WO2011071893A1 - Bio-marqueur de diagnostic pour identifier des femmes qui présentent un risque d'accouchement avant terme - Google Patents

Bio-marqueur de diagnostic pour identifier des femmes qui présentent un risque d'accouchement avant terme Download PDF

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Publication number
WO2011071893A1
WO2011071893A1 PCT/US2010/059248 US2010059248W WO2011071893A1 WO 2011071893 A1 WO2011071893 A1 WO 2011071893A1 US 2010059248 W US2010059248 W US 2010059248W WO 2011071893 A1 WO2011071893 A1 WO 2011071893A1
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WO
WIPO (PCT)
Prior art keywords
biomarkers
sample
women
expression
preterm
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Application number
PCT/US2010/059248
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English (en)
Inventor
Ozlem Equils
Charles F. Simmons Jr.
Original Assignee
Cedars-Sinai Medical Center
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Publication date
Application filed by Cedars-Sinai Medical Center filed Critical Cedars-Sinai Medical Center
Priority to CA2807050A priority Critical patent/CA2807050C/fr
Priority to US13/512,212 priority patent/US20120238469A1/en
Priority to EP10836527.1A priority patent/EP2510356B1/fr
Publication of WO2011071893A1 publication Critical patent/WO2011071893A1/fr
Priority to US14/836,013 priority patent/US20160069891A1/en
Priority to US15/683,487 priority patent/US20180038871A1/en

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/689—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to pregnancy or the gonads
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6869—Interleukin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G01N2333/5428—IL-10
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G01N2333/5437—IL-13
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/54—Interleukins [IL]
    • G01N2333/545—IL-1
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/36—Gynecology or obstetrics
    • G01N2800/368—Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour

