PLX3397, a CSF1 receptor inhibitor, limits allotransplantation-induced vascular remodeling

2021 ◽  
Author(s):  
Vanessa M Almonte ◽  
Unimunkh Uriyanghai ◽  
Lander Egaña-Gorroño ◽  
Dippal Parikh ◽  
Gustavo H Oliveira-Paula ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Disease ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Kinase Inhibitor ◽  
Treatment Options ◽  
Solid Organ Transplantation ◽  
Preclinical Model ◽  
Solid Organ ◽  
Pharmacologic Inhibition

Abstract Aims Graft vascular disease (GVD), a clinically important and highly complex vascular occlusive disease, arises from the interplay of multiple cellular and molecular pathways. While occlusive intimal lesions are composed predominantly of smooth muscle-like cells (SMLCs), the origin of these cells and the stimuli leading to their accumulation in GVD are uncertain. Macrophages have recently been identified as both potential drivers of intimal hyperplasia and as precursors that undergo transdifferentiation to become SMLCs in non-transplant settings. Colony stimulating factor-1 (CSF1) is a well-known regulator of macrophage development and differentiation, and prior preclinical studies have shown that lack of CSF1 limits GVD. We sought to identify the origins of SMLCs and of cells expressing the CSF1 receptor (CSF1R) in GVD, and to test the hypothesis that pharmacologic inhibition of CSF1 signaling would curtail both macrophage and SMLC activities and decrease vascular occlusion. Methods and Results We used genetically modified mice and a vascular transplant model with minor antigen mismatch to assess cell origins. We found that neointimal SMLCs derive from both donor and recipient, and that transdifferentiation of macrophages to SMLC phenotype is minimal in this model. Cells expressing CSF1R in grafts were identified as recipient-derived myeloid cells of Cx3cr1-lineage, and these cells rarely expressed smooth muscle marker proteins. Blockade of CSF1R activity using the tyrosine kinase inhibitor PLX3397 limited the expression of genes associated with innate immunity and decreased levels of circulating monocytes and intimal macrophages. Importantly, PLX3397 attenuated the development of GVD in arterial allografts. Conclusion These studies provide proof of concept for pharmacologic inhibition of the CSF1/CSF1R signaling pathway as a therapeutic strategy in GVD. Further preclinical testing of this pathway in GVD is warranted. Translational Perspective Graft vascular disease is a major limitation to the long-term success of clinical solid organ transplantation. Currently, there are no effective treatment options to prevent the development of neointimal lesions that obstruct blood flow to the graft. In this study we found that PLX3397, a selective inhibitor of CSF1R signaling, reduced the accumulation of macrophages and ACTA2+ cells within neointimal lesions in a preclinical model of graft vascular disease. Our study highlights a promising role for the pharmacologic targeting of CSF1R signaling to further study the molecular mechanisms that regulate allotransplantation-induced vascular remodeling.

scholarly journals How I treat EBV lymphoproliferation

Blood ◽  
2009 ◽  
Vol 114 (19) ◽  
pp. 4002-4008 ◽  
Author(s):  
Helen E. Heslop
Keyword(s):  
B Cells ◽  
Cytotoxic T Lymphocyte ◽  
Cytotoxic T Cells ◽  
Treatment Options ◽  
Solid Organ Transplantation ◽  
Solid Organ ◽  
Hematopoietic Stem ◽  
Serial Measurement ◽  
Barr Virus ◽  
Epstein Barr

Abstract Epstein-Barr virus (EBV)–associated B-cell lymphoproliferation is a life-threatening complication after hematopoietic stem cell or solid organ transplantation resulting from outgrowth of EBV-infected B cells that would normally be controlled by EBV-cytotoxic T cells. During the past decade, early detection strategies, such as serial measurement of EBV-DNA load in peripheral blood samples, have helped to identify high-risk patients and to diagnose early lymphoproliferation. Treatment options include manipulation of the balance between outgrowing EBV-infected B cells and the EBV cytotoxic T lymphocyte response and targeting the B cells with monoclonal antibodies or chemotherapy. Major challenges remain for defining indications for preemptive therapies and integrating novel and conventional therapies.

