Methods of diagnosing and treating complications of pregnancy

Abstract

Disclosed herein are methods for diagnosing a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder by measuring the level or biological activity of soluble endoglin. Also disclosed herein are methods for treating a a pregnancy related hypertensive disorder, such as pre-eclampsia and eclampsia, using compounds that alter soluble endoglin levels or biological activity.

1 . A method of treating or preventing a pregnancy related hypertensive disorder in a subject, said method comprising the step of administering to said subject a compound capable of specifically binding to soluble endoglin, wherein said administering is for a time and in an amount sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         2 . The method of  claim 1 , wherein said pregnancy related hypertensive disorder is selected from the group consisting of pre-eclampsia, eclampsia, gestational hypertension, chronic hypertension, HELLP syndrome, and pregnancy with a small for gestational age (SGA) infant.  
     
     
         3 . The method of  claim 2 , wherein said pregnancy related hypertensive disorder is pre-eclampsia or eclampsia.  
     
     
         4 . The method  claim 1 , wherein said compound is a purified soluble endoglin antibody or a soluble endoglin antigen-binding fragment.  
     
     
         5 . The method of  claim 1 , wherein said compound is a growth factor.  
     
     
         6 . The method of  claim 5 , wherein said growth factor is selected from the group consisting of: transforming growth factor (TGF)-β1 or TGF-β3, activin A, Bone Morphogenic Protein (BMP)-2 or BMP-7.  
     
     
         7 . The method of  claim 1 , further comprising administering a compound selected from the group consisting of a purified sFlt-1 antibody, an sFlt antigen-binding fragment, nicotine, theophylline, adenosine, nifedipine, minoxidil, magnesium sulfate, vascular endothelial growth factor (VEGF), and placental growth factor (PlGF).  
     
     
         8 . The method of  claim 7 , wherein said VEGF is VEGF121 or VEGF 165.  
     
     
         9 . The method of  claim 1 , further comprising the step of administering to a subject an anti-hypertensive compound.  
     
     
         10 . The method of  claim 1 , wherein said subject is a pregnant human.  
     
     
         11 . The method of  claim 1 , wherein said subject is a post-partum human.  
     
     
         12 . The method of  claim 1 , wherein said subject is a non-human.  
     
     
         13 . The method of  claim 12 , wherein said subject is selected from the group consisting of a cow, a horse, a sheep, a pig, a goat, a dog, and a cat.  
     
     
         14 . A method of treating or preventing a pregnancy related hypertensive disorder in a subject, said method comprising the step of administering to said subject a compound that increases the level of a growth factor capable of binding to soluble endoglin, wherein said administering is in an amount and for a time sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         15 . The method of  claim 14 , wherein said growth factor is selected from the group consisting of TGF-β1, TGF-3β, activin A, BMP-2, and BMP-7.  
     
     
         16 . The method of  claim 14 , wherein said compound is selected from the group consisting of cyclosporine, alpha tocopherol, methysergide, bromocriptine, and aldomet.  
     
     
         17 . A method of treating or preventing a pregnancy related hypertensive disorder in a subject, said method comprising administering to said subject a compound that inhibits growth factor binding to a soluble endoglin polypeptide, wherein said administering is sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         18 . The method of  claim 17 , wherein said compound binds to soluble endoglin and blocks growth factor binding.  
     
     
         19 . A method of treating or preventing a pregnancy related hypertensive disorder in a subject, said method comprising administering to said subject a compound capable of reducing soluble endoglin expression or biological activity, wherein said administering is sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         20 . The method of  claim 19 , wherein said compound is a compound that inhibits a proteolytic enzyme selected from the group consisting of a matrix metalloproteinase (MMP), cathepsin, and elastase.  
     
     
         21 . The method of  claim 20 , wherein said MMP is MMP9.  
     
     
         22 . The method of  claim 20 , wherein MMP is membrane type matrix metalloproteinase-1.  
     
     
         23 . The method of  claim 19 , wherein said compound comprises an antisense nucleobase oligomer that is at least 95% complementary to at least a portion of an endoglin nucleic acid sequence, wherein said administering is sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         24 . The method of  claim 23 , wherein said antisense nucleobase oligomer is 8 to 30 nucleotides in length.  
     