Definitions

  • This invention relates to preterm delivery. More specifically, the invention provides biomarkers and methods of using biomarkers for determining preterm delivery risk.
  • Preterm delivery is one of the most important fetal health problems in the United States today. Approximately one in eight newborns is delivered preterm and the incidence of prematurity has not decreased in the last 20 years. Most preterm babies, if they survive, often have cardiac, neurologic, ophthalmic, and gastrointestinal problems that can extend even beyond childhood, and perhaps lead to adult diseases such as atherosclerosis. Currently, there are few, if any diagnostic biomarkers available that effectively identify women who are going to deliver preterm. Biomarkers that are able to identify these women at risk would be useful in the deployment of prevention strategies. Thus, there is a need to develop novel diagnostics that may identify women who will deliver preterm. Especially, diagnostic biomarkers that may be detected in non-pregnant women or in women during the first trimester.
  • the invention includes methods of identifying a non-pregnant woman at risk for preterm delivery, comprising: obtaining a sample from the non-pregnant woman, determining the expression level of one or more biomarkers in the sample, and comparing the expression level of the one or more biomarkers with the expression level of biomarkers from those observed in women who delivered fullterm (controls). Since there is individual variation, the control values will constitute a range. If the level of expression of the one or more biomarkers in the sample is higher or lower than the level of expression of the same biomarkers in women who delivered fullterm (outside the range), then it is indicative that the non-pregnant woman is at risk for preterm delivery.
  • the risk of preterm delivery may be determined as "low, medium or high.”
  • the biomarkers include but not limited toIL-10, IL-13 and/or IL-1RA.
  • the sample can comprise stimulated PBMC/whole blood supernatant (in the presence or absence of Cortisol). Alternatively, the sample can comprise serum.
  • the invention includes methods of diagnosing susceptibility of preterm delivery in a woman, comprising: obtaining a sample from the woman, determining the expression level of one or more diagnostic biomarkers in the sample, and comparing the expression level of the one or more biomarkers with the expression level of biomarkers from women who delivered fullterm (normal range). If the level of expression of the one or more diagnostic biomarkers in the sample is higher or lower than the level of expression of the same diagnostic biomarkers in the women who delivered fullterm (normal range), then it is indicative that the woman is at risk for preterm delivery.
  • the diagnostic biomarkers can be inflammatory and/or anti-inflammatory cytokines.
  • the diagnostic biomarkers selected include but not limited to IL-10, IL-13 and/or IL-1RA.
  • Biomarker expression levels that are at least one to three times less than the biomarker expression levels of a control are indicative of preterm delivery.
  • the sample can comprise stimulated PBMC/whole blood supernatant (in the presence or absence of Cortisol).
  • the sample can comprise serum.
  • the women may be non-pregnant or in the first-trimester of pregnancy.
  • the present invention is also directed to a kit for preterm delivery.
  • the kit is useful for practicing the inventive method of determining the risk of preterm delivery of non-pregnant women.
  • the kit is an assemblage of materials or components, including a diagnostic bioassay of the present invention.
  • kits configured for the purpose of diagnosing susceptibility of women for preterm delivery.
  • the kit is configured particularly for the purpose of determining the risk of preterm delivery of non-pregnant women.
  • the kit is configured particularly for the purpose of diagnosing susceptibility of women for preterm delivery of pregnant or non-pregnant women.
  • Instructions for use may be included in the kit. "Instructions for use” typically include a tangible expression describing the technique to be employed in using the components of the kit to effect a desired outcome, such as to treat ischemia.
  • the kit also contains other useful components, such as, diluents, buffers, pharmaceutically acceptable carriers, syringes, catheters, applicators, pipetting or measuring tools, bandaging materials or other useful paraphernalia as will be readily recognized by those of skill in the art.
  • useful components such as, diluents, buffers, pharmaceutically acceptable carriers, syringes, catheters, applicators, pipetting or measuring tools, bandaging materials or other useful paraphernalia as will be readily recognized by those of skill in the art.
  • the materials or components assembled in the kit can be provided to the practitioner stored in any convenient and suitable ways that preserve their operability and utility.
  • the components can be in dissolved, dehydrated, or lyophilized form; they can be provided at room, refrigerated or frozen temperatures.
  • the components are typically contained in suitable packaging material(s).
  • packaging material refers to one or more physical structures used to house the contents of the kit, such as inventive compositions and the like.
  • the packaging material is constructed by well known methods, preferably to provide a sterile, contaminant-free environment.
  • package refers to a suitable solid matrix or material such as glass, plastic, paper, foil, and the like, capable of holding the individual kit components.
  • a package can be a glass vial used to contain suitable quantities of an inventive composition containing a polyphenol analog.
  • the packaging material generally has an external label which indicates the contents and/or purpose of the kit and/or its components.
  • Figure 1 Graphical representation of expression levels of various biomarkers of preterm individuals as compared to full term individuals. Empirical means and standard errors for each cytokine and treatment group are presented.
  • the inventors are the first to have discovered a biomarker assay that provides physicians with a tool to identify women who are at risk for preterm delivery, even before they become pregnant or while in the first trimester. Identification of women at risk allows the physician to better focus on preventative strategies in these women and improve pregnancy outcome.
  • the test may be done on small amounts of blood sample obtained from the subject, thus being minimally invasive to the fetus. Furthermore, the test may be repeated multiple times during the course of pregnancy. Thus, it provides dynamic assessment of the preterm delivery risk under the influence of changing environmental/physiologic factors, as wells as requires minimal skills to draw blood as opposed to obtaining amniotic fluid.
  • premature delivery refers to a premature birth or conditions associated with a premature birth, including for example, a child delivered before 34 weeks of amenorrhea.
  • Biomarker refers to a molecular indicator that is associated with a particular pathological or physiological state.
  • the "biomarker” as used herein is an indicator for preterm delivery.
