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Placenta-derived exosomes continuously increase in maternal circulation over the first trimester of pregnancy.

Sarker S, Scholz-Romero K, Perez A, Illanes SE, Mitchell MD, Rice GE, Salomon C - J Transl Med (2014)

Bottom Line: A time-series experimental design was used to establish pregnancy-associated changes in maternal plasma exosome concentrations during the first trimester.There is no significant decline in their yield with the freeze/thawing processes or change in their EM morphology.During normal healthy pregnancy, the number of exosomes present in the maternal plasma increased significantly with gestational age across the first trimester of pregnancy.

View Article: PubMed Central - PubMed

Affiliation: UQ Centre for Clinical Research, Centre for Clinical Diagnostics, Royal Brisbane and Women's Hospital, University of Queensland, Building 71/918, Herston QLD 4029, Queensland, Australia. c.salomongallo@uq.edu.au.

ABSTRACT

Background: Human placenta releases specific nanovesicles (i.e. exosomes) into the maternal circulation during pregnancy, however, the presence of placenta-derived exosomes in maternal blood during early pregnancy remains to be established. The aim of this study was to characterise gestational age related changes in the concentration of placenta-derived exosomes during the first trimester of pregnancy (i.e. from 6 to 12 weeks) in plasma from women with normal pregnancies.

Methods: A time-series experimental design was used to establish pregnancy-associated changes in maternal plasma exosome concentrations during the first trimester. A series of plasma were collected from normal healthy women (10 patients) at 6, 7, 8, 9, 10, 11 and 12 weeks of gestation (n = 70). We measured the stability of these vesicles by quantifying and observing their protein and miRNA contents after the freeze/thawing processes. Exosomes were isolated by differential and buoyant density centrifugation using a sucrose continuous gradient and characterised by their size distribution and morphology using the nanoparticles tracking analysis (NTA; Nanosight™) and electron microscopy (EM), respectively. The total number of exosomes and placenta-derived exosomes were determined by quantifying the immunoreactive exosomal marker, CD63 and a placenta-specific marker (Placental Alkaline Phosphatase PLAP).

Results: These nanoparticles are extraordinarily stable. There is no significant decline in their yield with the freeze/thawing processes or change in their EM morphology. NTA identified the presence of 50-150 nm spherical vesicles in maternal plasma as early as 6 weeks of pregnancy. The number of exosomes in maternal circulation increased significantly (ANOVA, p = 0.002) with the progression of pregnancy (from 6 to 12 weeks). The concentration of placenta-derived exosomes in maternal plasma (i.e. PLAP+) increased progressively with gestational age, from 6 weeks 70.6 ± 5.7 pg/ml to 12 weeks 117.5 ± 13.4 pg/ml. Regression analysis showed that weeks is a factor that explains for >70% of the observed variation in plasma exosomal PLAP concentration while the total exosome number only explains 20%.

Conclusions: During normal healthy pregnancy, the number of exosomes present in the maternal plasma increased significantly with gestational age across the first trimester of pregnancy. This study is a baseline that provides an ideal starting point for developing early detection method for women who subsequently develop pregnancy complications, clinically detected during the second trimester. Early detection of women at risk of pregnancy complications would provide an opportunity to develop and evaluate appropriate intervention strategies to limit acute adverse sequel.

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Related in: MedlinePlus

Exosome profiling across first trimester pregnancy. Enriched exosomal population (i.e. number of exosome particles) and placenta-derived exosomes (i.e. exosomal PLAP) were quantified in in peripheral plasma of women in the first trimester of pregnancy by ELISA. (A) exosomes as particles per ml plasma. (B) individual variation in exosome number for each week (C) exosomal PLAP during first trimester of pregnancy (i.e. 6–12 weeks). (D) individual variation in exosomal PLAP for each week. Data are presented as aligned dot plot and values are mean ± SEM. In A, two-way ANOVA **p = 0.0048, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 7 weeks and †p < 0.005: 6 vs. 12 weeks. In C, two-way ANOVA ***p < 0.0001, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 9 and 10 weeks, †p < 0.005: 6 vs. 11 and 12 weeks, and ‡p < 0.005: 8 vs. 11 and 12 weeks.
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Fig3: Exosome profiling across first trimester pregnancy. Enriched exosomal population (i.e. number of exosome particles) and placenta-derived exosomes (i.e. exosomal PLAP) were quantified in in peripheral plasma of women in the first trimester of pregnancy by ELISA. (A) exosomes as particles per ml plasma. (B) individual variation in exosome number for each week (C) exosomal PLAP during first trimester of pregnancy (i.e. 6–12 weeks). (D) individual variation in exosomal PLAP for each week. Data are presented as aligned dot plot and values are mean ± SEM. In A, two-way ANOVA **p = 0.0048, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 7 weeks and †p < 0.005: 6 vs. 12 weeks. In C, two-way ANOVA ***p < 0.0001, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 9 and 10 weeks, †p < 0.005: 6 vs. 11 and 12 weeks, and ‡p < 0.005: 8 vs. 11 and 12 weeks.

Mentions: The gestational age variation in plasma exosome number was analysed by two-way ANOVA with the variance partitioned between gestational age and subject. A significantly effect of gestational age was identified (n = 69, one missing value, p < 0.005). A post-hoc multiple range test was used to identify statistically significant (p <0.05) differences between pairwise comparisons (Figure 3A). In addition, a significant effect of subject was identified (n = 69, one missing value, p < 0.05) (Figure 3B). In addition, NEP and gestational age (i.e. 6–12 weeks) displayed a significant positive linear relationship (r2 = 0.202, p < 0.001, n = 69, one missing value).Figure 3


Placenta-derived exosomes continuously increase in maternal circulation over the first trimester of pregnancy.

