0E−13 PC PC 37:3 −30.4 7.4E−09 PC PC 37:6 −20.3 8.9E−03 PC PC 38:0 −29.1 5.0E−11 PC PC 38:1 −9.8 2.8E−02 PC PC 38:2 −27.9 3.7E−08 PC PC 38:3 −30.6 1.5E−09 PC PC 38:4 −24.1 6.4E−06 PC PC 38:5 −15.7 6.6E−05 PC PC 38:6 −35.0 2.6E−10 PC PC 38:7 −30.9 7.8E−05 PC PC 39:0 −18.7 9.9E−07 PC PC 39:4 −19.5 5.6E−06 PC PC 39:5 −12.0 3.5E−02 PC PC 40:1 −22.9 3.6E−08 PC PC 40:3 −15.8 1.6E−04 PC PC 40:4 −29.9 9.0E−12 PC PC 40:5 −14.8 1.4E−03 PC PC 40:8 −33.4 6.8E−10 PC PC O-32:0 −24.5 2.7E−08 PC O/P PC O-32:1 −31.3 7.1E−10 PC O/P PC O-34:0 −20.9 1.1E−06 PC O/P PC O-34:1 −22.2 1.2E−08 PC O/P PC O-34:2 −39.7 6.7E−17 PC O/P PC O-36:1 −27.6 1.5E−07 PC O/P PC O-36:2 −36.3 4.2E−14 PC O/P PC O-36:3 −38.7 4.8E−16 PC O/P PC O-36:4 −23.1 2.4E−09 PC O/P PC O-38:0 −21.3 8.0E−09 PC O/P PC O-38:1 −27.9 5.8E−12 PC O/P PC O-38:2 −28.3 4.8E−09 PC O/P PC O-38:3 −31.7 2.9E−11 PC O/P PC O-38:4 −31.5 7.2E−12 PC O/P PC O-38:5 −16.0 3.4E−05 PC O/P PC O-38:6 −13.5 7.2E−03 PC O/P PC O-40:1 −23.6 4.1E−06 PC O/P PC O-40:3 −37.5 2.5E−10 PC O/P PC O-40:4 −18.1 8.5E−04 PC O/P PC O-40:5 −20.3 1.3E−04 PC O/P PC P-36:2 −26.8 6.2E−08 PC O/P PC P-32:0 −20.2 8.1E−07 PC O/P PC P-32:1 −21.4 1.6E−05 PC O/P PC P-34:1 −13.9 3.6E−04 PC O/P PC P-34:2 −33.3 2.7E−13 PC O/P PC P-36:3 −17.8 2.4E−06 PC O/P PC P-36:4 −20.0 8.1E−07 PC O/P PC P-38:1 −16.5 4.7E−04 PC O/P PC P-38:2 −17.0 2.7E−03 PC O/P PC P-38:4 −21.4 2.1E−05 PC O/P PC P-38:5 −11.3 1.5E−03 PC O/P PC P-38:6 −22.9 5.9E−04 PC O/P PC P-40:1 −15.9 5.1E−04 PC O/P PC P-40:2 −21.6 1.3E−05 PC O/P PC P-40:3 −23.1 3.4E−07 PC O/P PC P-40:4 −21.8 2.8E−07 PC O/P PC P-40:5 −17.3 2.4E−05 PC O/P PC P-40:6 −6.2 3.7E−02 PC O/P PE 32:1 −39.1 2.3E−04 PE PE 34:2 −39.2 1.0E−06 PE PE 34:3 −49.7 8.1E−07 PE PE 36:3 −53.8 2.0E−07 PE PE 36:4 −30.7 2.9E−04 PE PE 36:5 −26.5 2.3E−02 PE PE 38:1 −37.4 7.9E−05 PE PE 38:2 −29.4 2.0E−02 PE PE 38:3 −40.4 6.8E−07 PE PE 38:4 −47.6 7.3E−07 PE PE 38:5 −33.5 5.1E−06 PE PE 40:4 −34.8 1.2E−03 PE PE 40:5 −21.8 8.3E−04 PE PE 40:7 −42.4 1.3E−03 PE PE O-34:1 −12.4 4.7E−02 PE O II PE O-36:4 −49.1 4.9E−11 PE O II PE O-38:4 −48.9 1.5E−12 PE O II PE O-38:5 −38.5 1.3E−09 PE O II PE O-38:6 −16.2 4.4E−02 PE O II PE P-34:2 −20.6 1.4E−03 PE P PE P-36:2 −41.7 7.0E−10 PE P PE P-36:4 −31.6 1.0E−06 PE P PE P-38:4 −36.4 7.4E−10 PE P PE P-38:6 −31.2 1.4E−07 PE P PE P-40:6 −20.6 1.6E−03 PE P PG 34:1 −22.9 1.1E−02 PG PG 36:2 −25.6 3.5E−04 PG PI 32:0 −49.6 3.0E−04 PI PI 34:0 −45.7 5.7E−04 PI PI 34:1 −31.3 4.7E−04 PI PI 34:2 −31.3 6.4E−06 PI PI 36:1 −40.1 3.8E−07 PI PI 36:2 −27.9 5.2E−06 PI PI 36:3 −51.5 9.9E−10 PI PI 36:4 −24.3 2.7E−03 PI PI 38:2 −40.5 3.5E−07 PI PI 38:3 −33.8 1.6E−09 PI PI 38:5 −24.2 4.3E−05 PI PI 40:4 −19.9 1.1E−03 PI S1P d16:1 −20.0 1.4E−08 S1P/SA1P S1P d18:1 −19.3 1.3E−07 S1P/SA1P S1P d18:2 −29.3 1.2E−13 S1P/SA1P SA1P d18:0 −24.8 5.3E−13 S1P/SA1P SM 30:2 −35.8 2.2E−11 SM SM 31:0 −12.2 4.2E−02 SM SM 31:1 −24.7 1.2E−07 SM SM 31:2 −12.3 7.5E−03 SM SM 32:0 −24.2 8.6E−08 SM SM 32:2 −28.0 3.3E−11 SM SM 33:1 −11.6 4.7E−03 SM SM 33:2 −9.2 4.4E−02 SM SM 34:0 −15.1 5.0E−04 SM SM 34:1 −13.2 4.6E−04 SM SM 34:2 −15.8 2.3E−05 SM SM 35:0 −25.3 3.2E−06 SM SM 35:1 −10.3 9.7E−03 SM SM 35:2 −9.3 9.1E−03 SM SM 36:3 −14.6 2.7E−03 SM SM 37:0 −10.1 1.6E−02 SM SM 37:1 −14.8 1.9E−04 SM SM 37:2 −34.8 1.4E−13 SM SM 38:0 −22.5 2.0E−05 SM SM 38:1 −20.4 1.0E−06 SM SM 38:3 −20.2 8.5E−05 SM SM 39:0 −18.0 7.1E−03 SM SM 39:2 −21.6 1.5E−03 SM SM 40:0 −27.0 5.9E−07 SM SM 40:3 −19.8 2.6E−06 SM SM 41:0 −32.5 1.3E−09 SM SM 41:2 −17.0 7.1E−05 SM SM 42:0 −30.9 3.8E−10 SM SM 42:3 −15.8 3.9E−05 SM SM 44:1 −23.7 1.1E−08 SM SM 44:3 −16.6 1.3E−04 SM

