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1.
Biomed Pharmacother ; 147: 112664, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35131655

RESUMEN

The lymphatic vascular system is crucial for maintaining tissue fluid homeostasis and immune surveillance. Promoting lymphatic function represents a new strategy to treat several diseases including lymphedema, chronic inflammation and impaired wound healing. By screening a plant extract library, a petroleum ether extract from the aerial parts of Eupatorium perfoliatum (E. perfoliatum) was found to possess lymphangiogenic properties. With the aid of HPLC activity profiling the active compound was identified as pheophorbide a. Both plant extract and pheophorbide a induced the sprouting and tube formation of human primary lymphatic endothelial cells (LECs). The proliferation of the LECs was increased upon treatment with pheophorbide a but not the E. perfoliatum extract. Treatment with the MEK1/2 inhibitor U0126 reduced the LEC sprouting activity, indicating a potential mechanism of action. These studies suggest that pheophorbide a could represent novel natural therapeutic agent to treat human lymphatic vascular insufficiencies.


Asunto(s)
Clorofila/análogos & derivados , Células Endoteliales/efectos de los fármacos , Eupatorium , Linfangiogénesis/efectos de los fármacos , Extractos Vegetales/farmacología , Butadienos/farmacología , Línea Celular , Clorofila/farmacología , Humanos , Vasos Linfáticos/efectos de los fármacos , MAP Quinasa Quinasa 1/antagonistas & inhibidores , MAP Quinasa Quinasa 2/antagonistas & inhibidores , Nitrilos/farmacología
2.
Microvasc Res ; 139: 104274, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34717967

RESUMEN

OBJECTIVE: Besides actions including their venotonic, anti-inflammatory, and anti-oxidant effects, venoactive drugs are expected to act on edema via their action on lymphatics. The objective of this study was to evaluate the effect of the combination of Ruscus, hesperidin methyl chalcone and Vitamin C (Ruscus/HMC/Vit C) on intracellular calcium mobilization and contraction of human lymphatic smooth muscle cells (LSMCs) to better characterize the mechanism of its lymphotonic activity. METHODS: Calcium mobilization was evidenced by videomicroscopy analysis of the fluorescence emitted by a specific Ca2+ sensitive dye and measured after injection of Ruscus/HMC/Vit C at 0.1, 0.3, 1.0, and 3.0 mg/mL into LSMCs. RESULTS: Ruscus/HMC/Vit C induced a strong and reproducible concentration-dependent calcium mobilization in LSMCs. On the contrary, another venoactive drug used as comparator, micronized purified flavonoid fraction (MPFF), did not induce calcium mobilization whatever the tested concentration. CONCLUSION: Although alternative mechanisms of action may result in potential lymphotonic effects, the efficacy of lymphotonic products is nonetheless related to their stimulating effect on the contractile activity of the smooth muscle cells surrounding lymphatic vessels. In the light of the results obtained in this study, the direct effect of Ruscus/HMC/Vit C on LSMC contraction may partially explain its clinical efficacy on lymphotonic activity, as has been observed in terms of objective signs of edema as reported in the recent guidelines on chronic venous disease.


Asunto(s)
Ácido Ascórbico/farmacología , Chalconas/farmacología , Hesperidina/análogos & derivados , Vasos Linfáticos/efectos de los fármacos , Contracción Muscular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Extractos Vegetales/farmacología , Ruscus , Calcio/metabolismo , Señalización del Calcio , Células Cultivadas , Diosmina/farmacología , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Regulación de la Expresión Génica , Hesperidina/farmacología , Humanos , Vasos Linfáticos/metabolismo , Masculino , Persona de Mediana Edad , Miocitos del Músculo Liso/metabolismo , Extractos Vegetales/aislamiento & purificación , Ruscus/química , Factores de Tiempo
3.
Microvasc Res ; 139: 104273, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34699844

