Functional interaction between BMPR-II and Tctex-1, a light chain of Dynein, is isoform-specific and disrupted by mutations underlying primary pulmonary hypertension

Diverse heterozygous mutations of bone morphogenetic receptor type II (BMPR-II) underlie the inherited form of the vascular disorder primary pulmonary hypertension (PPH). As yet, the molecular detail of how such defects contribute to the pathogenesis of PPH remains unclear. BMPR-II is a member of the transforming growth factor-beta cell signalling superfamily. Ligand binding induces cell surface receptor complex formation and activates a cascade of phosphorylation events of intracellular intermediaries termed Smads, which initiate transcriptional regulation. Some 30% of PPH-causing mutations localize to exon 12, which may be spliced out forming an isoform depleted of the unusually long BMPR-II cytoplasmic tail. To further elucidate the consequences of BMPR2 mutation, we sought to characterize aspects of the cytoplasmic domain function by seeking intracellular binding partners. We now report that Tctex-1, a light chain of the motor complex dynein, interacts with the cytoplasmic domain of BMPR-II and demonstrate that Tctex-1 is phosphorylated by BMPR-II, a function disrupted by PPH disease causing mutations within exon 12. Finally we show that BMPR-II and Tctex-1 co-localize to endothelium and smooth muscle within the media of pulmonary arterioles, key sites of vascular remodelling in PPH. Taken together, these data demonstrate a discrete function for the cytoplasmic domain of BMPR-II and justify further investigation of whether the interaction with and phosphorylation of Tctex-1 contributes to the pathogenesis of PPH

Primary pulmonary hypertension is associated with reduced pulmonary vascular expression of type II bone morphogenetic protein receptor

BACKGROUND: Mutations in the type II receptor for bone morphogenetic protein (BMPR-II), a receptor member of the transforming growth factor-beta (TGF-beta) superfamily, underlie many familial and sporadic cases of primary pulmonary hypertension (PPH). METHODS AND RESULTS: Because the sites of expression of BMPR-II in the normal and hypertensive lung are unknown, we studied the cellular localization of BMPR-II and the related type I and II receptors for TGF-beta by immunohistochemistry in lung sections from patients undergoing heart-lung transplantation for PPH (n=11, including 3 familial cases) or secondary pulmonary hypertension (n=6) and from unused donor lungs (n=4). In situ hybridization was performed for BMPR-II mRNA. Patients were screened for the presence of mutations in BMPR2. In normal lungs, BMPR-II expression was prominent on vascular endothelium, with minimal expression in airway and arterial smooth muscle. In pulmonary hypertension cases, the intensity of BMPR-II immunostaining varied between lesions but involved endothelial and myofibroblast components. Image analysis confirmed that expression of BMPR-II was markedly reduced in the peripheral lung of PPH patients, especially in those harboring heterozygous BMPR2 mutations. A less marked reduction was also observed in patients with secondary pulmonary hypertension. In contrast, there was no difference in level of staining for TGF-betaRII or the endothelial marker CD31. CONCLUSIONS: The cellular localization of BMPR-II is consistent with a role in the formation of pulmonary vascular lesions in PPH, and reduced BMPR-II expression may contribute to the process of vascular obliteration in severe pulmonary hypertension

Probability of Physical Association of 104 Blended Companions to Kepler Objects of Interest Using Visible and Near-Infrared Adaptive Optics Photometry

We determine probabilities of physical association for stars in blended Kepler Objects of Interest (KOIs), and find that 14.5%_(-3.4%)^(+3.8%) of companions within ~ 4" are consistent with being physically unassociated with their primary. This produces a better understanding of potential false positives in the Kepler catalog and will guide models of planet formation in binary systems. Physical association is determined through two methods of calculating multi-band photometric parallax using visible and near-infrared adaptive optics observations of 84 KOI systems with 104 contaminating companions within ~ 4". We find no evidence that KOI companions with separations of less than 1" are more likely to be physically associated than KOI companions generally. We also reinterpret transit depths for 94 planet candidates, and calculate that 2.6% ± 0.4% of transits have R > 15R_⊕, which is consistent with prior modeling work

The Robo-AO KOI survey: laser adaptive optics imaging of every Kepler exoplanet candidate

