Free DNA and Nucleosome Concentrations in Pathological Pregnancies

Study Overview
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Brief Summary
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The primary objective of this study is to demonstrate that plasma concentrations of nucleosomes and free DNA differ between three groups: pregnant patients with complications typical of placental insufficiency or venous thrombosis (group P), healthy women (Group T1) and healthy pregnant women (Group T2).

Detailed Description
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Our secondary objectives include the following: To describe, in 15 healthy, non-pregnant women changes in plasma concentrations of nucleosomes and free DNA over 3 months. To describe, in 15 pregnant women (without complications), changes in plasma concentrations of nucleosomes and free DNA over the last 7 months of pregnancy To show that plasma concentrations of nucleosomes and free DNA, in patients with complicated pregnancies differ according to the nature of the complication To show that a relationship exists between the concentrations of nucleosomes, free DNA, and total granulocyte microparticles (and trophoblast particles for pregnant women) To evaluate the relationship between nucleosome concentrations, free DNA concentrations and circulating leukocyte populations To evaluate the relationship between nucleosome concentrations, free DNA concentrations and hemostasis markers To describe changes in hemostasis markers throughout pregnancy To evaluate the relationship between nucleosome concentrations, free DNA concentrations and the angiogenic marker CD146 To add to the Nîmes University Hospital biological collections.

Official Title
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Comparative Study of Plasma Free DNA and Nucleosome Concentrations: Pathological Versus Normal Pregnancies

Conditions
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Pregnancy, Venous Thrombosis, Pulmonary Embolism, Hypertension, Pregnancy-Induced, Eclampsia, HELLP Syndrome, Pre-Eclampsia, Fetal Death, Placental Insufficiency

Intervention / Treatment
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* Biological: Bloodwork, baseline
* Biological: Blood work, Months 1 & 2
* Biological: Bloodwork, Months -1 to -6


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria for all patients: The patient must have given her informed and signed consent The patient must be insured or beneficiary of a health insurance plan Inclusion Criteria for patients in group P: The patient is pregnant and has complications typical of placental vascular disease (preeclampsia, eclampsia, HELLP syndrome, retro-placental hematoma, in utero fetal death) or venous thromboembolism (deep vein thrombosis, pulmonary embolism) Inclusion Criteria for patients in group T1: The patient is available for 3 months of follow-up The patient is a non-pregnant healthy volunteer No identifiable chronic pathologies No history of neoplastic disease No history of chronic infectious disease No acute disease (such as benign infection), now or within the past two weeks Inclusion Criteria for patients in group T2: The patient is available for 7 months of follow-up The patient is pregnant, with no identifiable pregnancy complications No identifiable chronic pathologies No history of neoplastic disease No history of chronic infectious disease No acute disease (such as benign infection), now or within the past two weeks Exclusion Criteria for all patients: The patient is participating in another study (with the exception of the following studies: PAPILLO-PMA (2013-A00538-37), ElastoMAP (2013-A01148-37), ElastoDéclenche (2014-A00828-39), LXRs (2009-A00968-49), Bakri (2013-A00914-41), OASIS 2 (2013-A00022-43)). The patient is in an exclusion period determined by a previous study The patient is under judicial protection, under tutorship or curatorship The patient refuses to sign the consent It is impossible to correctly inform the patient The patient cannot read French The patient is receiving hormonal ovarian stimulation in the context of medically assisted procreation Impossible to perform venipuncture under good conditions New complication or pathology during the study (except for pregnancy complications in group P) Exclusion Criteria for group P: Twin or multiple pregnancy The patient is participating in another study (with the exception of those mentioned in the exclusion criteria for all patients and the DG Postpartum study (2013-A00277-38) Exclusion Criteria for group T1: The patient is pregnant The patient is breast feeding The patient has given birth within the last 3 months Known history of chronic disease History of treated neoplastic disease Acute disease within the past two weeks (includes benign disease) Exclusion Criteria for group T2: Pregnancy with complications Known history of chronic disease History of treated neoplastic disease Acute disease within the past two weeks (includes benign disease) Twin or multiple pregnancy