    [0215] Table 4 shows statistically significantly (p<0.05) increased or decreased lipid biomarkers in ovarian cancer patients as compared to control group according to some embodiments of all aspects of the present disclosure. The results were derived from cohort I.

    TABLE-US-00004 TABLE 4 Increased (group A) (a) and decreased (group B) (b) lipid biomarkers in ovarian cancer patients according to some embodiments of the present disclosure. Abbreviations are described in the detailed description of the present disclosure. CHANGE (%) P-VALUE CLASS Table 4a. Group A molecules Cer(d16:1/18:0) 24.6 1.3E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/16:0) 27.0 3.2E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/18:0) 49.7 5.7E−05 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/20:0) 22.0 1.8E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:0/24:1) 16.4 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/18:0) 71.4 7.2E−12 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/20:0) 39.7 9.9E−08 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/22:0) 11.9 2.1E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:1/24:1) 30.5 1.1E−06 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/16:0) 15.2 1.2E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/18:0) 26.2 5.2E−04 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d18:2/24:1) 13.5 1.5E−02 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/22:0) 21.6 2.8E−03 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 Cer(d20:1/24:1) 43.9 2.1E−07 Ceramides with FA 16:0, 18:0, 20:0, 22:0, 24:1 LPC 20:1_sn2 20.6 3.8E−02 LPL I LPC 20:5_sn1 18.6 2.6E−02 LPL I LPC O-18:1 35.9 1.9E−02 LPL I LPC P-18:1 29.2 4.4E−02 LPL I LPE 18:0_sn1 35.5 1.4E−03 LPL I LPE 18:0_sn2 33.5 2.6E−03 LPL I PE O-36:1 62.4 1.9E−05 PE O I PE O-36:2 93.3 6.5E−08 PE O I PE O-36:3 66.0 9.3E−05 PE O I Table 4b. Group B molecules CE 14:0 −22.3 4.1E−06 CE CE 14:1 −22.0 4.7E−05 CE CE 15:0 −16.8 2.0E−04 CE CE 16:0 −12.5 1.4E−04 CE CE 16:2 −21.1 2.0E−07 CE CE 17:0 −16.5 1.2E−04 CE CE 17:1 −10.1 2.1E−02 CE CE 18:0 −26.7 4.9E−10 CE CE 18:1 −12.7 1.9E−04 CE CE 18:2 −21.9 7.2E−10 CE CE 19:1 −0.9 4.6E−02 CE CE 20:2 −23.3 2.6E−07 CE CE 20:3 −21.3 5.8E−06 CE CE 20:4 −7.4 5.0E−02 CE CE 22:3 −15.8 3.9E−02 CE CE 22:4 −14.9 1.6E−03 CE CE 22:5 −35.3 2.0E−08 CE CE 24:6 −22.7 1.2E−04 CE Cer(d16:1/23:0) −24.9 5.7E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/24:0) −29.3 6.1E−08 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d16:1/26:0) −17.1 4.0E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/23:0) −17.4 3.8E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:0/24:0) −14.8 2.5E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/14:0) −10.3 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:1/24:0) −11.9 8.6E−03 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/23:0) −18.5 5.6E−04 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/24:0) −22.4 