Abstract 512: Collagen Homologous Sequence R1R2 Mediates Vascular Remodeling by Decreasing Inflammation and Smooth Muscle Proliferation

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Ting-Hein Lee ◽  
Hou-Yu Chiang
Keyword(s):  
Smooth Muscle ◽  
Vascular Disease ◽  
Vascular Remodeling ◽  
Vessel Wall ◽  
Inflammatory Cell ◽  
Vascular Diseases ◽  
Inflammatory Cell Infiltration ◽  
Collagen I ◽  
Cell Infiltration ◽  
Ecm Proteins

The extracellular matrix (ECM) is a major constituent of the vessel wall. Except for providing a structural scaffold for cells, ECM controls numerous cellular functions like adhesion, growth, migration and differentiation. The components of ECM are mediated by the interplay between ECM synthesis, deposition, degradation and the interaction between ECM proteins. Vascular remodeling occurs in the vascular diseases and is characterized by endothelial cell activation, inflammatory cell infiltration, smooth muscle cell (SMC) proliferation/migration, and augmented deposition of ECM proteins. Collagen I is the major ECM component in the arterial wall, excess collagen I accumulation may exacerbate the vascular disease by further facilitating SMC proliferation/migration. Therefore, treatments to inhibit excess collagen deposition could provide a remedy for vascular disease. R1R2, a peptide derived from the bacterial adhesin SFS with sequence homology to collagen, is known to inhibit collagen I deposition by inhibiting the binding of fibronectin to collagen. Studies have revealed that R1R2 affects collagen I-dependent cell growth and migration in vitro. However, the in vivo functions of R1R2 during vascular remodeling remain unknown. We hypothesized that R1R2 prevents excess collagen I accumulation and SMC proliferation, resulting in decreased neointimal formation. We induced vascular remodeling by ligating the carotid artery on mice. Delivery of R1R2 was periadventially applied using pluronic gel and evaluated its effects on vascular remodeling, ECM deposition, SMC proliferation and differentiation. Morphometric analysis demonstrated that R1R2 reduced intima-media thickening by 50% compared to the control group. R1R2 treatment also decreased collagen I deposition in the vessel wall and maintained SMC in the contractile phenotype. Interestingly, R1R2 dramatically reduced inflammatory cell infiltration into the vessel by 80% accompanied with decreased VCAM-1 and ICAM-1. In conclusion, our data showed that R1R2 attenuates the vascular remodeling response by decreasing inflammation and SMC proliferation/migration. These studies provide a therapeutic potential of periadventitially delivering R1R2 in treating vascular diseases.

scholarly journals Activation of Protein Kinases in Canine Basilar Artery in Vasospasm

1999 ◽  
Vol 19 (1) ◽  
pp. 44-52 ◽  
Author(s):  
Hirokazu Fujikawa ◽  
Eiichi Tani ◽  
Ikuya Yamaura ◽  
Isao Ozaki ◽  
Katsuya Miyaji ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Basilar Artery ◽  
Molecular Mechanisms ◽  
Kinase Inhibitor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Intracellular Ca ◽  
Canine Basilar Artery ◽  
Kinase Pathway

