Methods of diagnosis and treatment of pancreatic cancer

Abstract

According to the present disclosure, novel proteomic profiles for pancreatic cancer are used to treat and / or ameliorate pancreatic cancer. In certain embodiments, the treatment includes removal of cancerous tissue after obtaining biopsy tissue from the patient. Optionally, the treatment comprises the administration of an anti-cancer therapeutic agent for reducing the progression of pancreatic cancer. In additional or alternative embodiments, novel proteomic profiles are used to monitor disease progression or remission.

Description

Technical Field

[0001] This disclosure generally pertains to the field of diagnosis and treatment of pancreatic cancer.

[0002] background

[0003] Pancreatic cancer has a poor prognosis. This is often because it is difficult to detect early. Patients usually have no symptoms until the cancer is untreatable and / or has spread throughout the body. Furthermore, because the tumor is located deep within the body, it is difficult to observe.

[0004] To facilitate early diagnosis, genetic testing is performed on patients with a family history of the disease. This testing detects gene changes that cause the genetic condition. However, these tests have limitations in their ability to predict pancreatic cancer risk. For individuals identified as having a risk of developing pancreatic cancer through genetic testing, endoscopic ultrasound or magnetic resonance imaging (MRI) is still used to assess the likelihood of pancreatic cancer. Although physicians can use genetic analysis to detect treatable pancreatic cancer early in members of the susceptible public, this testing is not used to screen the general population. (See American Cancer Society: www.cancer.org / cancer / pancreatic-cancer / detection-diagnosis-staging / detect.html).

[0005] Furthermore, very few drugs are available for treating pancreatic cancer. In recent years, numerous targeted agents have been tested, such as ECFR inhibitors (erlotinib), VEGF and VEGFR inhibitors, phosphoinositol-3-kinase-mTOR pathway inhibitors, Janus kinase inhibitors, and Ras pathway inhibitors. Unfortunately, most of these agents have proven ineffective in treating the disease (Zhu et al., 2018, “Pancreatic cancer: Challenge and opportunities”, BMC Medicine 16(214)).

[0006] Therefore, pancreatic cancer is not only difficult to detect in its early stages, but it also remains one of the most untreatable forms of cancer in humans.

[0007] Therefore, there is an urgent need for improved methods for diagnosing pancreatic cancer and / or determining an individual's predisposition to develop pancreatic cancer and its treatment. There is also an urgent need for screening and treatment of individuals with early-stage (e.g., stage 1 or 2) pancreatic cancer.

[0008] Overview

[0009] This disclosure provides methods for diagnosing and treating pancreatic cancer or for treating pancreatic cancer.

[0010] According to this disclosure, novel proteomic profiles of pancreatic cancer are used for the diagnosis, treatment, and / or improvement of pancreatic cancer. In some embodiments, the treatment includes surgery to remove the tumor or optionally administration of anticancer therapeutic agents such as those used to reduce the progression of pancreatic cancer. In other or alternative embodiments, the novel proteomic profiles are used to monitor disease progression and / or remission.

[0011] The proteomic profile of pancreatic cancer includes at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin. In one embodiment, pancreatic cancer is diagnosed by determining whether one, two, three, four, five, six, seven, or more proteins are elevated, for example, by an elevation of at least 1.2, 1.4, 1.6, 1.8, 2.0, or more times relative to levels considered normal in individuals without pancreatic cancer (e.g., baseline).

[0012] Surprisingly, it has been found that proteomic profiles containing multiple of the aforementioned proteins or their peptide fragments are elevated in individuals with early-stage pancreatic cancer (stage 1) compared to individuals with negative pancreatic cancer results. In some embodiments, the levels of pancreatic cancer-associated proteins are elevated in the circulation of individuals with pancreatic cancer compared to individuals without pancreatic cancer. In other embodiments, the levels of pancreatic cancer-associated proteins in the circulation of an individual are monitored as described herein, and if changes over time are detected, the individual is identified for further evaluation. In some embodiments, the levels of pancreatic cancer-associated proteins are altered in the blood (e.g., serum, plasma), body fluids (e.g., cerebrospinal fluid, pleural fluid, amniotic fluid, semen, or saliva), urine, and / or feces of individuals with pancreatic cancer. Not wishing to be bound by theory, novel combinations of pancreatic cancer-associated proteins are believed to play a role in the development of pancreatic cancer. Treatment may also include obtaining biopsy tissue to definitively confirm the presence of pancreatic cancer, followed by surgery and / or administration of pancreatic cancer drugs. Typically, this treatment comprises a combination of biopsy and surgery following a positive diagnosis.

[0013] According to one aspect of this disclosure, a method for treating pancreatic cancer in an individual is provided, the method comprising: (i) receiving a proteomic profile from the individual, the proteomic profile having previously been obtained by: (a) providing a biological sample obtained from the individual; (b) measuring a concentration level from the obtained sample of one or a combination of at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; and (c) comparing the concentration levels of the pancreatic cancer-associated proteins or peptide fragments thereof from the obtained sample with a negative reference value for pancreatic cancer and / or a reference value from a sample obtained from the individual at an earlier time point. (ii) Compare the concentration levels of pancreatic cancer-associated proteins or their peptide fragments to the obtained sample; (ii) if the concentration level of pancreatic cancer-associated proteins or their peptide fragments from the obtained sample is compared with the concentration level of pancreatic cancer-negative reference values ​​and / or reference concentration levels of pancreatic cancer-associated proteins or their peptide fragments from samples obtained at an earlier time point from the individual, the individual is identified as being at risk of pancreatic cancer; (iii) optionally obtain biopsy tissue from the individual identified as being at risk of pancreatic cancer in step (ii); (iv) analyze the biopsy tissue to determine the presence of cancer cells; (v) if the individual is identified as being at risk of pancreatic cancer based on the proteomic profile in step (ii) and if cancer cells are present in the biopsy tissue, the individual is identified as having pancreatic cancer; and (vi) if the individual is identified as having pancreatic cancer, optionally remove cancerous tissue from the pancreas and / or bile duct of the individual.

[0014] According to another aspect of this disclosure, a method for diagnosing and treating pancreatic cancer in an individual is provided, the method comprising: (a) providing a biological sample obtained from said individual; (b) measuring the concentration level of one or a combination of at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from said obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; and (c) dispensing the pancreatic cancer-associated protein or peptide fragments thereof from said obtained sample. (d) If the concentration level of the pancreatic cancer-related protein or its peptide fragment from the obtained sample differs from the concentration level of the pancreatic cancer negative reference value and / or from the concentration level of the reference pancreatic cancer-related protein or its peptide fragment from the sample obtained at an earlier time point from the individual, then the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer; and (e) optionally, treatment is administered using a pancreatic cancer treatment regimen or treatment is initiated for the individual identified as having pancreatic cancer, optionally including surgery and / or administration of chemotherapy agents or radiation therapy.

[0015] According to another aspect of this disclosure, a method for diagnosing and treating pancreatic cancer in an individual is provided, the method comprising: (a) providing a biological sample obtained from said individual; (b) measuring, or having measured, in a spectral unit a concentration level of a combination of pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; and (c) comparing, or having compared, the concentration level of the pancreatic cancer-associated protein or peptide fragment thereof determined in the spectral unit with that obtained from said individual. (d) If the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the concentration level of pancreatic cancer-negative sample from the individual and / or the concentration level of reference pancreatic cancer-associated protein or its peptide fragment from the obtained sample, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer; and (e) optionally, treatment with a pancreatic cancer treatment regimen or inducing treatment in an individual identified as having pancreatic cancer, optionally including surgery and / or administration of chemotherapy or radiation therapy.

[0016] In one implementation, pancreatic cancer treatment includes reducing the blood levels of one or more pancreatic cancer-related proteins or peptide fragments thereof in an individual diagnosed with pancreatic cancer.

[0017] According to any of the foregoing aspects or implementation methods, the blood levels of one or more pancreatic cancer-associated proteins or peptide fragments thereof in an individual are adjusted until the pancreatic cancer-associated proteins or peptide fragments thereof in the individual are reduced to a predetermined level.

[0018] According to any of the foregoing aspects or embodiments, the identification step occurs when determining the concentration level of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof from the obtained sample relative to the concentration level of the reference pancreatic cancer-associated protein or peptide fragment from a negative reference value for pancreatic cancer and / or relative to the concentration level in a previously obtained sample from the individual that is increased by about 10% or more, 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more.

[0019] According to any of the foregoing aspects or implementation methods, measuring the concentration level of pancreatic cancer-related proteins or peptide fragments includes measuring at least one, at least two, at least three, or each of L-selectin, tetraconnectin, phospholipid transferin, and fibronectin.

[0020] According to any of the foregoing aspects or implementation methods, the concentration level of at least four linkers is measured.

[0021] According to any of the foregoing aspects or implementation methods, the obtained sample is blood or urine.

[0022] According to any of the foregoing aspects or embodiments, the obtained sample is serum or plasma.

[0023] According to any of the foregoing aspects or implementation methods, the obtained sample is urine.

[0024] According to any of the foregoing aspects or embodiments, pancreatic cancer-related proteins or their peptide fragments are measured by spectroscopic techniques, wherein the spectroscopic techniques are selected from liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, high performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectroscopy (NMR), Raman spectroscopy, and infrared spectroscopy.

[0025] According to any of the foregoing aspects or implementation methods, the spectroscopic techniques include mass spectrometry.

[0026] According to any of the foregoing aspects or implementation methods, the comparison of the concentration levels of pancreatic cancer-related proteins or their peptide fragments from the obtained samples with the concentration levels of reference values ​​includes the use of multivariate statistical analysis.

[0027] According to any of the foregoing aspects or embodiments, the multivariate statistical analysis is selected from autonomous component analysis (PCA) or partial least squares discriminant analysis of latent structures (PLS-DA).

[0028] According to another aspect of this disclosure, a method is provided for monitoring and treating pancreatic cancer in an individual, the method comprising: (a) providing a first biological sample obtained from said individual at a first time point; (b) assessing a first pancreatic cancer-related proteome profile by measuring at least one, at least two, at least three, at least four, or at least five pancreatic cancer-related proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin from the first biological sample; (c) comparing the first pancreatic cancer-related proteome profile with a reference pancreatic cancer-related proteome profile from a pancreatic cancer-negative sample; and (d) determining the concentration levels of the first pancreatic cancer-related proteome profile and the reference pancreatic cancer-negative sample. (e) A first difference exists between the reference pancreatic cancer-associated proteome profile and the first difference indicates pancreatic cancer; (f) A second biological sample obtained from the individual at a second time point after the first time point is provided; (g) The second pancreatic cancer-associated proteome profile is assessed by measuring the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from the second biological sample; (h) The second pancreatic cancer-associated proteome profile is compared with a reference pancreatic cancer-associated proteome profile from a pancreatic cancer-negative sample; (h) A second difference exists between the first pancreatic cancer-associated proteome profile and the reference pancreatic cancer-associated proteome profile from the pancreatic cancer-negative sample and the second difference indicates pancreatic cancer; (i) The risk of pancreatic cancer progression or having pancreatic cancer is determined at least in part based on the first and second differences; and (j) If pancreatic cancer is identified, the individual is optionally treated with a pancreatic cancer treatment regimen or given treatment, optionally including surgery and / or administration of anticancer agents or radiation therapy.

