Vascular TSP1-CD47 signaling promotes sickle cell-associated arterial vasculopathy and pulmonary hypertension in mice

Novelli, Enrico M.; Little-Ihrig, Lynda; Knupp, Heather E.; Rogers, Natasha M.; Yao, Mingyi; Baust, Jeffrey; Meijles, Daniel N.; Croix, Claudette M. St.; Ross, Mark A.; Pagano, Patrick J.; DeVallance, Evan; Miles, George; Potoka, Karin P; Isenberg, Jeffrey S.; Gladwin, Mark T.

doi:10.1152/ajplung.00302.2018

Cited by 42 publications

(38 citation statements)

References 81 publications

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“…Direct confirmation of vasoocclusion in sickle mice is challenging, and typically limited to the finding of vascular congestion with large numbers of sickled RBCs. 51 In a postmortem study of a patient with SCD who died during a VOE, the bone marrow histology showed dilated and engorged small vessels which were not completely occluded by sickled RBCs, a possible fixation artifact. 52 In our experiments, it is likely that restoration of vascular patency and blood flow may have occurred by the time the organs were harvested 24 hours after LPS challenge, a prespecified time point required by our imaging protocol, thus precluding us from detecting earlier microvascular obstructions and their attendant ischemic changes.…”

Section: Discussionmentioning

confidence: 99%

Integrin VLA-4 as a PET imaging biomarker of hyper-adhesion in transgenic sickle mice

Perkins

Nyiranshuti²,

Little-Ihrig

et al. 2020

Blood Advances

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In sickle cell disease (SCD), very late antigen-4 (VLA-4 or integrin α4β1) mediates the adhesion of reticulocytes to inflamed, proinflammatory endothelium, a key process in promoting vaso-occlusive episodes (VOEs). We hypothesized that a radionuclide tracer targeting VLA-4 could be harnessed as a positron emission tomography (PET) imaging biomarker of VOEs. We tested the VLA-4 peptidomimetic PET tracer 64Cu-CB-TE1A1P-PEG4-LLP2A (64Cu-LLP2A) for imaging hyper-adhesion–associated VOEs in the SCD Townes mouse model. With lipopolysaccharide (LPS)-induced VOEs, 64Cu-LLP2A uptake was increased in the bone marrow of the humeri and femurs, common sites of VOEs in SCD mice compared with non-SCD mice. Treatment with a proven inhibitor of VOEs (the anti-mouse anti-P-selectin monoclonal antibody [mAb] RB40.34) during LPS stimulation led to a reduction in the uptake of 64Cu-LLP2A in the humeri and femurs to baseline levels, implying blockade of VOE hyper-adhesion. Flow cytometry with Cy3-LLP2A demonstrated an increased percentage of VLA-4–positive reticulocytes in SCD vs non-SCD mice in the bone and peripheral blood after treatment with LPS, which was abrogated by anti-P-selectin mAb treatment. These data, for the first time, show in vivo imaging of VLA-4–mediated hyper-adhesion, primarily of SCD reticulocytes, during VOEs. PET imaging with 64Cu-LLP2A may serve as a valuable, noninvasive method for identifying sites of vaso-occlusion and may provide an objective biomarker of disease severity and anti-P-selectin treatment efficacy in patients with SCD.

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Section: Discussionmentioning

confidence: 99%

Integrin VLA-4 as a PET imaging biomarker of hyper-adhesion in transgenic sickle mice

Perkins

Nyiranshuti²,

Little-Ihrig

et al. 2020

Blood Advances

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“…157 Finally, in a model of sickle cell disease-associated PH severity of pulmonary vascular disease correlated with levels of CD47. 158 This study also demonstrated that mice with myeloid cell deficiency of CD47 were conferred protection against development of PH, associated with decrease ROS generation and a decreased DNA damage response. Thus, CD47 regulation through cytosolic DNA sensing in innate immune cells remains a plausible—and thought-provoking—novel method of treating the underlying pathophysiology predisposing to pulmonary vascular disease.…”

Section: Exposures and Therapymentioning

confidence: 62%

“…Supporting empiric data demonstrated normalization of pulmonary pressures in CD47 -/mice, as well as ex vivo demonstration of decreased ET-1 production by human pulmonary artery endothelial cells treated with a neutralizing CD47 antibody 157 . Finally, in a model of sickle cell diseaseassociated PH severity of pulmonary vascular disease correlated with levels of CD47 158 . This study also demonstrated that mice with myeloid cell deficiency of CD47 were conferred protection against P e e r R e v i e w V e r s i o n development of PH, associated with decrease ROS generation and a decreased DNA damage response.…”