RESUMEN

The lymphatic system plays important roles in various physiological and pathological phenomena. As a bioactive phospholipid, lysophosphatidic acid (LPA) has been reported to function as a lymphangiogenic factor as well as some growth factors, yet the involvement of phospholipids including LPA and its derivatives in lymphangiogenesis is not fully understood. In the present study, we have developed an in-vitro lymphangiogenesis model (termed a collagen sandwich model) by utilizing type-I collagen, which exists around the lymphatic endothelial cells of lymphatic capillaries in vivo. The collagen sandwich model has revealed that cyclic phosphatidic acid (cPA), and not LPA, augmented the tube formation of human dermal lymphatic endothelial cells (HDLECs). Both cPA and LPA increased the migration of HDLECs cultured on the collagen. As the gene expression of LPA receptor 6 (LPA6) was predominantly expressed in HDLECs, a siRNA experiment against LPA6 attenuated the cPA-mediated tube formation. A synthetic LPA1/3 inhibitor, Ki16425, suppressed the cPA-augmented tube formation and migration of the HDLECs, and the LPA-induced migration. The activity of Rho-associated protein kinase (ROCK) located at the downstream of the LPA receptors was augmented in both the cPA- and LPA-treated cells. A potent ROCK inhibitor, Y-27632, suppressed the cPA-dependent tube formation but not the migration of the HDLECs. Furthermore, cPA, but not LPA, augmented the gene expression of VE-cadherin and ß-catenin in the HDLECs. These results provide novel evidence that cPA facilitates the capillary-like morphogenesis and the migration of HDLECs through LPA6/ROCK and LPA1/3 signaling pathways in concomitance with the augmentation of VE-cadherin and ß-catenin expression. Thus, cPA is likely to be a potent lymphangiogenic factor for the initial lymphatics adjacent to type I collagen under physiological conditions.


Asunto(s)
Células Endoteliales/efectos de los fármacos , Linfangiogénesis/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Lisofosfolípidos/farmacología , Ácidos Fosfatidicos/farmacología , Antígenos CD/genética , Antígenos CD/metabolismo , Cadherinas/genética , Cadherinas/metabolismo , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Colágeno Tipo I/metabolismo , Células Endoteliales/metabolismo , Humanos , Vasos Linfáticos/metabolismo , Masculino , Receptores del Ácido Lisofosfatídico/agonistas , Receptores del Ácido Lisofosfatídico/genética , Receptores del Ácido Lisofosfatídico/metabolismo , Transducción de Señal , beta Catenina/genética , beta Catenina/metabolismo , Quinasas Asociadas a rho/metabolismo
4.
Dev Cell ; 56(22): 3128-3145.e15, 2021 11 22.
Artículo en Inglés | MEDLINE | ID: mdl-34762852

RESUMEN

Identification of physiological modulators of nuclear hormone receptor (NHR) activity is paramount for understanding the link between metabolism and transcriptional networks that orchestrate development and cellular physiology. Using libraries of metabolic enzymes alongside their substrates and products, we identify 1-deoxysphingosines as modulators of the activity of NR2F1 and 2 (COUP-TFs), which are orphan NHRs that are critical for development of the nervous system, heart, veins, and lymphatic vessels. We show that these non-canonical alanine-based sphingolipids bind to the NR2F1/2 ligand-binding domains (LBDs) and modulate their transcriptional activity in cell-based assays at physiological concentrations. Furthermore, inhibition of sphingolipid biosynthesis phenocopies NR2F1/2 deficiency in endothelium and cardiomyocytes, and increases in 1-deoxysphingosine levels activate NR2F1/2-dependent differentiation programs. Our findings suggest that 1-deoxysphingosines are physiological regulators of NR2F1/2-mediated transcription.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Organogénesis/efectos de los fármacos , Esfingolípidos/farmacología , Animales , Diferenciación Celular/fisiología , Regulación de la Expresión Génica/fisiología , Humanos , Vasos Linfáticos/efectos de los fármacos , Ratones , Organogénesis/fisiología , Proteínas Represoras/fisiología
5.
Am J Physiol Renal Physiol ; 321(6): F675-F688, 2021 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-34658261

RESUMEN

Expansion of renal lymphatic networks, or lymphangiogenesis (LA), is well recognized during development and is now being implicated in kidney diseases. Although LA is associated with multiple pathological conditions, very little is known about its role in acute kidney injury. The purpose of this study was to evaluate the role of LA in a model of cisplatin-induced nephrotoxicity. LA is predominately regulated by vascular endothelial growth factor (VEGF)-C and VEGF-D, ligands that exert their function through their cognate receptor VEGF receptor 3 (VEGFR3). We demonstrated that use of MAZ51, a selective VEGFR3 inhibitor, caused significantly worse structural and functional kidney damage in cisplatin nephrotoxicity. Apoptotic cell death and inflammation were also increased in MAZ51-treated animals compared with vehicle-treated animals following cisplatin administration. Notably, MAZ51 caused significant upregulation of intrarenal phospho-NF-κB, phospho-JNK, and IL-6. Cisplatin nephrotoxicity is associated with vascular congestion due to endothelial dysfunction. Using three-dimensional tissue cytometry, a novel approach to explore lymphatics in the kidney, we detected significant vascular autofluorescence attributed to erythrocytes in cisplatin alone-treated animals. Interestingly, no such congestion was detected in MAZ51-treated animals. We found increased renal vascular damage in MAZ51-treated animals, whereby MAZ51 caused a modest decrease in the endothelial markers endomucin and von Willebrand factor, with a modest increase in VEGFR2. Our findings identify a protective role for de novo LA in cisplatin nephrotoxicity and provide a rationale for the development of therapeutic approaches targeting LA. Our study also suggests off-target effects of MAZ51 on the vasculature in the setting of cisplatin nephrotoxicity.NEW & NOTEWORTHY Little is known about injury-associated LA in the kidney and its role in the pathophysiology of acute kidney injury (AKI). Observed exacerbation of cisplatin-induced AKI after LA inhibition was accompanied by increased medullary damage and cell death in the kidney. LA inhibition also upregulated compensatory expression of LA regulatory proteins, including JNK and NF-κB. These data support the premise that LA is induced during AKI and lymphatic expansion is a protective mechanism in cisplatin nephrotoxicity.