[0029] According to the foregoing, in some implementations, the time period between the first time point and the second time point is at least 1 month, at least 2 months, at least 3 months, or at least 6 months.

[0030] According to the foregoing aspects or their implementation schemes, measuring the concentration level of pancreatic cancer-related proteins or their peptide fragments from the first and / or second biological samples includes measuring at least one, at least two, at least three, or each of L-selectin, tetraconnectin, phospholipid transferin, and fibronectin.

[0031] According to the foregoing or its implementation scheme, the concentration level of at least four linkers is measured.

[0032] According to the foregoing aspects or their implementation schemes, the first sample, the second sample, or both are blood or urine, or two of the samples are of the same sample type and are selected from serum, plasma, or urine.

[0033] According to another aspect of this disclosure, a kit for diagnosing pancreatic cancer is provided, comprising: (a) a detector configured to detect at least one, at least two, at least two, at least three, at least four, or at least five concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin obtained from a biological sample; and (b) a composition comprising, at control levels corresponding to control groups of pancreatic cancer-negative individuals, apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, and fibronectin. L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; (c) a multivariate analysis system configured to analyze the difference in concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof with control levels; and (d) optionally, providing a description of a method for diagnosing pancreatic cancer, wherein the method includes measuring the levels of pancreatic cancer-associated proteins or peptide fragments thereof from the obtained biological sample using the detector, and comparing the levels of the obtained pancreatic cancer-associated proteins or peptide fragments thereof with control levels of pancreatic cancer-associated proteins or peptide fragments thereof from pancreatic cancer-negative individuals and / or from samples obtained at spoofed time points from the individuals.

[0034] According to one embodiment of the foregoing aspects of this disclosure, the detector includes a multiproteome detector configured to measure the levels of pancreatic cancer-related proteins or peptide fragments thereof comprising apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

[0035] In some embodiments of the foregoing or of the embodiments thereof, the detector is configured to contain concentration levels of at least one, at least two, at least three, or each of the pancreatic cancer-related proteins or peptide fragments of L-selectin, tetraconnector, phospholipid transfer protein, and fibronectin.

[0036] In some embodiments of the foregoing or of its implementation, the detector is configured to measure at least four connectants.

[0037] According to another aspect of this disclosure, a computer-implemented method is provided for processing biological samples from an individual, diagnosing pancreatic cancer, and treating pancreatic cancer, the computer-implemented method comprising: (a) receiving a biological sample obtained from the individual; (b) processing the sample in a spectral unit directly or wirelessly connected to a processing device having a memory for storing measurement data from the spectral unit; (c) in the spectral unit, measuring at least one, at least two, at least three, at least four, or at least five pancreatic cancer-related proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin, and storing the measurement data in a processor; (d) comparing the stored measurement data with reference values ​​in a memory representing a pancreatic cancer-negative sample, optionally using multivariate statistical analysis; (e) storing the data on the processing device and the values ​​from the sample. The obtained sample corresponds to at least one, at least two, at least three, at least four, or at least five pancreatic cancer-related proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin. The results indicate that the measured data representing the level of pancreatic cancer-related proteins or peptide fragments thereof correspond to the results from pancreatic cancer negative... (f) If the concentration levels of a reference pancreatic cancer-associated protein or its peptide fragments differ in the sex sample, the results identify an individual as having pancreatic cancer or at risk of having pancreatic cancer; and (g) optionally administer or induce treatment to an individual identified as having pancreatic cancer, optionally including surgery and / or administration of anticancer agents or radiation therapy.

[0038] In some embodiments of the foregoing aspects or their implementations, the displayed results are included in the user interface, which serves as a dashboard.

[0039] In some implementations of the foregoing aspects or embodiments thereof, the result of the display is part of a plurality of sets of information displayed on the user interface.

[0040] In some embodiments of the foregoing or of the embodiments thereof, the spectral unit includes a detector or the spectral unit is operatively connected to a detector configured to measure the concentration level of pancreatic cancer-related proteins or peptide fragments, optionally configured to measure at least one, at least two, at least three, or each of L-selectin, tetraconnector, phospholipid transfer protein, and fibronectin.

[0041] In some embodiments of the foregoing or of its implementation, the detector is configured to measure at least four connectants.

[0042] According to another aspect of this disclosure, a method for diagnosing and treating pancreatic cancer in an individual is provided, the method comprising: (a) providing a biological sample obtained from said individual; (b) measuring the concentration level of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetrahydropalmatine, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin; and (c) taking a pancreatic cancer phase from the obtained sample. (d) The concentration level of the pancreatic cancer-associated protein or its peptide fragment is compared with the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample and / or from a sample obtained at an earlier time point from the individual; and (e) if the concentration level of the pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample and / or from a sample obtained at an earlier time point from the individual, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer; and (e) optionally, the individual is treated with a pancreatic cancer treatment regimen or radiation therapy.

[0043] In some embodiments of the foregoing or of the embodiments thereof, measuring the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from the first and / or second biological samples includes measuring at least one, at least two, at least three, or each of L-selectin, tetraconnectin, phospholipid transferin, and fibronectin.

[0044] In some embodiments of the foregoing or of its implementation, the concentration levels of at least four linkers are measured.

[0045] According to another aspect of this disclosure, a method for diagnosing and treating pancreatic cancer in an individual is provided, the method comprising: (a) providing a first biological sample obtained from the individual; (b) at a first time point, measuring the concentration level of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-related proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor and fibronectin; (c) comparing the concentration level of pancreatic cancer-related proteins or peptide fragments thereof from the obtained sample with the concentration level of reference pancreatic cancer-related proteins or peptide fragments thereof from a pancreatic cancer-negative sample and / or from a sample obtained at an earlier time point from the individual; and (d) at a second time point, measuring the concentration level of a second biological sample obtained from the individual ... tetraconnector, tetraconnector, tetraconnector, tetraconnector, tetraconnector, tetraconnector, tetraconnector, tetraconnector, tetraconnector, - Levels of at least one, at least two, at least three, at least four, or at least five concentrations of pancreatic cancer-associated proteins or peptide fragments thereof, including selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulant protein, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin; (e) comparing the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from said obtained samples at a second time point with the concentration levels of reference pancreatic cancer-associated proteins or peptide fragments thereof from pancreatic cancer-negative samples and / or from samples obtained at a first time point or from an earlier time point of the individual; (f) if the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from the first and second obtained samples differ from the concentration levels of reference pancreatic cancer-associated proteins or peptide fragments thereof from pancreatic cancer-negative samples and / or from samples obtained at the first time point, then identifying the individual as having pancreatic cancer or at risk of having pancreatic cancer; and (g) optionally, if so identified as having pancreatic cancer, treating or inducing treatment with a pancreatic cancer treatment regimen.

[0046] In certain embodiments of the foregoing aspects or embodiments thereof, the identification step (f) occurs when the concentration level of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof from the obtained sample is increased by about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more, relative to the concentration level of the reference pancreatic cancer-associated protein or peptide fragment from the pancreatic cancer-negative sample.

[0047] In certain embodiments of the foregoing or of the embodiments thereof, the identification step (f) occurs when the concentration levels of at least three, at least four, or at least five pancreatic cancer-associated proteins or their peptide fragments from the obtained sample are increased by about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more relative to the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample.

[0048] In some embodiments of the foregoing or of the embodiments thereof, measuring the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from the first and / or second biological samples includes measuring at least one, at least two, at least three, or each of L-selectin, tetraconnectin, phospholipid transferin, and fibronectin.

[0049] In some embodiments of the foregoing or of its implementation, the concentration levels of at least four linkers are measured.

[0050] In some embodiments of the foregoing or of its implementation, the obtained sample is blood or urine, or serum, plasma or urine.

[0051] In some embodiments of the foregoing or of its implementation, pancreatic cancer-associated proteins or peptide fragments thereof are measured by spectroscopic techniques selected from liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, high performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectroscopy (NMR), Raman spectroscopy, and infrared spectroscopy.

[0052] In some embodiments of the foregoing or its implementation, the comparison of the concentration levels of pancreatic cancer-associated proteins or peptide fragments from the obtained sample with the concentration levels of reference pancreatic cancer-associated proteins or peptide fragments from a pancreatic cancer-negative sample includes multivariate statistical analysis.

[0053] In certain embodiments of the foregoing or of the embodiments thereof, measuring the concentration level of pancreatic cancer-related proteins or peptide fragments thereof from a second biological sample includes measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, and phospholipid transfer protein, and wherein if the measured levels of L-selectin, tetraconnector, and / or phospholipid transfer protein in the second biological sample are increased by at least 10%, 15%, 20%, or 25% compared to the corresponding concentration levels measured at a first time point, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer.

[0054] In some embodiments of the foregoing or of its implementation, at least the concentration level of fibronectin is measured, and wherein if the level of fibronectin in the measured second biological sample increases or decreases by at least 10%, 15%, 20%, or 25% compared to the corresponding concentration level measured at the first time point, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer.

[0055] According to another aspect, a proteolytic sample for mass spectrometry to diagnose pancreatic cancer in an individual is provided, comprising one or a combination of peptide fragments of apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

[0056] According to another aspect, a proteolytic sample containing peptide fragments of L-selectin, tetraconnector, phospholipid transfer protein and / or fibronectin is provided for mass spectrometry to diagnose pancreatic cancer in individuals.

[0057] In some embodiments of any of the foregoing aspects or of any of the implementations thereof, the method further includes obtaining a biopsy tissue sample from an individual identified as having a risk of pancreatic cancer after measuring pancreatic cancer protein or fragments thereof, and surgically removing the cancerous tissue from the pancreas and / or bile duct of the individual if cancer cells are identified in the sample.

