Introduction Tumor necrosis factor alpha (TNF-α) is elevated 2-fold in women with preeclampsia. Preclinical investigation shows TNF-α blockade reduces maternal blood pressure in reduced uterine perfusion pressure (RUPP) rats and attenuates the reduction in fetal weight. However, the benefits versus harms of this therapy on fetal growth and underlying pathogenesis are unknown. Thus, this study tested the hypothesis that maternal treatment with the soluble TNF-α inhibitor Etanercept (Etan) during late gestation improves placental perfusion, nutrient transport, and morphology, thereby improving fetal growth in the RUPP model of preeclampsia compared with Sham controls. We further hypothesized that maternal Etan treatment is associated with improved blood pressure and inflammatory profiles in offspring. Methods Sham or RUPP surgery was performed at gestational day (GD) 14, with vehicle or Etan (0.4 mg/kg, s.c.) administered at GD18. Results Fetal weight (p = 0.0365) and survival (p = 0.0002) were reduced in RUPP (p = 0.0365) at GD20; only fetal weight was improved in Etan-RUPP (p = 0.0480). At GD20, uterine artery resistance index (UARI) was increased in RUPP (p = 0.0094), but attenuated in Etan-RUPP, indicating improved placental perfusion. Impaired placental transport and morphology were evident in RUPP, with no improvement in Etan-RUPP. Birthweight was improved in Etan-RUPP (p = 0.0312), although total NK cells (p = 0.0376) were increased in female Etan-RUPP offspring. Circulating AT1-AA activity was elevated in adult male and female RUPP offspring (p = 0.0013, p = 0.0006), but attenuated in female Etan-RUPP offspring. Discussion: These results suggest improved fetal growth in Etan-RUPP, independent of placental morphology or nutrient transporter expression, with sex-specific effects on inflammation in adult RUPP offspring.

Preeclampsia (PE), new-onset hypertension during pregnancy, is accompanied by organ dysfunction, in addition to placental dysfunction, and is associated with chronic inflammation and fetal growth restriction. Agonistic autoantibodies against the angiotensin II type 1 receptor (AT1-AA) are produced in PE women and induce a PE-like phenotype when infused into pregnant rats. PE offspring are at higher risk for cardiovascular disease and hypertension; however, AT1-AA's role in these risks is unknown. Therefore, we hypothesized that AT1-AA exposure during pregnancy contributes to hypertension in adult offspring. We infused AT1-AA (1:40) starting on gestational day (GD) 14, allowed the dams to give birth, and birth weight was measured within 12 h. Offspring were aged to 4 months, one male and one female per litter were randomly selected to undergo carotid catheterization, mean arterial pressure (MAP) measurement, and blood and tissue collection. Female AT1-AA offspring had elevated MAP compared to control female offspring. Female AT1-AA offspring also had elevated progesterone, measured by mass spectroscopy, and renal endothelin-1, measured by RT-PCR and ELISA, compared to female control offspring. Male and female AT1-AA offspring had increased circulating AT1-AA, measured by cardiomyocyte bioassay, compared to male and female control offspring. These data demonstrate that AT1-AA infusion during pregnancy can predispose female offspring to have elevated blood pressure and persistently increased renal ET-1 and AT1-AA in adulthood.