Asunto(s)
Indoles/toxicidad , Enfermedades Renales/inducido químicamente , Riñón/efectos de los fármacos , Linfangiogénesis/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Naftalenos/toxicidad , Inhibidores de Proteínas Quinasas/toxicidad , Receptor 3 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Animales , Apoptosis/efectos de los fármacos , Cisplatino , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Riñón/enzimología , Riñón/patología , Riñón/fisiopatología , Enfermedades Renales/enzimología , Enfermedades Renales/patología , Enfermedades Renales/fisiopatología , Vasos Linfáticos/enzimología , Vasos Linfáticos/patología , Vasos Linfáticos/fisiopatología , Masculino , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Fosforilación , Transducción de Señal , Receptor 3 de Factores de Crecimiento Endotelial Vascular/metabolismo
6.
PLoS One ; 16(9): e0257175, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34516572

RESUMEN

BACKGROUNDS: We demonstrated that coronary adventitial inflammation plays important roles in the pathogenesis of drug-eluting stent (DES)-induced coronary hyperconstricting responses in pigs in vivo. However, no therapy is yet available to treat coronary adventitial inflammation. We thus developed the low-intensity pulsed ultrasound (LIPUS) therapy that ameliorates myocardial ischemia by enhancing angiogenesis. AIMS: We aimed to examine whether our LIPUS therapy suppresses DES-induced coronary hyperconstricting responses in pigs in vivo, and if so, what mechanisms are involved. METHODS: Sixteen normal male pigs were randomly assigned to the LIPUS or the sham therapy groups after DES implantation into the left anterior descending (LAD) coronary artery. In the LIPUS group, LIPUS (32 cycles, 193 mW/cm2) was applied to the heart at 3 different levels (segments proximal and distal to the stent edges and middle of the stent) for 20 min at each level for every other day for 2 weeks. The sham therapy group was treated in the same manner but without LIPUS. At 4 weeks after stent implantation, we performed coronary angiography, followed by immunohistological analysis. RESULTS: Coronary vasoconstricting responses to serotonin in LAD at DES edges were significantly suppressed in the LIPUS group compared with the sham group. Furthermore, lymph transport speed in vivo was significantly faster in the LIPUS group than in the sham group. Histological analysis at DES edges showed that inflammatory changes and Rho-kinase activity were significantly suppressed in the LIPUS group, associated with eNOS up-regulation and enhanced lymph-angiogenesis. CONCLUSIONS: These results suggest that our non-invasive LIPUS therapy is useful to treat coronary functional abnormalities caused by coronary adventitial inflammation, indicating its potential for the novel and safe therapeutic approach of coronary artery disease.


Asunto(s)
Adventicia/patología , Implantación de Prótesis Vascular , Vasos Coronarios/patología , Vasos Coronarios/fisiopatología , Stents Liberadores de Fármacos , Inflamación/terapia , Ondas Ultrasónicas , Vasoconstricción , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , Adventicia/efectos de los fármacos , Adventicia/fisiopatología , Animales , Vasos Coronarios/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Inflamación/patología , Linfangiogénesis/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/fisiopatología , Modelos Biológicos , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Serotonina/metabolismo , Porcinos , Vasoconstricción/efectos de los fármacos , Quinasas Asociadas a rho/metabolismo
7.
Mar Drugs ; 19(9)2021 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-34564157