1.1E−05 Ceramides with FA 14:0, 23:0, 24:0, 26:0 Cer(d18:2/26:0) −14.2 4.0E−02 Ceramides with FA 14:0, 23:0, 24:0, 26:0 DAG(14:0/18:1) −39.1 6.9E−04 DAG DAG(14:0/18:2) −38.3 1.6E−03 DAG DAG(18:1/20:3) −31.3 1.1E−02 DAG DAG(16:0/16:1) −27.6 1.3E−02 DAG DAG(18:0/20:4) −22.0 3.9E−02 DAG Gb3(d18:1/16:0) −14.8 3.0E−04 Gb3 Gb3(d18:1/18:0) −13.4 2.2E−02 Gb3 Gb3(d18:1/22:0) −19.5 4.5E−04 Gb3 Gb3(d18:1/23:0) −16.2 5.6E−03 Gb3 Gb3(d18:1/24:0) −23.1 8.6E−05 Gb3 Glc/GalCer(d16:1/20:0) −16.8 9.7E−04 Glc/GalCer Glc/GalCer(d16:1/22:0) −23.4 1.5E−05 Glc/GalCer Glc/GalCer(d16:1/23:0) −19.3 3.8E−04 Glc/GalCer Glc/GalCer(d16:1/24:0) −23.1 5.0E−06 Glc/GalCer Glc/GalCer(d18:1/16:0) −13.3 1.1E−02 Glc/GalCer Glc/GalCer(d18:1/18:0) −16.1 7.5E−04 Glc/GalCer Glc/GalCer(d18:1/20:0) −19.7 9.2E−05 Glc/GalCer Glc/GalCer(d18:1/22:0) −25.9 2.6E−07 Glc/GalCer Glc/GalCer(d18:1/23:0) −23.8 7.8E−07 Glc/GalCer Glc/GalCer(d18:1/24:0) −24.2 4.7E−07 Glc/GalCer Glc/GalCer(d18:1/26:0) −21.0 9.4E−05 Glc/GalCer Glc/GalCer(d18:2/20:0) −23.3 1.7E−04 Glc/GalCer Glc/GalCer(d18:2/22:0) −17.9 5.0E−04 Glc/GalCer Glc/GalCer(d18:2/23:0) −21.6 5.2E−05 Glc/GalCer Glc/GalCer(d18:2/24:0) −23.9 4.8E−08 Glc/GalCer LacCer(d16:1/16:0) −21.1 5.3E−05 LacCer LacCer(d18:1/22:0) −27.7 7.6E−10 LacCer LacCer(d18:1/23:0) −26.0 1.7E−07 LacCer LacCer(d18:1/24:0) −23.2 1.5E−08 LacCer LacCer(d18:1/24:1) −21.0 2.8E−06 LacCer LacCer(d18:2/16:0) −25.2 5.5E−08 LacCer LacCer(d18:2/22:0) −18.2 3.9E−02 LacCer LacCer(d18:2/24:0) −19.7 4.0E−03 LacCer LacCer(d18:2/24:1) −10.7 2.6E−02 LacCer LPC 20:0_sn1 −16.6 3.4E−03 LPL II LPC 20:0_sn2 −14.6 1.1E−02 LPL II LPC 20:2_sn1 −5.7 3.7E−02 LPL II LPC 20:2_sn2 −16.4 3.2E−05 LPL II LPC 22:0_sn1 −28.7 6.3E−10 LPL II LPC 22:4_sn1 −11.1 1.7E−02 LPL II LPC 22:4_sn2 −19.6 4.9E−05 LPL II LPC 24:0_sn1 −28.7 1.5E−12 LPL II LPC 24:0_sn2 −28.9 2.3E−13 LPL II LPC O-20:0 −15.5 7.1E−04 LPL II LPC O-22:0 −25.3 4.8E−11 LPL II LPC O-22:1 −17.5 1.9E−03 LPL II LPC O-24:0 −26.6 2.9E−10 LPL II LPC O-24:1 −17.8 8.2E−05 LPL II LPC O-24:2 −33.0 1.2E−08 LPL II PC 28:0 −59.0 1.5E−15 PC PC 30:0 −43.4 4.0E−15 PC PC 30:1 −52.1 2.4E−13 PC PC 30:2 −61.4 2.1E−14 PC PC 31:0 −26.0 1.6E−07 PC PC 31:1 −36.9 1.9E−08 PC PC 32:0 −15.5 4.3E−05 PC PC 32:1 −29.0 1.9E−06 PC PC 32:2 −52.5 4.0E−19 PC PC 32:3 −58.5 4.2E−17 PC PC 33:0 −17.1 2.0E−03 PC PC 33:1 −25.2 5.2E−06 PC PC 33:2 −35.6 6.9E−13 PC PC 33:3 −42.0 4.2E−11 PC PC 34:0 −11.9 3.6E−03 PC PC 34:3 −37.9 9.7E−14 PC PC 34:5 −41.3 1.4E−05 PC PC 35:0 −21.3 5.5E−04 PC PC 35:1 −15.5 4.0E−03 PC PC 35:2 −26.3 2.6E−06 PC PC 35:3 −39.8 3.1E−15 PC PC 35:4 −25.8 1.5E−07 PC PC 36:0 −21.7 1.3E−07 PC PC 36:5 −32.1 1.4E−07 PC PC 36:6 −46.4 2.0E−09 PC PC 36:7 −27.4 2.1E−05 PC PC 37:1 −20.5 4.6E−06 PC PC 37:2 −36.3 6.0E−13 PC PC 37:3 −30.4 7.4E−09 PC PC 37:6 −20.3 8.9E−03 PC PC 38:0 −29.1 5.0E−11 PC PC 38:1 −9.8 2.8E−02 PC PC 38:7 −30.9 7.8E−05 PC PC 39:0 −18.7 9.9E−07 PC PC 39:4 −19.5 5.6E−06 PC PC 39:5 −12.0 3.5E−02 PC