[0058] In certain alternative embodiments of any of the foregoing aspects or embodiments thereof, an individual is identified as having pancreatic cancer (e.g., stage 1) if the level of tetraconnector increases over time when measured at two or more time points within a 1-month to 3-year time period, or if it is higher than the control at a single time point. Such embodiments may also include measuring the level of at least one of L-selectin and phospholipid transfer protein, wherein the level of at least one of L-selectin and phospholipid transfer protein increases over time when measured at two or more time points within a 1-month to 3-year time period, or if it is higher than the control at a single time point. In some embodiments, the method or kit further includes measuring fibronectin. In some embodiments, the method further includes measuring at least one of apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, or combinations thereof.

[0059] All features of the exemplary embodiments described in this disclosure, which are not mutually exclusive, can be combined with each other. Elements of one embodiment can be used in other embodiments without further mention. Other aspects and features of this disclosure will become apparent to those skilled in the art upon reading the following description of specific embodiments in conjunction with the accompanying drawings.

[0060] Detailed Explanation

[0061] The following provides a detailed description of one or more embodiments of the present invention. The invention has been described in conjunction with these embodiments, but is not limited to any particular embodiment described herein. The scope of the invention is defined only by the claims and their equivalents. Numerous specific details are set forth in the following description in order to provide a thorough understanding of the invention. These details are provided to provide non-limiting examples, and the invention may be practiced without some or all of these specific details, according to the claims.

[0062] definition

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. As used herein, and unless otherwise stated or required by context, each of the following terms shall have the definition set forth below.

[0064] When used in claims, articles such as “a” and “an” are understood to refer to one or more of the claimed or described content.

[0065] The term "pancreatic cancer negative" generally refers to biological samples from individuals who do not have pancreatic cancer or are unlikely to develop it.

[0066] The terms “baseline” or “control” generally refer to the concentration of a protein or its peptide fragments in individuals who do not have pancreatic cancer or are unlikely to develop pancreatic cancer, or values ​​derived from a population of such individuals and / or from publicly available data.

[0067] The term "pancreatic cancer treatment regimen" generally refers to an intervention for an individual with pancreatic cancer. The objectives of this regimen may include, but are not limited to, relieving or preventing symptoms, slowing or stopping the progression or worsening of pancreatic cancer, and inducing remission of pancreatic cancer.

[0068] In some implementations, a “pancreatic cancer treatment regimen” refers to treatment by administering one or more appropriate therapeutic agents, such as chemotherapeutic agents (e.g., altering pancreatic cancer-related proteome levels).

[0069] The terms “comprises,” “comprising,” “include,” “includes,” “including,” “contains,” and “containing” are non-restrictive, meaning that additional steps and other parts that do not affect the conclusion of the result can be added. The above terms include the terms “composed of” and “substantially composed of.”

[0070] The term "pancreatic cancer-associated proteome" or "proteomic spectrum" generally refers to a spectrum of proteins associated with pancreatic cancer, including at least one or a combination of apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

[0071] The phrase “reference value obtained”, which relates to comparing protein levels in a sample to reference values, refers to values ​​obtained from the sample or from multiple individuals who do not have pancreatic cancer. For example, reference values ​​can be obtained from statistical data previously collected from individuals who do not have pancreatic cancer.

[0072] The terms “preventing” and “prevention” are used interchangeably and generally refer to any activity that reduces the risk of developing pancreatic cancer in an individual.

[0073] The term "individual" generally refers to a vertebrate, such as a mammal. The term "mammal" is defined as an individual belonging to the class Mammalia. In some implementations, the individual is a human.

[0074] The term "treatment" generally refers to an intervention for pancreatic cancer or its related symptoms. The goals of treatment may include, but are not limited to, alleviating or preventing pancreatic cancer, slowing or stopping the progression or worsening of pancreatic cancer, and achieving remission of pancreatic cancer. In some embodiments, "treatment" refers to surgical procedures and / or the administration of therapeutic agents for the treatment of pancreatic cancer. Such treatment may also include radiation therapy.

[0075] The term "pancreatic cancer drug" refers to any therapeutic agent or prodrug used to treat or reduce the progression of pancreatic cancer. Drugs may be contained in pharmaceutical preparations and optionally include excipients.

[0076] As used herein in determining whether an individual has pancreatic cancer, the term "biopsy tissue" refers to, but is not limited to, a composition of cells and / or fluids from one or more of the individual's duodenum, bile duct, pancreas, and / or pancreatic duct.

[0077] The terms “preferred” or “preferred” refer to non-limiting examples of this disclosure and should not be construed as limiting.

[0078] The term “computer-implemented” for a method or process means that all or most of the steps of the method are performed by an electronic data processor and / or distributed computing such as cloud computing.

[0079] In all embodiments of this disclosure, unless otherwise stated, all percentages, concentrations, parts, and ratios are based on the total weight of the compositions of this disclosure. Unless otherwise stated, all weights relating to the listed ingredients are based on activity levels and therefore do not include solvents or byproducts that may be included in commercially available materials.

[0080] Modeling data

[0081] In one implementation, 1 to 50, 2 to 40, or 5 to 30 biomarkers are evaluated based on pancreatic cancer modeling data. This data is used to assess whether a given biomarker predicts pancreatic cancer.

[0082] In one embodiment, the modeling data is obtained from pancreatic cancer test and control group data. Each group of data undergoes computer-implemented calculations and includes at least one of the following: computer-generated receiver operating characteristic (ROC) curve analysis, principal component analysis (PCA) plots, and partial least squares discriminant analysis (PLS-DA) models of latent structures and / or projected variable importance (VIP) plots, thereby obtaining a set of at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more biomarkers identified as contributing to the diagnosis, development, or regression of pancreatic cancer relative to other measured biomarkers. In some embodiments, this disclosure is based on identifying an individual at risk of pancreatic cancer by determining whether a combination of novel proteins or fragments thereof identified by the inventors as predicting the presence of pancreatic cancer in a biological sample from the patient. The identified individual is then subjected to one or more steps, including obtaining biopsy tissue (e.g., cells and / or fluid from the duodenum, bile duct, pancreas, and / or pancreatic duct) and analyzing the presence of cancer cells in the biopsy tissue.

[0083] In some embodiments, the biomarkers are previously selected from proteomic profiles obtained from computer modeling. In some non-limiting instances, at least 2, 3, 4, or 5 proteomic biomarkers are measured. In other embodiments, up to 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, or 20 proteomic biomarkers are identified based on computer modeling. Optionally, this includes selecting metabolomic biomarkers in addition to proteomic biomarkers.

[0084] In some embodiments, the biomarkers are selected based on computer modeling that assigns weights to biomarkers in a proteomic profile and optionally further based on a metabolomic profile, the weights being based on the biomarkers’ ability to diagnose and / or assess pancreatic cancer progression.

[0085] Methods for diagnosing and treating pancreatic cancer

[0086] On one hand, this disclosure relates to methods for the diagnosis and treatment of pancreatic cancer and any related symptoms in an individual (e.g., at an early stage). In some embodiments, this disclosure provides at least in part the identification of proteins or peptide fragments thereof, which provide etiological data associated with pancreatic cancer and offer the opportunity for a protein- or peptide-based objective diagnosis of pancreatic cancer that can lead to more effective treatment. Given the complexity of the interactions between genetics and environment, proteomic profiling, optionally combined with other profiles (e.g., genomic profiles and / or metabolomics profiles), can provide methods for better understanding pancreatic cancer and developing diagnostic tests that contribute to individualized treatment decisions. In one embodiment, the method includes proteomic analysis within a multi-omics analysis, comprising proteomic and genomic and / or metabolomics analyses. Multi-omics-based analysis has the advantage of identifying a biomarker profile of genes derived from an individual and capturing the interactions of an individual's current lifestyle behaviors (e.g., smoking, alcohol consumption, sleep behavior, physical activity, etc.), gut microbiota, diet, and environmental factors that contribute to the unique proteomic profile of an individual with pancreatic cancer. Another advantage of combining early diagnosis with pancreatic cancer treatment regimens is improved treatment outcomes. This document describes a method for identifying novel proteomic profiles in individuals with pancreatic cancer for the diagnosis and treatment of these individuals. Therefore, this disclosure provides an advancement in the art.

[0087] According to this disclosure, a novel proteomic profile of pancreatic cancer has been identified in individuals with pancreatic cancer. The proteomic profile of pancreatic cancer includes at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin.

[0088] Apolipoprotein A1 is identified as a biomarker for pancreatic cancer in this study, and in some embodiments, its level is increased to approximately 1.2 to 10 times or more compared to the median level of reference apolipoprotein A1 in pancreatic cancer-negative individuals. In some embodiments, a 1.2-fold or higher level of apolipoprotein A1 identifies an individual with pancreatic cancer, such as stage 1 or 2 pancreatic cancer. Apolipoprotein A1 is currently considered a tumor suppressor gene. Apolipoprotein A1 (ApoA1) is a protein involved in lipid metabolism, and its potential role in pancreatic cancer has been investigated.

[0089] Alternatively, apolipoprotein A-II is identified herein as a biomarker for pancreatic cancer, and in some embodiments, is elevated to approximately 1.2 to 10 times or higher than the median level of apolipoprotein A-II in pancreatic cancer-negative individuals. In some embodiments, a 1.2-fold or higher level of apolipoprotein A-II identifies an individual with pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0090] Alternatively, plasma protease C1 inhibitors, which serve as biomarkers for pancreatic cancer, can be measured. Plasma protease C1 inhibitor levels can be elevated to approximately 1.2 to 10 times or higher compared to the median level of a reference plasma protease C1 inhibitor in individuals negative for pancreatic cancer. In some embodiments, an elevation of 1.2 times or more in plasma protease C1 inhibitors identifies an individual as having pancreatic cancer, such as stage 1 or 2. Plasma protease inhibitor 1 (C1 inhibitor) is a protein involved in the regulation of the complement system, a component of the immune system.

[0091] Alternatively, the protein L-selectin, a biomarker for pancreatic cancer, can be measured. L-selectin is a cell adhesion molecule expressed on the surface of immune cells and involved in the migration of these cells to sites of inflammation.