Section: Cd47 Blockadementioning

confidence: 92%

The Third Man: DNA sensing as espionage in pulmonary vascular health and disease

et al. 2021

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For as long as nucleic acids have been utilized to vertically and horizontally transfer genetic ma-terial, living organisms have had to develop methods of recognizing cytosolic DNA as either pathogenic (microbial invasion) or physiologic (mitosis and cellular proliferation). Derangement in key signaling molecules involved in these pathways of nucleotide sensing result in a family of diseases labeled interferonopathies. An interferonopathy, characterized by constitutive expres-sion of type I interferons, ultimately manifests as severe autoimmune disease at a young age. Afflicted patients present with a constellation of immune-mediated conditions, including primary lung manifestations such as pulmonary fibrosis and pulmonary hypertension. The latter condi-tion is especially interesting in light of the known role that DNA damage plays in a variety of types of inherited and induced pulmonary hypertension, with free DNA detection elevated in the circula-tion of affected individuals. While little is known regarding the role of cytosolic DNA sensing in development of pulmonary vascular disease, exciting new research in the related fields of immu-nology and oncology potentially sheds light on future areas of fruitful exploration. As such, the goal of this review is to summarize the state of the field of nucleic acid sensing, extrapolating common shared pathways that parallel our knowledge of pulmonary hypertension, in a molecular and cell-specific manner. Principles of DNA sensing related to known pulmonary injury inducing stimuli are also evaluated, in addition to potential therapeutic targets. Finally, future directions in pulmonary hypertension research and treatments will be briefly discussed.

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“…TSP-1 has previously been implicated in the pathophysiology of vascular occlusion and pulmonary hypertension associated with sickle cell disease (SCD) [ 53 , 54 ]. Increased prothrombotic risk in SCD is linked to elevated TSP-1 levels, which not only inhibit ADAMTS13 proteolysis of von Willebrand Factor [ 55 ], but also provoke RBC MP shedding; this process, in turn, favors RBC adhesion to endothelial cells as well as stimulation of endothelial cell apoptosis [ 56 ].…”

Section: Discussionmentioning

confidence: 99%

Thrombospondin-1/CD47 signaling modulates transmembrane cation conductance, survival, and deformability of human red blood cells

et al. 2020

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Background Thrombospondin-1 (TSP-1), a Ca2+-binding trimeric glycoprotein secreted by multiple cell types, has been implicated in the pathophysiology of several clinical conditions. Signaling involving TSP-1, through its cognate receptor CD47, orchestrates a wide array of cellular functions including cytoskeletal organization, migration, cell-cell interaction, cell proliferation, autophagy, and apoptosis. In the present study, we investigated the impact of TSP-1/CD47 signaling on Ca2+ dynamics, survival, and deformability of human red blood cells (RBCs). Methods Whole-cell patch-clamp was employed to examine transmembrane cation conductance. RBC intracellular Ca2+ levels and multiple indices of RBC cell death were determined using cytofluorometry analysis. RBC morphology and microvesiculation were examined using imaging flow cytometry. RBC deformability was measured using laser-assisted optical rotational cell analyzer. Results Exposure of RBCs to recombinant human TSP-1 significantly increased RBC intracellular Ca2+ levels. As judged by electrophysiology experiments, TSP-1 treatment elicited an amiloride-sensitive inward current alluding to a possible Ca2+ influx via non-selective cation channels. Exogenous TSP-1 promoted microparticle shedding as well as enhancing Ca2+- and nitric oxide-mediated RBC cell death. Monoclonal (mouse IgG1) antibody-mediated CD47 ligation using 1F7 recapitulated the cell death-inducing effects of TSP-1. Furthermore, TSP-1 treatment altered RBC cell shape and stiffness (maximum elongation index). Conclusions Taken together, our data unravel a new role for TSP-1/CD47 signaling in mediating Ca2+ influx into RBCs, a mechanism potentially contributing to their dysfunction in a variety of systemic diseases.

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Vascular TSP1-CD47 signaling promotes sickle cell-associated arterial vasculopathy and pulmonary hypertension in mice

Cited by 42 publications

References 81 publications

Integrin VLA-4 as a PET imaging biomarker of hyper-adhesion in transgenic sickle mice

Integrin VLA-4 as a PET imaging biomarker of hyper-adhesion in transgenic sickle mice

The Third Man: DNA sensing as espionage in pulmonary vascular health and disease

Thrombospondin-1/CD47 signaling modulates transmembrane cation conductance, survival, and deformability of human red blood cells

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