ABSTRACT
Adult stem cells (ASCs) can be cultured with difficulty from most tissues, often requiring chemical or transgenic modification to achieve adequate quantities. We show here that mouse primary fibroblasts, grown in suspension, change from the elongated and flattened morphology observed under standard adherent culture conditions of generating rounded cells with large nuclei and scant cytoplasm and expressing the mesenchymal stem cell (MSC) marker (Sca1; Ly6A) within 24 h. Based on this initial observation, we describe here a suspension culture method that, irrespective of the lineage used, mouse fibroblast or primary human somatic cells (fibroblasts, hepatocytes and keratinocytes), is capable of generating a high yield of cells in spheroid form which display the expression of ASC surface markers, circumventing the anoikis which often occurs at this stage. Moreover, mouse fibroblast-derived spheroids can be differentiated into adipogenic and osteogenic lineages. An analysis of single-cell RNA sequence data in mouse fibroblasts identified eight distinct cell clusters with one in particular comprising approximately 10% of the cells showing high levels of proliferative capacity expressing high levels of genes related to MSCs and self-renewal as well as the extracellular matrix (ECM). We believe the rapid, high-yield generation of proliferative, multi-potent ASC-like cells via the process we term suspension-induced stem cell transition (SIST) could have significant implications for regenerative medicine.
Subject(s)
Adult Stem Cells , Mesenchymal Stem Cells , Adult , Humans , Animals , Mice , Stem Cells , Cell Differentiation , Mesenchymal Stem Cells/metabolism , Animals, Genetically ModifiedSubject(s)
Cardiovascular System , Hypertension, Pulmonary , Humans , Caveolin 1 , Cell Line, Tumor , LungABSTRACT
Rationale: Extremely preterm infants develop bronchopulmonary dysplasia (BPD), a chronic lung injury that lacks effective treatment. TSP-1 (thrombospondin-1) is an antiangiogenic protein that activates TGF-ß1 (transforming growth factor-ß1), a cytokine strongly linked to both experimental and human BPD. Objectives:1) To examine effects of inhibiting TSP-1-mediated TGF-ß1 activation (LSKL [leucine-serine-lysine-leucine]) in neonatal rats with bleomycin-induced lung injury; 2) to examine effects of a TSP-1 mimic (ABT-510) on lung morphology; and 3) to determine whether TSP-1 and related signaling peptides are increased in lungs of human preterm infants at risk for BPD. Methods: From Postnatal Days 1 to 14, rat pups received daily intraperitoneal bleomycin (1 mg/kg) or vehicle and were treated with daily subcutaneous LSKL (20 mg/kg) or vehicle alone. Separate animals were treated with vehicle or ABT-510 (30 mg/kg/d). Paraffin-embedded lung tissues from 47 autopsies (controls; death <28 d, n = 30 and BPD at risk; death ⩾28 d, n = 17) performed on infants born <29 completed weeks' gestation were semiquantified for injury markers (collagen, macrophages, and 3-nitrotyrosine), TSP-1, and TGF-ß1. Measurements and Main Results: Bleomycin or ABT-510 increased lung TGF-ß1 activity and macrophage influx, caused pulmonary hypertension, and led to alveolar and microvascular hypoplasia. Treatment with LSKL partially prevented abnormal lung morphology secondary to bleomycin. Lungs from human infants at risk for BPD had increased contents of TSP-1 and TGF-ß1 when compared with controls. TGF-ß1 content correlated with markers of lung injury. Conclusions: TSP-1 inhibits alveologenesis in neonatal rats, in part via the upregulated activity of TGF-ß1. Observations in human lungs suggest a similar pathogenic role for TSP-1 in infants at risk for BPD.
Subject(s)
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Bronchopulmonary Dysplasia , Lung Injury , Animals , Bleomycin , Humans , Infant, Newborn , Infant, Premature , Leucine , Rats , Thrombospondin 1/metabolism , Thrombospondin 1/pharmacology , Transforming Growth Factor beta1/metabolismABSTRACT
Circulatory transition after birth presents a critical period whereby the pulmonary vascular bed and right ventricle must adapt to rapidly changing loading conditions. Failure of postnatal transition may present as hypoxemic respiratory failure, with disordered pulmonary and systemic blood flow. In this review, we present the biological and clinical contributors to pathophysiology and present a management framework.