[0092] In some implementations, L-selectin is elevated to approximately 1.2 to 10 times or more than the median level of reference L-selectin in individuals negative for pancreatic cancer. In some implementations, an elevated L-selectin level of 1.2 times or more identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0093] Alternatively, tetrajoint, a protein that serves as a biomarker for pancreatic cancer, can be measured. Tetrajoint, also known as plasminogen activator inhibitor-tissue factor pathway inhibitor-2, is a protein involved in blood clotting and has also been shown to possess antitumor properties. Tetrajoint is produced by various cell types in the body, including endothelial cells, platelets, and monocytes. It has also been found in various tissues, including the liver, lungs, and placenta. In some embodiments, tetrajoint is elevated to approximately 1.2 to 10 times or more than the median level of a reference tetrajoint in individuals negative for pancreatic cancer. In some embodiments, an elevation of 1.2 times or more in tetrajoint levels identifies an individual with pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0094] Alternatively, phospholipid transfer protein, a biomarker for pancreatic cancer, can be measured. In some embodiments, the level of phospholipid transfer protein is increased to approximately 1.2 to 10 times or more compared to the median level of a reference phospholipid transfer protein in individuals negative for pancreatic cancer. In some embodiments, a phospholipid transfer protein level of 1.2 times or more identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0095] Alternatively, apolipoprotein M, a protein that serves as a biomarker for pancreatic cancer, can be measured, and in some embodiments, its level is increased to approximately 1.2 to 10 times or more compared to the median level of a reference apolipoprotein M in individuals negative for pancreatic cancer. In some embodiments, a level of 1.2 times or more of apolipoprotein M identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0096] Alternatively, C1s complement factor, a biomarker for pancreatic cancer, can be measured, and in some embodiments, the C1s complement factor is elevated to approximately 1.2 to 10 times or more than the median level of a reference C1s complement factor in pancreatic cancer-negative individuals. In some embodiments, an elevation of 1.2 times or more in C1s complement factor levels identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0097] Alternatively, C4b complement factor, a biomarker for pancreatic cancer, can be measured, and in some embodiments, it is elevated to approximately 1.2 to 10 times or higher than the median level of a reference C4b complement factor in pancreatic cancer-negative individuals. In some embodiments, a C4b complement factor level of 1.2 times or higher identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0098] Alternatively, C6 complement factor, a biomarker for pancreatic cancer, can be measured, and in some embodiments, it is elevated to approximately 1.2 to 10 times or more than the median level of a reference C6 complement factor in pancreatic cancer-negative individuals. In some embodiments, an elevated level of 1.2 times or more of C6 complement factor identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0099] Alternatively, the protein coagulant protein can be measured as a biomarker for pancreatic cancer. Coagulant protein has been established to play a role in cell motility, and in some embodiments, coagulant protein levels are elevated to approximately 1.2 to 10 times or higher compared to the median level of a reference coagulant protein in pancreatic cancer-negative individuals. In some embodiments, a coagulant protein level of 1.2 times or higher identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0100] Alternatively, adipocyte membrane-associated proteins (AMAs), which serve as biomarkers for pancreatic cancer, can be measured. AMAs, also known as lipophilic proteins, are proteins expressed in lipid droplets, organelles that store lipids such as fat. Lipophiles are expressed in adipose (adipose) tissue, and therefore, weight loss can lead to a decrease in their levels in the blood. Regular physical activity has been shown to reduce adipose tissue and thus lower blood lipophilic protein levels. Consuming a low-fat and high-fiber diet rich in fruits and vegetables can help lower blood lipophilic protein levels. Certain medications used to lower cholesterol levels, such as statins, have been shown to reduce blood lipophilic protein levels, and in some embodiments, to increase them by approximately 1.2 to 10 times or more compared to the median level of AMAs in reference adipocyte membrane-associated proteins in pancreatic cancer-negative individuals. In some embodiments, a 1.2-fold or higher level of AMAs identifies an individual with pancreatic cancer, such as stage 1 or 2.

[0101] Alternatively, fibronectin, a protein that serves as a biomarker for pancreatic cancer, can be measured, and in some embodiments, its level is increased to approximately 1.2 to 10 times or more compared to the median level of a reference fibronectin in individuals negative for pancreatic cancer. In some embodiments, a 1.2-fold or higher level of fibronectin identifies an individual as having pancreatic cancer, such as stage 1 or 2 pancreatic cancer.

[0102] On one hand, this disclosure provides a method for diagnosing and treating pancreatic cancer in an individual. The method includes step (a) providing a biological sample obtained from an individual, such as a human. According to the method disclosed herein, any type of biological sample derived from anywhere in an individual's body can be tested, including but not limited to blood (including but not limited to serum or plasma), cerebrospinal fluid (“CSF”), pleural fluid, urine, feces, sweat, tears, breath condensate, salivary vitreous fluid, tissue samples, amniotic fluid, chorionic villus sampling, brain tissue, biopsy tissue of any solid tissue including tumors, adjacent normal tissue, smooth and skeletal muscle, adipose tissue, liver, skin, hair, brain, kidney, pancreas, lungs, etc. In one embodiment, the biological sample is obtained from blood. Pancreatic cancer-related proteins can be extracted from the biological source using any number of extraction / purification methods commonly used in quantitative analytical chemistry.

[0103] The method further includes step (b), which comprises measuring the concentration level of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-related proteins or peptide fragments thereof selected from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin. In some embodiments, the method includes measuring at least 8, 9, 10, 11, 12, 13, 14, or 15 pancreatic cancer-related proteins or fragments thereof from the obtained sample.

[0104] In some embodiments, the method may further include measuring the concentration levels of one or more other pancreatic cancer-associated proteins or peptide fragments thereof, in addition to the proteins described above, including but not limited to any proteins known in the art. Such biomarkers include carbohydrate antigen 19-9 (CA19-9) (see Goonetilleke and Siriwardena, 2007, Eur J SurgOncol. 33(3):266-70, which is incorporated herein by reference), CA242, hCGβ, CA72-4, and carcinoembryonic antigen (CEA) (see Louhimo et al., 2004, Oncology, 66(2):125-31, which is incorporated herein by reference).

[0105] The method described herein further includes step (c), comparing the concentration level of a pancreatic cancer-associated protein or its peptide fragment from the obtained sample with the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample, or a reference value derived therefrom. Those skilled in the art will understand that a reference can be determined as a value representing the level of a pancreatic cancer-associated protein or its peptide fragment in a population without pancreatic cancer for comparison. Various criteria can be used to determine the inclusion and / or exclusion of a particular individual in the reference population, including the individual's age (e.g., the reference individual can be within the same age group as the individual requiring treatment) and the individual's sex (e.g., the reference individual can be of the same sex as the individual requiring treatment). In some embodiments, the reference is an average of pancreatic cancer-negative samples or multiple samples. In another example, the reference concentration level of the pancreatic cancer-associated protein or its peptide fragment is derived from a patient population.

[0106] The method described herein further includes step (d), in which an individual is identified as having pancreatic cancer if the concentration level of a pancreatic cancer-associated protein or a peptide fragment thereof from the obtained sample differs from the concentration level of a reference pancreatic cancer-associated protein or a peptide fragment thereof from a pancreatic cancer-negative sample or a reference value derived therefrom. In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least one pancreatic cancer-associated protein or a peptide fragment thereof from the obtained sample differs from the concentration level of at least one reference pancreatic cancer-associated protein or a reference value derived therefrom from a pancreatic cancer-negative sample by about 10% or more, 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more. In some embodiments, the identification step (d) occurs when the concentration levels of at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or their peptide fragments from the obtained sample differ from the concentration levels of a reference pancreatic cancer-associated protein or fragment from a pancreatic cancer-negative sample by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more.

[0107] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least apolipoprotein A1 from the obtained sample is lower than the concentration level of a reference apolipoprotein A1 from or derived from a pancreatic cancer negative sample. In some aspects, the concentration level of apolipoprotein A1 from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein A1 from or derived from a pancreatic cancer negative sample. In some aspects, the concentration level of apolipoprotein A1 from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein A1 from or derived from a pancreatic cancer negative sample.

[0108] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least apolipoprotein A-II from the obtained sample is lower than the concentration level of a reference apolipoprotein A-II from a pancreatic cancer-negative sample or a reference value derived therefrom. In some aspects, the concentration level of apolipoprotein A-II from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein A-II from a pancreatic cancer-negative sample or a reference value derived therefrom. In some aspects, the concentration level of apolipoprotein A-II from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein A-II from a pancreatic cancer-negative sample.

[0109] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least one plasma proteinase C1 inhibitor from the obtained sample is lower than the concentration level of a reference plasma proteinase C1 inhibitor from a pancreatic cancer negative sample or a reference value derived therefrom. In some aspects, the concentration level of the plasma proteinase C1 inhibitor from the obtained sample is about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more higher than the concentration level of a reference plasma proteinase C1 inhibitor from a pancreatic cancer negative sample or a reference value derived therefrom. In some aspects, the concentration level of the plasma proteinase C1 inhibitor from the obtained sample is less than about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more lower than the concentration level of a reference plasma proteinase C1 inhibitor from a pancreatic cancer negative sample or a reference value derived therefrom.

[0110] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least L-selectin from the obtained sample is lower than the concentration level of a reference L-selectin from a pancreatic cancer-negative sample. In some aspects, the concentration level of L-selectin from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference L-selectin from a pancreatic cancer-negative sample. In other aspects, the concentration level of L-selectin from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference L-selectin from a pancreatic cancer-negative sample.

[0111] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least tetrajoint from the obtained sample is lower than the concentration level of a reference tetrajoint from a pancreatic cancer-negative sample. In some aspects, the concentration level of tetrajoint from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference tetrajoint from a pancreatic cancer-negative sample. In other aspects, the concentration level of tetrajoint from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference tetrajoint from a pancreatic cancer-negative sample.

[0112] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least phospholipid transfer protein from the obtained sample is lower than the concentration level of a reference phospholipid transfer protein from a pancreatic cancer-negative sample. In some aspects, the concentration level of phospholipid transfer protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference phospholipid transfer protein from a pancreatic cancer-negative sample. In other aspects, the concentration level of phospholipid transfer protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference phospholipid transfer protein from a pancreatic cancer-negative sample.

[0113] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least apolipoprotein M from the obtained sample is lower than the concentration level of a reference apolipoprotein M from a pancreatic cancer-negative sample. In some aspects, the concentration level of apolipoprotein M from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein M from a pancreatic cancer-negative sample. In other aspects, the concentration level of apolipoprotein M from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference apolipoprotein M from a pancreatic cancer-negative sample.

[0114] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least one coagulant protein from the obtained sample is lower than the concentration level of a reference coagulant protein from a pancreatic cancer-negative sample. In some aspects, the concentration level of the coagulant protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference coagulant protein from a pancreatic cancer-negative sample. In other aspects, the concentration level of the coagulant protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference coagulant protein from a pancreatic cancer-negative sample.

[0115] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least adipocyte membrane-associated protein from the obtained sample is lower than the concentration level of a reference adipocyte membrane-associated protein from a pancreatic cancer-negative sample. In some aspects, the concentration level of adipocyte membrane-associated protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference adipocyte membrane-associated protein from a pancreatic cancer-negative sample. In other aspects, the concentration level of adipocyte membrane-associated protein from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference adipocyte membrane-associated protein from a pancreatic cancer-negative sample.

