BMPR2-expressing bone marrow-derived endothelial-like progenitor cells alleviate pulmonary arterial hypertension in vivo.

BACKGROUND AND OBJECTIVE: Pulmonary arterial hypertension (PAH) is characterized by increased resistance in the distal pulmonary arteries, ultimately leading to right heart failure and, despite the available therapeutics, survival remains poor. Reduced expression of bone morphogenetic protein receptor type 2 (BMPR2) is strongly associated with PAH. Cell therapies are of interest in PAH, but whether this approach can upregulate BMPR2 is not known. Our objective was to evaluate a preclinical cell therapy approach based on upregulation of BMPR2. METHODS: We assessed the therapeutic effect of intravenously injected BMPR2-augmented rat bone marrow-derived endothelial-like progenitor cells (BMPR2-BM-ELPC) on PAH in the rat monocrotaline (MCT) model. RESULTS: The cells accumulate in the lungs with negligible systemic distribution, but the vast majority are lost from the lungs by 24 h. Lungs from rats treated with BMPR2-BM-ELPC exhibited an immediate increase in BMPR2 and related intracellular signalling proteins. Treatment with BMPR2-BM-ELPC attenuated PAH as demonstrated by a reduction in right ventricular hypertrophy as well as right ventricular systolic and mean pulmonary arterial pressures. In addition, this treatment reversed PAH-induced vascular remodelling with a significant reduction in vessel thickness and muscularization. In view of the short retention time of injected cells in the lungs, the mechanism for the effects seen may be intracellular communication via exosomes. In support of this hypothesis, we demonstrate that BMPR2-transduced outgrowth endothelial progenitor cells (OECs) release BMPR2-expressing exosomes. CONCLUSION: BMPR2-augmented ELPC demonstrate therapeutic benefits in the rat model and may have clinical translation potential.

Anogenital distance reflects the sex ratio of a gilt's birth litter and predicts her reproductive success1.

Anogenital distance (AGD) has been used to reflect masculinization in litter-bearing species. As masculinization affects behavior and reproduction, AGD could be measured to assist in selecting gilts with a temperament more suited to commercial production and greater reproductive potential. We hypothesized that gilts from a male-biased litter would have a longer AGD and poorer reproductive performance. In Exp. 1, AGD and weight were measured at day 1, day 21, and week 16 of age for gilts from male-biased litters (≥60% males; n = 51) and female-biased litters (≥60% females; n = 51). Sow AGD was measured 3 d after farrowing. In Exp. 2, AGD was measured at gilt selection at approximately 24 wk of age and gilts followed to second parity. Litter sex ratio affected AGD at 16 wk of age, with gilts from female-biased litters having longer AGD (mean ± SEM, 9.1 ± 0.7 mm vs. 11.0 ± 0.6 mm, P = 0.013). Anogenital distance was not different on day 1 or day 21. There was no effect of sex ratio on weight at any time, and sow AGD was not associated with the sex ratio of her litter. Gilts with an AGD longer than the mean of 11.55 mm were heavier (mean ± SEM, 118.8 ± 0.4 kg vs. 117.7 ± 0.4 kg, P = 0.023), were achieved puberty earlier (179.6 ± 0.6d vs. 182.2 ± 0.6 d, P = 0.001), were mated younger (200.6 ± 0.6 d vs. 203.2 ± 0.6 d, P = 0.001), and were more likely to be mated (91% vs. 83%, P = 0.005) than gilts with an AGD shorter than the mean. Gilts with an AGD greater than 11.55 mm had a greater born alive litter size (11.79 ± 0.20 vs. 11.20 ± 0.19, P = 0.018) compared with gilts with an AGD shorter than 11.55 mm. At 16 wk, AGD was associated with sex bias and could be used as a selection tool to predict reproductive success of the first parity, with a longer AGD being associated with gilts that had been born into a female-biased litter and that had better reproductive performance.

Sex bias of the birth litter affects surge but not tonic LH secretion in gilts.