Sarker S, Scholz-Romero K, Perez A, Illanes SE, Mitchell MD, Rice GE, Salomon C - J Transl Med (2014)

Exosome profiling across first trimester pregnancy. Enriched exosomal population (i.e. number of exosome particles) and placenta-derived exosomes (i.e. exosomal PLAP) were quantified in in peripheral plasma of women in the first trimester of pregnancy by ELISA. (A) exosomes as particles per ml plasma. (B) individual variation in exosome number for each week (C) exosomal PLAP during first trimester of pregnancy (i.e. 6–12 weeks). (D) individual variation in exosomal PLAP for each week. Data are presented as aligned dot plot and values are mean ± SEM. In A, two-way ANOVA **p = 0.0048, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 7 weeks and †p < 0.005: 6 vs. 12 weeks. In C, two-way ANOVA ***p < 0.0001, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 9 and 10 weeks, †p < 0.005: 6 vs. 11 and 12 weeks, and ‡p < 0.005: 8 vs. 11 and 12 weeks.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4283151&req=5

Fig3: Exosome profiling across first trimester pregnancy. Enriched exosomal population (i.e. number of exosome particles) and placenta-derived exosomes (i.e. exosomal PLAP) were quantified in in peripheral plasma of women in the first trimester of pregnancy by ELISA. (A) exosomes as particles per ml plasma. (B) individual variation in exosome number for each week (C) exosomal PLAP during first trimester of pregnancy (i.e. 6–12 weeks). (D) individual variation in exosomal PLAP for each week. Data are presented as aligned dot plot and values are mean ± SEM. In A, two-way ANOVA **p = 0.0048, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 7 weeks and †p < 0.005: 6 vs. 12 weeks. In C, two-way ANOVA ***p < 0.0001, Dunn’s post-hoc test analysis = *p < 0.05 6 vs. 9 and 10 weeks, †p < 0.005: 6 vs. 11 and 12 weeks, and ‡p < 0.005: 8 vs. 11 and 12 weeks.
Mentions: The gestational age variation in plasma exosome number was analysed by two-way ANOVA with the variance partitioned between gestational age and subject. A significantly effect of gestational age was identified (n = 69, one missing value, p < 0.005). A post-hoc multiple range test was used to identify statistically significant (p <0.05) differences between pairwise comparisons (Figure 3A). In addition, a significant effect of subject was identified (n = 69, one missing value, p < 0.05) (Figure 3B). In addition, NEP and gestational age (i.e. 6–12 weeks) displayed a significant positive linear relationship (r2 = 0.202, p < 0.001, n = 69, one missing value).Figure 3

Bottom Line: A time-series experimental design was used to establish pregnancy-associated changes in maternal plasma exosome concentrations during the first trimester.There is no significant decline in their yield with the freeze/thawing processes or change in their EM morphology.During normal healthy pregnancy, the number of exosomes present in the maternal plasma increased significantly with gestational age across the first trimester of pregnancy.

View Article: PubMed Central - PubMed

Affiliation: UQ Centre for Clinical Research, Centre for Clinical Diagnostics, Royal Brisbane and Women's Hospital, University of Queensland, Building 71/918, Herston QLD 4029, Queensland, Australia. c.salomongallo@uq.edu.au.

ABSTRACT

Background: Human placenta releases specific nanovesicles (i.e. exosomes) into the maternal circulation during pregnancy, however, the presence of placenta-derived exosomes in maternal blood during early pregnancy remains to be established. The aim of this study was to characterise gestational age related changes in the concentration of placenta-derived exosomes during the first trimester of pregnancy (i.e. from 6 to 12 weeks) in plasma from women with normal pregnancies.

Methods: A time-series experimental design was used to establish pregnancy-associated changes in maternal plasma exosome concentrations during the first trimester. A series of plasma were collected from normal healthy women (10 patients) at 6, 7, 8, 9, 10, 11 and 12 weeks of gestation (n = 70). We measured the stability of these vesicles by quantifying and observing their protein and miRNA contents after the freeze/thawing processes. Exosomes were isolated by differential and buoyant density centrifugation using a sucrose continuous gradient and characterised by their size distribution and morphology using the nanoparticles tracking analysis (NTA; Nanosight™) and electron microscopy (EM), respectively. The total number of exosomes and placenta-derived exosomes were determined by quantifying the immunoreactive exosomal marker, CD63 and a placenta-specific marker (Placental Alkaline Phosphatase PLAP).

Results: These nanoparticles are extraordinarily stable. There is no significant decline in their yield with the freeze/thawing processes or change in their EM morphology. NTA identified the presence of 50-150 nm spherical vesicles in maternal plasma as early as 6 weeks of pregnancy. The number of exosomes in maternal circulation increased significantly (ANOVA, p = 0.002) with the progression of pregnancy (from 6 to 12 weeks). The concentration of placenta-derived exosomes in maternal plasma (i.e. PLAP+) increased progressively with gestational age, from 6 weeks 70.6 ± 5.7 pg/ml to 12 weeks 117.5 ± 13.4 pg/ml. Regression analysis showed that weeks is a factor that explains for >70% of the observed variation in plasma exosomal PLAP concentration while the total exosome number only explains 20%.

Conclusions: During normal healthy pregnancy, the number of exosomes present in the maternal plasma increased significantly with gestational age across the first trimester of pregnancy. This study is a baseline that provides an ideal starting point for developing early detection method for women who subsequently develop pregnancy complications, clinically detected during the second trimester. Early detection of women at risk of pregnancy complications would provide an opportunity to develop and evaluate appropriate intervention strategies to limit acute adverse sequel.

Show MeSH
Related in: MedlinePlus