RESUMEN

Increased inflammation is the main pathophysiology of nonalcoholic fatty liver disease (NAFLD). Inflammation affects lymphatic vessel function that contributes to the removal of immune cells or macromolecules. Dysfunctional lymphatic vessels with decreased permeability are present in NAFLD. High-fat diet (HFD) is known to increase body weight, food intake, and inflammation in the liver. Previously, it was reported that Ecklonia cava extracts (ECE) decreased food intake or weight gain, and low-calorie diet and weight loss is known as a treatment for NAFLD. In this study, the effects of ECE and dieckol (DK)-which is one component of ECE that decreases inflammation and increases lymphangiogenesis and lymphatic drainage by controlling lymphatic permeability in high-fat diet (HFD)-fed mice-on weight gain and food intake were investigated. ECE and DK decreased weight gain and food intake in the HFD-fed mice. NAFLD activities such as steatosis, lobular inflammation, and ballooning were increased by HFD and attenuated by ECE and DK. The expression of inflammatory cytokines such as IL-6 and TNF-α and infiltration of M1 macrophages were increased by HFD, and they were decreased by ECE or DK. The signaling pathways of lymphangiogenesis, VEGFR-3, PI3K/pAKT, and pERK were decreased by HFD, and they were restored by either ECE or DK. The expression of VE-cadherin (which represents lymphatic junctional function) was increased by HFD, although it was restored by either ECE or DK. In conclusion, ECE and DK attenuated NAFLD by decreasing weight gain and food intake, decreasing inflammation, and increasing lymphangiogenesis, as well as modulating lymphatic vessel permeability.


Asunto(s)
Antiinflamatorios/uso terapéutico , Benzofuranos/uso terapéutico , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Phaeophyceae , Extractos Vegetales/uso terapéutico , Administración Oral , Animales , Antiinflamatorios/administración & dosificación , Antiinflamatorios/farmacología , Organismos Acuáticos , Benzofuranos/administración & dosificación , Benzofuranos/farmacología , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Ingestión de Energía/efectos de los fármacos , Hígado/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología
8.
Sci Rep ; 11(1): 16801, 2021 08 19.
Artículo en Inglés | MEDLINE | ID: mdl-34413352

RESUMEN

The lymphatic network of mammalian heart is an important regulator of interstitial fluid compartment and immune cell trafficking. We observed a remodeling of the cardiac lymphatic vessels and a reduced lymphatic efficiency during heart hypertrophy and failure induced by transverse aortic constriction. The lymphatic endothelial cell number of the failing hearts was positively correlated with cardiac function and with a subset of cardiac macrophages. This macrophage population distinguished by LYVE-1 (Lymphatic vessel endothelial hyaluronic acid receptor-1) and by resident macrophage gene expression signature, appeared not replenished by CCR2 mediated monocyte infiltration during pressure overload. Isolation of macrophage subpopulations showed that the LYVE-1 positive subset sustained in vitro and in vivo lymphangiogenesis through the expression of pro-lymphangiogenic factors. In contrast, the LYVE-1 negative macrophage subset strongly expressed MMP12 and decreased the endothelial LYVE-1 receptors in lymphatic endothelial cells, a feature of cardiac lymphatic remodeling in failing hearts. The treatment of mice with a CCR2 antagonist during pressure overload modified the proportion of macrophage subsets within the pathological heart and preserved lymphatic network from remodeling. This study reports unknown and differential functions of macrophage subpopulations in the regulation of cardiac lymphatic during pathological hypertrophy and may constitute a key mechanism underlying the progression of heart failure.


Asunto(s)
Vasos Linfáticos/metabolismo , Macrófagos/metabolismo , Miocardio/patología , Presión , Animales , Benzoxazinas/farmacología , Células CHO , Polaridad Celular/efectos de los fármacos , Cricetulus , Electrocardiografía , Células Endoteliales/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Linfangiogénesis/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Masculino , Ratones Endogámicos C57BL , Monocitos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores CCR2/metabolismo , Compuestos de Espiro/farmacología , Transcriptoma , Proteínas de Transporte Vesicular/metabolismo
9.
Bull Exp Biol Med ; 171(2): 281-285, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-34173919

RESUMEN

We used specific histochemical fluorescence-microscopic method of visualization of catecholamines to study adrenergic innervation of the thyroid gland tissue, blood vessels of the thyroid gland, cervical lymphatic vessel and lymph nodes in rats during correction of hypothyroidism with a bioactive formulation (Vozrozhdenie Plus balm with Potentilla alba L.). In experimental hypothyroidism, adrenergic innervation of the thyroid gland and the wall of the cervical lymph node, concentrated mainly along the arterial vessels and the cervical lymphatic vessel, retained its structural formations (plexuses and varicosities), but diffusion of catecholamines outside these formations was observed. Correction with the bioactive formulation restored of the contours of the nerve plexuses and varicosities and their brighter fluorescence in the thyroid gland and cervical lymphatic vessel and node. During correction of hypothyroidism with the bioactive formulation, reorganization of regional lymphatic vessels and nodes was more pronounced than reorganization of the thyroid gland.


