Hanna Tinel

Transcriptome Characterization of Estrogen-Treated Human Myocardium Identifies Myosin Regulatory Light Chain Interacting Protein as a Sex-Specific Element Influencing Contractile Function

OBJECTIVES This study investigated the effects of 17β-estradiol (E2) on gene regulation in human cardiac tissues. We hypothesized that a candidate E2 effect is cardiomyocyte (CM)- and sex-specific, conserved between humans and mice, and that E2 impairs contractile function in male CMs only. BACKGROUND Both men and women produce E2 locally from androgenic precursors. E2 regulates cardiovascular function, but specific mechanisms, protective or harmful, are not fully understood. METHODS We performed genome-wide expression profiling of E2-treated cardiac tissues from men and women, and studied gene expression and function in CMs from hearts of male and female E2-treated mice. RESULTS We found 36 E2-dependent genes regulated in a sex-specific manner. Of these, after E2 exposure, the myosin regulatory light chain interacting protein (MYLIP) gene was induced in tissues of men only. Focusing on Mylip and employing isolated mouse CMs, we confirmed our hypotheses that the E2 effect is CM- and sex-specific and conserved between humans and mice. The E2-treatment led to impaired contractile function in male CMs only, which was characterized by increased Mylip mRNA and protein levels, and decreased myosin regulatory light chain (Mrlc) protein. Our report is the first to our knowledge to show that cardiac Mrlc is an in vivo substrate for Mylip, leading to augmented Mrlc ubiquitination. Of relevance, we found that MYLIP expression levels rise with increasing age in hearts of men. CONCLUSIONS E2 directly influences cardiac gene regulation, and E2 actions may be different between the sexes. Since E2 levels rise in older and/or obese men, pharmacological targeting of MYLIP in men with elevated E2 levels could possibly decrease their risk for the development or progression of cardiovascular disease.

Synergistic Effects of BAY 60-4552 and Vardenafil on Relaxation of Corpus Cavernosum Tissue of Patients with Erectile Dysfunction and Clinical Phosphodiesterase Type 5 Inhibitor Failure

INTRODUCTION Overall efficacy rates of phosphodiesterase type 5 inhibitors (PDE5-i) for erectile dysfunction (ED) are 60-70%. PDE5-i treatment failures currently have to resort to invasive treatment options for restoration of erectile function. AIMS.: To assess changes in the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP)/protein kinase (PKG) pathway in human corpus cavernosum (HCC) of PDE5-i nonresponders compared with healthy controls. To evaluate the effects of BAY 60-4552, a stimulator of soluble guanylate cyclase (sGC), and vardenafil on relaxation of HCC strips from PDE5-i nonresponders. MAIN OUTCOME MEASURES mRNA expression, morphological localization of the NO/cGMP/PKG pathway, and relaxant capacity of both compounds alone or combined. Analysis of variance, t-test or Mann-Whitney test based upon number of groups and normality of data. METHODS HCC tissues were harvested after consent from individuals undergoing penile prosthesis implantation (patients) and potent patients undergoing transurethral surgery (healthy controls, needle biopsy). HCC tissues of patients were compared with those of healthy controls for the expression of mRNA coding for PDE5A, eNOS, PKGα1, PKG2, sGCα1, sGCα2, sGCβ1, sGCβ2, α-smooth muscle actin (aSMA) and β-actin by quantitative polymerase chain reaction (qPCR). The respective proteins were localized using immunofluorescence. Tissue strips of patients were precontracted with phenylepinephrine followed by incubation with 1 μM of either vardenafil or BAY 60-4552, or both simultaneously. RESULTS The main targets in the NO/cGMP/sGC pathway were downregulated in PDE5-i nonresponders. The pathway was morphologically located to HCC smooth muscle, of which the overall content was preserved in ED patients based on aSMA expression. BAY 60-4552 and vardenafil have synergistic effects on relaxation of HCC of PDE5-i nonresponders. The main limitation is the small amount of control tissue precluding functional testing on these samples. CONCLUSION Despite downregulation of the NO/cGMP/PKG pathway, combining BAY 60-4552 with vardenafil significantly enhanced relaxation HCC strips of PDE5-i nonresponders.

