Circle Formation and Detection Biogenesis Large-scale RNA profiling has indicated that approximately 75% of the human genome is transcribed into RNA and gives rise to millions of RNA transcripts. 15,16 Retention or skipping of single exons can generate multiple distinct mRNAs from one pre-mRNA, known as alternative splicing. The formation of covalently closed exon circRNAs occurs when a 3 0 end of an exon (splice donor site) is joined to a 5 0 end of the same (single-exon circRNA) or

Background: Myocardial fibrosis is a hallmark of cardiac remodeling and functionally involved in heart failure development, a leading cause of deaths worldwide. Clinically, no therapeutic strategy is available that specifically attenuates maladaptive responses of cardiac fibroblasts, the effector cells of fibrosis in the heart. Therefore, our aim was to develop novel antifibrotic therapeutics based on naturally derived substance library screens for the treatment of cardiac fibrosis. Methods: Antifibrotic drug candidates were identified by functional screening of 480 chemically diverse natural compounds in primary human cardiac fibroblasts, subsequent validation, and mechanistic in vitro and in vivo studies. Hits were analyzed for dose-dependent inhibition of proliferation of human cardiac fibroblasts, modulation of apoptosis, and extracellular matrix expression. In vitro findings were confirmed in vivo with an angiotensin II–mediated murine model of cardiac fibrosis in both preventive and therapeutic settings, as well as in the Dahl salt-sensitive rat model. To investigate the mechanism underlying the antifibrotic potential of the lead compounds, treatment-dependent changes in the noncoding RNAome in primary human cardiac fibroblasts were analyzed by RNA deep sequencing. Results: High-throughput natural compound library screening identified 15 substances with antiproliferative effects in human cardiac fibroblasts. Using multiple in vitro fibrosis assays and stringent selection algorithms, we identified the steroid bufalin (from Chinese toad venom) and the alkaloid lycorine (from Amaryllidaceae species) to be effective antifibrotic molecules both in vitro and in vivo, leading to improvement in diastolic function in 2 hypertension-dependent rodent models of cardiac fibrosis. Administration at effective doses did not change plasma damage markers or the morphology of kidney and liver, providing the first toxicological safety data. Using next-generation sequencing, we identified the conserved microRNA 671-5p and downstream the antifibrotic selenoprotein P1 as common effectors of the antifibrotic compounds. Conclusions: We identified the molecules bufalin and lycorine as drug candidates for therapeutic applications in cardiac fibrosis and diastolic dysfunction.

The plasma levels of long noncoding RNA LIPCAR are elevated in heart failure (HF) patients with reduced ejection fraction and associated with left ventricular remodeling and poor outcomes. We studied whether the presence of chronic kidney disease (CKD), as defined by an estimated glomerular filtration rate value <60mL/(min·1.73m2) modified the associations of plasma LIPCAR with left ventricular remodeling and outcomes in HF patients. Two hundred and thirty-four patients (mean age 74 [9.14] years, 50% male) were enrolled and followed for 4.73 (0.24–7.25) years. Plasma LIPCAR was detected by real-time quantitative polymerase chain reaction. LIPCAR was increased ( P =0.005) in patients compared with 17 age- and sex-matched controls, directly correlated with age ( P =0.001) and with the maximal early transmitral flow velocity to the mean peak early diastolic velocity of the mitral annulus displacement ratio ( P =0.001) and inversely correlated with estimated glomerular filtration rate ( P <0.001). LIPCAR was associated with hospitalization for HF, cardiovascular death, and a composite of hospitalization for HF or cardiovascular death ( P ≤0.010), these associations being dependent of estimated glomerular filtration rate. The interactions between estimated glomerular filtration rate and LIPCAR with respect to these outcomes were statistically significant or of borderline significance ( P ≤0.060). LIPCAR was increased in CKD patients compared with non-CKD patients ( P =0.021). LIPCAR was independently associated with hospitalization for HF ( P ≤0.039) only in non-CKD patients, but its addition to traditional risk factors did not improve risk prediction in these patients. In conclusion, plasma LIPCAR prognosticates outcomes in elderly HF patients without CKD. Thus, there is an effect modification of CKD on the association of circulating LIPCAR with outcomes in HF patients.

Fabry disease is an X-linked deficiency of the lysosomal hydrolase alpha-galactosidase A (alpha-Gal).This results in an accumulation of globotriaosylceramide (GL-3/Gb3) in a variety of cells with subsequent functional impairment. The continuous progress of FD often leads to decreased quality of life and premature death caused by multi-organic complications. The overall aim of our study was to determine the amount of circulating miRNAs in Fabry patients and to test whether ERT would alter the level of individual circulating miRNAs. We used miRNA sequencing by the HTG EdgeSeq System to identify the circulating miRNA pool from Fabry patients with and without enzyme replacement therapy (n = 6). In total, 296 miRNAs in serum of patients were identified. Among them 9 miRNAs were further evaluated in extra serum samples (n = 31) using real-time qPCR and 6 of them showed significant differential expression. The resulting miRNA pattern may help to better understand mechanisms involved in the beneficial effects of ERT and these new miRNA markers could help to estimate the efficacy of ERT or to identify Fabry patients with specific need for ERT.Fabry disease (FD) is an X-chromosome linked disorder caused by mutations in gene GLA coding for alpha-galactosidase-A enzyme (alpha-Gal). The enzyme activity deficiency that results in an accumulation of globotriaosylceramide (GL-3/Gb3) in a variety of cells often leads to subsequent functional impairment 1 . The initial manifestations of Fabry disease usually start in adolescence stage of life, including neuropathic pain (acroparesthesia) and abdominal discomfort 2 . The continuous progress of FD results in decreased quality of life and premature death caused by multi-organic complications 3,4 . As a specific treatment, Enzyme replacement therapy (ERT) has been shown to stabilize and reduce many signs and symptoms of Fabry disease 5-7 . More recently, oral chaperone therapy was shown to be also effective in selected Fabry patients depending on the underlying gene mutation 8 . Of clinical importance is the fact that early diagnosis and treatment in the disease course may delay or prevent the progression towards irreversible organ dysfunction and the consequent life-threatening complications. This is sometimes difficult due to the high variability of the severity and multi-organ system involvement in Fabry disease 9 . Next to the clinical features, enzyme activity tests and DNA sequencing are available to confirm the diagnosis 10 . Globotriaosylsphingosine (LysoGb3) serves as a useful biomarker to improve the diagnosis of heterozygous Fabry disease for therapeutic evaluation and monitoring 11 . In addition, circulating serum proteins in the blood of Fabry patients may help to get more information about the underlying pathophysiological mechanisms 12 .Recently, a group of small RNA molecules known as microRNAs (miRNAs) have been proved to play essential roles in the cardiac function 13,14 . Moreover, the expression levels of miRNAs that present in circulating fluid usually differ betw...

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