Subject(s)
Hypertension, Pulmonary , Respiratory Insufficiency , Consensus , Critical Illness/therapy , Hemodynamics/physiology , Humans , Hypertension, Pulmonary/therapy , Infant, Newborn , Respiratory Insufficiency/therapyABSTRACT
Premature infants often require mechanical ventilation and oxygen therapy, which can result in bronchopulmonary dysplasia (BPD), characterized by developmental arrest and impaired lung function. Conventional clinical methods for assessing the prenatal lung are not adequate for the detection and assessment of long-term health risks in infants with BPD, highlighting the need for a noninvasive tool for the characterization of lung microstructure and function. Theoretical diffusion models, like the model of xenon exchange (MOXE), interrogate alveolar gas exchange by predicting the uptake of inert hyperpolarized (HP) 129Xe gas measured with HP 129Xe magnetic resonance spectroscopy (MRS). To investigate HP 129Xe MRS as a tool for noninvasive characterization of pulmonary microstructural and functional changes in vivo, HP 129Xe gas exchange data were acquired in an oxygen exposure rat model of BPD that recapitulates the fewer and larger distal airways and pulmonary vascular stunting characteristics of BPD. Gas exchange parameters from MOXE, including airspace mean chord length (Lm), apparent hematocrit in the pulmonary capillaries (HCT), and pulmonary capillary transit time (tx), were compared with airspace mean axis length and area density (MAL and ρA) and percentage area of tissue and air (PTA and PAA) from histology. Lm was significantly larger in the exposed rats (P = 0.003) and correlated with MAL, ρA, PTA, and PAA (0.59<|ρ|<0.66 and P < 0.05). Observed increase in HCT (P = 0.012) and changes in tx are also discussed. These findings support the use of HP 129Xe MRS for detecting fewer, enlarged distal airways in this rat model of BPD, and potentially in humans.
Subject(s)
Bronchopulmonary Dysplasia/metabolism , Capillaries/metabolism , Lung/metabolism , Magnetic Resonance Spectroscopy , Pulmonary Gas Exchange , Animals , Animals, Newborn , Bronchopulmonary Dysplasia/chemically induced , Bronchopulmonary Dysplasia/pathology , Capillaries/pathology , Disease Models, Animal , Female , Humans , Lung/blood supply , Lung/pathology , Male , Rats , Rats, Sprague-Dawley , Xenon IsotopesABSTRACT
PURPOSE: To investigate the dependence of dissolved 129 Xe chemical shift on the fraction of inhaled oxygen, Fi O2 , in the lungs of healthy rats. METHODS: The chemical shifts of 129 Xe dissolved in red blood cells, δRBC , and blood plasma and/or tissue, δPlasma , were measured using MRS in 12 Sprague Dawley rats mechanically ventilated at Fi O2 values of 0.14, 0.19, and 0.22. Regional effects on the chemical shifts were controlled using a chemical shift saturation recovery sequence with a fixed delay time. MRS was also performed at an Fi CO2 value of 0.085 to investigate the potential effect of the vascular response on δRBC and δPlasma . RESULTS: δRBC increased with decreasing Fi O2 (P = .0002), and δPlasma showed no dependence on Fi O2 (P = .23). δRBC at Fi CO2 = 0 (210.7 ppm ± 0.1) and at Fi CO2 = 0.085 (210.6 ppm ± 0.2) were not significantly different (P = .67). δPlasma at Fi CO2 = 0 (196.9 ppm ± 0.3) and at Fi CO2 = 0.085 (197.0 ppm ± 0.1) were also not significantly different (P = .81). CONCLUSION: Rat lung δRBC showed an inverse relationship to Fi O2 , opposite to the relationship previously demonstrated for in vitro human blood. Rat lung δRBC did not depend on Fi CO2 .