PC 40:1 −22.9 3.6E−08 PC PC 40:3 −15.8 1.6E−04 PC PC 40:4 −29.9 9.0E−12 PC PC 40:8 −33.4 6.8E−10 PC PC O-32:0 −24.5 2.7E−08 PC O/P PC O-32:1 −31.3 7.1E−10 PC O/P PC O-34:0 −20.9 1.1E−06 PC O/P PC O-34:1 −22.2 1.2E−08 PC O/P PC O-34:2 −39.7 6.7E−17 PC O/P PC O-36:1 −27.6 1.5E−07 PC O/P PC O-36:2 −36.3 4.2E−14 PC O/P PC O-36:3 −38.7 4.8E−16 PC O/P PC O-36:4 −23.1 2.4E−09 PC O/P PC O-38:0 −21.3 8.0E−09 PC O/P PC O-38:1 −27.9 5.8E−12 PC O/P PC O-38:2 −28.3 4.8E−09 PC O/P PC O-38:3 −31.7 2.9E−11 PC O/P PC O-38:4 −31.5 7.2E−12 PC O/P PC O-38:5 −16.0 3.4E−05 PC O/P PC O-38:6 −13.5 7.2E−03 PC O/P PC O-40:1 −23.6 4.1E−06 PC O/P PC O-40:3 −37.5 2.5E−10 PC O/P PC O-40:4 −18.1 8.5E−04 PC O/P PC O-40:5 −20.3 1.3E−04 PC O/P PC P-36:2 −26.8 6.2E−08 PC O/P PC P-32:0 −20.2 8.1E−07 PC O/P PC P-32:1 −21.4 1.6E−05 PC O/P PC P-34:1 −13.9 3.6E−04 PC O/P PC P-34:2 −33.3 2.7E−13 PC O/P PC P-36:4 −20.0 8.1E−07 PC O/P PC P-38:1 −16.5 4.7E−04 PC O/P PC P-38:2 −17.0 2.7E−03 PC O/P PC P-38:4 −21.4 2.1E−05 PC O/P PC P-38:5 −11.3 1.5E−03 PC O/P PC P-38:6 −22.9 5.9E−04 PC O/P PC P-40:1 −15.9 5.1E−04 PC O/P PC P-40:2 −21.6 1.3E−05 PC O/P PC P-40:3 −23.1 3.4E−07 PC O/P PC P-40:4 −21.8 2.8E−07 PC O/P PC P-40:5 −17.3 2.4E−05 PC O/P PE 32:1 −39.1 2.3E−04 PE PE 34:2 −39.2 1.0E−06 PE PE 34:3 −49.7 8.1E−07 PE PE 36:2 −37.9 1.5E−06 PE PE 36:3 −53.8 2.0E−07 PE PE 36:4 −30.7 2.9E−04 PE PE 36:5 −26.5 2.3E−02 PE PE 38:1 −37.4 7.9E−05 PE PE 38:2 −29.4 2.0E−02 PE PE 38:3 −40.4 6.8E−07 PE PE 38:5 −33.5 5.1E−06 PE PE 40:4 −34.8 1.2E−03 PE PE 40:5 −21.8 8.3E−04 PE PE 40:7 −42.4 1.3E−03 PE PE O-34:1 −12.4 4.7E−02 PE O II PE O-36:4 −49.1 4.9E−11 PE O II PE O-38:4 −48.9 1.5E−12 PE O II PE O-38:5 −38.5 1.3E−09 PE O II PE O-38:6 −16.2 4.4E−02 PE O II PE P-34:2 −20.6 1.4E−03 PE P PE P-36:2 −41.7 7.0E−10 PE P PE P-36:4 −31.6 1.0E−06 PE P PE P-38:4 −36.4 7.4E−10 PE P PE P-38:6 −31.2 1.4E−07 PE P PG 34:1 −22.9 1.1E−02 PG PG 36:2 −25.6 3.5E−04 PG PI 32:0 −49.6 3.0E−04 PI PI 34:0 −45.7 5.7E−04 PI PI 34:1 −31.3 4.7E−04 PI PI 34:2 −31.3 6.4E−06 PI PI 36:1 −40.1 3.8E−07 PI PI 36:2 −27.9 5.2E−06 PI PI 36:3 −51.5 9.9E−10 PI PI 36:4 −24.3 2.7E−03 PI PI 38:2 −40.5 3.5E−07 PI PI 38:3 −33.8 1.6E−09 PI PI 38:5 −24.2 4.3E−05 PI PI 40:4 −19.9 1.1E−03 PI SM 30:2 −35.8 2.2E−11 SM SM 31:0 −12.2 4.2E−02 SM SM 31:1 −24.7 1.2E−07 SM SM 31:2 −12.3 7.5E−03 SM SM 32:0 −24.2 8.6E−08 SM SM 33:1 −11.6 4.7E−03 SM SM 33:2 −9.2 4.4E−02 SM SM 34:0 −15.1 5.0E−04 SM SM 34:2 −15.8 2.3E−05 SM SM 35:0 −25.3 3.2E−06 SM SM 35:1 −10.3 9.7E−03 SM SM 35:2 −9.3 9.1E−03 SM SM 36:3 −14.6 2.7E−03 SM SM 37:0 −10.1 1.6E−02 SM SM 37:1 −14.8 1.9E−04 SM SM 37:2 −34.8 1.4E−13 SM SM 38:0 −22.5 2.0E−05 SM SM 38:1 −20.4 1.0E−06 SM SM 38:3 −20.2 8.5E−05 SM SM 39:0 −18.0 7.1E−03 SM SM 39:2 −21.6 1.5E−03 SM SM 40:0 −27.0 5.9E−07 SM SM 40:3 −19.8 2.6E−06 SM SM 41:0 −32.5 1.3E−09 SM SM 41:2 −17.0 7.1E−05 SM SM 42:0 −30.9 3.8E−10 SM SM 42:3 −15.8 3.9E−05 SM SM 44:1 −23.7 1.1E−08 SM SM 44:3 −16.6 1.3E−04 SM