Preeclampsia (PE), new-onset hypertension during pregnancy in association with other-organ dysfunction including the placenta. It is associated with inflammatory cytokines and fetal growth restriction (FGR) which itself is associated with increased risk for hypertension and cardiovascular disease in adult offspring. PE women have increased tumor necrosis factor alpha (TNF-α) and agonistic autoantibodies to the angiotensin II type 1 receptor (AT1-AA). Moreover, female PE children are at elevated risk for PE during pregnancy indicating a familial connection with PE. The reduced uterine perfusion pressure (RUPP) model of PE recaptures many of the characteristics of PE including hypertension, inflammatory activation, and fetal growth restriction. Growth-restricted RUPP offspring develop hypertension as adults and we have recently shown that adult RUPP offspring also produce AT1-AA. Importantly, RUPP rats have increased TNF-α and we have shown that TNF-α blockade with etanercept improves maternal blood pressure. The use of etanercept has not resulted in negative effects for mothers with autoimmune disorders that are kept on their treatment throughout pregnancy, however, its effect on fetal weight and long-term health is still being investigated. We hypothesize that etanercept could improve cardiovascular and chronic inflammatory outcomes in adult RUPP offspring. Pregnant rats were divided into the following groups: Sham, Sham + Etanercept, RUPP, RUPP + Etanercept. RUPP surgeries were performed on gestational day 14 and Etanercept (400 mcg/kg) was injected i.p. on gestational day 18. The dams were allowed to deliver naturally and birth weights were recorded within 12 hours. At 4 months of age mean arterial pressure (MAP) was recorded via carotid catheters, and blood and tissues were collected. Immune cells were measured by flow cytometry. AT1-AA was measured using a cardiomyocyte bioassay. A two-way ANOVA was used for statistical analysis. Etanercept improves birth weight in male RUPP offspring (5.2±0.2 g, n=7 (RUPP) vs 6.0±0.3g, n=9, p<0.05 (RUPP+Etanercept)), but did not affect adult blood pressure or inflammatory factors. Although Etanercept did not improve female birth weight, Etanercept trended to improve MAP in RUPP female offspring (130±5 mmHg, n=6, (RUPP) vs 123±4 mmHg, n=7 (RUPP+Etanercept)). In addition, there was a trending decrease in circulating B cells in the RUPP female offspring compared to sham female offspring which was normalized in the RUPP + Etanercept female offspring. (4.24±0.81 % gated, n=5 (RUPP); vs 9.56±2.07 % gated, n=7 (Sham), vs 9.04±1.42 % gated, n=6, (RUPP+Etanercept)). RUPP + Etanercept female offspring also had decreased in circulating AT1-AA which was elevated in RUPP female offspring (16±3 ΔBPM (RUPP), n=5, 7±2 ΔBPM, n=4 (RUPP+Etanercept), -1±2 ΔBPM, n=5, (Sham)). Overall, perinatal Etanercept treatment did not worsen fetal outcomes. Importantly, Etanercept may have a more positive effect to improve overall health outcomes of female offspring as seen by the trend to reduce blood pressure, normalize immune cells, and reduce circulating AT1-AA in adult females. Blocking TNF-α during PE may be a potential therapeutic strategy to improve both maternal and female fetal outcomes. This study was supported by NIH grants R01HL170622 (BL), R01HL143459 (BTA), F31HD110230 (NC), P20GM104357 (BTA), P30GM149404 (BTA), and P20GM121334 (BL, BTA). This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

Background Preeclampsia, new‐onset hypertension during pregnancy alongside other organ dysfunction, is the leading cause of mortality for the mother and low birth weight for the baby. Low birth weight contributes to high risk of cardiovascular disorders later in life. Women with preeclampsia have activated B cells producing agonistic autoantibodies to AT1‐AA (angiotensin II type I receptor). We hypothesize that rituximab, a B cell‐depleting chemotherapeutic, will deplete maternal B cells in reduced uterine perfusion pressure (RUPP) rats without worsening the effect of placental ischemia on pup growth and survival. Methods and Results To test this hypothesis, the RUPP procedure was performed, and rituximab was continuously infused via miniosmotic pump. Maternal blood and tissues were collected. A separate group of dams were allowed to deliver, pup weights were recorded, and at 4 months of age, tissues were collected from offspring. Immune cells were measured via flow cytometry, and AT1‐AA was quantified using a contraction bioassay. Blood pressure increased in RUPP rats and was normalized with rituximab treatment. RUPP offspring also had increased circulating B cells, cytolytic natural killer cells, and increased circulating AT1‐AA, which were normalized with maternal rituximab treatment. This is the first study to analyze the AT1‐AA in RUPP offspring, which was normalized with rituximab. Conclusions Our findings indicate that perinatal rituximab lowers maternal mean arterial pressure in RUPP rats and improves birth weight, circulating AT1‐AA, and circulating natural killer cells, indicating that rituximab improves adverse fetal outcomes in response to placental ischemia.