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Group P: pregnancy w/complications<br>The patient is pregnant and has complications typical of placental vascular disease (preeclampsia, eclampsia, HELLP syndrome, retro-placental hematoma, in utero fetal death) or venous thromboembolism (deep vein thrombosis, pulmonary embolism). 100 patients will be included. Interventions to be administered: Bloodwork, baseline | Biological: Bloodwork, baseline<br>* 36 ml of blood are drawn at baseline (last month of pregnancy for groups P and T2, inclusion for group T1) in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>|
| Group T1: Healthy volunteers<br>Healthy volunteers with no history of chronic or neoplastic disease. 30 healthy volunteers will be included. Interventions to be administered: Bloodwork, baseline | Biological: Bloodwork, baseline<br>* 36 ml of blood are drawn at baseline (last month of pregnancy for groups P and T2, inclusion for group T1) in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>|
| Group T2: Pregnancy, no complications<br>Pregnant patients with no identifiable pregnancy complications, and no history of chronic or neoplastic disease. 50 pregnant volunteers will be included. Interventions to be administered: Bloodwork, baseline | Biological: Bloodwork, baseline<br>* 36 ml of blood are drawn at baseline (last month of pregnancy for groups P and T2, inclusion for group T1) in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>|
| Group T1x: 15 Healthy volunteers<br>15 Healthy volunteers selected from group T1 (the first 15). These patients will have 2 additional months of follow up. Interventions to be administered: Blood work, Months 1 & 2 | Biological: Bloodwork, baseline<br>* 36 ml of blood are drawn at baseline (last month of pregnancy for groups P and T2, inclusion for group T1) in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>Biological: Blood work, Months 1 & 2<br>* 36 ml of blood are drawn at 1 & 2 months after inclusion in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>|
| Group T2x: 15 Pregnancy, no complications<br>15 patients selected from group T2 (the first 15); these patients will have 7 months of follow up during pregnancy. Interventions to be administered: Bloodwork, Months -1 to -6 | Biological: Bloodwork, baseline<br>* 36 ml of blood are drawn at baseline (last month of pregnancy for groups P and T2, inclusion for group T1) in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>Biological: Bloodwork, Months -1 to -6<br>* 36 ml of blood are drawn at the third, fourth, fifth, sixth, seventh and eight months of normal pregnancy (corresponding to months -1 to -6 before comparative baseline; this group is included in the study early during pregnancy)in order to quantify the following: plasma free DNA concentration, plasma nucleosome concentration, hemoglobin, platelets, leukocytes, polynuclear neutrophils, mean corpuscular volume, monocytes, mean corpuscular hemoglobin, lymphocytes, polynuclear eosinophils, polynuclear basophils, D-Dimers, Fibrin monomers, Trophoblast microparticles, Angiogenic marker CD146.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Total plasma concentration of free DNA (ng/ml) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | Base line (day 0) | 
| Total plasma concentration of nucleosomes (AU) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth | Base line (day 0) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Hemoglobin (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Platelets (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Leukocytes (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Polynuclear neutrophils(g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Mean corpuscular volume (fL) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Monocytes (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Mean corpuscular hemoglobin (pg/GR) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Lymphocytes (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Polynuclear eosinophils (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Polynuclear basophils (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| D-dimers (ng/ml) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Fibrin monomers (ng/ml) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Trophoblast microparticles (%) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Angiogenic marker CD146 (ng/ml) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | baseline (day 0) | 
| Hemoglobin (g/l) | For group T1x only | 1 month | 
| Platelets (g/l) | For group T1x only | 1 month | 
| Leukocytes (g/l) | For group T1x only | 1 month | 
| Polynuclear neutrophils(g/l) | For group T1x only | 1 month | 
| Mean corpuscular volume (fL) | For group T1x only | 1 month | 
| Monocytes (g/l) | For group T1x only | 1 month | 
| Mean corpuscular hemoglobin (pg/GR) | For group T1x only | 1 month | 
| Lymphocytes (g/l) | For group T1x only | 1 month | 
| Polynuclear eosinophils (g/l) | For group T1x only | 1 month | 
| Polynuclear basophils (g/l) | For group T1x only | 1 month | 
| D-dimers (ng/ml) | For group T1x only | 1 month | 
| Fibrin monomers (ng/ml) | For group T1x only | 1 month | 
| Trophoblast microparticles (%) | For group T1x only | 1 month | 
| Angiogenic marker CD146 (ng/ml) | For group T1x only | 1 month | 
| Total plasma concentration of free DNA (ng/ml) | For group T1x only | 1 month | 
| Total plasma concentration of nucleosomes (AU) | For group T1x only | 1 month | 
| Hemoglobin (g/l) | For group T1x only | 2 months | 
| Platelets (g/l) | For group T1x only | 2 months | 
| Leukocytes (g/l) | For group T1x only | 2 months | 
| Polynuclear neutrophils(g/l) | For group T1x only | 2 months | 
| Mean corpuscular volume (fL) | For group T1x only | 2 months | 
| Monocytes (g/l) | For group T1x only | 2 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T1x only | 2 months | 
| Lymphocytes (g/l) | For group T1x only | 2 months | 
| Polynuclear eosinophils (g/l) | For group T1x only | 2 months | 
| Polynuclear basophils (g/l) | For group T1x only | 2 months | 
| D-dimers (ng/ml) | For group T1x only | 2 months | 
| Fibrin monomers (ng/ml) | For group T1x only | 2 months | 
| Trophoblast microparticles (%) | For group T1x only | 2 months | 
| Angiogenic marker CD146 (ng/ml) | For group T1x only | 2 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T1x only | 2 months | 