Asunto(s)
Hipotiroidismo , Ganglios Linfáticos/patología , Vasos Linfáticos/patología , Glándula Tiroides/irrigación sanguínea , Glándula Tiroides/inervación , Fibras Adrenérgicas/efectos de los fármacos , Fibras Adrenérgicas/patología , Fibras Adrenérgicas/ultraestructura , Animales , Vasos Sanguíneos/diagnóstico por imagen , Vasos Sanguíneos/efectos de los fármacos , Vasos Sanguíneos/patología , Hipotiroidismo/diagnóstico por imagen , Hipotiroidismo/tratamiento farmacológico , Hipotiroidismo/patología , Ganglios Linfáticos/diagnóstico por imagen , Ganglios Linfáticos/efectos de los fármacos , Vasos Linfáticos/diagnóstico por imagen , Vasos Linfáticos/efectos de los fármacos , Masculino , Microscopía Fluorescente , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Yoduro de Potasio/farmacología , Yoduro de Potasio/uso terapéutico , Ratas , Glándula Tiroides/diagnóstico por imagen , Glándula Tiroides/efectos de los fármacos , Hormonas Tiroideas/farmacología , Hormonas Tiroideas/uso terapéutico
10.
Cells ; 10(5)2021 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-34067671

RESUMEN

Despite improvement during the last ten years in the longevity of patients with metastatic clear cell renal cell carcinoma (mccRCC) the disease remains incurable. Hence, new therapeutic strategies are urgently needed. Relapse following anti-angiogenic treatment depends on the over-expression of vascular endothelial growth factor C (VEGFC), one of the main drivers of lymphangiogenesis. Therefore, we developed specific mouse monoclonal antibodies and evaluated their therapeutic efficacy in vitro and in vivo. Immunization of mice with the domain of VEGFC that stimulates the VEGF receptor 3 (VEGFR3) led to the selection of one hybridoma producing specific anti-VEGFC monoclonal antibodies. The selected 1E9 antibodies were sequenced, and the corresponding variable light and heavy chains were subcloned into expression vectors in frame with sequences encoding the human IgG1 constant heavy and light chains. CHO cells were stably transfected and cloned to produce chimeric antibodies. These antibodies inhibited the activation of VEGFR3 signaling, and therefore the proliferation and migration of VEGFC-stimulated endothelial cells. Moreover, they inhibited the proliferation of VEGFC-expressing renal cancer cells through NRP2 signaling. 1E9 antibodies inhibited the growth of experimental RCC, and their therapeutic efficacy was enhanced by the anti-VEGF antibody bevacizumab. Hence, our results suggest that targeting VEGFC could have a relevant therapeutic impact on mccRCC that relapse following anti-angiogenic treatment.


Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Antineoplásicos Inmunológicos/farmacología , Carcinoma de Células Renales/tratamiento farmacológico , Células Endoteliales/efectos de los fármacos , Neoplasias Renales/tratamiento farmacológico , Neovascularización Patológica , Factor C de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Animales , Carcinoma de Células Renales/metabolismo , Carcinoma de Células Renales/patología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Endoteliales/metabolismo , Células Endoteliales/patología , Femenino , Humanos , Neoplasias Renales/metabolismo , Neoplasias Renales/patología , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/metabolismo , Vasos Linfáticos/patología , Ratones Desnudos , Transducción de Señal , Carga Tumoral/efectos de los fármacos , Factor C de Crecimiento Endotelial Vascular/inmunología , Factor C de Crecimiento Endotelial Vascular/metabolismo , Receptor 3 de Factores de Crecimiento Endotelial Vascular/metabolismo , Ensayos Antitumor por Modelo de Xenoinjerto
11.
Nat Commun ; 12(1): 3460, 2021 06 08.
Artículo en Inglés | MEDLINE | ID: mdl-34103491

RESUMEN

Lack or dysfunction of the lymphatics leads to secondary lymphedema formation that seriously reduces the function of the affected organs and results in degradation of quality of life. Currently, there is no definitive treatment option for lymphedema. Here, we utilized nucleoside-modified mRNA encapsulated in lipid nanoparticles (LNPs) encoding murine Vascular Endothelial Growth Factor C (VEGFC) to stimulate lymphatic growth and function and reduce experimental lymphedema in mouse models. We demonstrated that administration of a single low-dose of VEGFC mRNA-LNPs induced durable, organ-specific lymphatic growth and formation of a functional lymphatic network. Importantly, VEGFC mRNA-LNP treatment reversed experimental lymphedema by restoring lymphatic function without inducing any obvious adverse events. Collectively, we present a novel application of the nucleoside-modified mRNA-LNP platform, describe a model for identifying the organ-specific physiological and pathophysiological roles of the lymphatics, and propose an efficient and safe treatment option that may serve as a novel therapeutic tool to reduce lymphedema.