    [0216] Tables 5-9 show examples of combinations of increasing and decreasing small molecule biomarkers from Group A and Group B, respectively, and are provided to illustrate various aspects of the present disclosure. They are not intended to limit the present disclosure, which is defined by the accompanying claims.

    Combination of Small Molecule Biomarkers Improves Diagnostic Performance

    [0217] The embodiments of the present disclosure, i.e. combining increasing and decreasing small molecule biomarkers, improves diagnostic performance. This is illustrated in Table 5 by ratios of small molecule biomarkers. In Table 5 the results are shown for patients with malignant tumors vs. controls comparison in cohort I. When taking the ratio of two small molecule biomarkers, the performance measured by AUC as well as sum of sensitivity and specificity is improved when comparing the performance to single small molecule biomarker components of the ratios.

    TABLE-US-00005 TABLE 5 Performance of small molecule ratios (a) and single small molecule biomarkers (b) in ovarian cancer detection. Abbreviations are described in the detailed description of the present disclosure. AUC SE + SP SE SP Table 5a. Small molecule ratios 2-Hydroxybutyric acid|Ketoleucine 0.895 1.66 0.92 0.73 3,4-Dihydroxybutyric acid|Alanine 0.910 1.68 0.97 0.71 3,4-Dihydroxybutyric acid|PC 30:0 0.885 1.62 0.88 0.74 3,4-Dihydroxybutyric acid|PC 32:2 0.886 1.62 0.74 0.89 3,4-Dihydroxybutyric acid|PC 32:3 0.888 1.62 0.77 0.85 3,4-Dihydroxybutyric acid|PC 34:4 0.870 1.57 0.73 0.84 3,4-Dihydroxybutyric acid|PC 36:3 0.873 1.61 0.81 0.80 3,4-Dihydroxybutyric acid|PC O-34:2 0.876 1.61 0.81 0.80 3,4-Dihydroxybutyric acid|PC O-36:2 0.870 1.59 0.80 0.79 3,4-Dihydroxybutyric acid|PC O-36:3 0.877 1.60 0.86 0.74 3,4-Dihydroxybutyric acid|S1P d18:1 0.874 1.61 0.87 0.74 3,4-Dihydroxybutyric acid|S1P d18:2 0.885 1.63 0.89 0.73 3-Hydroxybutyric acid|Ketoleucine 0.887 1.63 0.87 0.77 3-Hydroxybutyric acid|PC O-34:2 0.875 1.60 0.82 0.79 3-Hydroxybutyric acid|PC O-36:3 0.870 1.63 0.81 0.82 3-Hydroxybutyric acid|PE O-38:4 0.871 1.58 0.86 0.71 3-Hydroxybutyric acid|PE P-34:2 0.870 1.66 0.98 0.68 3-Hydroxybutyric acid|SM 37:2 0.874 1.64 0.83 0.81 Adipic acid|Ketoleucine 0.857 1.63 0.89 0.73 Cer(d18:1/18:0)|Alanine 0.859 1.61 0.90 0.70 Cer(d18:1/24:1)|PC O-36:3 0.858 1.62 0.78 0.85 LPE 18:0_sn1|LPC 18:2_sn2 0.869 1.62 0.84 0.79 Table 5b. Single small molecule biomarkers 2-Hydroxybutyric acid 0.724 1.38 0.82 0.56 3,4-Dihydroxybutyric acid 0.853 1.63 0.73 0.90 3-Hydroxybutyric acid 0.832 1.54 0.93 0.61 Adipic acid 0.712 1.39 0.68 0.70 Alanine 0.842 1.58 0.91 0.67 Cer(d18:1/18:0) 0.740 1.35 0.58 0.78 Cer(d18:1/24:1) 0.675 1.28 0.55 0.73 Ketoleucine 0.808 1.51 0.78 0.72 LPC 18:2_sn2 0.755 1.43 0.72 0.70 LPE 18:0_sn1 0.595 1.25 0.31 0.94 PC 30:0 0.784 1.46 0.82 0.64 PC 32:2 0.818 1.51 0.80 0.71 PC 32:3 0.801 1.46 0.74 0.72 PC 34:4 0.789 1.43 0.78 0.64 PC 36:3 0.772 1.42 0.61 0.81 PC O-34:2 0.795 1.45 0.59 0.87 PC O-36:2 0.782 1.46 0.74 0.71 PC O-36:3 0.801 1.52 0.70 0.82 PE O-38:4 0.746 1.34 0.71 0.63 PE P-34:2 0.658 1.28 0.50 0.78 S1P d18:1 0.698 1.32 0.58 0.74 S1P d18:2 0.788 1.50 0.88 0.62 SM 37:2 0.777 1.45 0.66 0.79

    Combination of Small Molecule Biomarkers Improves Diagnostic Performance of Early Stage Ovarian Cancer Patients

    [0218] Combination of increasing and decreasing small molecule biomarkers improves diagnostic performance also in the detection of early stage (stage I or II) ovarian cancer patients. This is illustrated in Table 6 by ratios of small molecule biomarkers. In Table 6 the results are shown for patients with stage I or II ovarian cancer vs. controls in cohort II. When taking the ratio of two small molecule biomarkers, the performance measured by AUC as well as sum of sensitivity and specificity is improved when comparing the performance to single small molecule biomarker components of the ratios.

    TABLE-US-00006 TABLE 6 Performance of small molecule ratios (a) and single small molecule biomarkers (b) in early-stage ovarian cancer detection. Abbreviations are described in the detailed description of the present disclosure. AUC SE + SP SE SP Table 6a. Small molecule ratios Acetoacetic acid|Gb3(d18:1/22:0) 0.833 1.55 0.69 0.86 Acetoacetic acid|Glc/GalCer(d16:1/20:0) 0.826 1.51 0.54 0.97 Acetoacetic acid|Glc/GalCer(d16:1/23:0) 0.833 1.53 0.58 0.95 Acetoacetic acid|Glc/GalCer(d16:1/24:0) 0.827 1.52 0.85 0.67 Acetoacetic acid|Ketoleucine 0.828 1.53 0.77 0.76 Acetoacetic acid|LacCer(d18:1/23:0) 0.844 1.57 0.77 0.80 Acetoacetic acid|PC 34:3 0.828 1.51 0.58 0.94 Acetoacetic acid|PC O-36:1 0.833 1.55 0.96 0.59 Acetoacetic acid|PC P-38:2 0.839 1.57 0.88 0.69 Acetoacetic acid|SM 31:2 0.832 1.55 0.65 0.89 Cer(d18:0/18:0)|LacCer(d18:1/23:0) 0.755 1.46 0.71 0.75 Cer(d18:1/24:1)|LPC 20:2_sn2 0.785 1.49 0.81 0.69 Cer(d18:1/24:1)|PI 36:3 0.777 1.47 0.73 0.74 Table 6b. Single small molecule biomarkers Acetoacetic acid 0.810 1.49 0.54 0.95 Cer(d18:0/18:0) 0.607 1.23 0.50 0.73 Cer(d18:1/24:1) 0.535 1.17 0.73 0.44 Gb3(d18:1/22:0) 0.668 1.29 0.46 0.83 Glc/GalCer(d16:1/20:0) 0.688 1.40 0.85 0.56 Glc/GalCer(d16:1/23:0) 0.640 1.25 0.62 0.63 Glc/GalCer(d16:1/24:0) 0.689 1.39 0.81 0.58 Ketoleucine 0.652 1.33 0.69 0.64 LacCer(d18:1/23:0) 0.713 1.46 0.73 0.73 LPC 20:2_sn2 0.750 1.43 0.58 0.85 PC 34:3 0.624 1.24 0.81 0.43 PC O-36:1 0.673 1.44 0.73 0.71 PC P-38:2 0.611 1.22 0.35 0.87 PI 36:3 0.754 1.42 0.65 0.76 SM 31:2 0.631 1.33 0.52 0.81
    Combination of Small Molecule Biomarkers with CA-125 Improves Diagnostic Performance of Early Stage Ovarian Cancer Patients