The Robo-AO Kepler Planetary Candidate Survey is observing every Kepler planet candidate host star (KOI) with laser adaptive optics imaging to hunt for blended nearby stars which may be physically associated companions. With the unparalleled efficiency provided by the first fully robotic adaptive optics system, we perform the critical search for nearby stars (0.15" to 4.0" separation with contrasts up to 6 magnitudes) that dilute the observed planetary transit signal, contributing to inaccurate planetary characteristics or astrophysical false positives. We present 3313 high resolution observations of Kepler planetary hosts from 2012-2015, discovering 479 nearby stars. We measure an overall nearby star probability rate of 14.5±0.8%. With this large data set, we are uniquely able to explore broad correlations between multiple star systems and the properties of the planets which they host, providing insight into the formation and evolution of planetary systems in our galaxy. Several KOIs of particular interest will be discussed, including possible quadruple star systems hosting planets and updated properties for possible rocky planets orbiting with in their star's habitable zone

Investigation of chlorine radical chemistry in the Eyjafjallajkull volcanic plume using observed depletions in non-methane hydrocarbons

As part of the effort to understand volcanic plume composition and chemistry during the eruption of the Icelandic volcano Eyjafjallajkull, the CARIBIC atmospheric observatory was deployed for three special science flights aboard a Lufthansa passenger aircraft. Measurements made during these flights included the collection of whole air samples, which were analyzed for non-methane hydrocarbons (NMHCs). Hydrocarbon concentrations in plume samples were found to be reduced to levels below background, with relative depletions characteristic of reaction with chlorine radicals (Cl). Recent observations of halogen oxides in volcanic plumes provide evidence for halogen radical chemistry, but quantitative data for free halogen radical concentrations in volcanic plumes were absent. Here we present the first observation-based calculations of Cl radical concentrations in volcanic plumes, estimated from observed NMHC depletions. Inferred Cl concentrations were between 1.3 × 10 and 6.6 × 10 Cl cm. The relationship between NMHC variability and local lifetimes was used to investigate the ratio between OH and Cl within the plume, with [OH]/[Cl] estimated to be ∼37. Copyright 2011 by the American Geophysical Union

ROBO-AO KEPLER PLANETARY CANDIDATE SURVEY. II. ADAPTIVE OPTICS IMAGING OF 969 KEPLER EXOPLANET CANDIDATE HOST STARS

We initiated the Robo-AO Kepler Planetary Candidate Survey in 2012 to observe each Kepler exoplanet candidate host star with high angular resolution, visible light, laser adaptive optics (AOs) imaging. Our goal is to find nearby stars lying in Kepler's photometric apertures that are responsible for the relatively high probability of false-positive exoplanet detections and that cause underestimates of the size of transit radii. Our comprehensive survey will also shed light on the effects of stellar multiplicity on exoplanet properties and will identify rare exoplanetary architectures. In this second part of our ongoing survey, we observed an additional 969 Kepler planet candidate hosts and we report blended stellar companions up to that contribute to Kepler's measured light curves. We found 203 companions within ~4'' of 181 of the Kepler stars, of which 141 are new discoveries. We measure the nearby star probability for this sample of Kepler planet candidate host stars to be 10.6% ± 1.1% at angular separations up to 25, significantly higher than the 7.4% ± 1.0% probability discovered in our initial sample of 715 stars; we find the probability increases to 17.6% ± 1.5% out to a separation of 40. The median position of Kepler Objects of Interest (KOIs) observed in this survey are 11 closer to the galactic plane, which may account for some of the nearby star probability enhancement. We additionally detail 50 Keck AO images of Robo-AO observed KOIs in order to confirm 37 companions detected at a <5σ significance level and to obtain additional infrared photometry on higher significance detected companions

