Children and adolescents with pulmonary arterial hypertension : baseline and follow-up data from the polish registry of pulmonary hypertension (BNP-PL)

We present the results from the pediatric arm of the Polish Registry of Pulmonary Hypertension. We prospectively enrolled all pulmonary arterial hypertension (PAH) patients, between the ages of 3 months and 18 years, who had been under the care of each PAH center in Poland between 1 March 2018 and 30 September 2018. The mean prevalence of PAH was 11.6 per million, and the estimated incidence rate was 2.4 per million/year, but it was geographically heterogeneous. Among 80 enrolled children (females, n = 40; 50%), 54 (67.5%) had PAH associated with congenital heart disease (CHD-PAH), 25 (31.25%) had idiopathic PAH (IPAH), and 1 (1.25%) had portopulmonary PAH. At the time of enrolment, 31% of the patients had significant impairment of physical capacity (WHO-FC III). The most frequent comorbidities included shortage of growth (n = 20; 25%), mental retardation (n = 32; 40%), hypothyroidism (n = 19; 23.8%) and Down syndrome (n = 24; 30%). The majority of children were treated with PAH-specific medications, but only half of them with double combination therapy, which improved after changing the reimbursement policy. The underrepresentation of PAH classes other than IPAH and CHD-PAH, and the geographically heterogeneous distribution of PAH prevalence, indicate the need for building awareness of PAH among pediatricians, while a frequent coexistence of PAH with other comorbidities calls for a multidisciplinary approach to the management of PAH children

Seismic constraints from a Mars impact experiment using InSight and Perseverance

NASA’s InSight (Interior Exploration using Seismic Investigations, Geodesy and Heat Transport) mission has operated a sophisticated suite of seismology and geophysics instruments on the surface of Mars since its arrival in 2018. On 18 February 2021, we attempted to detect the seismic and acoustic waves produced by the entry, descent and landing of the Perseverance rover using the sensors onboard the InSight lander. Similar observations have been made on Earth using data from both crewed1,2 and uncrewed3,4 spacecraft, and on the Moon during the Apollo era5, but never before on Mars or another planet. This was the only seismic event to occur on Mars since InSight began operations that had an a priori known and independently constrained timing and location. It therefore had the potential to be used as a calibration for other marsquakes recorded by InSight. Here we report that no signal from Perseverance’s entry, descent and landing is identifiable in the InSight data. Nonetheless, measurements made during the landing window enable us to place constraints on the distance–amplitude relationships used to predict the amplitude of seismic waves produced by planetary impacts and place in situ constraints on Martian impact seismic efficiency (the fraction of the impactor kinetic energy converted into seismic energy)

Ultra-Rare Mutation in Long-Range Enhancer Predisposes to Thyroid Carcinoma with High Penetrance

Peer reviewe

Disturbed Expression of Splicing Factors in Renal Cancer Affects Alternative Splicing of Apoptosis Regulators, Oncogenes, and Tumor Suppressors

BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is the most common type of renal cancer. One of the processes disturbed in this cancer type is alternative splicing, although phenomena underlying these disturbances remain unknown. Alternative splicing consists of selective removal of introns and joining of residual exons of the primary transcript, to produce mRNA molecules of different sequence. Splicing aberrations may lead to tumoral transformation due to synthesis of impaired splice variants with oncogenic potential. In this paper we hypothesized that disturbed alternative splicing in ccRCC may result from improper expression of splicing factors, mediators of splicing reactions. METHODOLOGY/PRINCIPAL FINDINGS: Using real-time PCR and Western-blot analysis we analyzed expression of seven splicing factors belonging to SR proteins family (SF2/ASF, SC35, SRp20, SRp75, SRp40, SRp55 and 9G8), and one non-SR factor, hnRNP A1 (heterogeneous nuclear ribonucleoprotein A1) in 38 pairs of tumor-control ccRCC samples. Moreover, we analyzed splicing patterns of five genes involved in carcinogenesis and partially regulated by analyzed splicing factors: RON, CEACAM1, Rac1, Caspase-9, and GLI1. CONCLUSIONS/SIGNIFICANCE: We found that the mRNA expression of splicing factors was disturbed in tumors when compared to paired controls, similarly as levels of SF2/ASF and hnRNP A1 proteins. The correlation coefficients between expression levels of specific splicing factors were increased in tumor samples. Moreover, alternative splicing of five analyzed genes was also disturbed in ccRCC samples and splicing pattern of two of them, Caspase-9 and CEACAM1 correlated with expression of SF2/ASF in tumors. We conclude that disturbed expression of splicing factors in ccRCC may possibly lead to impaired alternative splicing of genes regulating tumor growth and this way contribute to the process of carcinogenesis