    [0219] Combination of increasing and decreasing small molecule biomarkers improves diagnostic performance also when combined together with protein biomarker CA-125. This is illustrated in Table 7 by ratios of small molecule biomarkers. In Table 7 the results are shown for patients with stage I or II ovarian cancer vs. controls in cohort II. When constructing a logistic regression model taking the ratio of two small molecule biomarkers and CA-125, the performance measured by AUC is improved when comparing the performance to a logistic regression model incorporating a single small molecule biomarker component and CA-125. Moreover, the AUC values are higher than those obtained for CA-125 alone.

    TABLE-US-00007 TABLE 7 Performance of small molecule ratios (a) and single small molecule biomarkers (b) in early-stage ovarian cancer detection when combined with CA-125. (c) Performance of CA-125 alone. Abbreviations are described in the detailed description of the present disclosure. AUC Table 7a. Small molecule ratios 2-Hydroxybutyric acid|SM 39:0 0.870 3-Hydroxybutyric acid|SM 39:0 0.882 Acetoacetic acid|Cer(d16:1/26:0) 0.914 Acetoacetic acid|Gb3(d18:1/22:0) 0.921 Acetoacetic acid|Glc/GalCer(d16:1/22:0) 0.913 Acetoacetic acid|Glc/GalCer(d16:1/23:0) 0.914 Acetoacetic acid|LacCer(d16:1/16:0) 0.915 Acetoacetic acid|PC 37:3 0.914 Acetoacetic acid|PC O-40:5 0.913 Acetoacetic acid|PC P-34:2 0.913 Acetoacetic acid|PC P-38:2 0.914 Acetoacetic acid|SM 31:2 0.920 Acetoacetic acid|SM 39:0 0.913 Acetoacetic acid|SM 39:1 0.913 Cer(d18:1/24:1)|LPC 14:0_sn1 0.874 Cer(d18:1/24:1)|PI 36:1 0.874 Cer(d18:1/24:1)|PI 36:3a 0.882 TAG(18:2/18:2/18:2)|SM 39:0 0.880 Table 7b. Single small molecule biomarkers 2-Hydroxybutyric acid 0.807 3-Hydroxybutyric acid 0.827 Acetoacetic acid 0.903 Cer(d16:1/26:0) 0.821 Cer(d18:1/24:1) 0.800 Gb3(d18:1/22:0) 0.839 Glc/GalCer(d16:1/22:0) 0.830 Glc/GalCer(d16:1/23:0) 0.816 LacCer(d16:1/16:0) 0.833 LPC 14:0_sn1 0.853 PC 37:3 0.869 PC O-40:5 0.832 PC P-34:2 0.835 PC P-38:2 0.802 PI 36:1 0.844 PI 36:3a 0.854 SM 31:2 0.827 SM 39:0 0.864 SM 39:1 0.836 TAG(18:2/18:2/18:2) 0.816 Table 7c. CA-125 value CA-125 0.806

    Combination of Small Molecule Biomarkers Improves Diagnostic Performance in Premenopausal Women

    [0220] Combination of increasing and decreasing small molecule biomarkers improves diagnostic performance especially in premenopausal women suffering from ovarian cancer. This is illustrated in Table 8 by ratios of small molecule biomarkers. In Table 8 the results are shown for premenopausal patients with ovarian cancer vs. premenopausal controls in cohort II. When taking the ratio of two small molecule biomarkers, the performance measured by AUC as well as sum of sensitivity and specificity is improved when comparing the performance to single small molecule biomarker components of the ratios. The performance is improved also when compared to the clinically used protein biomarker CA-125.

    TABLE-US-00008 TABLE 8 Performance of small molecule ratios (a), single small molecule biomarkers (b) and CA-125 in detection of ovarian cancer in premenopausal women. Abbreviations are described in the detailed description of the present disclosure. AUC SE + SP SE SP Table 8a. Small molecule ratios 2-Hydroxybutyric acid|PC O-36:1 0.824 1.66 0.94 0.72 Acetoacetic acid|PC 30:2 0.825 1.51 0.65 0.87 Cer(d18:1/24:1)|LPC 14:0_sn1 0.832 1.53 0.76 0.77 Cer(d20:1/24:1)|LPC 14:0_sn1 0.830 1.56 0.94 0.62 PE O-36:2|PE P-36:2 0.839 1.64 0.94 0.69 TAG(18:1/18:1/22:6)|LPC 14:0_sn1 0.840 1.60 0.76 0.83 TAG(18:1/18:1/22:6)|LPC 14:0_sn2 0.839 1.58 0.76 0.82 TAG(18:1/18:1/22:6)|PC 36:6 0.855 1.59 0.82 0.77 TAG(18:1/18:1/22:6)|PC O-36:1 0.837 1.61 0.94 0.67 TAG(18:1/18:1/22:6)|TAG(14:0/18:2/18:2) 0.861 1.72 0.88 0.83 Table 8b. Single small molecule biomarkers 2-Hydroxybutyric acid 0.700 1.42 0.82 0.60 Acetoacetic acid 0.788 1.45 0.88 0.57 Cer(d18:1/24:1) 0.615 1.37 0.82 0.55 Cer(d20:1/24:1) 0.638 1.32 0.59 0.73 LPC 14:0_sn1 0.818 1.57 0.88 0.68 LPC 14:0_sn2 0.790 1.51 0.71 0.80 PC 30:2 0.802 1.51 0.82 0.68 PC 36:6 0.748 1.51 0.65 0.87 PC O-36:1 0.790 1.62 0.88 0.73 PE O-36:2 0.520 1.16 0.65 0.52 PE P-36:2 0.810 1.56 0.88 0.68 TAG(14:0/18:2/18:2) 0.744 1.50 0.76 0.73 TAG(18:1/18:1/22:6) 0.680 1.34 0.82 0.52 Table 8c. Result for CA-125 CA125 0.777 1.51 0.94 0.57
    Combination of Small Molecule Biomarkers Improves Prediction of Overall Survival in Patients with Ovarian Cancer