Discovery of the Soluble Guanylate Cyclase Stimulator Vericiguat (BAY 1021189) for the Treatment of Chronic Heart Failure

The first-in-class soluble guanylate cyclase (sGC) stimulator riociguat was recently introduced as a novel treatment option for pulmonary hypertension. Despite its outstanding pharmacological profile, application of riociguat in other cardiovascular indications is limited by its short half-life, necessitating a three times daily dosing regimen. In our efforts to further optimize the compound class, we have uncovered interesting structure-activity relationships and were able to decrease oxidative metabolism significantly. These studies resulting in the discovery of once daily sGC stimulator vericiguat (compound 24, BAY 1021189), currently in phase 3 trials for chronic heart failure, are now reported.

Phosphodiesterase 5 inhibitors and erectile dysfunction

Background: The introduction of phosphodiesterase 5 (PDE5) inhibitors for the treatment of erectile dysfunction was a landmark in the therapy of this highly prevalent male disorder. After the launch of sildenafil in 1998, which was followed by vardenafil and tadalafil in 2003, > 40 million men worldwide have been treated with these compounds. It turned out that erectile dysfunction treatment with PDE5 inhibitors is not only very effective, but also well tolerated and safe. Objective: As the erectile dysfunction market is still growing, new developments have been initiated, addressing mainly pharmacokinetic parameters by new compounds, reformulations of the available drugs and combinations of PDE5 inhibitors with other treatment principles. This review summarises and updates the patent information and the present status of developments within the world of PDE5 inhibitors for the treatment of erectile dysfunction. Methods: The information was collected from various scientific databases (e.g., Dolphin, IDDB, Integrity Prous Science, Medline, chemical abstracts, ClinicalTrials.gov) focused on the literature from 2002 onwards. Results/conclusion: Because of the high standard in erectile dysfunction therapy with existing PDE5 inhibitors, positioning of new PDE5 inhibitors for the treatment of erectile dysfunction will become very difficult. Combinations with new drugs having a different mode of action could be successful in patients presently resistant to the therapy with PDE5 inhibitors. The use of PDE5 inhibitors within other indications beyond erectile dysfunction has become an important new aspect in the development of these drugs and will have a substantial influence on research and development within this unique class of drugs.

Additive effects of the Rho kinase inhibitor Y‐27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure

To evaluate the expression of the Rho/Rho‐associated protein kinase (ROCK) pathway in the corpus cavernosum of patients with severe erectile dysfunction (ED) compared with healthy human corpus cavernosum, and to test the functional effects of two Rho kinase inhibitors (RKIs) on erectile tissue of patients with severe ED, which did not respond to phosphodiesterase type 5 inhibitors (PDE5Is).

PDE5 GENE EXPRESSION AND RELAXANT EFFECTS OF VARDENAFIL IN MALE AND FEMALE HUMAN AND RAT DETRUSOR MUSCLE

1. J Urol. 2007 Apr;177(4):1401-7. Tadalafil relieves lower urinary tract symptoms secondary to benign prostatic hyperplasia. McVary KT, Roehrborn CG, Kaminetsky JC, Auerbach SM, Wachs B, Young JM, Esler A, Sides GD, Denes BS. 2. J Urol. 2007 Mar;177(3):1071-7. Sildenafil citrate improves erectile function and urinary symptoms in men with erectile dysfunction and lower urinary tract symptoms associated with benign prostatic hyperplasia: a randomized, double-blind trial. McVary KT, Monnig W, Camps JL Jr, Young JM, Tseng LJ, van den Ende G. 3. J Urol 2008 (soon to be published). A randomised, placebo-controlled study to assess the efficacy of twice-daily vardenafil in the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia. Stief CG; Porst H; Neuser D; Beneke M; Ulbrich E.

The use of PDE5 inhibitors in the treatment of benign prostate hyperplasia and lower urinary tract symptoms: preclinical evidences

Material-methods The mRNA expression of PDE5 was determined in rat tissues of the lower urinary tract. To test the functional relevance of the PDE5 expression, sildenafil, vardenafil, and tadalafil were tested on pre-contracted isolated rat bladder, prostate and urethra strips. The in vivo efficacy was tested in a partial bladder outlet obstruction (BOO) model after acute and chronic treatment.