Objective: Preeclampsia (PE), new-onset hypertension during the third trimester of pregnancy alongside other organ dysfunction, is associated with chronic inflammation and fetal growth restriction. Agonistic autoantibodies against the angiotensin II type 1 receptor (AT1-AA) are produced in PE women and induce a PE-like phenotype when infused into pregnant rats. AT1-AA is still present in maternal circulation up to eight years postpartum. We have shown that AT1-AA exposure during pregnancy contributes to impairment of cerebral blood flow in the postpartum period. Moreover, PE offspring are at higher risk for cardiovascular disease and hypertension but AT1-AA’s role in this elevated risk is unknown. The objective of this study was to determine if AT1-AA contributes to the risk for hypertension in PE offspring. Therefore, we hypothesized that AT1-AA infusion during pregnancy would cause offspring to have low birth weight and hypertension in adulthood. Methods: We infused AT1-AA (1:40) starting on gestational day (GD) 14 and allowed the dams to give birth on GD21. Birth weight was measured within 12 hours. Offspring were aged to 4 months. At that time, one male and female per litter were randomly selected to undergo carotid catheterization. The following day mean arterial pressure (MAP) was measured and blood and tissues were collected. Immune cells were measured by flow cytometry. Sex hormones were measured by mass spectroscopy. AT1-AA was measured by cardiomyocyte bioassay. Renal endothelin was measured by ELISA and RT-PCR. A two-way ANOVA was used for statistical analysis. Results: AT1-AA male and female offspring (6.1±0.1g, n=19, p<0.05; 5.8±0.1g, n=19, p<0.05) were larger at birth than NP male and female offspring (5.8±0.1g, n=11; 5.5±0.1g, n=11). Female AT1-AA offspring had elevated MAP (124±2 mmHg, n=14, p<0.05) compared to NP female offspring (110±5 mmHg, n=8) while male AT1-AA offspring had similar MAP to male NP offspring. Circulating cytolytic NK cells trended higher in female AT1-AA offspring (2.73±0.37 % gated, n=9, p=0.0949) compared to female NP offspring (1.66±0.31 % gated, n=6). Female AT1-AA offspring also had elevated progesterone (41.3±6.0 pg/μL, n=8, p=0.0556) compared to female NP offspring (26.9±4.9 pg/μL, n=7). Renal PPET expression was increased in female AT1-AA offspring (2.6±0.69 fold increase, n=4, p<0.05) compared to female NP offspring. Renal endothelin-1 was increased in female AT1-AA offspring (1.02±0.23 pg/mg, n=7, p=0.062) compared to female NP offspring (0.59±0.06 pg/mg, n=6). Male and female AT1-AA offspring had increased circulating AT1-AA (9.5±2.2 ΔBPM, n=6, p<0.0001; 7.9±0.9 ΔBPM, n=8, p<0.001) compared to male and female NP offspring (-0.4±.4 ΔBPM, n=7; -0.8±0.4 ΔBPM, n=4). Conclusion: These data demonstrate that AT1-AA infusion during pregnancy can predispose female offspring to have elevated blood pressure in adulthood. This study was supported by NIH grants RO1HD067541 (BL), F31HD110230 (NC), P20GM121334 (BL, LMA), HL151407 (DCC) and American heart association (AHA) early career award 19CDA34670055 (LMA). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Preeclampsia (PE), new onset hypertension during pregnancy, is associated with increased tumor necrosis factor alpha (TNF-α), agonistic autoantibodies to the angiotensin II type 1 receptor (AT1-AA), and low birth weight babies. We have previously shown that Etanercept ( E ), a TNF-α inhibitor, improves PE features in the reduced uterine perfusion pressure (RUPP) rat. However, the long-term effect of E on offspring health and immune competency has not been studied. Therefore, we hypothesize that E will improve maternal health without affecting offspring outcomes in a rat model of PE. To test this hypothesis, E (0.4 mg/kg) was injected on gestation day 18 into sham and RUPP dams which were allowed to deliver. Weights were recorded at birth and 16 weeks of age when mean arterial pressure (MAP) was measured. Whole blood was collected to measure immune cells by flow cytometry. AT1-AA was measured using a cardiomyocyte bioassay. A student’s t-test was used to compare differences among male and female RUPP offspring with and without E . RUPP male and female offspring were smaller (5.21±0.19 g, n=7; 5.24±0.10 g, n=7) than sham offspring (5.94±0.14 g, n=7, p<0.05; 5.86±0.16 g, n=9, p<0.01) while RUPP male offspring were larger and female offspring were unchanged following perinatal E (6.01±0.29 g, n=9, p<0.05; 5.28±0.14, n=9). MAP was comparable between sham (133±4 mmHg) and RUPP (132±4 mmHg) male offspring with no change seen following perinatal E (133±4 mmHg). Perinatal E did not impact adult male circulating B cells. Circulating AT1-AA was elevated in male RUPP offspring (16±2 ΔBPM, p<0.01) compared to sham and was not improved with E (13±2 ΔBPM). MAP was comparable in sham (131±5 mmHg) and RUPP (130±5 mmHg) female offspring but trended downward with perinatal E (123±4 mmHg). RUPP female offspring had reduced circulating B cells (4.24±0.81 % gated, p=0.066) compared to sham females (9.56±2.07 % gated) which was normalized following perinatal E (9.04±1.42 % gated, p<0.05). AT1-AA trended lower following perinatal E (7±2 ΔBPM, p=0.0888) compared to RUPP female offspring (16±3 ΔBPM). Our findings indicate that E does not worsen fetal outcomes. Moreover, E improves male birth weight and reduces adult female AT1-AA without compromising offspring immune competency.

