Vicky K. Yang

Mammalian heart renewal by pre-existing cardiomyocytes

Although recent studies have revealed that heart cells are generated in adult mammals, the frequency of generation and the source of new heart cells are not yet known. Some studies suggest a high rate of stem cell activity with differentiation of progenitors to cardiomyocytes. Other studies suggest that new cardiomyocytes are born at a very low rate, and that they may be derived from the division of pre-existing cardiomyocytes. Here we show, by combining two different pulse–chase approaches—genetic fate-mapping with stable isotope labelling, and multi-isotope imaging mass spectrometry—that the genesis of cardiomyocytes occurs at a low rate by the division of pre-existing cardiomyocytes during normal ageing, a process that increases adjacent to areas of myocardial injury. We found that cell cycle activity during normal ageing and after injury led to polyploidy and multinucleation, but also to new diploid, mononucleate cardiomyocytes. These data reveal pre-existing cardiomyocytes as the dominant source of cardiomyocyte replacement in normal mammalian myocardial homeostasis as well as after myocardial injury.

Concise Review: Developing Best-Practice Models for the Therapeutic Use of Extracellular Vesicles

Growing interest in extracellular vesicles (EVs, including exosomes and microvesicles) as therapeutic entities, particularly in stem cell‐related approaches, has underlined the need for standardization and coordination of development efforts. Members of the International Society for Extracellular Vesicles and the Society for Clinical Research and Translation of Extracellular Vesicles Singapore convened a Workshop on this topic to discuss the opportunities and challenges associated with development of EV‐based therapeutics at the preclinical and clinical levels. This review outlines topic‐specific action items that, if addressed, will enhance the development of best‐practice models for EV therapies. Stem Cells Translational Medicine 2017;6:1730–1739

Comparisons of morphometric measurements and serum insulin-like growth factor concentration in healthy cats and cats with hypertrophic cardiomyopathy.

OBJECTIVE To compare morphometric measurements and serum insulin-like growth factor (IGF-1) concentration in cats with and without hypertrophic cardiomyopathy (HCM), and assess the hypothesis that cats with HCM have larger body size and skeletal features and higher serum IGF-1 concentrations than healthy cats. ANIMALS 25 cats with HCM and 22 healthy control cats. PROCEDURES Physical examination and echocardiography were performed to classify cats into the HCM and control groups. Data collected from each cat included diet history, body weight, body condition score, lengths of the humerus and 4th and 12th thoracic vertebrae, heart size, head length and width, and abdominal circumferences. Comparisons of these variables were made between groups. RESULTS Body condition score in HCM-affected and control cats did not differ significantly. However, median head width; lengths of the head, 4th and 12th thoracic vertebrae, and humerus; and body weight in the HCM-affected group were significantly greater than values in the control group. Median serum concentration of IGF-1 was not significantly different between groups. CONCLUSIONS AND CLINICAL RELEVANCE These data suggested that among the study cats, those with HCM were skeletally larger, but not more obese, than healthy cats. Whether this was attributable to differences in early growth or other causes requires additional investigation.

Circulating exosome microRNA associated with heart failure secondary to myxomatous mitral valve disease in a naturally occurring canine model

ABSTRACT Myxomatous mitral valve disease (MMVD) is functionally and histologically identical to mitral valve prolapse (MVP) in humans. Currently, there are no medical treatments that can delay the progression of this valvular disease or associated cardiac remodelling. Therefore, there is a need to understand the molecular pathology associated with MMVD and MVP better, and thus identify potential therapeutic targets. Circulating exosomes contain small RNA, including miRNA, which reflect cell physiology and pathology. This study explored the association between circulating exosomal miRNA (ex-miRNA) content and MMVD, heart failure due to MMVD (MMVD-CHF) and ageing, which is strongly associated with MMVD. Ex-miRNA was isolated from old normal/healthy dogs (n = 6), young normal dogs (n = 7), dogs with MMVD (n = 7) and dogs with MMVD-CHF (n = 7). Separately, total plasma miRNA was isolated from normal dogs (n = 8), dogs with MMVD (n = 8) and dogs with MMVD-CHF (n = 11). Using reverse transcription quantitative polymerase chain reaction, exosomal miR-181c (p = 0.003) and miR-495 (p = 0.0001) significantly increased in dogs with MMVD-CHF compared to the other three groups. Exosomal miR-9 (p = 0.002) increased in dogs with MMVD and MMVD-CHF compared to age-matched (old) normal dogs. Exosomal miR-599 (p = 0.002) decreased in dogs with MMVD compared to old normal dogs. In total plasma, 58 miRNA were deemed significantly different (p < 0.04) between normal dogs, dogs with MMVD and dogs with MMVD-CHF. However, in contrast to ex-miRNA, none of the miRNA in total plasma remained statistically significant if the false discovery rate was <15%. Changes in ex-miRNA are observed in dogs as they age (miR-9, miR-495 and miR-599), develop MMVD (miR-9 and miR-599) and progress from MMVD to CHF (miR-181c and miR-495). Ex-miRNA expression-level changes appear to be more specific to disease states than total plasma miRNA. RESPONSIBLE EDITOR Elena Aikawa, Harvard Medical School, USA