[0116] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least C1s complement factor from the obtained sample is lower than the concentration level of a reference C1s complement factor from a pancreatic cancer-negative sample. In some aspects, the concentration level of C1s complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference C1s complement factor from a pancreatic cancer-negative sample. In other aspects, the concentration level of C1s complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference C1s complement factor from a pancreatic cancer-negative sample.

[0117] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least C4b complement factor from the obtained sample is lower than the concentration level of a reference C4b complement factor from a pancreatic cancer-negative sample. In some aspects, the concentration level of C4b complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference C4b complement factor from a pancreatic cancer-negative sample. In other aspects, the concentration level of C4b complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of a reference C4b complement factor from a pancreatic cancer-negative sample.

[0118] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least C6 complement factor from the obtained sample is lower than the concentration level of reference C6 complement factor from a pancreatic cancer-negative sample. In some aspects, the concentration level of C6 complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of reference C6 complement factor from a pancreatic cancer-negative sample. In other aspects, the concentration level of C6 complement factor from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of reference C6 complement factor from a pancreatic cancer-negative sample.

[0119] In some embodiments, the identification step (d) occurs when it is determined that the concentration level of at least fibronectin from the obtained sample is lower than the concentration level of reference fibronectin from a pancreatic cancer-negative sample. In some aspects, the concentration level of fibronectin from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of reference fibronectin from a pancreatic cancer-negative sample. In other aspects, the concentration level of fibronectin from the obtained sample is about 20% or higher, about 30% or higher, about 40% or higher, about 50% or higher, about 60% or higher, or about 70% or higher than the concentration level of reference fibronectin from a pancreatic cancer-negative sample.

[0120] The method described herein also includes the step of treating an individual thus identified as having pancreatic cancer using a pancreatic cancer treatment regimen (e).

[0121] In some embodiments of any of the methods described herein, comparing the concentration level of at least one pancreatic cancer-associated protein from the obtained sample, or a value derived therefrom, with the concentration level of a reference pancreatic cancer-associated protein from a pancreatic cancer-negative sample includes the use of multivariate statistical analysis. In one embodiment, the multivariate statistical analysis is selected from autonomous component analysis (“PCA”) or partial least squares discriminant analysis of latent structures (PLS-DA). In some embodiments, a computer is used for statistical analysis. Software capable of using statistical methods known in the art can extract data for statistical analysis from chromatograms (i.e., the spectrum of the mass signal).

[0122] In some aspects, this disclosure relates to methods for monitoring pancreatic cancer progression and treating pancreatic cancer in an individual. In one embodiment, the method includes quantifying pancreatic cancer-related proteins or peptide fragments thereof at one or more time points after the initiation of treatment to monitor pancreatic cancer progression or regression in an individual. Therefore, the method includes: (a) providing a first biological sample obtained from an individual at a first time; (b) assessing a first pancreatic cancer-associated proteome profile by measuring the concentration levels of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin from a previously obtained sample; (c) comparing the first pancreatic cancer-associated proteome profile with a reference pancreatic cancer-associated proteome profile from a pancreatic cancer-negative sample; and (d) determining the first pancreatic cancer-associated proteome profile compared with a reference pancreatic cancer-associated proteome profile from a pancreatic cancer-negative sample. (e) providing a second biological sample obtained from the individual at a second time after the first time; (f) assessing the second pancreatic cancer-related proteome profile by measuring the concentration levels of pancreatic cancer-related proteins or peptide fragments thereof from the second obtained sample; (g) comparing the second pancreatic cancer-related proteome profile with a reference pancreatic cancer-related proteome profile from the pancreatic cancer-negative sample; (h) determining that a second difference exists between the first pancreatic cancer-related proteome profile from the pancreatic cancer-negative sample and the reference pancreatic cancer-related proteome profile, the second difference indicating pancreatic cancer; (i) determining pancreatic cancer progression or regression based at least in part on the first and second differences; and (j) treating the identified individual with a pancreatic cancer treatment regimen.

[0123] In some embodiments of the above method, the time between the first and second times is at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 9 months, or at least 12 months, preferably at least 3 months. In some embodiments, the individual has been treated before the first two biological samples have been obtained. In other embodiments, the individual has been treated during the interval between the collection of biological samples. In some embodiments, the first biological sample, the second biological sample, or both are blood or urine, preferably serum, plasma, or urine.

[0124] This disclosure also provides methods for diagnosing and treating pancreatic cancer in individuals. The method includes: (a) providing a biological sample obtained from an individual; (b) measuring the concentration levels of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or protein fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin pancreatic cancer-associated protein or peptide fragments thereof; (c) comparing the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from said obtained sample with the concentration levels of reference pancreatic cancer-associated proteins or peptide fragments thereof from a pancreatic cancer-negative sample; (d) identifying the individual as having pancreatic cancer if the concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof from said obtained sample differ from the concentration levels of reference pancreatic cancer-associated proteins or peptide fragments thereof from a pancreatic cancer-negative sample; and (e) treating the so-identified individual using a pancreatic cancer treatment regimen.

[0125] This disclosure also provides a method for diagnosing and treating pancreatic cancer in an individual, the method comprising: (a) providing a biological sample obtained from said individual; (b) measuring the concentration level of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin from the obtained sample; (c) comparing the concentration level of pancreatic cancer-associated proteins or fragments thereof from said obtained sample with the concentration level of a reference pancreatic cancer-associated protein or fragments thereof or a baseline from a pancreatic cancer-negative sample; and (d) measuring the concentration level of apolipoprotein A1, apolipoprotein A-II, plasma protein C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, and fibronectin from the obtained sample. (e) The concentration levels of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-related proteins or fragments of collagen, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; (f) The concentration levels of pancreatic cancer-related proteins or fragments thereof from the obtained sample are compared with the concentration levels of reference pancreatic cancer-related proteins or fragments thereof from a pancreatic cancer-negative sample or a baseline; (g) If the concentration levels of the pancreatic cancer-related proteins or fragments thereof from the obtained sample differ from the concentration levels of the reference pancreatic cancer-related proteins or fragments thereof from the pancreatic cancer-negative sample, and the concentration levels of the pancreatic cancer-related proteins or fragments thereof from the obtained sample differ from the concentration levels of the reference pancreatic cancer-related proteins or fragments thereof from the pancreatic cancer-negative sample or a baseline, the individual is identified as having pancreatic cancer; and (g) The identified individual is treated with a pancreatic cancer treatment regimen.

[0126] In some embodiments of the above method, the identification step (f) occurs when the concentration level of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or fragments thereof from the obtained sample differs from the concentration level of a reference pancreatic cancer-associated protein or fragment thereof or a baseline by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more.

[0127] Methods for quantifying pancreatic cancer-related proteins or their peptide fragments.

[0128] In one embodiment, pancreatic cancer-related proteins or their peptide fragments are measured by spectroscopic techniques selected from liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, high-performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectroscopy (NMR), Raman spectroscopy, and infrared spectroscopy.

[0129] In some embodiments, the concentration levels of pancreatic cancer-associated proteins or their peptide fragments can be measured by methods including mass spectrometry, including but not limited to gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (e.g., LC-MS, LC-MS-MS, LC-MRM, LC-SIM, and LC-SRM). In some embodiments, the mass spectrometry method is multiple reaction monitoring mass spectrometry, parallel reaction monitoring mass spectrometry, matrix-assisted laser desorption / ionization (MALDI) mass spectrometry, or data-independent acquisition mass spectrometry. In another embodiment, the multiple reaction monitoring mass spectrometry method is immuno-multiple reaction monitoring mass spectrometry, immuno-parallel reaction monitoring, immuno-MALDI, or immuno-data-independent acquisition mass spectrometry.

[0130] In one embodiment, the method including mass spectrometry includes introducing a protein sample into a mass spectrometry analysis unit, subsequently fragmenting it within the mass spectrometry analysis unit, and recording and providing the mass of the fragments for analysis.

[0131] In some embodiments, mass spectrometry methods for determining whether pancreatic-associated proteins are elevated include enzymatic or chemical digestion of proteins or peptide fragments of a sample obtained from an individual into peptide fragments. Optionally, the peptide fragments are isolated and / or ionized and captured by mass spectrometry. Digestion may include proteolytic digestion, involving treatment of a preparation containing pancreatic cancer-associated proteins with acid, base, or enzymes such as trypsin or other proteolytic enzymes. One embodiment includes shotgun proteomics quantification, in which all proteins in a complex mixture such as serum, urine, and cell lysates are hydrolyzed or cleaved into peptides, followed by multidimensional HPLC-MS, with the aim of producing a holistic spectrum of the protein mixture as a genome “shotgun” sequence.

[0132] Therefore, according to one aspect of this disclosure, a method is provided for determining whether pancreatic-associated proteins or peptide fragments thereof are elevated in a sample obtained from an individual, said pancreatic-associated proteins or peptide fragments thereof being selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, and C1s complement factor. The method comprises at least one pancreatic cancer-associated protein or peptide fragment thereof, including C4b complement factor, C6 complement factor, and fibronectin, wherein the protein or peptide fragment thereof from a sample obtained from an individual is enzymatically or chemically digested into peptide fragments to produce the peptide fragments; a solution containing the peptide fragments is introduced into a mass spectrometer, optionally followed by one or more treatments including liquid chromatography or other treatments, to quantify the peptide fragments; the concentration of the peptide fragments relative to a baseline, such as a standard (e.g., a peptide standard); and an assessment of whether the fragments are elevated relative to the baseline or standard; if one or more peptides are elevated relative to the baseline or standard, the individual is identified as having pancreatic cancer or predisposing to develop pancreatic cancer; and treatment or inducing treatment of the individual using pancreatic cancer treatments, optionally including chemotherapeutic agents approved for the treatment of pancreatic cancer.

[0133] Proteins or their peptide fragments can be fragmented within a mass spectrometry unit, and the quality of the fragments can be recorded and provided for analysis.

[0134] The baseline is a normal level of a protein or peptide derived from an individual or a population of individuals without pancreatic cancer. A standard may contain one or more pancreatic cancer-associated proteins or peptide fragments thereof, at concentrations corresponding to such normal levels. This standard may be derived from a sample or values ​​previously obtained from samples from individuals without pancreatic cancer. The standard may contain one or more of the following: apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin or fragments thereof.