     
         25 . The method of  claim 19 , wherein said compound comprises a double stranded RNA having one strand that is at least 95% complementary to at least a portion of an endoglin nucleic acid sequence, wherein said administering is sufficient to treat or prevent said pregnancy related hypertensive disorder in said subject.  
     
     
         26 . The method of  claim 25 , wherein said double stranded RNA is a small interfering RNA (siRNAs) 19 to 25 nucleotides in length.  
     
     
         27 . The method of  claim 19 , said compound comprising a purified antibody or antigen-binding fragment that specifically binds soluble endoglin.  
     
     
         28 . The method of  claim 14 ,  17 , or  19 , wherein said pregnancy related hypertensive disorder is selected from the group consisting of pre-eclampsia, eclampsia, gestational hypertension, chronic hypertension, HELLP syndrome, and pregnancy with a small for gestational age (SGA) infant.  
     
     
         29 . The method of  claim 28 , wherein said pregnancy related hypertensive disorder is pre-eclampsia or eclampsia.  
     
     
         30 . The method of  claim 1  or  19 , wherein said method further comprises monitoring said pregnancy related hypertensive disorder in said subject, wherein said monitoring comprises measuring the level of soluble endoglin polypeptide in a sample from said subject.  
     
     
         31 . The method of  claim 30 , wherein said sample is serum or plasma and a level of soluble endoglin polypeptide less than 25 ng/ml indicates an improvement in said pregnancy related hypertensive disorder.  
     
     
         32 . The method of  claim 30 , wherein said measuring of levels is done on two or more occasions and a decrease in said levels between measurements is an indicator of an improvement in said pregnancy related hypertensive disorder.  
     
     
         33 . The method of  claim 30 , wherein said measuring of levels is compared to a positive reference sample and a decrease in the level of soluble endoglin relative to said positive reference sample indicates an improvement in said pregnancy related hypertensive disorder in said subject.  
     
     
         34 . The method of  claim 30 , wherein said monitoring is used to determine the therapeutic dosage of the compound.  
     
     
         35 . The method of  claim 30 , wherein said measuring is done using an immunological assay.  
     
     
         36 . The method of  claim 30 , wherein the level of soluble endoglin is the level of free, bound, or total soluble endoglin.  
     
     
         37 . The method of  claim 30 , wherein the level of soluble endoglin is the level of an endoglin polypeptide resulting from degradation or enzymatic cleavage.  
     
     
         38 . The method of  claim 30 , wherein said monitoring comprises measuring the level of at least one of sFlt-1, VEGF, or PlGF polypeptide in a sample from said subject.  
     
     
         39 . The method of  claim 38 , further comprising calculating the relationship between said levels of soluble endoglin, sFlt-1, VEGF, or PlGF using a metric, wherein an alteration in the relationship between said levels in the subject sample relative to a reference sample, is a diagnostic indicator of a pregnancy related hypertensive disorder in said subject.  
     
     
         40 . The method of  claim 39 , wherein said metric is a pre-eclampsia anti-angiogenic index (PAAI): [sFlt-1/VEGF+PlGF].  
     
     
         41 . The method of  claim 40 , wherein a PAAI value less than 20 indicates an improvement in said pre-eclampsia or eclampsia.  
     
     
         42 . The method of  claim 39 , wherein said metric is a soluble endoglin anti-angiogenic index: [(sFlt-1+0.25 soluble endoglin)/PlGF] or [(soluble endoglin+sFlt-1)/PlGF].  
     
     
         43 . The method of  claim 39 , wherein said metric is used to determine the dosage of the therapeutic compound.  
     
     
         44 . The method of  claim 43 , wherein the therapeutic compound is administered in a dose such that the PAAI is less than 20.  
     
     
         45 . The method of  claim 1  or  19 , wherein said method further comprises monitoring the pregnancy related hypertensive disorder in said subject, wherein said monitoring comprises measuring the level of an endoglin nucleic acid molecule in a sample from said subject.  
     