The physiology and behavior of gilts that develop in a male-biased litter can differ from gilts that develop in a female-biased litter. We hypothesized that gilts from male-biased litters will have a delayed and attenuated luteinizing hormone (LH) surge, and reduced LH pulse frequency and amplitude compared to gilts from female-biased litters. Gilts were selected at birth from male-biased (>60% males n = 10) or female-biased (>60% females n = 9) litters. From 18 wk of age, detection of puberty using daily boar contact began and their subsequent estrous periods were synchronized with oral progestogen (altrenogest). On day 3 after altrenogest withdrawal, blood samples were obtained from 6 gilts per sex bias group at 10 min intervals from 0900 to 2100 h to determine LH pulse amplitude and frequency. From 0900 on day 4, all 19 gilts were sampled every 4 h until the end of estrus to characterize LH surge dynamics. There were no differences between groups in LH pulse characteristics. Compared to gilts from female-biased litters, the LH surge in gilts from male-biased litters was delayed [56.00 ± 3.32 h vs. 43.11 ± 3.76 h (mean ± standard error of the mean (SEM)), P < 0.05], the duration was decreased [29.78 ± 2.12 h vs. 37.71 ± 1.19 h (mean ± SEM), P < 0.05] and the total secretion as measured by area under the curve was decreased (91.42 ± 9.52 ng/mL vs. 120.28 ± 9.48 ng/mL, P < 0.05). Our results indicate that a male-biased uterine environment has different effects on the tonic secretion of LH than the LH surge, with only some elements of the LH surge being affected.

Response to gonadotrophins differs for gilts from female- and male-biased litters.

In several species, females masculinised by abnormal androgen exposure in utero have poor reproductive performance and gilts born into litters with a male bias are likely exposed to greater androgen concentrations prenatally than gilts born into female-biased litters. At 24h of age, piglet plasma testosterone concentrations in gilts from male-biased litters (>60% male; n=22) or female-biased litters (>60% female; n=27) were not different. At 18 wks of age, all gilts received an injection of 400IU equine chorionic gonadotrophin plus 200IU human chorionic gonadotrophin to stimulate oestrus. Two weeks after the injection gilts were slaughtered and ovaries collected for determination of numbers of corpora lutea (CL). Compared to gilts from female-biased litters, gilts from male-biased litters were more likely to ovulate (86.0% vs 59.5%, P=0.047) and had more CL (13.1±1.5 vs 7.2±1.7, P=0.015). The present data indicate an effect of birth litter sex-bias on pre-pubertal physiological development, possibly involving organisational effects at the ovarian cellular level impacting on future ovarian function. Potential impacts on subsequent fertility remain to be determined.

Human chorionic gonadotrophin in early gestation induces growth of estrogenic ovarian follicles and improves primiparous sow fertility during summer.

Reduced summer farrowing rates may be due to inadequate corpora luteal (CL) support. Porcine CL become dependent on LH from 12 d of pregnancy and the embryonic estrogen signal for maternal recognition of pregnancy (MRP) is initiated at about 11-12 d after insemination. We hypothesised that injection of the LH analogue human chorionic gonadotropin (hCG) would induce growth of estrogenic follicles and, by mimicking the signal for MRP and stimulating progesterone secretion, increase primiparous sow fertility. In Experiment 1, during a 28 d lactation 53 mixed parity sows were full-fed either throughout lactation (n=16) or until 18 d and then feed restricted during the last 10 d of lactation (n=36). At 12 d after mating restrict-fed sows were injected with 1000IU hCG (n=17) or were not injected (n=19); the full-fed sows acted as non-treated positive controls. Transrectal ovarian ultrasound exams were performed on days 12, 16, 20, 24, and 28; blood samples were obtained on days 12, 14, and 15 for estradiol and progesterone assay. For Experiment 2, during the summer months primiparous sows received 1000IU hCG 12 d after mating (n=28) or were non-injected controls (n=27). Pregnancy status was determined at 28 d and sows allowed to go to term to determine farrowing rates and litter sizes. In Experiment 1, injection of hCG increased (P<0.001) follicle diameter and serum concentrations of estradiol (P<0.01) and progesterone (P<0.05). There were no effects of lactation feeding level on wean-estrus interval, farrowing rate or subsequent litter size. In Experiment 2, hCG injection was associated with a higher pregnancy rate (P<0.05) and farrowing rate (P<0.08). There was no effect on litter size. These data confirm that hCG stimulates growth of estrogenic follicles and CL function, and improves primiparous sow fertility during the summer months.