The use of rivaroxaban for the treatment of thrombotic complications in four dogs

OBJECTIVE To describe the clinical use of rivaroxaban in the treatment of 4 dogs with vascular thrombosis, 2 with pulmonary thromboembolism and 2 with systemic thrombosis. CASE SERIES SUMMARY This report describes the use of a direct factor Xa anticoagulant newly approved in human patients for the treatment or prevention of arterial or venous thrombosis. The use of this medication in a clinical setting for canine patients with thromboembolism has not been described before. Two patients were treated with rivaroxaban for pulmonary thromboembolism. Decreases in thrombus size were seen in both patients, but one patient suffered acute respiratory distress and was euthanized while the other continued to do well at the time of this writing. The other 2 patients were treated for systemic thrombosis. Decreases in thrombus size were also noted. One patient later suffered hematochezia of unknown cause, and the other continued to do well at the time of this writing. NEW OR UNIQUE INFORMATION PROVIDED This is the first published report of the use of a new oral direct factor Xa anticoagulant in dogs in a clinical setting for the treatment of both pulmonary and systemic thrombosis. In this case series, we share our limited experience in the use of this new medication, our strategy in determining appropriate dosages, and our monitoring protocol.Objective To describe the clinical use of rivaroxaban in the treatment of 4 dogs with vascular thrombosis, 2 with pulmonary thromboembolism and 2 with systemic thrombosis. Case Series Summary This report describes the use of a direct factor Xa anticoagulant newly approved in human patients for the treatment or prevention of arterial or venous thrombosis. The use of this medication in a clinical setting for canine patients with thromboembolism has not been described before. Two patients were treated with rivaroxaban for pulmonary thromboembolism. Decreases in thrombus size were seen in both patients, but one patient suffered acute respiratory distress and was euthanized while the other continued to do well at the time of this writing. The other 2 patients were treated for systemic thrombosis. Decreases in thrombus size were also noted. One patient later suffered hematochezia of unknown cause, and the other continued to do well at the time of this writing. New or Unique Information Provided This is the first published report of the use of a new oral direct factor Xa anticoagulant in dogs in a clinical setting for the treatment of both pulmonary and systemic thrombosis. In this case series, we share our limited experience in the use of this new medication, our strategy in determining appropriate dosages, and our monitoring protocol.

Association between Survival Time and Changes in NT-proBNP in Cats Treated for Congestive Heart Failure

Background Reductions in N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) concentrations after treatment have been associated with improved survival in people with congestive heart failure (CHF), but have not been reported in cats with CHF. Objectives To evaluate changes in NT‐proBNP concentrations in cats with CHF after treatment and determine whether serial NT‐proBNP measurements provide prognostic information. Animals Thirty‐one client‐owned cats. Methods Prospective, observational study in cats with new onset CHF secondary to cardiomyopathy. Concentrations of NT‐proBNP were measured within 4 hours of admission to the hospital, on the day of discharge, and at re‐evaluation 7–10 days later. Results Median NT‐proBNP concentrations decreased significantly from admission (1,713 pmol/L [range, 160–3,784 pmol/L]) to discharge (902 pmol/L [range, 147–3,223 pmol/L]); P = .005) and from admission to re‐evaluation (1,124 pmol/L [range, 111–2,727 pmol/L]; P = .024). Median survival time was 109 days (range, 1–709 days), with 5 cats still alive at the time of analysis. Cats with a larger percent decrease in NT‐proBNP from admission to discharge had a longer survival time (P = .048). Cats with evidence of active CHF at the time of re‐evaluation (P = .010) and cats whose owners had difficulty administering medications (P = .045) had shorter survival times. Conclusions and clinical importance Cats with a larger percent decrease in NT‐proBNP during hospitalization and no evidence of CHF at the time of re‐evaluation had longer survival times. Additional studies are needed to determine whether NT‐proBNP can help guide treatment in cats with CHF.