    [0221] Combination of increasing and decreasing small molecule biomarkers improves the prediction of overall survival in patients suffering from ovarian cancer. This is illustrated in Table 9 by ratios of small molecule biomarkers. When taking the ratio of two small molecule biomarkers, the performance measured by hazard ratios and p-values of the cox regression models are improved when comparing the performance to single small molecule biomarker components of the ratios. The performance is improved also when compared to the clinically used protein biomarker CA-125.

    TABLE-US-00009 TABLE 9 Performance of small molecule ratios (a), single small molecule biomarkers (b) and CA-125 in predicting the overall survival of ovarian cancer patients. Abbreviations are described in the detailed description of the present disclosure. HR (95% CI) P-value Table 9a. Small molecule ratios 3,4-Dihydroxybutyric acid|PC 1.49 (1.19, 1.87) 5.0E−04 37:6 Adipic acid|PC O-38:2 1.57 (1.26, 1.96) 5.0E−05 Cer(d18:1/16:0)|PC O-38:2 1.81 (1.42, 2.32) 2.4E−06 Cer(d18:1/16:0)|SM 41:1 1.72 (1.36, 2.18) 4.8E−06 Cer(d20:1/24:1)|CE 20:3 1.80 (1.40, 2.31) 3.8E−06 Cer(d20:1/24:1)|PC 37:3 1.90 (1.45, 2.49) 3.0E−06 Cer(d20:1/24:1)|PC 37:6 1.84 (1.44, 2.36) 1.4E−06 Cer(d20:1/24:1)|PC 40:8 2.05 (1.51, 2.77) 3.4E−06 Cer(d20:1/24:1)|PC O-38:1 1.81 (1.41, 2.33) 3.1E−06 Cer(d20:1/24:1)|PC O-38:2 1.86 (1.45, 2.40) 1.4E−06 Cer(d20:1/24:1)|SM 41:1 1.82 (1.42, 2.34) 2.7E−06 TAG(18:1/18:1/20:4)|PC 37:3 1.51 (1.19, 1.91) 5.7E−04 Table 9b. Single small molecule biomarkers 3,4-Dihydroxybutyric acid 1.38 (1.10, 1.73) 6.0E−03 Adipic acid 1.37 (1.09, 1.70) 5.8E−03 CE 20:3 0.75 (0.61, 0.93) 7.7E−03 Cer(d18:1/16:0) 1.42 (1.12, 1.80) 3.4E−03 Cer(d20:1/24:1) 1.53 (1.20, 1.94) 5.3E−04 PC 37:3 0.74 (0.60, 0.92) 5.6E−03 PC 37:6 0.72 (0.57, 0.90) 3.4E−03 PC 40:8 0.75 (0.59, 0.94) 1.5E−02 PC O-38:1 0.72 (0.58, 0.88) 1.9E−03 PC O-38:2 0.65 (0.52, 0.82) 2.1E−04 SM 41:1 0.72 (0.59, 0.87) 9.5E−04 TAG(18:1/18:1/20:4) 1.24 (0.98, 1.55) 6.8E−02 Table 9c. Clinically used CA125 marker CA125 1.15 (0.85, 1.57) 3.6E−01
    Combination of Small Molecule Biomarkers with CA-125 Improves Diagnostic Performance of Early Stage Ovarian Cancer Patients

    [0222] Combination of cancer antigen 125 (CA-125) and decreasing small molecule biomarkers (group B) improves diagnostic performance also in the detection of early stage (stage I or II) ovarian cancer patients. This is illustrated in Table 10 by ratios of CA-125 and small molecule biomarkers. In Table 10 the results are shown for patients with stage I or II ovarian cancer vs. controls in cohort II. When taking the ratio of CA-125 and a small molecule biomarker, the performance measured by AUC as well as sum of sensitivity and specificity is improved when comparing the performance to CA-125 or single small molecule biomarker alone.