ROBO-AO KEPLER PLANETARY CANDIDATE SURVEY. II. ADAPTIVE OPTICS IMAGING OF 969 KEPLER EXOPLANET CANDIDATE HOST STARS

We initiated the Robo-AO Kepler Planetary Candidate Survey in 2012 to observe each Kepler exoplanet candidate host star with high angular resolution, visible light, laser adaptive optics (AOs) imaging. Our goal is to find nearby stars lying in Kepler's photometric apertures that are responsible for the relatively high probability of false-positive exoplanet detections and that cause underestimates of the size of transit radii. Our comprehensive survey will also shed light on the effects of stellar multiplicity on exoplanet properties and will identify rare exoplanetary architectures. In this second part of our ongoing survey, we observed an additional 969 Kepler planet candidate hosts and we report blended stellar companions up to that contribute to Kepler's measured light curves. We found 203 companions within ~4'' of 181 of the Kepler stars, of which 141 are new discoveries. We measure the nearby star probability for this sample of Kepler planet candidate host stars to be 10.6% ± 1.1% at angular separations up to 25, significantly higher than the 7.4% ± 1.0% probability discovered in our initial sample of 715 stars; we find the probability increases to 17.6% ± 1.5% out to a separation of 40. The median position of Kepler Objects of Interest (KOIs) observed in this survey are 11 closer to the galactic plane, which may account for some of the nearby star probability enhancement. We additionally detail 50 Keck AO images of Robo-AO observed KOIs in order to confirm 37 companions detected at a <5σ significance level and to obtain additional infrared photometry on higher significance detected companions

Genome-Wide RNA Sequencing of Human Trabecular Meshwork Cells Treated with TGF-β1: Relevance to Pseudoexfoliation Glaucoma

Pseudoexfoliation glaucoma (XFG) is an aggressive form of secondary open angle glaucoma, characterised by the production of exfoliation material and is estimated to affect 30 million people worldwide. Activation of the TGF-&beta; pathway by TGF-&beta;1 has been implicated in the pathogenesis of pseudoexfoliation glaucoma. To further investigate the role of TGF-&beta;1 in glaucomatous changes in the trabecular meshwork (TM), we used RNA-Seq to determine TGF-&beta;1 induced changes in the transcriptome of normal human trabecular meshwork (HTM) cells. The main purpose of this study was to perform a hypothesis-independent RNA sequencing analysis to investigate genome-wide alterations in the transcriptome of normal HTMs stimulated with TGF-&beta;1 and investigate possible pathophysiological mechanisms driving XFG. Our results identified multiple differentially expressed genes including several genes known to be present in exfoliation material. Significantly altered pathways, biological processes and molecular functions included extracellular matrix remodelling, Hippo and Wnt pathways, the unfolded protein response, oxidative stress, and the antioxidant system. This cellular model of pseudoexfoliation glaucoma can provide insight into disease pathogenesis and support the development of novel therapeutic interventions

Clinical and molecular genetic features of pulmonary hypertension in patients with hereditary hemorrhagic telangiectasia

BACKGROUND: Most patients with familial primary pulmonary hypertension have defects in the gene for bone morphogenetic protein receptor II (BMPR2), a member of the transforming growth factor beta (TGF-beta) superfamily of receptors. Because patients with hereditary hemorrhagic telangiectasia may have lung disease that is indistinguishable from primary pulmonary hypertension, we investigated the genetic basis of lung disease in these patients. METHODS: We evaluated members of five kindreds plus one individual patient with hereditary hemorrhagic telangiectasia and identified 10 cases of pulmonary hypertension. In the two largest families, we used microsatellite markers to test for linkage to genes encoding TGF-beta-receptor proteins, including endoglin and activin-receptor-like kinase 1 (ALK1), and BMPR2. In subjects with hereditary hemorrhagic telangiectasia and pulmonary hypertension, we also scanned ALK1 and BMPR2 for mutations. RESULTS: We identified suggestive linkage of pulmonary hypertension with hereditary hemorrhagic telangiectasia on chromosome 12q13, a region that includes ALK1. We identified amino acid changes in activin-receptor-like kinase 1 that were inherited in subjects who had a disorder with clinical and histologic features indistinguishable from those of primary pulmonary hypertension. Immunohistochemical analysis in four subjects and one control showed pulmonary vascular endothelial expression of activin-receptor-like kinase 1 in normal and diseased pulmonary arteries. CONCLUSIONS: Pulmonary hypertension in association with hereditary hemorrhagic telangiectasia can involve mutations in ALK1. These mutations are associated with diverse effects, including the vascular dilatation characteristic of hereditary hemorrhagic telangiectasia and the occlusion of small pulmonary arteries that is typical of primary pulmonary hypertension