Subarachnoid hemorrhage (SAH) often leads to a long-term narrowing of cerebral artery called vasospasm. To understand the molecular mechanisms in vasospasm, signal transduction of tyrosine kinase pathway and phosphorylation of myosin light chain (MLC) and calponin (CaP) in the basilar artery were studied. Vasospasm was produced in the canine basilar artery by a two-hemorrhage method, and vasocontraction was induced by a local application of KCl or serotonin to the basilar artery after a transclival exposure. Intracellular substrates of tyrosine kinase pathway, including Shc, Raf1, and extracellular-regulated kinases in the basilar artery, were activated after SAH, and the activation of Shc suggests stimulation of signal transductions from tyrosine kinase receptors, G-coupled receptors, or both. The activation of tyrosine kinase pathway in vasospasm also was supported by dose-dependent dilation of the spastic basilar artery on days 0 and 7 by topical application of genistein, a tyrosine kinase inhibitor, and associated marked inhibition of tyrosine phosphorylation of intracellular substrates, including Shc. In addition, the generation of protein kinase M, catalytic fragment of protein kinase Cα (PKCα), in vasospasm on days 0 and 7 was inhibited in response to genistein, indicating an inactivation of μ-calpain. It is suggested, therefore, that the reversal of vasospasm by genistein is closely associated with the restoration of intracellular Ca2+ levels. However, the increased activities of Raf1 and extracellular-regulated kinases in vasospasm were declined on day 7 compared with those on day 0 or 2, suggesting that the activation of tyrosine kinase pathway is more closely associated with the early stage of vasospasm than with the late stage of vasospasm. The analysis by pyrophosphate polyacrylamide gel electrophoresis (PPi-PAGE) demonstrated three MLC bands in vasospasm on days 2 and 7, as well as in KCl- and serotonin-induced vasocontraction. Since PPi-PAGE resolves smooth muscle MLC into three bands in the MLC kinase (MLCK)-mediated phosphorylation and into a single band in the PKC-mediated phosphorylation based on the phosphorylation state, the current results suggest that MLC in vasospasm is phosphorylated by MLCK but not by PKC. In basilar artery, CaP was significantly down-regulated, and in addition, significantly phosphorylated on serine and threonine residues only in vasospasm on days 2 and 7. Although the significance of CaP phosphorylations in vivo still is controversial, CaP down-regulation and phosphorylation may attenuate the inhibition of Mg2+-ATPase activity by CaP and induce a potential enhancement of smooth muscle contractility in delayed vasospasm. Since CaP is phosphorylated vivo by PKC, activated PKC in vasospasm may phosphorylate CaP. Thus, SAH stimulates tyrosine kinase pathway to increase intracellular Ca2+ and activate PKC, and the former activates MLCK to phosphorylate MLC, whereas the latter phosphorylates CaP but not MLC.

Epstein-Barr virus-associated smooth muscle tumors in children following solid organ transplantation: A review

2015 ◽  
Vol 19 (2) ◽  
pp. 235-243 ◽  
Author(s):  
Jacqueline Jossen ◽  
Jaime Chu ◽  
Hilary Hotchkiss ◽  
Birte Wistinghausen ◽  
Kishore Iyer ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Organ Transplantation ◽  
Epstein Barr Virus ◽  
Solid Organ Transplantation ◽  
Solid Organ ◽  
Barr Virus ◽  
Smooth Muscle Tumors ◽  
Epstein Barr

Fungal infections in solid organ transplantation

2017 ◽  
Author(s):  
Darius Armstrong James ◽  
Anand Shah ◽  
Anna Reed
Keyword(s):  
Organ Transplantation ◽  
Fungal Infections ◽  
Wide Spectrum ◽  
Treatment Options ◽  
Solid Organ Transplantation ◽  
Invasive Disease ◽  
Global Scale ◽  
Molecular Diagnostic ◽  
Individual Risk ◽  
Solid Organ

Fungal infections are a significant and life-threatening complication of organ transplantation, on a global scale. Risk varies according to transplant type, with liver, lung, and small bowel transplant recipients being at particular risk. Whilst invasive candidiasis is the most common fungal infection in organ transplantation overall, aspergillosis is a particular problem in lung transplantation. In addition, a wide spectrum of fungi may cause invasive disease in organ transplantation, consequently diagnosis and treatment can be challenging. Key challenges are to understand individual risk for infection, appropriate prophylactic strategies, and molecular diagnostic approaches. Treatment options are complicated by drug–drug interactions with transplant therapy, as well as intrinsic allograft dysfunction seen in many patients. In this chapter, we review the epidemiology, risk factors, diagnosis, and management of fungal infections in solid organ transplantation.