A poor uterine environment causes changes in fetal development that affect the health of offspring long-term. Although there are multiple pathways that contribute to the development of cardiovascular and neurological disease, low birth weight or fetal growth restriction (FGR) predisposes offspring to these diseases. There is a link between fetal exposure to adverse influences and hypertension later in life. Many epidemiological studies support the link between fetal life and the risk of disease later in life. Experimental models have sought to provide mechanistic proof of this link while simultaneously investigating potential therapeutics or treatment pathways. Preeclampsia (PE), one of several hypertensive disorders in pregnancy, is a leading cause of morbidity and mortality for both the mother and fetus. Studies have shown that PE is a state of chronic inflammation and there is an imbalance between pro-inflammatory and regulatory immune cells and mediators. There is no cure for PE beyond the delivery of the fetal-placental unit, and many PE pregnancies result in FGR and preterm birth. Epidemiological data demonstrate that the sex of the offspring is correlated with the degree of cardiovascular disease that develops with the age of the offspring yet few studies examine the effect of sex on the development of neurological disorders. Even fewer studies examine the effects of therapeutics on offspring of different genders following a PE pregnancy. Moreover, there remain significant gaps in knowledge concerning the role the immune system plays in FGR offspring developing hypertension or neurovascular disorders later in life. Therefore, the purpose of this review is to highlight current research on sex differences in the developmental programming of hypertension and neurological disorders following a PE pregnancy.

BACKGROUND: Preeclampsia, a new-onset hypertension with end -organ damage in pregnancy, is associated with maternal death and mor-bidity, low birthweight, and B cells producing agonistic autoantibodies to the angiotensin II type 1 receptor. Angiotensin II type 1 receptor agonistic autoantibodies are produced during pregnancy and after delivery and are in the fetal circulation of women with preeclampsia. Angiotensin II type 1 receptor agonistic autoantibodies are shown to contribute to endothelial dysfunction, renal dysfunction, hypertension, fetal growth restriction, and chronic inflammation in women with preeclampsia. The reduced uterine perfusion pressure rat model of preeclampsia exhibits these features. In addition, we have shown that the administration of a 'n7AAc', which blocks the actions of the angiotensin II type 1 receptor autoantibodies, improves preeclamptic features in the rat with reduced uterine perfusion pressure. However, the effect of a 'n7AAc' on the long-term health of the offspring of rats with reduced uterine perfusion pressure is unknown.OBJECTIVE: This study aimed to test the hypothesis that inhibition of angiotensin II type 1 receptor autoantibodies during pregnancy will improve offspring birthweight and prevent increased cardiovascular risk in offspring in adulthood.STUDY DESIGN: To test our hypothesis, a 'n7AAc' (24 mg/d) or vehi-cle (saline) was given on gestation day 14 via miniosmotic pumps to sham-operated (sham) and Sprague-Dawley rat dams with reduced uter-ine perfusion pressure. Dams were allowed to deliver naturally, and pup weights were recorded within 12 hours after birth. Pups were aged to 16 weeks, at which time mean arterial pressure was measured and whole blood was collected to measure immune cells by flow cytometry, cytokines by enzyme-linked immunosorbent assay, and angiotensin II type 1 recep-tor autoantibodies by bioassay. A 2-way analysis of variance with the Bon-ferroni multiple comparison posthoc test was used for statistical analysis.RESULTS: There was no significant change in offspring birthweight of 'n7AAc'-treated male (5.63 +/- 0.09 g) or female (5.66 +/- 0.14 g) offspringfrom reduced uterine