Asunto(s)
Linfangiogénesis/genética , Vasos Linfáticos/patología , Linfedema/patología , Nucleósidos/metabolismo , Factor C de Crecimiento Endotelial Vascular/genética , Animales , Vasos Sanguíneos/patología , Proliferación Celular/efectos de los fármacos , Toxina Diftérica/farmacología , Modelos Animales de Enfermedad , Células HEK293 , Humanos , Inmunidad/efectos de los fármacos , Inyecciones Intradérmicas , Lípidos/administración & dosificación , Lípidos/química , Vasos Linfáticos/efectos de los fármacos , Ratones Endogámicos C57BL , Nanopartículas/administración & dosificación , Nanopartículas/química , Especificidad de Órganos , Poli C/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Tamoxifeno/farmacología , Factor C de Crecimiento Endotelial Vascular/administración & dosificación , Factor C de Crecimiento Endotelial Vascular/metabolismo
12.
Adv Drug Deliv Rev ; 175: 113833, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34147531

RESUMEN

Imaging of the lymphatic vasculature has gained great attention in various fields, not only because lymphatic vessels act as a key draining system in the body, but also for their implication in autoimmune diseases, organ transplant, inflammation and cancer. Thus, neolymphangiogenesis, or the generation of new lymphatics, is typically an early event in the development of multiple tumor types, particularly in aggressive ones such as malignant melanoma. Still, the understanding of how lymphatic endothelial cells get activated at distal (pre)metastatic niches and their impact on therapy is still unclear. Addressing these questions is of particular interest in the case of immune modulators, because endothelial cells may favor or halt inflammatory processes depending on the cellular context. Therefore, there is great interest in visualizing the lymphatic vasculature in vivo. Here, we review imaging tools and mouse models used to analyze the lymphatic vasculature during tumor progression. We also discuss therapeutic approaches based on nanomedicines to target the lymphatic system and the potential use of extracellular vesicles to track and target sentinel lymph nodes. Finally, we summarize main pre-clinical models developed to visualize the lymphatic vasculature in vivo, discussing their applications with a particular focus in metastatic melanoma.


Asunto(s)
Vesículas Extracelulares/metabolismo , Sistema Linfático/diagnóstico por imagen , Sistema de Administración de Fármacos con Nanopartículas , Animales , Vesículas Extracelulares/patología , Humanos , Ganglios Linfáticos/diagnóstico por imagen , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/patología , Sistema Linfático/efectos de los fármacos , Sistema Linfático/patología , Vasos Linfáticos/diagnóstico por imagen , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/patología , Ganglio Linfático Centinela/diagnóstico por imagen , Ganglio Linfático Centinela/patología
14.
Int J Mol Sci ; 22(9)2021 Apr 24.
Artículo en Inglés | MEDLINE | ID: mdl-33923272

RESUMEN

The mechanisms of lymphedema development are not well understood, but emerging evidence highlights the crucial role the immune system plays in driving its progression. It is well known that lymphatic function deteriorates as lymphedema progresses; however, the connection between this progressive loss of function and the immune-driven changes that characterize the disease has not been well established. In this study, we assess changes in leukocyte populations in lymph nodes within the lymphatic drainage basin of the tissue injury site (draining lymph nodes, dLNs) using a mouse tail model of lymphedema in which a pair of draining collecting vessels are left intact. We additionally quantify lymphatic pump function using established near infrared (NIR) lymphatic imaging methods and lymph-draining nanoparticles (NPs) synthesized and employed by our team for lymphatic tissue drug delivery applications to measure lymphatic transport to and resulting NP accumulation within dLNs associated with swelling following surgery. When applied to assess the effects of the anti-inflammatory drug bestatin, which has been previously shown to be a possible treatment for lymphedema, we find lymph-draining NP accumulation within dLNs and lymphatic function to increase as lymphedema progresses, but no significant effect on leukocyte populations in dLNs or tail swelling. These results suggest that ameliorating this loss of lymphatic function is not sufficient to reverse swelling in this surgically induced disease model that better recapitulates the extent of lymphatic injury seen in human lymphedema. It also suggests that loss of lymphatic function during lymphedema may be driven by immune-mediated mechanisms coordinated in dLNs. Our work indicates that addressing both lymphatic vessel dysfunction and immune cell expansion within dLNs may be required to prevent or reverse lymphedema when partial lymphatic function is sustained.