KLF5 mediates vascular remodeling via HIF-1α in hypoxic pulmonary hypertension

2016 ◽  
Vol 310 (4) ◽  
pp. L299-L310 ◽  
Author(s):  
Xiaochen Li ◽  
Yuanzhou He ◽  
Yongjian Xu ◽  
Xiaomin Huang ◽  
Jin Liu ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Smooth Muscle ◽  
Right Ventricular ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Systolic Pressure ◽  
Cyclin B1 ◽  
Dependent Manner ◽  
Hypoxic Pulmonary Hypertension ◽  
Pulmonary Arterial

Hypoxic pulmonary hypertension (HPH) is characterized by active vasoconstriction and profound vascular remodeling. KLF5, a zinc-finger transcription factor, is involved in the excessive proliferation and apoptotic resistance phenotype associated with monocrotaline-induced pulmonary hypertension. However, the molecular mechanisms of KLF5-mediated pathogenesis of HPH are largely undefined. Adult male Sprague-Dawley rats were exposed to normoxia or hypoxia (10% O2) for 4 wk. Hypoxic rats developed pulmonary arterial remodeling and right ventricular hypertrophy with significantly increased right ventricular systolic pressure. The levels of KLF5 and hypoxia-inducible factor-1α (HIF-1α) were upregulated in distal pulmonary arterial smooth muscle from hypoxic rats. The knockdown of KLF5 via short-hairpin RNA attenuated chronic hypoxia-induced hemodynamic and histological changes in rats. The silencing of either KLF5 or HIF-1α prevented hypoxia-induced (5%) proliferation and migration and promoted apoptosis in human pulmonary artery smooth muscle cells. KLF5 was immunoprecipitated with HIF-1α under hypoxia and acted as an upstream regulator of HIF-1α. The cell cycle regulators cyclin B1 and cyclin D1 and apoptosis-related proteins including bax, bcl-2, survivin, caspase-3, and caspase-9, were involved in the regulation of KLF5/HIF-1α-mediated cell survival. This study demonstrated that KLF5 plays a crucial role in hypoxia-induced vascular remodeling in an HIF-1α-dependent manner and provided a better understanding of the pathogenesis of HPH.

In situ Evidence of Collagen V and Interleukin-6/Interleukin-17 Activation in Vascular Remodeling of Experimental Pulmonary Hypertension

Pathobiology ◽  
2020 ◽  
Vol 87 (6) ◽  
pp. 356-366
Author(s):  
Sabrina Setembre Batah ◽  
Maiara Almeida Alda ◽  
Rebeca Rodrigues Lopes Roslindo Figueira ◽  
Heloisa R. Cruvinel ◽  
Luis Perdoná Rodrigues da Silva ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cell ◽  
Muscle Cell ◽  
Interleukin 6 ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Yield Potential ◽  
Control Group ◽  
Interleukin 17

Several studies have reported the pathophysiologic and molecular mechanisms responsible for pulmonary arterial hypertension (PAH). However, the in situ evidence of collagen V (Col V) and interleukin-17 (IL-17)/interleukin-6 (IL-6) activation in PAH has not been fully elucidated. We analyzed the effects of collagen I (Col I), Col V, IL-6, and IL-17 on vascular remodeling and hemodynamics and its possible mechanisms of action in monocrotaline (MCT)-induced PAH. Twenty male Wistar rats were randomly divided into two groups. In the PAH group, animals received MCT 60 mg/kg intraperitoneally, whereas the control group (CTRL) received saline. On day 21, the pulmonary blood pressure (PAP) and right ventricular systolic pressure (RVSP) were determined. Lung histology (smooth muscle cell proliferation [α-smooth muscle actin; α-SMA] and periadventitial fibrosis), immunofluorescence (Col I, Col V, and α-SMA), immunohistochemistry (IL-6, IL-17, and transforming growth factor-beta [TGF-β]), and transmission electron microscopy to detect fibronexus were evaluated. The RVSP (40 ± 2 vs. 24 ± 1 mm Hg, respectively; <i>p</i> &#x3c; 0.0001), right ventricle hypertrophy index (65 ± 9 and 25 ± 5%, respectively; <i>p</i> &#x3c; 0.0001), vascular periadventitial Col I and Col V, smooth muscle cell α-SMA+, fibronexus, IL-6, IL-17, and TGF-β were higher in the MCT group than in the CTRL group. In conclusion, our findings indicate in situ evidence of Col V and IL-6/IL-17 activation in vascular remodeling and suggest that increase of Col V may yield potential therapeutic targets for treating patients with PAH.