  • the indicator can be a cytokine or an immuno modulating agent, including interleukins and interferons.
  • biomarkers include but are not limited IL-2, IL-4, IL-5, IL-8, IL-10, IL-12, IL-13, GM-CSF, IFN-g, and TNF-a.
  • biomarkers of the present invention include IL-10, IL-13 and/or IL-lPvA.
  • a "biomarker” of the present invention may be detected in a sample.
  • sample refers to cells or component parts, or a fraction or portion thereof of body fluids, including but not limited to blood or amniotic cord blood.
  • sample or “biological sample” further refers to plasma, serum, and/or peripheral blood mononuclear cells (PMBC).
  • PMBC peripheral blood mononuclear cells
  • the present invention provides a method of diagnosing susceptibility for preterm delivery in a non-pregnant woman comprising obtaining a sample from the woman and assaying the sample for the presence or absence of one or more diagnostic biomarkers, where the presence or absence of one or more diagnostic biomarkers is indicative of susceptibility for preterm delivery in the woman.
  • the sample comprises PMBC supernatant.
  • the sample comprises whole blood.
  • the sample comprises serum.
  • the one or more diagnostic biomarkers comprise a microbial component lipopolysacharide (LPS) or another immune stimulant induced cytokine expression profile.
  • the one or more diagnostic biomarkers comprise inflammation and/or anti-inflammatory cytokines.
  • the diagnostic biomarkers are analyzed in the presence of Cortisol.
  • the Cortisol concentration is in the range of 1 ug/ml to 500 ug/ml, with a preferred range of 1 ug/ml to 150 ug/ml.
  • the one or more diagnostic biomarkers comprise a low expression of IL-10, IL-13 and/or IL-1RA.
  • the low expression of IL-10, IL-13 and/or IL-1RA comprises a 1 to 3 fold decrease in expression compared to levels ordinarily found in a healthy individual.
  • cytokines or other biomarkers there are many techniques readily available in the field for detecting the presence or absence of cytokines or other biomarkers, including protein microarrays.
  • some of the detection paradigms that can be employed to this end include optical methods, electrochemical methods (voltametry and amperometry techniques), atomic force microscopy, and radio frequency methods, e.g., multipolar resonance spectroscopy.
  • Illustrative of optical methods in addition to microscopy, both confocal and non-confocal, are detection of fluorescence, luminescence, chemiluminescence, absorbance, reflectance, transmittance, and birefringence or refractive index (e.g., surface plasmon resonance, ellipsometry, a resonant mirror method, a grating coupler waveguide method or interferometry).
  • detection of fluorescence, luminescence, chemiluminescence, absorbance, reflectance, transmittance, and birefringence or refractive index e.g., surface plasmon resonance, ellipsometry, a resonant mirror method, a grating coupler waveguide method or interferometry.
  • cytokine may be captured using biospecific capture reagents, such as antibodies, aptamers or antibodies that recognize the biomarker and modified forms of it.
  • biospecific capture reagents such as antibodies, aptamers or antibodies that recognize the biomarker and modified forms of it.
  • This method could also result in the capture of protein interactors that are bound to the proteins or that are otherwise recognized by antibodies and that, themselves, can be biomarkers.
  • the biospecific capture reagents may also be bound to a solid phase. Then, the captured proteins can be detected by SELDI mass spectrometry or by eluting the proteins from the capture reagent and detecting the eluted proteins by traditional MALDI or by SELDI.
  • SELDI affinity capture mass spectrometry
  • SEAC Surface-Enhanced Affinity Capture
  • mass spectrometers are time-of-flight, magnetic sector, quadrupole filter, ion trap, ion cyclotron resonance, electrostatic sector analyzer and hybrids of these.
  • the presence of biomarkers such as cytokines may be detected using traditional immunoassay techniques.
  • Immunoassay requires biospecific capture reagents, such as antibodies, to capture the analytes.
  • the assay may also be designed to specifically distinguish protein and modified forms of protein, which can be done by employing a sandwich assay in which one antibody captures more than one form and second, distinctly labeled antibodies, specifically bind, and provide distinct detection of, the various forms.
  • Antibodies can be produced by immunizing animals with the biomolecules.
  • Traditional immunoassays may also include sandwich immunoassays including ELISA or fluorescence-based immunoassays, as well as other enzyme immunoassays.
  • cytokines Prior to detection, cytokines may also be fractionated to isolate them from other components in a solution or of blood that may interfere with detection. Fractionation may include platelet isolation from other blood components, sub-cellular fractionation of platelet components and/or fractionation of the desired cytokine from other biomolecules found in platelets using techniques such as chromatography, affinity purification, ID and 2D mapping, and other methodologies for purification known to those of skill in the art.
  • a sample is analyzed by means of a biochip.
  • Biochips generally comprise solid substrates and have a generally planar surface, to which a capture reagent (also called an adsorbent or affinity reagent) is attached.
  • the surface of a biochip comprises a plurality of addressable locations, each of which has the capture reagent bound there.
  • the presence of biomarkers such as cytokines may be detected using PCR techniques or flow cytometry. All of the available techniques for cytokine and chemokine detection may be used.
  • PBMC Peripheral blood mononuclear cells
  • LPS lipopolysaccharide
  • Empirical means and standard errors for each cytokine and treatment group are presented herein. Here, the averages of replicate measures were calculated first for each subject. The mean and SEM was then taken across each treatment group.
  • the inventors focused on the data with 0 or 300 ug/ml of Cortisol pretreatment crossed with 0 or 1 ug/ml LPS. This results in 4 samples per subject, each analyzed twice (with exception of the samples from the full term subject who only had one analysis of each sample). With 8 measurements per subject (with the exception of the full term subject who only has 4 measurements) this results in a data set with 44 measurements for each of the cytokines.
  • Cortisol did suppress secretion of IL-13 and IL-lra; LPS increased secretion of IL-4, IL-10, IL-13, TNF-a, and IL-lra; and women who had preterm deliveries had overall lower secretion levels of IL-10, IL-13, and IL-lra (Table 2).
  • Table 2 Global effects of Cortisol, LPS, and premature delivery on cytokine secretion levels in PBMC based on results of mixed modeling. Cytokine Cortisol Effect LPS Effect Pre-term Effect
  • Luu TM Ment LR, Schneider KC, Katz KH, Allan WC, Vohr BR. Lasting effects of preterm birth and neonatal brain hemorrhage at 12 years of age. Pediatrics. 2009 Mar; 123(3): 1037-44. -Morse SB, Zheng H, Tang Y, Roth J. Early school-age outcomes of late preterm infants.
  • Peltier MR Immunology of term and preterm labor. Reprod Biol Endocrinol. 2003 Dec 2; 1 : 122.
  • Kalish RB Vardhana S, Gupta M, Perni SC, Witkin SS, Interleukin-4 and -10 gene