Asunto(s)
Modelos Animales de Enfermedad , Leucina/análogos & derivados , Leucocitos/inmunología , Leucotrieno B4/antagonistas & inhibidores , Ganglios Linfáticos/inmunología , Vasos Linfáticos/inmunología , Linfedema/inmunología , Animales , Femenino , Cinética , Leucina/farmacología , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Leucocitos/patología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/patología , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/metabolismo , Vasos Linfáticos/patología , Linfedema/tratamiento farmacológico , Linfedema/metabolismo , Linfedema/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Inhibidores de Proteasas/farmacología
15.
Pediatr Hematol Oncol ; 38(4): 367-377, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33641614

RESUMEN

Multifocal lymphangioendotheliomatosis with thrombocytopenia (MLT) is a recently recognized disorder characterized by vascular lesions marked by distinct endothelial proliferation. Lesions affect multiple tissues, and MLT can be associated with refractory thrombocytopenia resulting in life-threatening bleeding. Diagnosing MLT may be challenging given its rarity and phenotypic variability. There is no consensus on the optimal management or treatment duration. We report a 4-month-old male who presented with multiple vascular malformations involving the gastrointestinal tract, lung, bones, choroid plexus, and spleen, with minimal cutaneous involvement and no thrombocytopenia. Wedge resection of a pulmonary nodule was strongly positive for lymphatic vessel endothelial hyaluronan receptor 1 favoring MLT despite the lack of thrombocytopenia. The patient's clinical symptoms and vascular lesions improved on sirolimus therapy. We review the literature to highlight the clinical variability of MLT and discuss the diagnostic and therapeutic options for MLT.


Asunto(s)
Angiomatosis/tratamiento farmacológico , Inmunosupresores/uso terapéutico , Vasos Linfáticos/patología , Sirolimus/uso terapéutico , Trombocitopenia/tratamiento farmacológico , Angiomatosis/complicaciones , Angiomatosis/patología , Endotelio Linfático/efectos de los fármacos , Endotelio Linfático/patología , Humanos , Lactante , Vasos Linfáticos/efectos de los fármacos , Masculino , Trombocitopenia/complicaciones , Trombocitopenia/patología
17.
J Pharmacol Exp Ther ; 376(1): 40-50, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33100270

RESUMEN

Pharmacological openers of ATP-sensitive potassium (KATP) channels are effective antihypertensive agents, but off-target effects, including severe peripheral edema, limit their clinical usefulness. It is presumed that the arterial dilation induced by KATP channel openers (KCOs) increases capillary pressure to promote filtration edema. However, KATP channels also are expressed by lymphatic muscle cells (LMCs), raising the possibility that KCOs also attenuate lymph flow to increase interstitial fluid. The present study explored the effect of KCOs on lymphatic contractile function and lymph flow. In isolated rat mesenteric lymph vessels (LVs), the prototypic KATP channel opener cromakalim (0.01-3 µmol/l) progressively inhibited rhythmic contractions and calculated intraluminal flow. Minoxidil sulfate and diazoxide (0.01-100 µmol/l) had similar effects at clinically relevant plasma concentrations. High-speed in vivo imaging of the rat mesenteric lymphatic circulation revealed that superfusion of LVs with cromakalim and minoxidil sulfate (0.01-10 µmol/l) maximally decreased lymph flow in vivo by 38.4% and 27.4%, respectively. Real-time polymerase chain reaction and flow cytometry identified the abundant KATP channel subunits in LMCs as the pore-forming Kir6.1/6.2 and regulatory sulfonylurea receptor 2 subunits. Patch-clamp studies detected cromakalim-elicited unitary K+ currents in cell-attached patches of LMCs with a single-channel conductance of 46.4 pS, which is a property consistent with Kir6.1/6.2 tetrameric channels. Addition of minoxidil sulfate and diazoxide elicited unitary currents of similar amplitude. Collectively, our findings indicate that KCOs attenuate lymph flow at clinically relevant plasma concentrations as a potential contributing mechanism to peripheral edema. SIGNIFICANCE STATEMENT: ATP-sensitive potassium (KATP) channel openers (KCOs) are potent antihypertensive medications, but off-target effects, including severe peripheral edema, limit their clinical use. Here, we demonstrate that KCOs impair the rhythmic contractions of lymph vessels and attenuate lymph flow, which may promote edema formation. Our finding that the KATP channels in lymphatic muscle cells may be unique from their counterparts in arterial muscle implies that designing arterial-selective KCOs may avoid activation of lymphatic KATP channels and peripheral edema.