Epigenetic Regulation of Thyroid Hormone Receptor Beta in Renal Cancer

<div><p>Abstract</p><p>Thyroid hormone receptor beta (<i>THRB</i>) gene is commonly deregulated in cancers and, as strengthened by animal models, postulated to play a tumor-suppressive role. Our previous studies revealed downregulation of <i>THRB</i> in clear cell renal cell carcinoma (ccRCC), but the culpable mechanisms have not been fully elucidated. Since epigenetic regulation is a common mechanism influencing the expression of tumor suppressors, we hypothesized that downregulation of <i>THRB</i> in renal cancer results from epigenetic aberrances, including CpG methylation and microRNA-dependent silencing. Our study revealed that ccRCC tumors exhibited a 56% decrease in <i>THRB</i> and a 37% increase in DNA methyltransferase 1 (DNMT1) expression when compared with paired non-neoplastic control samples. However, <i>THRB</i> CpG methylation analysis performed using BSP, SNaPshot and MSP-PCR consistently revealed no changes in methylation patterns between matched tumor and control samples. <i>In silico</i> analysis resulted in identification of four microRNAs (miR-155, miR-425, miR-592, and miR-599) as potentially targeting <i>THRB</i> transcript. Luciferase assay showed direct binding of miR-155 and miR-425 to 3′UTR of <i>THRB,</i> and subsequent in vivo analyses revealed that transfection of UOK171 cell line with synthetic miR-155 or miR-425 resulted in decreased expression of endogenous <i>TRHB</i> by 22% and 64%, respectively. Finally, real-time PCR analysis showed significant upregulation of miR-155 (354%) and miR-425 (162%) in ccRCC when compared with matched controls. Moreover, microRNA levels were negatively correlated with the amount of <i>THRB</i> transcript in tissue samples. We conclude that CpG methylation is not the major mechanism contributing to decreased <i>THRB</i> expression in ccRCC. In contrast, <i>THRB</i> is targeted by microRNAs miR-155 and miR-425, whose increased expression may be responsible for downregulation of <i>THRB</i> in ccRCC tumors.</p></div

Genetic evidence that thyroid hormone is indispensable for prepubertal insulin-like growth factor-I expression and bone acquisition in mice.

Understanding how bone growth is regulated by hormonal and mechanical factors during early growth periods is important for optimizing the attainment of peak bone mass to prevent or postpone the occurrence of fragility fractures later in life. Using genetic mouse models that are deficient in thyroid hormone (TH) (Tshr(-/-) and Duox2(-/-)), growth hormone (GH) (Ghrhr(lit/lit)), or both (Tshr(-/-); Ghrhr(lit/lit)), we demonstrate that there is an important period prior to puberty when the effects of GH are surprisingly small and TH plays a critical role in the regulation of skeletal growth. Daily administration of T3/T4 during days 5 to 14, the time when serum levels of T3 increase rapidly in mice, rescued the skeletal deficit in TH-deficient mice but not in mice lacking both TH and GH. However, treatment of double-mutant mice with both GH and T3/T4 rescued the bone density deficit. Increased body fat in the TH-deficient as well as TH/GH double-mutant mice was rescued by T3/T4 treatment during days 5 to 14. In vitro studies in osteoblasts revealed that T3 in the presence of TH receptor (TR) α1 bound to a TH response element in intron 1 of the IGF-I gene to stimulate transcription. In vivo studies using TRα and TRβ knockout mice revealed evidence for differential regulation of insulin-like growth factor (IGF)-I expression by the two receptors. Furthermore, blockade of IGF-I action partially inhibited the biological effects of TH, thus suggesting that both IGF-I-dependent and IGF-I-independent mechanisms contribute to TH effects on prepubertal bone acquisition

microRNAs targeting <i>THRB</i> transcript.

<p><b>A.</b> The effect of microRNA mimics on the activity of luciferase expressed from a reporter plasmid pGL3-luc-3′UTR-THRB. HeLa cells were transfected using the reporter plasmid and respective microRNA mimics: pre-miR-155, pre-miR-425, pre-miR-599, or pre-miR-221. The results are shown as percentage of control (Ctrl, cells transfected with non-targeting pre-miR). The plot shows results of three independent biological experiments, measured in triplicates. The relative activity of Firefly luciferase was normalized to Renilla luciferase activity. Statistical analysis was performed using ANOVA followed by Dunnett’s multiple comparison test. *** p<0.001. <b>B.</b> The effect of miR-155 and miR-425 on endogenous <i>THRB</i> expression in renal cancer cells. UOK171 cells were transfected using respective microRNA mimics (pre-miRs) or inhibitors (anti-miRs), and <i>THRB</i> mRNA expression was analyzed using real-time PCR. The results are shown as percent of control (the cells transfected with non-targeting pre-miR). The plots shows results of three independent biological experiments measured in triplicates. Statistical analysis was performed using ANOVA followed by Dunnett’s multiple comparison test. *p<0.05, *** p<0.001. <b>C.</b> The expression of miR-155 and miR-425 in nontumorous controls (C, n = 35) and paired ccRCC tissue samples (T, n = 35). The results are shown as percent of control. Real-time PCR for each sample was performed in triplicates. Statistical analysis was performed using Wilcoxon matched pair test. *p<0.05, **p<0.01.</p