perfusion pressure dams compared with vehicle male (5.51 +/- 0.17 g) or female (5.74 +/- 0.13 g) offspring from reduced uterine perfusion pressure dams. In addition, 'n7AAc' treatment did not affect the birthweight of sham male (5.83 +/- 0.11 g) or female (5.64 +/- 0.12) offspring compared with vehicle sham male (5.811 +/- 0.15 g) or female (5.40 +/- 0.24 g) offspring. At adulthood, mean arterial pressure was unchanged in 'n7AAc' treated-male (133 +/- 2 mm Hg) and female (127 +/- 3 mm Hg) offspring from reduced uterine perfusion pressure dams compared with vehicle male (142 +/- 3 mm Hg) and female (133 +/- 5 mm Hg) offspring from reduced uterine perfusion pressure dams, the 'n7AAc'-treated sham male (133 +/- 3 mm Hg) and female (135 +/- 3 mm Hg) offspring, and vehicle sham male (138 +/- 4 mm Hg) and female (130 +/- 5 mm Hg) offspring. The circulating angiotensin II type 1 receptor autoanti-bodies were increased in vehicle male (10 +/- 2 ABPM) and female (14 +/- 2 ABPM) offspring from reduced uterine perfusion pressure dams and 'n7AAc'-treated male (11 +/- 2 ABPM) and female (11 +/- 2 ABPM) off-spring from reduced uterine perfusion pressure dams compared with vehi-cle sham male (1 +/- 1 ABPM) and female (-1 +/- 1 ABPM) offspring and 'n7AAc'-treated sham male (-2 +/- 2 ABPM) and female (-2 +/- 2 ABPM) offspring. CONCLUSION: Our findings indicated that perinatal 7-amino acid sequence peptide treatment does not negatively impact offspring survival or weight at birth. Perinatal 'n7AAc' treatment did not prevent increased cardiovascular risk in offspring, but it also did not cause an increased car-diovascular risk in offspring with reduced uterine perfusion pressure com-pared with controls. Furthermore, perinatal 'n7AAc' treatment did not affect endogenous immunologic programming as observed by no change in circulating angiotensin II type 1 receptor autoantibodies in either sex of adult offspring from reduced uterine perfusion pressure dams.

Preeclampsia (PE), new onset hypertension with end-organ damage in pregnancy, is associated with maternal death and morbidity, increased uterine artery resistance (UARI), low birth weight, and B cells producing agonistic autoantibodies to the angiotensin II type 1 receptor (AT1-AA). AT1-AA is produced during pregnancy, postpartum, and is in fetal circulation of PE women. The Reduced Uterine Perfusion Pressure (RUPP) rat model of PE exhibits these features and we have shown that administration of a seven amino acid sequence peptide (7AA), which blocks the actions of AT1-AA, improves PE features in the RUPP rat. However, 7AA's effect on the long-term health of RUPP rat offspring is unknown. We hypothesize that 7AA will not worsen outcomes for offspring born to the RUPP rat model of PE. To test our hypothesis, 7AA (24 ug/day) was given on gestation day 14 via mini-osmotic pumps to control and RUPP dams. UARI was measured using Doppler ultrasound, pup weights were recorded within 12 hours of birth. Mean arterial pressure (MAP) was measured and whole blood was collected to measure immune cells by flow cytometry at 16 weeks. A two-way ANOVA was used for statistical analysis. UARI was increased in RUPP (0.70±0.02,n=10,p< 0.05) compared to controls (0.47±0.02,n=8) and improved with 7AA (0.49±0.02,n=10,p< 0.05). There was no significant change in offspring birth weight of RUPPs (6.0±0.21g,n=8) with or without 7AA (5.5±0.08g,n=7). At age 16 weeks there were no changes in weight (381±9g n=8 vs 374±12g n=6), MAP (138±4 vs 122±9mmHg), circulating B cells (8±4.2% vs 9±7.6%), or cytolytic NK cells (9±3.6% vs 2±1.1%) in male RUPP offspring with or without perinatal 7AA. There were no changes in weight (234±2g,n=7 vs 236±7g,n=5), MAP (123±7vs 132±10mmHg), circulating B cells (5±1.7% vs 4±2.2%), or NK cells (48±15.3% vs 22±15.5%) in female offspring of RUPP rats with or without 7AA. Our findings indicate that perinatal 7AA improves maternal PE features without negatively impacting fetal outcomes indicating promising potential to treat PE.