Congenital cardiac malformation with three-chambered right atrium and a persistent left cranial vena cava in a dog

This report describes an unusual congenital abnormality in a dog in which multiple distinct membranes were observed within the right atrium, creating obstruction to venous return from both the cranial vena cava and the caudal vena cava. A persistent left cranial vena cava was also identified. In addition to a membrane in the typical location for cor triatriatum dexter, the dog also had a perforated membrane separating the main right atrial body and tricuspid valve from a more cranial right atrial chamber and the right cranial vena cava. Balloon dilation was performed successfully to alleviate the obstruction to systemic venous return created by the two membranes. Due to the unusual anatomic features, angiography plus echocardiography was useful to completely characterize the congenital abnormality prior to intervention.

Dysregulation of valvular interstitial cell let-7c, miR-17, miR-20a, and miR-30d in naturally occurring canine myxomatous mitral valve disease

Canine myxomatous mitral valve disease (MMVD) resembles the early stages of myxomatous pathology seen in human non-syndromic mitral valve prolapse, a common valvular heart disease in the adult human population. Canine MMVD is seen in older subjects, suggesting age-related epigenetic dysregulation leading to derangements in valvular cell populations and matrix synthesis or degradation. We hypothesized that valvular interstitial cells (VICs) undergo disease-relevant changes in miRNA expression. In primary VIC lines from diseased and control valves, miRNA expression was profiled using RT-qPCR and next generation sequencing. VICs from diseased valves showed phenotypic changes consistent with myofibroblastic differentiation (vimentinlow+, α-SMAhigh+), increases in senescence markers (p21, SA-β-gαl), and decreased cell viability and proliferation potential. RT-qPCR and miRNA sequencing analyses both showed significant (p<0.05) downregulation of let-7c, miR-17, miR-20a, and miR-30d in VICs from diseased valves compared to controls. Decreased let-7c, miR-17, and miR-20a may contribute to myofibroblastic differentiation in addition to cell senescence, and decreased miR-30d may disinhibit cell apoptosis. These data support the hypothesis that epigenetic dysregulation plays an important role in age-related canine MMVD.

Clinical findings and survival time in dogs with advanced heart failure

Background Dogs with advanced heart failure are a clinical challenge for veterinarians but there are no studies reporting clinical features and outcome of this population. Hypothesis/Objectives To describe clinical findings and outcome of dogs with advanced heart failure caused by degenerative mitral valve disease (DMVD). Animals Fifty‐four dogs with advanced heart failure because of DMVD. Methods For study purposes, advanced heart failure was defined as recurrence of congestive heart failure signs despite receiving the initially prescribed dose of pimobendan, angiotensin‐converting‐enzyme inhibitor (ACEI), and furosemide >4 mg/kg/day. Data were collected for the time of diagnosis of Stage C heart failure and time of diagnosis of advanced heart failure. Date of death was recorded. Results At the diagnosis of advanced heart failure, doses of pimobendan (n = 30), furosemide (n = 28), ACEI (n = 13), and spironolactone (n = 4) were increased, with ≥1 new medications added in most dogs. After initial diagnosis of advanced heart failure, 38 (70%) dogs had additional medications adjustments (median = 2 [range, 0‐27]), with the final total medication number ranging from 2‐10 (median = 5). Median survival time after diagnosis of advanced heart failure was 281 days (range, 3‐885 days). Dogs receiving a furosemide dose >6.70 mg/kg/day had significantly longer median survival times (402 days [range, 3‐885 days] versus 129 days [range 9‐853 days]; P = .017). Conclusions and Clinical Importance Dogs with advanced heart failure can have relatively long survival times. Higher furosemide dose and non‐hospitalization were associated with longer survival.

Extracardiac intrapericardial myxosarcoma causing right ventricular outflow tract obstruction in a dog

A 13-year-old male castrated pomeranian cross was referred for evaluation of episodes of collapse and a suspected cardiac mass. The presence of a mass at the base of the heart within the pericardial space was confirmed by echocardiography. Additional diagnostics included computed tomography, ultrasound-guided fine-needle aspirate, and thoracic radiographs. The mass was surgically debulked and diagnosed as myxosarcoma via histopathology. This case report describes the diagnostic imaging, laboratory findings, and short-term positive clinical outcome of a dog with a myxosarcoma in a previously undescribed location.