[0135] In another embodiment, multiple standards comprising one or more pancreatic cancer-related peptide fragments are provided at concentrations corresponding to normal levels. The standards may include any combination of peptide fragments derived from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitors, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

[0136] Pancreatic cancer treatment

[0137] Before treatment for pancreatic cancer, an individual will typically undergo a biopsy to confirm its presence. Biopsies can be performed percutaneously, endoscopically, or surgically. A percutaneous biopsy involves inserting a hollow needle through the skin and into the pancreas to remove a sample of the tumor. Imaging, such as ultrasound or CT scans, can facilitate this procedure. An endoscopic biopsy involves inserting an endoscope through the throat and small intestine to the pancreas. Using endoscopic ultrasound, a needle is inserted into the tumor to obtain a sample from the bile duct or pancreatic duct, or a brush is used to remove cells from the bile duct or pancreatic duct. If it is desired to determine whether the pancreatic cancer has spread, a surgical biopsy can be performed. A laparoscopic (keyhole surgery) surgical biopsy can be performed. Optionally, additional tests are performed to assess whether the cancer cells obtained from the biopsy have mutations in certain genes. If gene mutations related to pancreatic cancer are found, targeted therapy may be part of the pancreatic treatment regimen. Data from proteomic profiling assessments can be used in conjunction with this information.

[0138] Imaging studies can be performed to aid in diagnosis. These studies can use X-rays, magnetic fields, sound waves, or radioactive materials to visualize tumors. Examples of imaging techniques include computed tomography (CT) scans, magnetic resonance imaging (MRI), ultrasound, cholangiopancreatography (CCP), such as magnetic resonance cholangiopancreatography (MRCP) or percutaneous transhepatic cholangiopancreatography (PTC), positron emission tomography (PET), or angiography.

[0139] Pancreatic treatment options may include surgery to remove the tumor.

[0140] Pancreatic cancer treatment regimens may alternatively or additionally include one or more therapeutic agents to treat pancreatic cancer. Examples of such therapeutic agents include: abuprofen (paclitaxel albumin-stabilized nanoparticle formulation); everolimus; capecitabine; erlotinib hydrochloride; everolimus; 5-FU (fluorouracil injection); fluorouracil; gemcitabine; MRTX1133 targeting the Kras G12D mutant; and / or olaparib. Pancreatic cancer treatment may additionally or alternatively include radiation therapy to reduce tumor size.

[0141] In one embodiment, one or more therapeutic agents are formulated in a delivery carrier such as lipid nanoparticles. The therapeutic agent formulated into the delivery carrier such as lipid nanoparticles may include one or more RNA sequences of the aforementioned pancreatic cancer biomarkers or RNA sequences of biomarkers associated with the aforementioned pancreatic biomarkers, which may be used to inhibit their disease effects or reduce their levels.

[0142] Reagent test kit

[0143] The proteomic profiles described herein can be used in tests, assays, methods, and kits for the diagnosis, prediction, regulation, or monitoring of pancreatic cancer, including ongoing assessments, monitoring, and / or susceptibility evaluations. This disclosure includes kits for diagnosing pancreatic cancer by measuring and identifying at least one or more pancreatic cancer-associated proteins or fragments thereof. Therefore, the kit may include (a) a detector configured to detect concentration levels of at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-associated proteins or fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin, which are control levels corresponding to a control group of pancreatic cancer-negative individuals; (c) a multivariate analysis system configured to analyze the difference between the concentration levels of pancreatic cancer-associated proteins or fragments thereof and control levels; and (d) optionally, instructions for a method of diagnosing pancreatic cancer, wherein the method includes measuring the level of pancreatic cancer-associated proteins or peptide fragments thereof from the obtained biological sample using the detector and comparing the level of the obtained pancreatic cancer-associated proteins or peptides thereof with the control level of pancreatic cancer-associated proteins or peptides thereof obtained from pancreatic cancer-negative individuals. In one embodiment, the pancreatic cancer diagnostic method includes a multi-metabolite detector configured to measure the levels of pancreatic cancer-related proteins or peptides thereof. The detector may be part of a mass spectrometry unit, optionally operatively connected to a chromatographic unit such as a liquid chromatography unit.

[0144] In some aspects, the kit can be used to measure pancreatic cancer-related proteins or fragments thereof using physical separation techniques (as described above). In other aspects, the kit can be used to measure pancreatic cancer-related proteins or peptides using methods other than physical separation methods, such as colorimetric, enzymatic, and immunological methods, as used in non-limiting examples. The kit may also include one or more suitable negative and / or positive controls. The kits disclosed herein may include other reagents such as buffers and solutions for performing the tests.

[0145] Computer implementation method

[0146] This disclosure also relates to computer-implemented methods for processing biological samples from individuals, diagnosing pancreatic cancer, and treating pancreatic cancer. The computer-implemented methods can also allow for monitoring pancreatic cancer progression across multiple time points to support more effective treatment options.

[0147] The computer-implemented method includes receiving a biological sample from the individual; processing the sample in a spectral unit directly or wirelessly connected to a processing device, or utilizing any suitable communication technology, the processing device having a memory for storing measurement data from the spectral unit; and in the spectral unit, measuring at least one, at least two, at least three, at least four, at least five, at least six, or at least seven levels of pancreatic cancer-associated proteins or fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin, and storing the measurement data in a processor. The processing device includes one or more data storage devices configured or adapted to store data related to the method. For example, the data storage device may be configured or adapted to store measurement data from the spectral unit. The data storage device may also include computer program code stored thereon. The program code of this embodiment may include program code for performing at least the method steps during execution.

[0148] The computer-implemented method further includes comparing the stored measurement data with values ​​in a memory representing a pancreatic cancer-negative sample using multivariate statistical analysis; storing in the processing device results corresponding to at least one, at least two, at least three, at least four, at least five, at least six, or at least seven pancreatic cancer-related proteins or fragments thereof from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin, wherein if the measurement data representing the level of the pancreatic cancer-related protein or fragment differs from the concentration value of a reference pancreatic cancer-related protein or fragment from a pancreatic cancer-negative sample, the result identifies the individual as having pancreatic cancer; and displaying the result or a pancreatic cancer treatment plan on an electronic display, the electronic display being directly or wirelessly connected to the processor, for identifying an individual as having pancreatic cancer or having a predisposition to develop pancreatic cancer.

[0149] The displayed results or treatment plans include electronic text, optionally with graphical icons. Optionally, the graphical icons are part of an electronic dashboard that displays the results in a format that simplifies the results and / or provides treatment recommendations based on the results.

[0150] As noted, computer-implemented methods could further allow for the monitoring of pancreatic cancer-related proteins across multiple time points, supporting more effective treatment options. Figure 1Non-limiting examples of this method are described herein. The method includes obtaining a biological sample (e.g., a serum sample) at time point 1 (T1). The sample is proteolytically hydrolyzed 10A to produce hydrolyzed proteins, the hydrolyzed proteins are analyzed by mass spectrometry 20A, and pancreatic cancer-associated proteins are measured 30A. The concentrations are compared with those of a pancreatic cancer-negative sample 40A, and it is determined whether the protein concentration is increased or decreased relative to a standard 50A. In this non-limiting example, the pancreatic cancer-associated protein is at least one of L-selectin, tetraconnector, phospholipid transfer protein, and / or fibronectin. In one example of this disclosure, at least one of L-selectin, tetraconnector, and / or phospholipid transfer protein is elevated relative to baseline (e.g., a pancreatic cancer-negative sample). In this case, the individual is identified as having a possible risk of having or developing pancreatic cancer (YES). If the concentrations of L-selectin, tetraconnector, phospholipid transfer protein, and / or fibronectin are not significantly different from those of a standard, the individual is identified as having a low risk of pancreatic cancer 60 (NO). It should be understood that the greater the elevation between the measured pancreatic cancer-associated protein and that from a pancreatic cancer-negative sample, the greater the individual's likelihood of developing or progressing to pancreatic cancer. It is desirable to obtain this information earlier in an individual's life (e.g., 40 years or younger, 35 years or younger, 30 years or younger, or 25 years or younger) to increase any benefit from delaying or offsetting the progression of pancreatic cancer.

[0151] In other embodiments, although a small increase in the measured pancreatic cancer-associated protein compared to that from a pancreatic cancer-negative sample reflects a low probability that the individual has or has developed pancreatic cancer up to that time point, there is no guarantee that the increase will remain small at later time points. Therefore, it is recommended that these individuals be monitored periodically for their risk of having or developing pancreatic cancer. For example, if an individual is identified as having a possible risk (YES) of having or developing pancreatic cancer, a second biological sample (e.g., a serum sample) is obtained at time point 2 (T2) after T1. In some embodiments, the individual's risk of having or developing pancreatic cancer is measured over their lifetime (or at least over an extended period, such as at least two months, at least four months, at least six months, at least one year, at least two years, at least five years, at least ten years, at least twenty years, or at least thirty years). The sample is subjected to proteolysis 10B to produce hydrolyzed proteins, the hydrolyzed proteins are analyzed by mass spectrometry 20B, and pancreatic cancer-associated proteins 30B are measured. The concentration is compared to T2 40B, and it is determined whether the protein concentration level is increased or decreased relative to T2 50B. If at least one pancreatic cancer-associated protein selected from L-selectin, tetraconnector, phospholipid transfer protein, and / or fibronectin increases by a threshold relative to T1 (e.g., at least 10%, at least 20%, at least 30%), the individual is identified as having a high risk of having or developing pancreatic cancer 70 (YES). An individual is typically identified as having pancreatic cancer after a biopsy and analysis for the presence of cancer cells (e.g., using a microscope). If the concentrations of L-selectin, tetraconnector, phospholipid transfer protein, and / or fibronectin do not differ significantly from T1, the individual is identified as having a low risk of pancreatic cancer 60 (NO). Pancreatic cancer-associated proteins can be measured at a third time point, i.e., time point 3 (T3), if necessary, after T2. The concentration of pancreatic cancer-associated proteins at T3 can be compared to the levels of one or both of T2 and T1 to identify an individual as having a high or low risk of having or developing pancreatic cancer. Therefore, the methods according to this disclosure allow for monitoring changes in the state of pancreatic cancer or the risk of developing (or redeveloping) pancreatic cancer over time, particularly after any treatment regimen. Treatment regimens typically involve surgery but may additionally or alternatively include the administration of chemotherapy and / or radiation therapy.

[0152] The following embodiments describe some exemplary patterns for implementing certain methods described herein. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the systems and methods described herein. Example

[0153] Example 1: Diagnosis and treatment of stage I pancreatic cancer patients determined by monitoring changes in pancreatic-associated protein concentration over time.