Subject(s)
Magnetic Resonance Imaging , Xenon Isotopes , Animals , Erythrocytes , Lung , Oxygen , Rats , Rats, Sprague-DawleyABSTRACT
INTRODUCTION: Although chronic pulmonary hypertension (cPH) secondary to chronic neonatal lung disease is associated with increased mortality and respiratory and neurodevelopmental morbidities, late diagnosis (typically ≥36 weeks postmenstrual age, PMA) and the use of qualitative echocardiographic diagnostic criterion (flat interventricular septum in systole) remain significant limitations in clinical care. Our objective in this study is to evaluate the utility of relevant quantitative echocardiographic indices to identify cPH in preterm neonates, early in postnatal course and to develop a diagnostic test based on the best combination of markers. METHODS AND ANALYSIS: In this ongoing international prospective multicentre observational diagnostic accuracy study, we aim to recruit 350 neonates born <27 weeks PMA and/or birth weight <1000 g and perform echocardiograms in the third week of age and at 32 weeks PMA (early diagnostic assessments, EDA) in addition to the standard diagnostic assessment (SDA) for cPH at 36 weeks PMA. Predefined echocardiographic markers under investigation will be measured at each EDA and examined to create a scoring system to identify neonates who subsequently meet the primary outcome of cPH/death at SDA. Diagnostic test characteristics will be defined for each EDA. Pulmonary artery acceleration time and tricuspid annular plane systolic excursion are the primary markers of interest. ETHICS AND DISSEMINATION: Ethics approval has been received by the Mount Sinai Hospital Research Ethics Board (REB) (#16-0111-E), Sunnybrook Health Sciences Centre REB (#228-2016), NHS Health Research Authority (IRAS 266498), University of Iowa Human Subjects Office/Institutional Review Board (201903736), Rotunda Hospital Research and Ethics Committee (REC-2019-008), and UBC Children's and Women's REB (H19-02738), and is under review at Boston Children's Hospital Institutional Review Board. Study results will be disseminated to participating families in lay format, presented to the scientific community at paediatric and critical care conferences and published in relevant peer-reviewed journals. TRAIL REGISTRATION NUMBER: NCT04402645.
Subject(s)
Hypertension, Pulmonary , Lung Diseases , Boston , Child , Echocardiography , Female , Humans , Hypertension, Pulmonary/diagnostic imaging , Hypertension, Pulmonary/etiology , Infant , Infant, Newborn , Prospective StudiesABSTRACT
During late lung development, alveolar and microvascular development is finalized to enable sufficient gas exchange. Impaired late lung development manifests as bronchopulmonary dysplasia (BPD) in preterm infants. Single-cell RNA sequencing (scRNA-seq) allows for assessment of complex cellular dynamics during biological processes, such as development. Here, we use MULTI-seq to generate scRNA-seq profiles of over 66,000 cells from 36 mice during normal or impaired lung development secondary to hyperoxia with validation of some of the findings in lungs from BPD patients. We observe dynamic populations of cells, including several rare cell types and putative progenitors. Hyperoxia exposure, which mimics the BPD phenotype, alters the composition of all cellular compartments, particularly alveolar epithelium, stromal fibroblasts, capillary endothelium and macrophage populations. Pathway analysis and predicted dynamic cellular crosstalk suggest inflammatory signaling as the main driver of hyperoxia-induced changes. Our data provides a single-cell view of cellular changes associated with late lung development in health and disease.