scholarly journals Activation of TFEB ameliorates dedifferentiation of arterial smooth muscle cells and neointima formation in mice with high-fat diet

2019 ◽  
Vol 10 (9) ◽  
Author(s):  
Yun-Ting Wang ◽  
Xiang Li ◽  
Jiajie Chen ◽  
Bradley K. McConnell ◽  
Li Chen ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
High Fat Diet ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Animal Experiments ◽  
Metabolic Stress ◽  
Neointima Formation ◽  
High Fat ◽  
Autophagy Pathway

Abstract Autophagy is recently implicated in regulating vascular smooth muscle cell (SMC) homeostasis and in the pathogenesis of vascular remodeling. Transcription factor EB (TFEB) is a master regulator of autophagy signaling pathways. However, the molecular mechanisms and functional roles of TFEB in SMC homeostasis have not been elucidated. Here, we surveyed the ability of TFEB to regulate autophagy pathway in SMCs, and whether pharmacological activation of TFEB favors SMC homeostasis preventing dedifferentiation and pathogenic vascular remodeling. In primary cultured SMCs, TFEB activator trehalose induced nuclear translocation of TFEB and upregulation of TFEB-controlled autophagy genes leading to enhanced autophagy signaling. Moreover, trehalose suppressed serum-induced SMC dedifferentiation to synthetic phenotypes as characterized by inhibited proliferation and migration. These effects of trehalose were mimicked by ectopic upregulation of TFEB and inhibited by TFEB gene silencing. In animal experiments, partial ligation of carotid arteries induced downregulation of TFEB pathway in the media layer of these arteries. Such TFEB suppression was correlated with increased SMC dedifferentiation and aggravated high-fat diet (HFD)-induced neointima formation. Treatment of mice with trehalose reversed this TFEB pathway suppression, and prevented SMC dedifferentiation and HFD-induced neointima formation. In conclusion, our findings have identified TFEB as a novel positive regulator for autophagy pathway and cellular homeostasis in SMCs. Our data suggest that suppression of TFEB may be an initiating mechanism that promotes SMC dedifferentiation leading to accelerated neointima formation in vascular disorders associated with metabolic stress, whereas trehalose reverses these changes. These findings warrant further evaluation of trehalose in the clinical settings.

scholarly journals miR-4632 mediates PDGF-BB-induced proliferation and antiapoptosis of human pulmonary artery smooth muscle cells via targeting cJUN

2017 ◽  
Vol 313 (4) ◽  
pp. C380-C391 ◽  
Author(s):  
Zhengjiang Qian ◽  
Yanjiao Li ◽  
Jidong Chen ◽  
Xiang Li ◽  
Deming Gou
Keyword(s):  
Smooth Muscle ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Muscle Cells ◽  
Pulmonary Vascular Remodeling ◽  
Proliferation And Apoptosis ◽  
Pulmonary Artery Smooth Muscle ◽  
Human Pulmonary Artery