[0154] Patients' proteomic profiles were monitored over a three-year period from 2019 to 2022 as part of a multi-assessment health assessment provided by the applicant. Each protein listed in Table 1 was identified as showing significant changes in blood concentration (nM) measured over the three-year period. Surprisingly, some proteins (L-selectin, tetraconnectin, and phospholipid transferin) also showed significant increases in concentration at time point 2 (T2) relative to time point 1 (T1). On the other hand, fibronectin decreased by 323.98% from time point 3 (T3) to time point 2 (T2).

[0155] Table 1: Changes in the concentration of pancreatic cancer-related proteins over time.

[0156]

[0157]

[0158] L-selectin, tetraconnector, and phospholipid transfer protein increased at T2 (time point 2), thus being identified as early indicators of pancreatic cancer. The concentrations of each of these proteins also increased at T3 (time point 3). Furthermore, fibronectin levels were low at T1 (242.0 nM) and T3 (295.7 nM), but increased at T2 (1254.2 nM). A -323.98% change in fibronectin was observed between T2 and T3.

[0159] The results were surprising, as previous studies had shown that tetraconnectin levels were reduced in pancreatic cancer (Caputo et al., 2022, Cancer (Basel), 14(19):4658; and Felix et al., 2013, PloS ONE, 8e82755). However, the inventors found that the protein increased over the time period measured.

[0160] The patient was evaluated and diagnosed with stage 1 pancreatic cancer by a licensed physician. A biopsy was obtained from the patient and examined under a microscope to confirm the diagnosis. The patient underwent surgery to remove the tumor. To the inventors' knowledge, this is a first-positive early diagnosis and treatment of pancreatic cancer utilizing the novel proteomic profile identified herein.

[0161] The measurement of changes in blood concentrations of one or a combination of pancreatic cancer-related proteins over time provides a tool for the early detection of pancreatic cancer, which is not possible using known diagnostic methods. Therefore, monitoring combinations of proteins identified as elevated in patients for the diagnosis and treatment of pancreatic cancer represents a significant advance in this field.

[0162] Other examples of implementation methods will become apparent from the teachings of this specification.

[0163] Note that, for the convenience of the reader, headings or subheadings may be used throughout this disclosure, but should in no way limit the scope of the invention. Furthermore, certain theories may be presented and disclosed herein; however, such theories should in no way limit the scope of the invention.

[0164] The elements of the methods and / or systems described in conjunction with the embodiments of this disclosure are applicable to other aspects of this disclosure as necessary. Therefore, it should be understood that the methods and / or systems of this disclosure encompass any method and / or system that includes any steps and / or components referenced herein, in any embodiment where each such step or component exists independently as defined herein. Many such methods and / or systems may be included, rather than those specifically set forth herein.

[0165] The dimensions and values ​​disclosed herein should not be construed as strictly limited to the exact numerical values ​​listed. Rather, unless otherwise stated, each such dimension refers to the stated value and the functionally equivalent range around that value. For example, a dimension disclosed as “40 mm” is intended to mean “approximately 40 mm”.

[0166] Unless expressly excluded or otherwise limited, every reference cited herein, including any cited or related patent or application and any patent application or patent claiming priority or benefit thereof, is incorporated herein in its entirety by reference. Reference to any document is not an admission that it is prior art relating to any disclosure disclosed or claimed herein, or that it, alone or in any combination with any other reference, teaches, suggests, or discloses any such disclosure. Furthermore, in the event of any conflict between the meaning or definition of any term in this document and any meaning or definition of the same term in any document incorporated by reference, the meaning or definition given to that term in this document shall prevail.

[0167] While specific embodiments of this disclosure have been described and illustrated, it will be apparent to those skilled in the art that various other changes and modifications can be made without departing from the scope of this disclosure. Therefore, the invention is intended to cover all such changes and modifications within the scope of this disclosure in the appended claims.

Claims

1. A method for treating pancreatic cancer in an individual, the method comprising: (i) Receive a proteomic profile from an individual, the proteomic profile previously obtained by: (a) Provide biological samples obtained from said individual; (b) Measure the concentration level of one or a combination of at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetralinkin, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin. (c) The concentration levels of pancreatic cancer-associated proteins or their peptide fragments from the obtained samples are compared with the concentration levels of pancreatic cancer-negative reference values ​​and / or reference pancreatic cancer-associated proteins or their peptide fragments from samples obtained at an earlier time point from the individual. (ii) If the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the negative reference value for pancreatic cancer and / or the reference concentration level of pancreatic cancer-associated protein or its peptide fragment from a sample obtained at an earlier time point from the individual, the individual is identified as being at risk of having pancreatic cancer. (iii) Optionally, obtain biopsy tissue from the individual identified in step (ii) as being at risk of having pancreatic cancer; (iv) Analyze the biopsy tissue to determine the presence of cancer cells; (v) If, based on the proteomic profile in step (ii), the individual is identified as being at risk of having pancreatic cancer and if cancer cells are present in the biopsy tissue, then the individual is identified as having pancreatic cancer; as well as (vi) If the individual is identified as having pancreatic cancer, cancerous tissue may optionally be removed from the pancreas and / or bile duct of the individual.

2. A method for diagnosing and treating pancreatic cancer in an individual, the method comprising: (a) Provide biological samples obtained from said individual; (b) Measure the concentration level of one or a combination of at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetralinkin, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin. (c) The concentration levels of pancreatic cancer-associated proteins or their peptide fragments from the obtained samples are compared with the concentration levels of pancreatic cancer-negative reference values ​​and / or reference pancreatic cancer-associated proteins or their peptide fragments from samples obtained at an earlier time point from the individual. (d) If the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the negative reference value for pancreatic cancer and / or the reference concentration level of pancreatic cancer-associated protein or its peptide fragment from the sample obtained at an earlier time point from the individual, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer. as well as (e) Optionally use pancreatic cancer treatment regimens to treat or cause treatment of an individual identified as having pancreatic cancer, optionally including surgery and / or administration of chemotherapy or radiation therapy.

3. A method for diagnosing and treating pancreatic cancer in an individual, the method comprising: (a) Provide biological samples obtained from said individual; (b) The concentration levels of pancreatic cancer-associated proteins or combinations thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor and fibronectin, or peptide fragments thereof, were measured or have been measured in a spectral unit. (c) The concentration levels of pancreatic cancer-associated proteins or their peptide fragments as determined in the spectral unit have been compared or have been compared with the concentration levels of pancreatic cancer-negative reference values ​​and / or reference pancreatic cancer-associated proteins or their peptide fragments as determined from samples obtained at an earlier time point from the individual. (d) If the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the concentration level of a pancreatic cancer-negative sample from the individual and / or a reference concentration level of pancreatic cancer-associated protein or its peptide fragment from a sample obtained at an earlier time point from the individual, then the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer. as well as (e) Optionally use pancreatic cancer treatment regimens to treat or cause treatment of an individual identified as having pancreatic cancer, optionally including surgery and / or administration of chemotherapy or radiation therapy.

4. The method of claim 1, 2 or 3, wherein the pancreatic cancer treatment comprises reducing the blood levels of one or more pancreatic cancer-related proteins or peptide fragments thereof in an individual diagnosed with pancreatic cancer.

5. The method of claim 4, wherein the blood levels of one or more pancreatic cancer-associated proteins or peptide fragments thereof in the individual are adjusted until the pancreatic cancer-associated proteins or peptide fragments thereof in the individual are reduced to a predetermined level.

6. The method of any one of claims 1 to 5, wherein the identification step occurs when determining that the concentration level of at least one, at least two, at least three, at least four, or at least five of the pancreatic cancer-associated proteins or peptide fragments thereof from the obtained sample is increased by about 10% or more, 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more, relative to the concentration level in a previously obtained sample from the individual.

7. The method according to any one of claims 1 to 6, wherein measuring the concentration level of pancreatic cancer-related proteins or peptide fragments comprises measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, phospholipid transferin, and fibronectin.

8. The method of any one of claims 1 to 7, wherein the concentration level of at least four linkers is measured.

9. The method according to any one of claims 1 to 8, wherein the obtained sample is blood or urine.

10. The method of any one of claims 1 to 8, wherein the obtained sample is serum or plasma.

11. The method of claim 9, wherein the obtained sample is urine.

12. The method of any one of claims 1 to 8, wherein the pancreatic cancer-associated protein or its peptide fragment is measured by a spectroscopic technique, wherein the spectroscopic technique is selected from liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, high performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectroscopy (NMR), Raman spectroscopy, and infrared spectroscopy.

13. The method of claim 12, wherein the spectroscopic technique comprises mass spectrometry.

14. The method of any one of claims 1 to 8, wherein the comparison of the concentration level of pancreatic cancer-associated protein or its peptide fragment or the peptide fragment thereof from the obtained sample with the concentration level of a reference value includes the use of multivariate statistical analysis.

15. The method of claim 14, wherein the multivariate statistical analysis is selected from autonomous component analysis (PCA) or partial least squares discriminant analysis of latent structures (PLS-DA).

16. A method for monitoring protein or peptide fragments in an individual and for treating pancreatic cancer in an individual, said method comprising: (a) Provide a first biological sample obtained from the individual at a first time point; (b) The first pancreatic cancer-associated proteome profile was assessed by measuring the concentration levels of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from the first biological sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin. (c) Compare the first pancreatic cancer-associated proteomic profile with a reference pancreatic cancer-associated proteomic profile from a pancreatic cancer-negative sample; (d) Determine that there is a first difference between the first pancreatic cancer-associated proteomic profile from the pancreatic cancer-negative sample and the reference pancreatic cancer-associated proteomic profile, the first difference indicating pancreatic cancer; (e) providing a second biological sample obtained from the individual at a second time point after the first time point; (f) Assess the second pancreatic cancer-related proteome profile by measuring the concentration levels of pancreatic cancer-related proteins or peptide fragments thereof from the second biological sample; (g) Compare the second pancreatic cancer-associated proteomic profile with a reference pancreatic cancer-associated proteomic profile from the pancreatic cancer-negative sample; (h) Determine that there is a second difference between a first pancreatic cancer-associated proteomic profile from the pancreatic cancer-negative sample and the reference pancreatic cancer-associated proteomic profile, the second difference indicating pancreatic cancer; (i) The risk of pancreatic cancer progression or having pancreatic cancer is determined at least in part based on the first and second differences; as well as (j) If diagnosed with pancreatic cancer, the individual may optionally be treated with or be treated with a pancreatic cancer treatment regimen, optionally including surgery and / or administration of anticancer agents or radiation therapy.

17. The method of claim 16, wherein the time period between the first time point and the second time point is at least one month, at least two months, at least three months, or at least six months.