Subject(s)
Hyperoxia/genetics , Hyperoxia/physiopathology , Lung/metabolism , Lung/pathology , Sequence Analysis, RNA/methods , Single-Cell Analysis/methods , Animals , Bronchopulmonary Dysplasia/genetics , Bronchopulmonary Dysplasia/pathology , Genotype , Male , MiceABSTRACT
OBJECTIVES: The aim of this study was to use a comprehensive imaging protocol to identify echocardiographic correlations of right and left ventricular size, function, and hemodynamics in neonates with persistent pulmonary hypertension of newborn and describe their relationship with key clinical variables. DESIGN: Retrospective case-control echocardiography-based study of persistent pulmonary hypertension of newborn. SETTING: A tertiary neonatal ICU in Canada. PATIENTS: Forty-nine neonates (gestational age ≥ 35 wk old) diagnosed with persistent pulmonary hypertension of newborn within first 3 days after birth and 50 age-matched controls. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The echocardiographic measurements comprised of right ventricular and left ventricular functional markers, including tricuspid annular plane systolic excursion, fractional area change, tissue Doppler imaging, and deformation imaging. Sample size was based on detecting an intergroup difference of 10% in tricuspid annular plane systolic excursion, which was considered the primary outcome. Linear correlations between the right and left ventricular indices, as well as their association with the outcome of death or extracorporeal membrane oxygenation were evaluated. Persistent pulmonary hypertension of newborn was associated with lower tricuspid annular plane systolic excursion (6.81 ± 1.92 vs 9.25 ± 1.30 mm), right-ventricular global longitudinal strain (16.9% ± 5.4% vs -21.6% ± 4.6%); left ventricular ejection fraction (49% ± 7% vs 55% ± 6%), left ventricular global longitudinal strain (-16.7% ± 3.3% vs -21.4% ± 2.0%) (all p < 0.01). Right and left ventricular diastolic and global function was also lower in persistent pulmonary hypertension of newborn, with more pronounced changes seen for the right ventricle. Moderate-to-strong linear correlations were observed between the right and left ventricular functional markers, with right ventricular global longitudinal strain and left ventricular global longitudinal strain being the strongest (r = 0.8). Within persistent pulmonary hypertension of newborn group, hypoxic ischemic encephalopathy was associated with lower right and left ventricular systolic and right ventricular diastolic performance. Tricuspid annular plane systolic excursion (p =0.08) and left ventricular systolic velocity (p = 0.09) tended to be lower in patients who subsequently died/needed extracorporeal membrane oxygenation. CONCLUSIONS: Persistent pulmonary hypertension of newborn is characterized by global cardiac dysfunction, involving both the right and left ventricles, with significant interventricular functional correlation. Cardiac dysfunction early in disease course may identify patients at highest risk of adverse outcome.
Subject(s)
Hypertension, Pulmonary , Ventricular Dysfunction, Right , Canada , Heart Ventricles/diagnostic imaging , Humans , Hypertension, Pulmonary/diagnostic imaging , Infant, Newborn , Retrospective Studies , Stroke Volume , Ventricular Dysfunction, Right/diagnostic imaging , Ventricular Dysfunction, Right/etiology , Ventricular Function, Left , Ventricular Function, RightABSTRACT
PURPOSE: To measure the chemical shift of hyperpolarized 129 Xe dissolved in the red blood cells(δRBC ) of a cohort of rats exposed to hyperoxia and intermittent hypoxia (IH) to mimic human bronchopulmonary dysplasia, and to investigate the effect of xenon-blood distribution time on δRBC . METHODS: δRBC was measured from spectra acquired using a chemical shift saturation recovery sequence from 15 Sprague-Dawley rats exposed to hyperoxia-IH and 10 age-matched control rats. Sensitization to the xenon-blood distribution time was achieved by varying the time between saturation pulses, τ. δRBC was compared with blood fraction measured by histology of the cohort and blood oxygenation measured directly using pulse oximetry following a hypoxic challenge in an identically exposed cohort. RESULTS: The mean δRBC in the hyperoxia-IH exposed rats was 0.55 ± 0.04 ppm lower than that of the healthy cohort (P = .0038), and this difference did not depend on τ (P = .996). The blood fraction of the exposed cohort was lower than that of the healthy cohort (P = .0397). Oximetry measurements showed that the baseline arterial oxygen saturation (Sa O2 ) of each cohort was not different (P = .72), but after a hypoxic challenge, the Sa O2 of the exposed cohort was lower than that of the healthy cohort (P = .003). CONCLUSION: δRBC is reduced in rats exposed to hyperoxia-IH compared with control rats. The change in δRBC is consistent with enhanced blood oxygen desaturation of the exposed cohort measured by pulse oximetry during a hypoxic challenge. This suggests that the observed change in δRBC reflects enhanced desaturation in the hyperoxia-IH exposed cohort compared with the healthy cohort.
