Sex, pregnancy and aortic disease in Marfan syndrome

Background : Sex-related differences as well as the adverse effect of pregnancy on aortic disease outcome are well-established phenomena in humans with Marfan syndrome (MFS). The underlying mechanisms of these observations are largely unknown. Objectives : In an initial (pilot) step we aimed to confirm the differences between male and female MFS patients as well as between females with and without previous pregnancy. We then sought to evaluate whether these findings are recapitulated in a pre-clinical model and performed in-depth cardiovascular phenotyping of mutant male and both nulliparous and multiparous female Marfan mice. The effect of 17 beta-estradiol on fibrillin-1 protein synthesis was compared in vitro using human aortic smooth muscle cells and fibroblasts. Results : Our small retrospective study of aortic dimensions in a cohort of 10 men and 20 women with MFS (10 pregnant and 10 non-pregnant) confirmed that aortic root growth was significantly increased in the pregnant group compared to the non-pregnant group (0.64mm/year vs. 0.12mm/year, p = 0.018). Male MFS patients had significantly larger aortic root diameters compared to the non-pregnant and pregnant females at baseline and follow-up (p = 0.002 and p = 0.007, respectively), but no significant increase in aortic root growth was observed compared to the females after follow-up (p = 0.559 and p = 0.352). In the GT-8/+ MFS mouse model, multiparous female Marfan mice showed increased aortic diameters when compared to nulliparous females. Aortic dilatation in multiparous females was comparable to Marfan male mice. Moreover, increased aortic diameters were associated with more severe fragmentation of the elastic lamellae. In addition, 17 beta-estradiol was found to promote fibrillin-1 production by human aortic smooth muscle cells. Conclusions : Pregnancy-related changes influence aortic disease severity in otherwise protected female MFS mice and patients. There may be a role for estrogen in the female sex protective effect

Hybrid Silica-Coated PLGA Nanoparticles for Enhanced Enzyme-Based Therapeutics

Some cancer cells rely heavily on non-essential biomolecules for survival, growth, and proliferation. Enzyme based therapeutics can eliminate these biomolecules, thus specifically targeting neoplastic cells; however, enzyme therapeutics are susceptible to immune clearance, exhibit short half-lives, and require frequent administration. Encapsulation of therapeutic cargo within biocompatible and biodegradable poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) is a strategy for controlled release. Unfortunately, PLGA NPs exhibit burst release of cargo shortly after delivery or upon introduction to aqueous environments where they decompose via hydrolysis. Here, we show the generation of hybrid silica-coated PLGA (SiLGA) NPs as viable drug delivery vehicles exhibiting sub-200 nm diameters, a metastable Zeta potential, and high loading efficiency and content. Compared to uncoated PLGA NPs, SiLGA NPs offer greater retention of enzymatic activity and slow the burst release of cargo. Thus, SiLGA encapsulation of therapeutic enzymes, such as asparaginase, could reduce frequency of administration, increase half-life, and improve efficacy for patients with a range of diseases

Correction: Sex, pregnancy and aortic disease in Marfan syndrome.

[This corrects the article DOI: 10.1371/journal.pone.0181166.]

Aortic root morphology in wild-type and heterozygous GT-8/+ mice at the age of 8 months.

<p>A. The aortic elastic lamellae of GT-8/+ mice show clear fragmentation compared to the wild-type controls. Major breaks spanning at least 3 consecutive layers of elastic lamellae can be found in male and multiparous female GT-8/+ mice. Scale bars = 100 μm. B. At the age of 8 months, GT-8/+ male mice show significantly more fragmentation of the aortic elastic lamellae (left graph) and an increased number of major breaks (right graph) compared to wild-type littermates. Similarly, nulliparous and multiparous female GT-8/+ mice both show significantly more fragmentation of the aortic elastic lamellae compared to wild-type (left graph). The number of major breaks in the aortic root of multiparous GT-8/+ female mice is, however, significantly higher than in nulliparous GT-8/+ female mice and multiparous wild-type female mice (right graph). Each symbol represents a single animal. Means with 95% confidence intervals are shown. F: female; M: male; np: nulliparous; mp: multiparous; p-values: * <0.05, ** <0.005, *** <0.001.</p

Aortic root characteristics in MFS patients.

<p>A. At baseline, aortic root diameters are higher in men compared to women. Non-pregnant women have higher aortic root diameters at baseline compared to pregnant women. After several years of follow-up, aortic root diameters increase in men and pregnant women, but these barely increase in non-pregnant women. Time interval between baseline and after pregnancy is 1.8 months, and between after pregnancy and follow-up is 5.5 years. * Values were only available for the pregnant women group at this time point. B. The growth rate of the aortic root, measured at baseline and last follow-up, was significantly higher in the pregnant group in comparison to the non-pregnant group. No significant difference in aortic growth rate was observed between the men and women. Results are shown as means ± standard deviation, unless the values were not normally distributed in which case median ± interquartile range are shown. ns: not significant; * p-value <0.05.</p

Characteristics of the aortic root of MFS patients at baseline and after several years of follow-up.

<p>Characteristics of the aortic root of MFS patients at baseline and after several years of follow-up.</p

Fibrillin-1 production by (A) human aortic smooth muscle cells (HASMC) and (B) fibroblasts.

<p>Cells were treated with 17β- estradiol and TGFβ-1 and compared to untreated control cells. Higher concentrations of 17β-estradiol (4–8 nmol/L) compared to control (ctl) (0, 1 or 2 nmol/L 17β-estradiol) promoted fibrillin-1 production by HASMC. In contrast, fibrillin-1 production by fibroblasts was stimulated by TGFβ-1 but not by 17β-estradiol. Results shown were obtained using the antibody pair B15-AP201. Error bars (standard deviation) are based on combining two separate experiments with HASMC and three separate experiments with fibroblasts. In each experiment, samples were tested in duplicate. Significant p-values are marked with an *.</p

Aortic dimensions of wild-type (+/+) and heterozygous GT-8/+ males (M) and nulliparous (np) and multiparous (mp) females (F) at age 8 (panel A) and 12 months (panel B).

<p>Male and multiparous female GT-8/+ mice show more severe aortic disease than wild-type mice and nulliparous GT-8/+ females at both time points. Nulliparous GT-8/+ females differed only significantly from the nulliparous wild-type females at the level of the aortic root at 12 months. In addition, nulliparous females have smaller aortic diameters than the GT-8/+ males. Black lines represent the mean ± standard deviation. Non-significant differences are not indicated in the figure. p-values: * <0.05, ** <0.005, *** <0.001, **** <0.0001.</p