18. The method of claim 16 or 17, wherein measuring the concentration level of pancreatic cancer-related proteins or peptide fragments thereof from the first and / or second biological samples comprises measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, phospholipid transferin, and fibronectin.

19. The method of claim 16, 17 or 18, wherein the concentration level of at least four linkers is measured.

20. The method of claim 17, wherein the first sample, the second sample, or both are blood or urine, or both samples are of the same sample type and are selected from serum, plasma, or urine.

21. A reagent kit for diagnosing pancreatic cancer, comprising: (a) A detector configured to detect concentration levels of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof obtained from a biological sample, selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin; from; (b) A composition comprising, at control levels corresponding to those of a control group of individuals with negative pancreatic cancer, apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin. (c) A multivariate analysis system configured to analyze the difference in concentration levels of pancreatic cancer-related proteins or their peptide fragments compared to control levels; and (d) Optionally, description of a method for diagnosing pancreatic cancer is provided, wherein the method includes measuring the level of pancreatic cancer-associated protein or a peptide fragment thereof from the obtained biological sample using the detector, and comparing the obtained level of pancreatic cancer-associated protein or a peptide fragment thereof with control levels from pancreatic cancer-negative individuals and / or from samples obtained from an earlier time point from the individual.

22. The kit of claim 21, wherein the detector comprises a multiproteomics detector configured to measure the levels of pancreatic cancer-related proteins or peptide fragments thereof, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, colloidin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

23. The kit of claim 21 or 22, wherein the detector is configured to measure the concentration level of at least one, at least two, at least three, or each of the pancreatic cancer-related protein or peptide fragments comprising L-selectin, tetraconnector, phospholipid transfer protein, and fibronectin.

24. The kit of claim 23, wherein the detector is configured to measure at least four connectives.

25. A computer-implemented method for processing biological samples of individuals, diagnosing pancreatic cancer, and treating pancreatic cancer, the computer-implemented method comprising: (a) Receiving biological samples obtained from said individual; (b) The sample is processed in a spectral unit that is directly or wirelessly connected to a processing device having a memory for storing measurement data from the spectral unit; (c) In the spectral unit, the levels of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-related proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor or C6 complement factor and fibronectin are measured, and the measurement data are stored in the processor; (d) Compare the stored measurement data with reference values ​​in a memory representing pancreatic cancer negative samples, optionally using multivariate statistical analysis; (e) Storing on the processing device results corresponding to at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, or C6 complement factor, and fibronectin, wherein if the measurement data representing the level of pancreatic cancer-associated protein or peptide fragments thereof differs from the concentration level of a reference pancreatic cancer-associated protein or peptide fragments thereof from a pancreatic cancer-negative sample, the results identify an individual having pancreatic cancer or a risk of having pancreatic cancer; (f) Displaying the results on an electronic display, directly or wirelessly connected to the processor, for identifying individuals who are diagnosed with pancreatic cancer, at risk of developing pancreatic cancer, or have a predisposition to develop pancreatic cancer; and (g) Optionally use pancreatic cancer treatment protocols or induce treatment in individuals identified as having pancreatic cancer, optionally including surgery and / or administration of anticancer agents or radiation therapy.

26. The method of claim 25, wherein the displayed results are included in the user interface serving as a dashboard.

27. The method of claim 25 or 26, wherein the result of the display is a portion of a plurality of sets of information displayed on the user interface.

28. The method of claim 25, 26 or 27, wherein the spectral unit includes a detector or the spectral unit is operatively connected to a detector, the detector being configured to measure the concentration level of the pancreatic cancer-related protein or peptide fragment, optionally configured to measure at least one, at least two, at least three or each of L-selectin, tetraconnector, phospholipid transferin and fibronectin.

29. The method of claim 28, wherein the detector is configured to measure at least four connectives.

30. A method for diagnosing and treating pancreatic cancer in an individual, the method comprising: (a) Provide biological samples obtained from said individual; (b) Measure the concentration levels of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor or C6 complement factor and fibronectin; (c) The concentration levels of pancreatic cancer-associated proteins or their peptide fragments from the obtained samples were compared with the concentration levels of reference pancreatic cancer-associated proteins or their peptide fragments from pancreatic cancer-negative samples and / or from samples obtained at an earlier time point from the individual. (d) If the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample differs from the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample and / or from a sample obtained at an earlier time point from the individual, then the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer; and (e) Optionally treat or treat pancreatic cancer using a treatment regimen or radiation therapy, or treat such individuals.

31. The method of claim 30, wherein measuring the concentration level of pancreatic cancer-associated proteins or peptide fragments thereof from the first and / or second biological samples comprises measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, phospholipid transferin, and fibronectin.

32. The method of claim 30 or 31, wherein the concentration level of at least four linkers is measured.

33. A method for diagnosing and treating pancreatic cancer in an individual, the method comprising: (a) Provide a first biological sample obtained from said individual; (b) At the first time point, measure at least one, at least two, at least three, at least four, or at least five concentration levels of pancreatic cancer-associated proteins or peptide fragments thereof selected from the obtained sample, including apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor or C6 complement factor and fibronectin; (c) The concentration levels of pancreatic cancer-associated proteins or their peptide fragments from the obtained samples were compared with the concentration levels of reference pancreatic cancer-associated proteins or their peptide fragments from pancreatic cancer-negative samples and / or from samples obtained at an earlier time point from the individual. (d) At a second time point, the concentration level of at least one, at least two, at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof selected from apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor or C6 complement factor and fibronectin in a second biological sample obtained from the individual is measured. (e) The concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample at a second time point is compared with the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample and / or from a sample obtained at a first time point or from an earlier time point from the individual. (f) If the concentration level of the pancreatic cancer-associated protein or its peptide fragment from the first and second obtained samples is different from the concentration level of the reference pancreatic cancer-associated protein or its peptide fragment from the pancreatic cancer-negative sample and / or different from the sample obtained at the first time point, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer. as well as (g) Optionally, if pancreatic cancer is thus diagnosed, the individual shall be treated with a pancreatic cancer treatment regimen or given treatment.

34. The method of claim 33, wherein the identification step (f) occurs when the concentration level of at least one, at least two, at least three, at least four, or at least five of the pancreatic cancer-associated proteins or peptide fragments thereof from the obtained sample is increased by about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more, relative to the concentration level of the reference pancreatic cancer-associated protein or peptide fragment from the pancreatic cancer-negative sample.

35. The method of claim 33, wherein the identification step (f) occurs when the concentration levels of at least three, at least four, or at least five pancreatic cancer-associated proteins or peptide fragments thereof in the obtained sample are increased by about 10% or more, about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, or about 70% or more relative to the concentration level of a reference pancreatic cancer-associated protein or peptide fragment from a pancreatic cancer-negative sample.

36. The method of claim 33, wherein measuring the concentration level of pancreatic cancer-related proteins or peptide fragments thereof from the first and / or second biological samples comprises measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, phospholipid transferin, and fibronectin.

37. The method of any one of claims 33 to 36, wherein the concentration level of at least four linkers is measured.

38. The method of any one of claims 33 to 37, wherein the obtained sample is blood or urine, or serum, plasma or urine.

39. The method of any one of claims 33 to 38, wherein the pancreatic cancer-associated protein or its peptide fragment is measured by a spectroscopic technique, wherein the spectroscopic technique is selected from liquid chromatography, gas chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, high performance liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectroscopy (NMR), Raman spectroscopy, and infrared spectroscopy.

40. The method of any one of claims 33 to 39, wherein the comparison of the concentration level of pancreatic cancer-associated protein or its peptide fragment from the obtained sample with the concentration level of a reference pancreatic cancer-associated protein or its peptide fragment from a pancreatic cancer-negative sample includes multivariate statistical analysis.

41. The method of any one of claims 33 to 40, wherein measuring the concentration level of the pancreatic cancer-related protein or its peptide fragment from the second biological sample comprises measuring at least one, at least two, at least three, or each of L-selectin, tetraconnector, and phospholipid transfer protein, and wherein if the measured levels of L-selectin, tetraconnector, and / or phospholipid transfer protein in the second biological sample are increased by at least 10%, 15%, 20%, or 25% compared to the corresponding concentration levels measured at the first time point, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer.

42. The method of any one of claims 33 to 41, wherein at least the concentration level of fibronectin is measured, and wherein if the measured level of fibronectin in the second biological sample increases or decreases by at least 10%, 15%, 20%, or 25% compared to the corresponding concentration level measured at the first time point, the individual is identified as having pancreatic cancer or at risk of having pancreatic cancer.

43. A proteolytic sample for mass spectrometry to diagnose pancreatic cancer in an individual, comprising one or a combination of peptide fragments of apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, L-selectin, tetraconnector, phospholipid transfer protein, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, and fibronectin.

44. A proteolytic sample used for mass spectrometry to diagnose pancreatic cancer in an individual, comprising peptide fragments of L-selectin, tetraconnector, phospholipid transfer protein and / or fibronectin.

45. The method of any of the preceding claims, further comprising obtaining a biopsy tissue sample from the individual identified as having a risk of pancreatic cancer after measuring the pancreatic cancer protein or a fragment thereof, and surgically removing the cancerous tissue from the pancreas and / or bile duct of the individual if cancer cells are identified in the sample.

46. ​​The method or kit according to any one of claims 1 to 45, wherein the level of the tetralinker increases over time when measured at two or more time points within a period of 1 month to 3 years, or is higher than the control at a single time point.

47. The method or kit of any one of claims 1 to 45, further comprising measuring the level of at least one of L-selectin and phospholipid transfer protein, wherein the level of at least one of L-selectin and phospholipid transfer protein increases over time or is higher than the control at a single time point when measured at two or more time points over a period of 1 month to 3 years.

48. The method of claim 46, further comprising measuring the level of at least one of L-selectin and phospholipid transfer protein, wherein when measured at two or more time points over a period of 1 month to 3 years, the level of at least one of L-selectin and phospholipid transfer protein increases over time, or is higher than the control at a single time point.

49. The method or kit of claim 46, 47 or 48, further comprising measuring the levels of L-selectin and phospholipid transfer protein, wherein the levels of L-selectin and phospholipid transfer protein increase over time when measured at two or more time points within a time period of 1 month to 3 years, or are higher than the control at a single time point.

50. The method or kit of any one of claims 46 to 49, further comprising measuring fibronectin.

51. The method or kit of any one of claims 46 to 50, further comprising measuring at least one of apolipoprotein A1, apolipoprotein A-II, plasma protease C1 inhibitor, apolipoprotein M, coagulin, adipocyte membrane-associated protein, C1s complement factor, C4b complement factor, C6 complement factor, or combinations thereof.

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