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  • Urology & Nephrology (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Gynecology & Obstetrics (AREA)
  • Pregnancy & Childbirth (AREA)
  • Reproductive Health (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne des bio-marqueurs associés à un accouchement avant terme. Plus spécifiquement, l'invention concerne des procédés pour mesurer des bio-marqueurs que l'on trouve chez des femmes qui présentent un risque d'accouchement avant terme.
PCT/US2010/059248 2009-12-08 2010-12-07 Bio-marqueur de diagnostic pour identifier des femmes qui présentent un risque d'accouchement avant terme WO2011071893A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
CA2807050A CA2807050C (fr) 2009-12-08 2010-12-07 Bio-marqueur de diagnostic pour identifier des femmes qui presentent un risque d'accouchement avant terme
US13/512,212 US20120238469A1 (en) 2009-12-08 2010-12-07 Diagnostic biomarker to identify women at risk for preterm delivery
EP10836527.1A EP2510356B1 (fr) 2009-12-08 2010-12-07 Procédé de diagnostic pour identifier des femmes qui présentent un risque d'accouchement avant terme
US14/836,013 US20160069891A1 (en) 2009-12-08 2015-08-26 Diagnostic biomarker to identify women at risk for preterm delivery
US15/683,487 US20180038871A1 (en) 2009-12-08 2017-08-22 Diagnostic biomarker to identify women at risk for preterm delivery