     
         46 . The method of  claim 45 , wherein said levels of an endoglin nucleic acidis compared to a reference sample and an alteration in said levels in the subject sample compared to said reference is a diagnostic indicator of a pregnancy related hypertensive disorder in said subject.  
     
     
         47 . An antibody or antigen-binding fragment thereof that specifically binds soluble endoglin.  
     
     
         48 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody or antigen-binding fragment prevents binding of a growth factor to soluble endoglin.  
     
     
         49 . The antibody or antigen-binding fragment thereof of  claim 48 , wherein said growth factor is selected from the group consisting of TGF-β1, TGF-3β, activin A, BMP-2, and BMP-7.  
     
     
         50 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody is a monoclonal antibody.  
     
     
         51 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody or antigen-binding fragment thereof is a human or humanized antibody.  
     
     
         52 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody lacks an Fc portion.  
     
     
         53 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody is an F(ab′) 2 , an Fab, or an Fv structure.  
     
     
         54 . The antibody or antigen-binding fragment thereof of  claim 47 , wherein said antibody or antigen-binding fragment thereof is present in a pharmaceutically acceptable carrier.  
     
     
         55 . A method of diagnosing a subject as having, or having a predisposition to, a pregnancy related hypertensive disorder, said method comprising measuring the level of a soluble endoglin polypeptide in a sample from said subject.  
     
     
         56 . The method of  claim 55 , wherein said pregnancy related hypertensive disorder is pre-eclampsia, eclampsia, gestational hypertension, chronic hypertension, HELLP syndrome, and pregnancy with a SGA infant.  
     
     
         57 . The method of  claim 56 , wherein said pregnancy related hypertensive disorder is pre-eclampsia or eclampsia.  
     
     
         58 . The method of  claim 55 , wherein the level of soluble endoglin is the level of free, bound, or total soluble endoglin.  
     
     
         59 . The method of  claim 55 , wherein a level of soluble endoglin greater than 20 ng/ml is a diagnostic indicator of a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         60 . The method of claims  55 , wherein said measuring is done using an immunological assay.  
     
     
         61 . The method of  claim 60 , wherein said immunological assay is an ELISA.  
     
     
         62 . The method of  claim 55 , wherein an increase in the level of said soluble endoglin polypeptide relative to a normal reference is a diagnostic indicator of pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         63 . The method of  claim 55  wherein said measuring of levels is done on two or more occasions and a change in said levels between measurements is a diagnostic indicator of said pregnancy related hypertensive disorder.  
     
     
         64 . The method of  claim 55 , wherein said monitoring further comprises measuring the level of at least one of sFlt-1, VEGF, or PlGF polypeptide in a sample from said subject.  
     
     
         65 . The method of  claim 64 , further comprising calculating the relationship between said levels of soluble endoglin, sFlt-1, VEGF, or PlGF using a metric, wherein an alteration in the relationship between said levels in the subject sample relative to a reference sample, is a diagnostic indicator of a pregnancy related hypertensive disorder in said subject.  
     
     
         66 . The method of  claim 65 , wherein said metric is a pre-eclampsia anti-angiogenic index (PAAI): [sFlt-1VEGF+PlGF].  
     
     
         67 . The method of  claim 66 , wherein a PAAI value greater than 10 is a diagnostic indicator of a pregnancy related hypertensive disorder in said subject.  
     
     
         68 . The method of  claim 65 , wherein said metric is a soluble endoglin anti-angiogenic index: [(sFlt-1+0.25 soluble endoglin)/PlGF].  
     
     
         69 . The method of  claim 68 , wherein a soluble endoglin anti-angiogenic index value greater than 100 is a diagnostic indicator of a pregnancy related hypertensive disorder in said subject.  
     
     
         70 . The method of  claim 65 , wherein said metric is [(soluble endoglin+sFlt-1)/PlGF].  
     
     
         71 . The method of  claim 66 ,  68 , or  70 , wherein said metric further comprises the body mass index of the mother or the gestational age of the fetus.  
     