Preeclampsia (PE), new onset hypertension during pregnancy, is the leading cause of death and morbidity for the mother and low birth weight in offspring. Low birth weight has been shown to cause offspring to be at high risk for cardiovascular and metabolic disorders later in life. PE has been associated with chronic immune activation including T helper cells and natural killer cells. PE women also have activated B cells producing agonistic autoantibodies to the angiotensin II type I receptor (AT1‐AA). AT1‐AA has been implicated in numerous pathways in the pathophysiology of PE. We have shown Rituximab (R), used clinically for B cell depletion, lowers mean arterial pressure (MAP), B cells, and AT1‐AA in the reduced uterine perfusion pressure (RUPP) rat model of PE. Clinical studies show no untoward effects on offspring of pregnant women maintained on R for treating lymphoma. R is not used during PE, therefore, effects of maternal B cell depletion on offspring survival and growth in response to placental ischemia is unknown. We hypothesize that R will deplete maternal B cells in RUPP rats without worsening the effect of placental ischemia on pup growth and survival. To test this hypothesis, the RUPP procedure was performed and R (250 mcg/kg) was given on gestation day (GD) 14 via mini‐osmotic pump. On GD 19, B cells were measured by flow cytometry, and MAP and pup weights were recorded. A separate group of dams were allowed to deliver, pup weights were recorded within 12 hours of birth and weekly until 16 weeks, and B cells were analyzed. A one‐way or two‐way ANOVA was used for statistical analysis. MAP increased in RUPP 123±2 (n=19, p<0.05) compared to NP controls 101±1 (n=18) and was 106±3 mmHg in RUPP+R (n=8, p<0.05). On GD19, maternal circulating B cells were 16±2 % (n=14) in RUPPs, 8±2 % in NP rats, (n=7, p<0.05), and 5.5±1% gate in RUPP+R (n=5, p<0.05). RUPP male and female offspring tended to be smaller (5.11±0.23 g, 5.19±0.14 g; n=4, n=4) at birth than NP offspring (6.09±0.15 g, 5.87±0.12 g; n=6, p<0.064; n=6, p<0.948) or RUPP+R offspring (5.75±0.24 g, 5.36±0.28 g; n=6, p=0.291; n=6, p>0.999). At 12 weeks, male and female RUPP offspring had elevated circulating B cells (21.24±2.92, 20.04±0.72 %; n=6; n=9) compared to NP (1.38±0.23, 1.63±0.06 %; n=4, p<0.05; n=3, p<0.04) which was normalized in RUPP+R offspring (0.40±0.10, 0.27±0.03 % gate; n=3, p<0.05; n=8, p<0.05). At 16 weeks, B cells were comparable in male and female offspring from NP (0.78±0.09 %, n=10; 1.06±0.21 % gate, n=6) and RUPP+R rats (1.21±0.34% gate, n=5; 1.62±0.34% gate, n=5), but were elevated in RUPP male and female offspring (2.45±0.27% gate, n=9; 1.68±0.45% gate, n=13) compared to NP (p<0.05; p>0.99) and RUPP+R (p<0.05; p>0.99). Our findings indicate that R lowers maternal circulating B cells and MAP in RUPP rats and improves fetal weight and circulating B cells, indicating that R does not worsen adverse fetal outcomes in response to placental ischemia.

The seven amino acid inhibitory peptide to the AT1-AA ('n7AAc') has limited transfer to the fetus at gestational day 20, improves uterine blood flow and fetal growth in the reduced uterine perfusion pressure model of preeclampsia (PE), and does not impair fetal survival during gestation in sham-operated or placental ischemic rats. Collectively, these findings suggest that maternal administration of 'n7AAc' as an effective strategy for the treatment of PE is associated with improved outcomes in the fetus.

Preeclampsia (PE), new onset hypertension during pregnancy, is a progesterone deficient state and is associated with an imbalance among CD4+ T cells, natural killer (NK) cells, and inflammatory cytokines. Progesterone induces progesterone induced blocking factor (PIBF) which stimulates IL-4/T helper 2 cells (TH2) as mechanisms to quell inflammation. UMMC PE patients have significantly lower progesterone compared to normotensive pregnant women (NP).This study was designed to test the hypothesis that progesterone, in the form of 17-hydroxyprogesterone caproate (17-OHPC), stimulates PIBF/IL-4/TH2, as a mechanism of reducing NK/TH1s and improves blood pressure in PE. PE participants (27-33 weeks of gestation) were recruited from University of Mississippi Medical Center (UMMC) and once enrolled the participant received 17-OHPC (250 mg, I.M.) weekly, with blood draws before and after 17-OHPC injection. Participants received 1-3 doses (average). Placentas were collected at delivery. Importantly, our data demonstrate that PIBF was 18.6 +/-1.0 pg/mL in NT (n=4), 14.53 +/- 1.0 pg/mL in PE (n=10, p< 0.05), and 15.78 +/-0.85 in PE+17-OHPC (n=6). Placental CD4+ T cells were 2.46+/- 0.9 % gate in NT, 6.5+/- 2.7 in PE (n=3), 4.6+/- 2.0 in PE +17-OHPC (n=4). Circulating CD4+ T cells were 18.23 +/-4.8 in PE (n=4), 14.4+/-1.2 % gate in PE+17-OHPC (n=5). Placental and circulating NK cells were 20.84 +/- 6.8, 7.2 +/- 2.0 in PE (n=5) which reduced to 4.7+/-1.03, 5.3 +/- 1.5 % gate in PE+17-OHPC (n=4, p< 0.05). Circulating TNF-alpha was 22.4 +/- 4.5 in NP pg/mL, 32.0 +/- 3.4 in PE (n=8), which decreased to 21.1+/-5.5 in PE+17OHPC (n=4, p< 0.05). Placental TH2 cells were 81.7+/-10.6 in PE and 84.7+/-11.0 in PE+17-OHPC. Circulating IL-4 was 4.0 +/- 0.75 pg/mL in PE (n=10) which increased to 5.4 +/- 0.5 in PE+17-OHPC (n=11). 17-OHPC prolonged time to delivery beyond 72h on average and average systolic blood pressure was 151 +/- 5 mmHg in PE (n=18) and 137+/-4 in PE+17-OHPC (n=13). Therefore, 17-OHPC reduced inflammation, lowered blood pressure and prolonged time do delivery during PE.