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US26769309P 2009-12-08 2009-12-08
US61/267,693 2009-12-08

Related Child Applications (3)

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US13/512,212 A-371-Of-International US20120238469A1 (en) 2009-12-08 2010-12-07 Diagnostic biomarker to identify women at risk for preterm delivery
AU2012230065A Division AU2012230065B2 (en) 2009-12-08 2012-03-13 Diagnostic biomarker to predict women at risk for preterm delivery
US14/836,013 Continuation US20160069891A1 (en) 2009-12-08 2015-08-26 Diagnostic biomarker to identify women at risk for preterm delivery

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RU2486519C1 (ru) * 2011-11-15 2013-06-27 Государственное бюджетное образовательное учреждение высшего профессионального образования "Российский национальный исследовательский медицинский университет им. Н.И. Пирогова" Министерства здравоохранения и социального развития Российской Федерации (ГБОУ ВПО РНИМУ им. Н.И. Пирогова Минздравсоцразви Способ прогнозирования преждевременных родов и внутриутробного инфицирования плода
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US10545156B2 (en) 2011-03-17 2020-01-28 RPI Consulting, LLC Diagnostic biomarker to predict women at risk for preterm delivery
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EP2912458B1 (fr) 2012-10-24 2018-07-18 NYU Winthrop Hospital Biomarqueurs non-invasifs pour identifier les sujets à risque d'accouchement prématuré
KR101578526B1 (ko) * 2015-07-14 2015-12-17 이화여자대학교 산학협력단 32주 미만의 조산 위험성을 예측하기 위한 마커 il-13 및 이의 이용
US11293931B2 (en) 2016-05-05 2022-04-05 Indiana University Research And Technology Corporation Quantitative profiling of progesterone metabolites for the prediction of spontaneous preterm delivery
CA3023242A1 (fr) 2016-05-05 2017-11-09 Indiana University Research & Technology Corporation Profilage quantitatif de metabolites de la progesterone pour la prediction de l'accouchement premature spontane
EP3682250A4 (fr) 2017-09-13 2021-03-03 Progenity, Inc. Biomarqueurs de pré-éclampsie ainsi que systèmes et procédés associés
WO2020215051A1 (fr) * 2019-04-19 2020-10-22 Rpi Consulting Llc Traitements de troubles liés à la grossesse
EP4070113A4 (fr) 2019-12-04 2023-12-20 Biora Therapeutics, Inc. Évaluation de la prééclampsie à l'aide de dosages du facteur de croissance placentaire libre et dissocié
US20220291243A1 (en) * 2019-12-04 2022-09-15 Garbha Health, Inc. Analyzing hormone panels for the prediction of spontaneous preterm delivery

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WETTA L ET AL., AMERICAN JOURNAL OF OBSTETRICS & GYNECOLOGY, vol. 199, no. 6, 1 December 2008 (2008-12-01), pages S59

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US10545156B2 (en) 2011-03-17 2020-01-28 RPI Consulting, LLC Diagnostic biomarker to predict women at risk for preterm delivery
RU2486519C1 (ru) * 2011-11-15 2013-06-27 Государственное бюджетное образовательное учреждение высшего профессионального образования "Российский национальный исследовательский медицинский университет им. Н.И. Пирогова" Министерства здравоохранения и социального развития Российской Федерации (ГБОУ ВПО РНИМУ им. Н.И. Пирогова Минздравсоцразви Способ прогнозирования преждевременных родов и внутриутробного инфицирования плода
EP2807277A4 (fr) * 2012-01-27 2016-02-17 Univ Leland Stanford Junior Procédés de profilage et de quantification d'arn acellulaire
US10155986B2 (en) 2012-01-27 2018-12-18 The Board Of Trustees Of The Leland Stanford Junior University Methods for profiling and quantitating cell-free RNA
US10240200B2 (en) 2012-01-27 2019-03-26 The Board Of Trustees Of The Leland Stanford Junior University Methods for profiling and quantitating cell-free RNA
US10240204B2 (en) 2012-01-27 2019-03-26 The Board Of Trustees Of The Leland Stanford Junior University Methods for profiling and quantitating cell-free RNA
US10287632B2 (en) 2012-01-27 2019-05-14 The Board Of Trustees Of The Leland Stanford Junior University Methods for profiling and quantitating cell-free RNA
US11845988B2 (en) 2019-02-14 2023-12-19 Mirvie, Inc. Methods and systems for determining a pregnancy-related state of a subject
US11851706B2 (en) 2019-02-14 2023-12-26 Mirvie, Inc. Methods and systems for determining a pregnancy-related state of a subject

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US20120238469A1 (en) 2012-09-20
EP2510356B1 (fr) 2018-10-31
US20160069891A1 (en) 2016-03-10
EP2510356A1 (fr) 2012-10-17
US20180038871A1 (en) 2018-02-08
EP2510356A4 (fr) 2013-05-08
CA2807050A1 (fr) 2011-06-16
CA2807050C (fr) 2019-06-18

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