MicroRNAs (miRNAs) can regulate the proliferative status of pulmonary artery smooth muscle cells (PASMCs), which is a core factor modulating pulmonary vascular remodeling diseases, such as atherosclerosis and pulmonary arterial hypertension (PAH). Our previous work has shown that miR-4632, a rarely reported miRNA, is significantly downregulated in platelet-derived growth factor (PDGF)-BB-stimulated human pulmonary artery smooth muscle cells (HPASMCs), yet its cell function and the underlying molecular mechanisms remain to be elucidated. Here, we find that miR-4632 is highly expressed in HPASMCs and its expression significantly decreased in response to different stimuli. Functional studies revealed that miR-4632 inhibited proliferation and promoted apoptosis of HPASMCs but had no effects on cell contraction and migration. Furthermore, the cJUN was identified as a direct target gene of miR-4632, while knockdown of cJUN was necessary for miR-4632-mediated HPASMC proliferation and apoptosis. In addition, the downregulation of miR-4632 by PDGF-BB was found to associate with histone deacetylation through the activation of PDGF receptor/phosphatidylinositol 3′-kinase/histone deacetylase 4 signaling. Finally, the expression of miR-4632 was reduced in the serum of patients with PAH. Overall, our results suggest that miR-4632 plays an important role in regulating HPASMC proliferation and apoptosis by suppression of cJUN, providing a novel therapeutic miRNA candidate for the treatment of pulmonary vascular remodeling diseases. It also implies that serum miR-4632 has the potential to serve as a circulating biomarker for PAH diagnosis.

Abstract 458: Role of Colony Stimulating Factor 1 Receptor in Graft Vascular Disease

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Vanessa Almonte ◽  
Oriyanhan Wunimenghe ◽  
Dario F Riascos-Bernal ◽  
Prameladevi Chinnasamy ◽  
Smitha Jayakumar ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Disease ◽  
Neointimal Hyperplasia ◽  
Lineage Tracing ◽  
Vascular Lesions ◽  
Solid Organ ◽  
Colony Stimulating Factor ◽  
Male Mice ◽  
The Media ◽  
Stimulating Factor

Heart and kidney transplants are effective treatments for end-stage organ failure, but their long-term success is limited by graft vascular disease (GVD), the leading cause of solid organ transplant failure. This process manifests as concentric thickening of vessel walls due to neointimal hyperplasia in the donor organ, characterized by expansion of cells, notably smooth muscle-like cells (SMLCs) and macrophages (MPs), which accumulate, proliferate, and eventually occlude the lumen of arteries. Our lab recently reported that loss of colony stimulating factor-1 (CSF1) expression in either donor or recipient mice limits GVD, and showed that SMLCs isolated from neointimal lesions express high levels of CSF1 and its receptor, CSF1R. While CSF1-mediated activation of CSF1R has been studied extensively in MP biology, its role in SMLCs and GVD has not been well characterized. We hypothesize that CSF1R activation in neointimal MPs and SMCs occurs after organ transplantation and promotes the development of GVD. To test this idea, carotid arteries from 8-12 week old C57/B6J male mice were transplanted orthotopically into female or male mice. At day 30 post transplantation, sex-mismatched transplants developed significant neointimal lesions not seen in sex-matched controls. Neointimal, adventitial, and to a lesser extent medial cells were positive for CSF1R and CD68, which were scarcely detected in control untransplanted arteries. Cells in the media of transplanted vessels co-stained for smooth muscle alpha actin (SMA) and calponin. SMA-positive cells were found in neointimal lesions, with few cells co-expressing calponin. Proliferation, assessed by Phospho-histone H3 staining, was evident in cells of uncertain origin in the media and neointima. In conclusion, H-Y antigen-driven histoincompatibility in this mouse transplant model yielded vascular lesions that resemble GVD, with significant neointima formation, preservation of medial cells, and evidence of CSF1R expression and of cell proliferation. Future studies will focus on lineage tracing of smooth muscle and myeloid cells to evaluate neointimal cell origins, plus genetic depletion of CSF1R in these cell lineages to determine the requirement for CSF1R expression in the development of GVD.

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