Subject(s)
Bronchopulmonary Dysplasia , Hyperoxia , Animals , Erythrocytes , Humans , Infant, Newborn , Lung , Rats , Rats, Sprague-Dawley , XenonSubject(s)
Fractals , Lung Injury , Female , Humans , Infant, Newborn , Observer Variation , Placenta , PregnancyABSTRACT
Deficient nitric oxide (NO) signaling plays a critical role in the pathogenesis of chronic neonatal pulmonary hypertension (PHT). Physiological NO signaling is regulated by S-nitrosothiols (SNOs), which act both as a reservoir for NO and as a reversible modulator of protein function. We have previously reported that therapy with inhaled NO (iNO) increased peroxynitrite-mediated nitration in the juvenile rat lung, although having minimal reversing effects on vascular remodeling. We hypothesized that sodium nitrite (NaNO2) would be superior to iNO in enhancing lung SNOs, thereby contributing to reversal of chronic hypoxic PHT. Rat pups were exposed to air or hypoxia (13% O2) from postnatal days 1 to 21. Dose-response prevention studies were conducted from days 1-21 to determine the optimal dose of NaNO2. Animals then received rescue therapy with daily subcutaneous NaNO2 (20 mg/kg), vehicle, or were continuously exposed to iNO (20 ppm) from days 14-21. Chronic PHT secondary to hypoxia was both prevented and reversed by treatment with NaNO2. Rescue NaNO2 increased lung NO and SNO contents to a greater extent than iNO, without causing nitration. Seven lung SNO proteins upregulated by treatment with NaNO2 were identified by multiplex tandem mass tag spectrometry, one of which was leukotriene A4 hydrolase (LTA4H). Rescue therapy with a LTA4H inhibitor, SC57461A (10 mg·kg-1·day-1 sc), partially reversed chronic hypoxic PHT. We conclude that NaNO2 was superior to iNO in increasing tissue NO and SNO generation and reversing chronic PHT, in part via upregulated SNO-LTA4H.
Subject(s)
Hypertension, Pulmonary/prevention & control , Hypertrophy, Right Ventricular/prevention & control , Hypoxia/complications , Indicators and Reagents/administration & dosage , Sodium Nitrite/administration & dosage , Vascular Remodeling/drug effects , Administration, Inhalation , Animals , Animals, Newborn , Chronic Disease , Female , Hypertension, Pulmonary/etiology , Hypertension, Pulmonary/pathology , Hypertrophy, Right Ventricular/etiology , Hypertrophy, Right Ventricular/pathology , Male , Nitric Oxide/metabolism , Peroxynitrous Acid/metabolism , Rats , Rats, Sprague-Dawley , Signal TransductionABSTRACT
OBJECTIVE: To characterize the natural history of cardiopulmonary physiology in the first 24 hours after birth. STUDY DESIGN: A prospective observational study of healthy newborns was conducted at a large tertiary perinatal center. Echocardiography was performed at <0.5, 2-3, 7-10, and 22-24 hours of age. Specifically, assessment of pulmonary vascular resistance (PVR) (pulmonary artery acceleration time [PAAT], right ventricular ejection time, right ventricular ejection time:PAAT [PVR index], and PAAT indexed to heart rate [PAATi]), ventricular outputs (right and left), and ventricular function (tricuspid annular planar excursion, right ventricular [RV] fractional area change [FAC], RV/left ventricular [LV] global peak longitudinal strain, and LV ejection fraction) were performed. One-way repeated-measures ANOVA analysis was performed for time-dependent variables. RESULTS: In total, 15 neonates (9 males), born at 40 ± 0.8 weeks and 3.5 ± 0.5 kg, respectively, were studied. We observed increased PAATi (P < .05) by 2-3 hours, followed by a subsequent decline in all indices of PVR (PVR index, PAATi, midsystolic notching, and right-to-left ductal flow [P < .0001]). Although right and left ventricular stroke volume increased over the study interval (P < .001), LV output remained stable. All indices of RV function (tricuspid annular planar excursion, RV fractional area change 4-chamber, and RV global peak longitudinal strain-3 chamber [P < .001]) increased during the study interval. CONCLUSION: The immediate transition after birth is characterized by lower PVR, reversal of the transductal shunt, and increased biventricular stroke volume. The differential adaptive response of the RV and LV is novel and may relate to loading conditions and patent ductus arteriosus closure.