Preeclampsia (PE), new onset hypertension during pregnancy, is associated with maternal death and morbidity, low birth weight in offspring, increased tumor necrosis factor alpha (TNF-α) and B cells producing autoantibodies. We have shown that Etanercept (E), a TNF-α inhibitor, improves blood pressure in the reduced uterine perfusion pressure (RUPP) rat model of PE and that Rituximab (R), used clinically for B cell depletion, lowered blood pressure and B cells in RUPP rats. Clinical use of R or E during pregnancy has not resulted in adverse outcomes for the mother, however, the effects of maternal B cell depletion or TNF-α blockade on offspring immune cells in response to placental ischemia is unknown. We hypothesize that E and R will not worsen outcomes on fetal immune function in a rat model of preeclampsia. To test this hypothesis, R (250 mcg/kg) was given on gestation day (GD) 14 via mini-osmotic pumps. E (400 mcg/kg)was i.p. injected on GD 18. After delivery, pup weights were recorded within 12 hours and weekly until 16 weeks. At 16 weeks whole blood was taken to measure B cells by flow cytometry. A one-way ANOVA was used for statistical analysis. 18 litters were examined. RUPP male and female offspring were smaller (5.11±0.23 g, n=4; 5.19±0.14 g, n=4) than NP (6.09±0.15 g, n=6, p< 0.05; 5.87±0.12 g, n=6, p< 0.05) which was not negatively impacted by E. RUPP+ R offspring (5.75±0.24 g, n=6; 5.36±0.28 g, n=6) tended to be larger than RUPP controls. At 12 weeks, male and female RUPP offspring had elevated circulating B cells (21±3%; 20±1 %,) compared to NP rats (1±0.23%, p< 0.05; 1.6±0.06 %, p< 0.04) which was normalized in RUPP+ R offspring (0.4±0.1%, p< 0.05; 0.3±0.03 %, p< 0.05). At 16 weeks, B cells were comparable in NP male and female offspring (0.78±0.09 %; 1.06±0.21 %) and RUPP+ R rats (0.80±0.04; 1.88±1.00 %) and RUPP + E (0.36±0.14, n=2; 6.3±2.79 % gate, n=2). Our findings indicate that neither E nor R worsen fetal outcomes while having positive maternal effects on blood pressure in response to placental ischemia.