    TABLE-US-00010 TABLE 10 (a) Performance of ratios of CA-125 and small molecule biomarkers from group B in early-stage ovarian cancer detection, (b) Performance of CA-125 and single small molecule biomarkers alone. Abbreviations are described in the detailed description of the present disclosure. AUC SE + SP SE SP Table 10a. CA-125/small molecule ratios CA-125|LPC 14:0_sn2 0.858 1.62 0.88 0.73 CA-125|PI 36:3a 0.855 1.61 0.85 0.77 CA-125|PC 37:3 0.854 1.59 0.88 0.70 CA-125|LPC 14:0_sn1 0.853 1.60 0.85 0.75 CA-125|LPC 20:3_sn1 0.847 1.61 0.88 0.72 CA-125|PI 36:3b 0.845 1.61 0.85 0.77 CA-125|PC 34:4 0.845 1.59 0.92 0.67 CA-125|PC 40:4 0.843 1.64 0.92 0.71 CA-125|SM 39:0 0.842 1.61 0.87 0.74 CA-125|LPC 20:2_sn2 0.842 1.60 0.88 0.72 CA-125|PI 36:1 0.842 1.60 0.88 0.71 CA-125|S1P d18:2 0.838 1.62 0.92 0.70 CA-125|SM 39:1 0.836 1.64 0.88 0.76 CA-125|PI 38:5b 0.836 1.62 0.92 0.69 CA-125|PC 36:3b + 1 0.834 1.65 0.81 0.84 CA-125|PC P-34:2 + 1 0.830 1.64 0.88 0.76 CA-125|LPC 24:0_sn1 0.830 1.62 0.88 0.74 CA-125|SM 31:2 0.826 1.63 0.78 0.85 CA-125|SM 31:1 0.824 1.63 0.85 0.79 CA-125|SM 32:1 0.821 1.63 0.85 0.79 CA-125|Gb3(d18:1/23:0) 0.819 1.62 0.91 0.71 Table 10b. CA-125 or single small molecule biomarkers CA-125 0.806 1.55 0.92 0.63 LPC 14:0_sn2 0.713 1.34 0.85 0.50 PI 36:3a 0.754 1.42 0.65 0.76 PC 37:3 0.737 1.47 0.73 0.74 LPC 14:0_sn1 0.731 1.43 0.85 0.59 LPC 20:3_sn1 0.722 1.39 0.65 0.73 PI 36:3b 0.691 1.36 0.50 0.86 PC 34:4 0.651 1.27 0.85 0.42 PC 40:4 0.715 1.39 0.73 0.66 SM 39:0 0.555 1.20 0.83 0.38 LPC 20:2_sn2 0.750 1.43 0.58 0.85 PI 36:1 0.715 1.38 0.73 0.65 S1P d18:2 0.695 1.35 0.92 0.43 SM 39:1 0.686 1.43 0.69 0.73 PI 38:5b 0.645 1.27 0.42 0.84 PC 36:3b + 1 0.658 1.30 0.81 0.50 PC P-34:2 + 1 0.742 1.41 0.88 0.52 LPC 20:4_sn1 0.685 1.34 1.00 0.34 SM 31:2 0.631 1.33 0.52 0.81 SM 31:1 0.642 1.35 0.50 0.85 SM 32:1 0.662 1.29 0.73 0.56 Gb3(d18:1/23:0) 0.600 1.29 0.59 0.70
    Combination of Small Molecule Biomarkers with CA-125 Improves Diagnostic Performance in Premenopausal Women

    [0223] Combination of cancer antigen 125 (CA-125) and decreasing small molecule biomarkers (group B) improves diagnostic performance especially in premenopausal women suffering from ovarian cancer. This is illustrated in Table 11 by ratios of CA-125 and small molecule biomarkers. In Table 11 the results are shown for premenopausal patients with ovarian cancer vs. premenopausal controls in cohort II. When taking the ratio of CA-125 and a small molecule biomarker, the performance measured by AUC as well as sum of sensitivity and specificity is improved when comparing the performance to CA-125 or a single small molecule biomarker alone.

    TABLE-US-00011 TABLE 11 (a) Performance of ratios of CA-125 and small molecule biomarkers from group B in detection of ovarian cancer in premenopausal patients. (b) Performance of CA-125 and single small molecule biomarkers alone. Abbreviations are described in the detailed description of the present disclosure. AUC SE + SP SE SP Table 11a. CA-125/small molecule ratios CA-125|PC 30:2 0.875 1.65 1.00 0.65 CA-125|PC 28:0 0.864 1.59 0.82 0.77 CA-125|PC 32:2 0.863 1.60 1.00 0.60 CA-125|PC 32:3 0.855 1.62 0.88 0.73 CA-125|LPC 20:0_sn1 0.851 1.62 0.82 0.80 CA-125|TAG(14:0/18:2/18:2) 0.850 1.58 0.88 0.70 CA-125|PC 36:2 + 2 0.846 1.61 0.94 0.67 CA-125|PC 35:3b 0.843 1.55 0.88 0.67 CA-125|PC 36:3a + 1 0.842 1.62 0.94 0.68 CA-125|LPC 22:0_sn1 0.841 1.59 0.82 0.76 CA-125|PC 37:2 0.841 1.54 0.82 0.72 CA-125|TAG(14:0/16:1/18:2) 0.839 1.53 0.76 0.77 CA-125|SM 31:1 0.828 1.57 0.88 0.68 CA-125|SM 37:2 0.828 1.59 0.94 0.65 CA-125|DAG(14:0/18:2) 0.827 1.54 0.86 0.68 CA-125|SM 39:1 0.847 1.61 0.94 0.67 CA-125|LPC 14:0_sn1 0.862 1.60 0.88 0.72 CA-125|SM 30:2 0.819 1.60 0.76 0.83 CA-125|LPC 20:3_sn2 0.837 1.59 0.94 0.65 Table 11b. CA-125 or single small molecule biomarkers CA-125 0.777 1.51 0.94 0.57 PC 30:2 0.802 1.51 0.82 0.68 PC 28:0 0.779 1.46 0.71 0.75 PC 32:2 0.782 1.52 0.71 0.82 PC 32:3 0.756 1.51 0.76 0.75 LPC 20:0_sn1 0.795 1.56 0.82 0.73 TAG(14:0/18:2/18:2) 0.744 1.50 0.76 0.73 PC 36:2 + 2 0.784 1.52 0.71 0.82 PC 35:3b 0.768 1.52 0.88 0.63 PC 36:3a +1 0.805 1.54 0.82 0.72 LPC 22:0_sn1 0.812 1.52 0.82 0.69 PC 37:2 0.797 1.51 0.71 0.80 TAG(14:0/16:1/18:2) 0.726 1.47 0.94 0.53 SM 31:1 0.750 1.51 0.82 0.68 SM 37:2 0.743 1.46 0.71 0.75 DAG(14:0/18:2) 0.707 1.39 0.71 0.68 SM 39:1 0.762 1.57 0.88 0.68 LPC 14:0_sn1 0.818 1.57 0.88 0.68 SM 30:2 0.683 1.37 0.88 0.48 LPC 20:3_sn2 0.768 1.50 0.76 0.73