Subject(s)
Adaptation, Physiological/physiology , Heart Ventricles/diagnostic imaging , Stroke Volume/physiology , Vascular Resistance/physiology , Ventricular Function, Left/physiology , Ventricular Function, Right/physiology , Echocardiography , Female , Humans , Infant, Newborn , Male , Prospective StudiesABSTRACT
PURPOSE: Inhaled nitric oxide (iNO) is a pulmonary vasodilator that is approved for use in term and near-term neonates with hypoxic respiratory failure associated with evidence of pulmonary hypertension. However, it is commonly used in infants and children to treat a variety of other cardiopulmonary diseases associated with pulmonary hypertension and hypoxic respiratory failure. In critically ill children, iNO therapy may be continued for a prolonged period, and this increases the risk for adverse consequences including toxicity and unnecessary costs. We implemented an iNO Stewardship Program with the aim of improving adherence to guidelines and reducing unnecessary iNO utilization. METHODS: Between April 1, 2011, and March 31, 2015, a before and after cohort study was conducted at The Hospital for Sick Children. Prospective iNO usage and outcome variables in the poststewardship period were examined. RESULTS: Patient characteristics and outcomes were similar before and after stewardship implementation. The number of iNO therapy courses were also similar in the before and after period. Inhaled nitric oxide utilization in the pediatric intensive care unit and the cardiac critical care unit decreased from 15 765 hours in the prestewardship period (April 2011 to March 2013) to 10 342 hours in the poststewardship period (April 2013 to March 2015), with significant improvement in adherence to the iNO guideline and a small decrease in expenditure (3%). CONCLUSION: Implementation of the iNO Stewardship was successful at reducing overall iNO utilization. This quality improvement initiative helped us optimize practice and subsequently expand the methodology to inform the clinical indication for iNO.
Subject(s)
Drug Utilization/standards , Hypertension, Pulmonary/drug therapy , Nitric Oxide/administration & dosage , Quality Improvement/organization & administration , Respiratory Insufficiency/drug therapy , Vasodilator Agents/therapeutic use , Administration, Inhalation , Adolescent , Advisory Committees/organization & administration , Child, Preschool , Critical Care/statistics & numerical data , Drug Utilization/statistics & numerical data , Female , Guideline Adherence , Humans , Infant , Infant, Newborn , Intensive Care Units, Pediatric/organization & administration , Intensive Care Units, Pediatric/statistics & numerical data , Male , Nitric Oxide/therapeutic use , Practice Guidelines as Topic , Prospective Studies , Quality Improvement/standards , Vasodilator Agents/administration & dosageABSTRACT
BACKGROUND/AIM: Mammalian target of rapamycin (mTOR) is a pivotal regulator of cell proliferation, survival, and autophagy. Autophagy is increased in adult experimental chronic pulmonary hypertension (PHT), but its contributory role to pulmonary vascular disease remains uncertain and has yet to be explored in the neonatal animal. Notch is a major pro-proliferative pathway activated by mTOR. A direct relationship between autophagy and Notch signaling has not been previously explored. Our aim was to examine changes in mTOR-, Notch-, and autophagy-related pathways and the therapeutic effects of autophagy modulators in experimental chronic neonatal PHT secondary to chronic hypoxia. METHODS: Rat pups were exposed to normoxia or hypoxia (13% O2 ) from postnatal days 1-21, while receiving treatment with temsirolimus (mTOR inhibitor), DAPT (Notch inhibitor), or chloroquine (inhibitor of autophagic flux). RESULTS: Exposure to hypoxia up-regulated autophagy and Notch3 signaling markers in lung, pulmonary artery (PA), and PA-derived smooth muscle cells (SMCs). Temsirolimus prevented chronic PHT and attenuated PA and SMC signaling secondary to hypoxia. These effects were replicated by DAPT. mTOR or Notch inhibition also down-regulated smooth muscle content of platelet-derived growth factor ß-receptor, a known contributor to vascular remodeling. In contrast, chloroquine had no modifying effects on markers of chronic PHT. Knockdown of Beclin-1 in SMCs had no effect on hypoxia-stimulated Notch3 signaling. CONCLUSIONS: mTOR-Notch3 signaling plays a critical role in experimental chronic neonatal PHT. Inhibition of autophagy did not suppress Notch signaling and had no effect on markers of chronic PHT.