Preeclampsia (PE), new onset hypertension during pregnancy, affects 5-7% of all pregnancies in the U.S. and is associated with reduced fetal weight, increased inflammation, vascular endothelial dysfunction (increased endothelin-1 (ET-1) and decreased nitric oxide (NO)) and hypertension. To date the best treatment remains early delivery of the feto-placental unit. Activated lymphocytes during normal pregnancy (NP) express progesterone receptors, which stimulate a protein called Progesterone Induced Blocking Factor (PIBF) that is reduces during hypertensive pregnancy disorders. Therefore, this study was designed to test the hypothesis that progesterone, in the form of 17-hydroxyprogesterone caproate (17-OHPC), reduces inflammation, markers of endothelial dysfunction while reducing blood pressure and prolong time to delivery in PE women. In our ongoing clinical trial, PE participants received 17-OHPC (250 mg, I.M.) with blood draws before and after injection. Placentas were collected at delivery. PIBF was 18.6 +/-1.0 pg/mL in NT (n=4), 14.53 +/- 1.0 pg/mL in PE (n=10, p<0.05), and 15.78 +/-0.85 in PE+17-OHPC (n=6). Placental CD4+ T cells were 6.5+/- 2.7 in PE (n=3), 4.6+/- 2.0 in PE +17-OHPC (n=4). Circulating CD4+ T cells were 18.23 +/-4.8 in PE (n=4), 14.4+/-1.2 % gate in PE+17-OHPC (n=5). Placental TH2 cells were 81.7+/-10.6% gate in PE and 84.7+/-11.0 in PE+17-OHPC. Placental and circulating NK cells were 20.84 +/- 6.8 % gate, 7.2 +/- 2.0 in PE which reduced to 4.7+/-1.03, 5.3 +/- 1.5 % gate in PE+17-OHPC. Circulating TNF-alpha was 32.0 +/- 3.4 pg/mL in PE (n=8), which decreased to 21.1+/-5.5 in PE+17OHPC (n=4). Circulating ET-1 was 2.53+/- 0.4 pg/mL in healthy normal pregnant (NP, n=5), 6.7 +/- 1.4 in PE (n=18, p<0.05) and 4.9 +/- 1.3 in PE+17-OHPC. Placenta preproendothelin -1 (PPET-1) increased 1.5 fold change in PE (n=4) compared to NP, which was reduced to 0.96 in PE+17-OHPC (n=4). Importantly, endotheling-1 measured in HUVECS media treated with PE sera was 68 +/- 22 pg/mg of protein in PE which reduced to 57 +/-19 in PE+17OHPC (n=6). Moreover, circulation nitrate-nitrite was 50 +/- 9 uM in PE and significantly increased to 94 +/-14 in PE+17-OHPC. 17-OHPC prolonged time of delivery beyond 72h on average and average systolic blood pressure was 151 +/- 5 mmHg in PE (n=18) and 137+/-4 in PE+17-OHPC (n=13). Our results suggest that 17-OHPC reduced inflammation, markers of endothelial dysfunction, lowered blood pressure and prolonged time do delivery in PE women.

Placenta ischemia, the initiating factor in preeclampsia (PE), is associated with intrauterine growth restriction (IUGR) and increased blood pressure (BP) in offspring. Yet, the only treatment for PE is delivery of the baby and placenta. The Reduced Uterine Perfusion Pressure (RUPP) rat model induced by placental ischemia at gestational day 14 (G14) mimics many facets of human PE including pregnancy-specific hypertension, an increase in the agonistic ANG II Type 1 receptor autoantibody (AT1-AA), IUGR and increased BP in the offspring. Inhibition of AT1-AA using an epitope-binding inhibitory peptide ('n7AAc') attenuates increased BP at gestational day 19 in the RUPP. Yet, whether use of ‘n7aac’ improves fetal growth and mitigates increased BP in the offspring is unknown. Thus, we tested the hypothesis that maternal administration of ‘n7aac’ improves fetal growth by attenuating reduced uterine blood flow and impaired placental remodeling. Sham or RUPP surgery was performed at G14 with administration of vehicle or ‘n7aac’ (144μg/day) via mini osmotic pump until gestational day 20 (G20). At G20 uterine artery resistance index was significantly elevated in vehicle RUPP (0.69±0.02 mm/s n=10) compared to vehicle Sham (0.48±0.02 mm/s n=8) (P<0.0001) and not increased in treated RUPP (0.49±0.02 mm/s n=10) or treated Sham (0.48±0.02 mm/s n=9). Fetal weight was significantly reduced in vehicle RUPP (3.24±0.2 g) compared to vehicle Sham (3.92±0.05 g) (P=0.013) and not decreased in treated RUPP (3.70±0.04 g) or Sham (3.98±0.10 g). Litter size of viable pups at G20 was only reduced in treated RUPP (5.3±1.4) compared to vehicle Sham (11.56±0.7) (P=0.003). Importantly, using in vivo imaging, little to no auto fluorescence of rhodamine-labeled peptide (480 μg/kg/day, n=4) was detectable in the pups at G20. Thus, our results demonstrate that maternal treatment with ‘n7aac’ in the RUPP rat model of PE improve UARI, which is associated with improved fetal weight at G20 in response to placental ischemia. Whether this benefit continues to birth and mitigates increased BP in IUGR offspring is unknown but is the focus of future studies. In conclusion, inhibition of the AT1AA during PE may not only provide benefit to the mother, but may also be associated with benefit in the offspring.