| Total plasma concentration of nucleosomes (AU) | For group T1x only | 2 months | 
| Hemoglobin (g/l) | For group T2x only | -1 months | 
| Hemoglobin (g/l) | For group T2x only | -2 months | 
| Hemoglobin (g/l) | For group T2x only | -3 months | 
| Hemoglobin (g/l) | For group T2x only | -4 months | 
| Hemoglobin (g/l) | For group T2x only | -5 months | 
| Hemoglobin (g/l) | For group T2x only | -6 months | 
| Platelets (g/l) | For group T2x only | -1 months | 
| Platelets (g/l) | For group T2x only | -2 months | 
| Platelets (g/l) | For group T2x only | -3 months | 
| Platelets (g/l) | For group T2x only | -4 months | 
| Platelets (g/l) | For group T2x only | -5 months | 
| Platelets (g/l) | For group T2x only | -6 months | 
| Leukocytes (g/l) | For group T2x only | -1 months | 
| Leukocytes (g/l) | For group T2x only | -2 months | 
| Leukocytes (g/l) | For group T2x only | -3 months | 
| Leukocytes (g/l) | For group T2x only | -4 months | 
| Leukocytes (g/l) | For group T2x only | -5 months | 
| Leukocytes (g/l) | For group T2x only | -6 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -1 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -2 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -3 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -4 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -5 months | 
| Polynuclear neutrophils(g/l) | For group T2x only | -6 months | 
| Mean corpuscular volume (fL) | For group T2x only | -1 months | 
| Mean corpuscular volume (fL) | For group T2x only | -2 months | 
| Mean corpuscular volume (fL) | For group T2x only | -3 months | 
| Mean corpuscular volume (fL) | For group T2x only | -4 months | 
| Mean corpuscular volume (fL) | For group T2x only | -5 months | 
| Mean corpuscular volume (fL) | For group T2x only | -6 months | 
| Monocytes (g/l) | For group T2x only. | -1 months | 
| Monocytes (g/l) | For group T2x only. | -2 months | 
| Monocytes (g/l) | For group T2x only. | -3 months | 
| Monocytes (g/l) | For group T2x only. | -4 months | 
| Monocytes (g/l) | For group T2x only. | -5 months | 
| Monocytes (g/l) | For group T2x only. | -6 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -1 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -2 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -3 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -4 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -5 months | 
| Mean corpuscular hemoglobin (pg/GR) | For group T2x only. | -6 months | 
| Lymphocytes (g/l) | For group T2x only. | -1 months | 
| Lymphocytes (g/l) | For group T2x only. | -2 months | 
| Lymphocytes (g/l) | For group T2x only. | -3 months | 
| Lymphocytes (g/l) | For group T2x only. | -4 months | 
| Lymphocytes (g/l) | For group T2x only. | -5 months | 
| Lymphocytes (g/l) | For group T2x only. | -6 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -1 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -2 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -3 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -4 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -5 months | 
| Polynuclear eosinophils (g/l) | For group T2x only. | -6 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -1 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -2 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -3 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -4 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -5 months | 
| Polynuclear basophils (g/l) | For group T2x only. | -6 months | 
| D-dimers (ng/ml) | For group T2x only. | -1 months | 
| D-dimers (ng/ml) | For group T2x only. | -2 months | 
| D-dimers (ng/ml) | For group T2x only. | -3 months | 
| D-dimers (ng/ml) | For group T2x only. | -4 months | 
| D-dimers (ng/ml) | For group T2x only. | -5 months | 
| D-dimers (ng/ml) | For group T2x only. | -6 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -1 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -2 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -3 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -4 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -5 months | 
| Fibrin monomers (ng/ml) | For group T2x only. | -6 months | 
| Trophoblast microparticles (%) | For group T2x only. | -1 months | 
| Trophoblast microparticles (%) | For group T2x only. | -2 months | 
| Trophoblast microparticles (%) | For group T2x only. | -3 months | 
| Trophoblast microparticles (%) | For group T2x only. | -4 months | 
| Trophoblast microparticles (%) | For group T2x only. | -5 months | 
| Trophoblast microparticles (%) | For group T2x only. | -6 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -1 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -2 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -3 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -4 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -5 months | 
| Angiogenic marker CD146 (ng/ml) | For group T2x only. | -6 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -1 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -2 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -3 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -4 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -5 months | 
| Total plasma concentration of nucleosomes (AU) | For group T2x only. | -6 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -1 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -2 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -3 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -4 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -5 months | 
| Total plasma concentration of free DNA (ng/ml) | For group T2x only. | -6 months | 
| Fibrinogen (g/l) | For group P, baseline occurs within the 30 days preceding birth. For group T1, baseline = time of inclusion. For group T2, baseline occurs within the 30 days preceding birth. | Base line (day 0) | 
| Fibrinogen (g/l) | For group T2x only. | -1 months | 
| Fibrinogen (g/l) | For group T2x only. | -2 months | 
| Fibrinogen (g/l) | For group T2x only. | -3 months | 
| Fibrinogen (g/l) | For group T2x only. | -4 months | 
| Fibrinogen (g/l) | For group T2x only. | -5 months | 
| Fibrinogen (g/l) | For group T2x only. | -6 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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plasma nucleosome concentration, plasma free DNA concentration