Subject(s)
Hypertension, Pulmonary/metabolism , Receptor, Notch3/metabolism , Receptor, Platelet-Derived Growth Factor beta/metabolism , TOR Serine-Threonine Kinases/metabolism , Animals , Animals, Newborn , Autophagy , Cell Proliferation/drug effects , Diamines/pharmacology , Female , Hypoxia/metabolism , Lung/blood supply , Lung/metabolism , Male , Myocytes, Smooth Muscle/metabolism , Pulmonary Artery/metabolism , Rats, Sprague-Dawley , Receptor, Notch3/antagonists & inhibitors , Signal Transduction , Sirolimus/analogs & derivatives , Sirolimus/pharmacology , TOR Serine-Threonine Kinases/antagonists & inhibitors , Thiazoles/pharmacologyABSTRACT
It is increasingly recognized that the abnormal physiologic consequences of pulmonary hypertension (PH) may contribute to poor cardiopulmonary health in premature babies. Conflicting literature has led to clinical uncertainty, pathological misinterpretation, and variability in treatment approaches among practitioners. There are several disorders with overlapping and interrelated presentations, and other disorders with a similar clinical phenotype but diverse pathophysiological contributors. In this review, we provide a diagnostic approach for acute hypoxemic respiratory failure in the preterm neonate, outline the pathophysiological conditions that may present as acute PH, and discuss the implications of high pulmonary vascular resistance (PVR) on the cardiovascular system. Although PVR and respiratory management are highly interrelated, there may be a population of preterm neonates in whom inhaled nitric oxide may improve illness severity and may relate to outcomes. A management approach based on physiology that considers common clinical conundrums is provided. A more comprehensive understanding of the physiology may help in informed decision-making in clinical situations where conclusive scientific evidence is lacking. Regardless, high-quality research is required, and appropriate definition of the target population is paramount. A thoughtful approach to cardiovascular therapy may also provide an avenue to improve neurodevelopmental outcomes while awaiting more clear answers.
Subject(s)
Hypertension, Pulmonary/drug therapy , Infant, Premature , Acute Disease , Diagnosis, Differential , Heart/physiology , Humans , Hypertension, Pulmonary/diagnosis , Hypoxia/diagnosis , Infant, Newborn , Infant, Premature, Diseases , Vasodilator Agents/administration & dosageABSTRACT
Emerging evidence suggests that long non-coding RNAs (lncRNAs) represent a cellular hub coordinating various cellular processes that are critical in health and disease. Mechanical stress triggers changes in vascular smooth muscle cells (VSMCs) that in turn contribute to pathophysiological changes within the vasculature. We sought to evaluate the role that lncRNAs play in mechanical stretch-induced alterations of human aortic smooth muscle cells (HASMCs). RNA (lncRNA and mRNA) samples isolated from HASMCs that had been subjected to 10 or 20% elongation (1 Hz) for 24 h were profiled with the Arraystar Human LncRNA Microarray V3.0. LncRNA expression was quantified in parallel via qRT-PCR. Of the 30,586 human lncRNAs screened, 580 were differentially expressed (DE, P < 0.05) in stretched HASMCs. Amongst the 26,109 protein-coding transcripts evaluated, 25 of those DE were associated with 25 of the aforementioned DE lncRNAs (P < 0.05). Subsequent Kyoto Encyclopedia of Genes and Genomes analysis revealed that the DE mRNAs were largely associated with the tumor necrosis factor signaling pathway and inflammation. Gene Ontology analysis indicated that the DE mRNAs were associated with cell differentiation, stress response, and response to external stimuli. We describe the first transcriptome profile of stretch-induced changes in HASMCs and provide novel insights into the regulatory switches that may be fundamental in governing aberrant VSMC remodeling.