     
         72 . A method of diagnosing a subject as having, or having a predisposition to, a pregnancy related hypertensive disorder, said method comprising measuring the level of an endoglin nucleic acid molecule in a sample from said subject and comparing it to a reference, wherein an alteration in said levels compared to a reference diagnoses a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         73 . The method of  claim 72 , wherein said pregnancy related hypertensive disorder is pre-eclampsia, eclampsia, gestational hypertension, HELLP syndrome, chronic hypertension, or pregnancy with a small for gestation age infant.  
     
     
         74 . The method of  claim 73 , wherein said pregnancy related hypertensive disorder is pre-eclampsia or eclampsia.  
     
     
         75 . The method of  claim 72 , wherein said alteration in said levels is an increase.  
     
     
         76 . The method of  claim 72 , wherein said endoglin nucleic acid is a soluble endoglin nucleic acid.  
     
     
         77 . The method of  claim 72 , further comprising measuring the level of at least one of sFlt-1, VEGF, or PlGF nucleic acid molecule in a sample from said subject and comparing it to a reference sample, wherein an alteration in said level diagnoses a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         78 . The method of  claim 72 , wherein said measuring of levels is done on two or more occasions and a change in said levels between measurements is a diagnostic indicator of said pregnancy related hypertensive disorder.  
     
     
         79 . A method of diagnosing a subject as having, or having a predisposition to a pregnancy related hypertensive disorder, said method comprising determining the nucleic acid sequence of an endoglin gene in a sample from a subject and comparing it to a reference sequence, wherein an alteration in the subject's nucleic acid sequence that is an alteration that changes the expression level of the gene product in said subject diagnoses the subject with a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         80 . The method of  claim 55 ,  72 , or  79 , wherein said sample is a bodily fluid of said subject in which said soluble endoglin is normally detectable.  
     
     
         81 . The method of  claim 80 , wherein said bodily fluid is selected from the group consisting of urine, amniotic fluid, blood, serum, plasma, and cerebrospinal fluid.  
     
     
         82 . The method of  claim 55 ,  72 , or  79 , wherein said sample is a cell or a tissue.  
     
     
         83 . The method of  claim 82 , wherein said cell is selected from the group consisting of an endothelial cell, a leukocyte, a monocyte, or a cell derived from the placenta.  
     
     
         84 . The method of  claim 82 , wherein said tissue is a placental tissue.  
     
     
         85 . The method of  claim 55 ,  72 , or  79 , wherein said subject is a non-pregnant human, a pregnant human, a post-partum human, or a non-human and said method diagnoses a predisposition to pre-eclampsia or eclampsia.  
     
     
         86 . The method of  claim 85 , wherein said non-human is selected from the group consisting of a cow, a horse, a sheep, a pig, a goat, a dog, or a cat.  
     
     
         87 . The method of  claim 55 ,  72 , or  79 , wherein said method is used to diagnose a pregnancy related hypertensive disorder, or a predisposition to a pregnancy related hypertensive disorder, at least 4 weeks prior to the onset of symptoms.  
     
     
         88 . A kit for the diagnosis of a pregnancy related hypertensive disorder, or a predisposition to a pregnancy related hypertensive disorder, in a subject comprising a nucleic acid molecule having an endoglin nucleic acid sequence or a sequence complementary thereto, or any combination thereof, and instructions for using said nucleic acid molecule to diagnose a pregnancy related hypertensive disorder or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         89 . The kit of  claim 88 , wherein said kit further comprises a nucleic acid sequence of VEGF, sFlt-1 or PlGF or a sequence complementary thereto, or any combination thereof.  
     
     
         90 . The kit of  claim 88 , wherein said endoglin nucleic acid is a soluble endoglin nucleic acid.  
     
     
         91 . A kit for the diagnosis of a pregnancy related hypertensive disorder, or a predisposition to a pregnancy related hypertensive disorder, in a subject comprising a soluble endoglin binding molecule and instructions for the use of said soluble endoglin binding molecule for the diagnosis of said pregnancy related hypertensive disorder, or a predisposition to a pregnancy related hypertensive disorder.  
     
     
         92 . The kit of  claim 91 , wherein said soluble endoglin binding molecule is an antibody, or antigen-binding fragment thereof, that specifically binds soluble endoglin.  
     
     
         93 . The kit of  claim 91 , wherein said kit further comprises a VEGF, sFlt-1 or PlGF binding molecule.