Subject(s)
Aorta/metabolism , Gene Expression Profiling , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , Oligonucleotide Array Sequence Analysis , RNA, Long Noncoding/biosynthesis , Stress, Mechanical , Aorta/cytology , Humans , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytologyABSTRACT
Bronchopulmonary dysplasia (BPD) is a chronic lung injury characterized by impaired alveologenesis that may persist into adulthood. Rat models of BPD using varying degrees of hyperoxia to produce injury either cause early mortality or spontaneously recover following removal of the inciting stimulus, thus limiting clinical relevance. We sought to refine an established rat model induced by exposure to 60% O2 from birth by following hyperoxia with intermittent hypoxia (IH). Rats exposed from birth to air or 60% O2 until day 14 were recovered in air with or without IH (FIO2 = 0.10 for 10 min every 6 h) until day 28 Animals exposed to 60% O2 and recovered in air had no evidence of abnormal lung morphology on day 28 or at 10-12 wk. In contrast, 60% O2-exposed animals recovered in IH had persistently increased mean chord length, more dysmorphic septal crests, and fewer peripheral arteries. Recovery in IH also increased pulmonary vascular resistance, Fulton index, and arterial wall thickness. IH-mediated abnormalities in lung structure (but not pulmonary hypertension) persisted when reexamined at 10-12 wk, accompanied by increased pulmonary vascular reactivity and decreased exercise tolerance. Increased mean chord length secondary to IH was prevented by treatment with a peroxynitrite decomposition catalyst [5,10,15,20-Tetrakis(4-sulfonatophenyl)-21H,23H-porphyrin iron (III) chloride, 30 mg/kg/day, days 14-28], an effect accompanied by fewer inflammatory cells. We conclude that IH during recovery from hyperoxia-induced injury prevents recovery of alveologenesis and leads to changes in lung and pulmonary vascular function lasting into adulthood, thus more closely mimicking contemporary BPD.
Subject(s)
Bronchopulmonary Dysplasia/complications , Bronchopulmonary Dysplasia/pathology , Hyperoxia/complications , Hypoxia/complications , Lung Injury/complications , Pulmonary Alveoli/growth & development , Pulmonary Alveoli/pathology , Animals , Animals, Newborn , Biomarkers/metabolism , Catalysis , Disease Models, Animal , Female , Hyperoxia/pathology , Hypertension, Pulmonary/complications , Hypoxia/pathology , Lung Injury/pathology , Male , Metalloporphyrins/pharmacology , Peroxynitrous Acid/metabolism , Physical Conditioning, Animal , Pneumonia/complications , Rats, Sprague-DawleyABSTRACT
OBJECTIVES: To evaluate whether incorporating conventional, tissue Doppler imaging and speckle tracking echocardiography are reliable and can characterize changes in left ventricular (LV) function properly in healthy neonates in the early transitional newborn period. STUDY DESIGN: A prospective observational study was conducted in 50 healthy term neonates with a mean ± SD gestational age and birth weight of 39.3 ± 1.2 weeks and 3.5 ± 0.44 kg, respectively. All infants underwent serial echocardiograms at 15 ± 2 (day 1) and 35 ± 2 hours (day 2) of age. The LV dimensions and various functional indices including tissue Doppler imaging velocities and speckle tracking echocardiography-derived peak longitudinal strain, and systolic and diastolic strain rate were acquired and compared between time points. RESULTS: All measurements were feasible from each scan except speckle tracking echocardiography in 10% and 20% of images on days 1 and 2 of age, respectively. LV dimensions, but not functional measures, demonstrated a small to moderate positive correlation with birth weight. On day 2, a small reduction was observed in LV basal diameter, mitral valve inflow velocity time integral, and systolic velocity of the lateral wall and septum. Other indices remained unchanged. Tissue Doppler imaging-derived functional and flow-derived hemodynamic measures demonstrated the least measurement bias, and strain measurements demonstrated better reliability than strain rate, fractional shortening, and ejection fraction. CONCLUSION: The relative reliability of various echocardiographic indices to quantify LV function in neonates establish a normative dataset and provide evidence for their validity during the first 2 days of life.