Asunto(s)
Edema/etiología , Canales KATP/metabolismo , Vasos Linfáticos/fisiología , Contracción Muscular , Potenciales de Acción , Animales , Células Cultivadas , Cromakalim/farmacología , Diazóxido/farmacología , Canales KATP/agonistas , Canales KATP/genética , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/metabolismo , Masculino , Minoxidil/análogos & derivados , Minoxidil/farmacología , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/fisiología , Potasio/metabolismo , Ratas , Ratas Sprague-Dawley
18.
Pediatr Res ; 89(1): 110-117, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32279070

RESUMEN

BACKGROUND: To investigate whether the YAP/TAZ (Yes-associated protein/transcriptional coactivator with PDZ binding motif) pathway contributes to the pathogenesis of lymphatic malformations (LMs). METHODS: YAP, TAZ, CTGF (connective tissue growth factor), and Ki-67 were detected in LMs by immunohistochemistry. The colocalization of YAP and Ki-67 was analyzed by double immunofluorescence. Pearson's correlation and cluster analyses were performed to analyze the relationships between these proteins. Human dermal lymphatic endothelial cells (HDLECs) were used for mechanistic investigation. Rat models of LMs were established to investigate the role of the YAP pathway in LM development. RESULTS: Compared with those in normal skin, the expression levels of YAP, TAZ, CTGF, and Ki-67 were significantly upregulated in lymphatic endothelial cells (LECs) of LMs. Interestingly, YAP and CTGF presented much higher expression levels in infected LMs. In experiments in vitro, lipopolysaccharide (LPS) enhanced the expression of YAP in a concentration- and time-dependent manner via the increased phosphorylation of Erk1/2 (extracellular signal-regulated kinase 1/2). Moreover, the proliferation, invasion, and tubule formation of HDLECs increased significantly in accordance with the activation of the YAP signaling pathway. Furthermore, LM rat models validated that LPS facilitated the development of LMs, which was dependent on the activation of YAP. CONCLUSIONS: The data reveal that activation of the YAP signaling pathway in LECs may play a crucial role in the progression of LMs. IMPACT: Compared with that in normal skin, the YAP signaling pathway was activated in LECs of LMs. Inhibiting the YAP signaling pathway attenuated the proliferation, invasion, and tubule formation of HDLECs. Additionally, the activation of the YAP signaling pathway could promote LM development in a rat model. Activation of the YAP signaling pathway in LECs may play a crucial role in the progression of LMs. The YAP signaling pathway was activated in LMs. Inhibition of the YAP signaling pathway could promote regression of the lesions.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Células Endoteliales/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Linfangiogénesis , Anomalías Linfáticas/metabolismo , Vasos Linfáticos/metabolismo , Factores de Transcripción/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Estudios de Casos y Controles , Movimiento Celular , Proliferación Celular , Células Cultivadas , Factor de Crecimiento del Tejido Conjuntivo/metabolismo , Modelos Animales de Enfermedad , Células Endoteliales/efectos de los fármacos , Células Endoteliales/patología , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Antígeno Ki-67/metabolismo , Linfangiogénesis/efectos de los fármacos , Anomalías Linfáticas/genética , Anomalías Linfáticas/patología , Anomalías Linfáticas/prevención & control , Vasos Linfáticos/anomalías , Vasos Linfáticos/efectos de los fármacos , Ratas , Transducción de Señal , Factores de Transcripción/genética , Proteínas Coactivadoras Transcripcionales con Motivo de Unión a PDZ , Verteporfina/farmacología , Proteínas Señalizadoras YAP
19.
Circ Res ; 128(1): 42-58, 2021 01 08.
Artículo en Inglés | MEDLINE | ID: mdl-33135960

RESUMEN

RATIONALE: The recent discovery of meningeal lymphatics in mammals is reshaping our understanding of fluid homeostasis and cellular waste management in the brain, but visualization and experimental analysis of these vessels is challenging in mammals. Although the optical clarity and experimental advantages of zebrafish have made this an essential model organism for studying lymphatic development, the existence of meningeal lymphatics has not yet been reported in this species. OBJECTIVE: Examine the intracranial space of larval, juvenile, and adult zebrafish to determine whether and where intracranial lymphatic vessels are present. METHODS AND RESULTS: Using high-resolution optical imaging of the meninges in living animals, we show that zebrafish possess a meningeal lymphatic network comparable to that found in mammals. We confirm that this network is separate from the blood vascular network and that it drains interstitial fluid from the brain. We document the developmental origins and growth of these vessels into a distinct network separated from the external lymphatics. Finally, we show that these vessels contain immune cells and perform live imaging of immune cell trafficking and transmigration in meningeal lymphatics. CONCLUSIONS: This discovery establishes the zebrafish as a important new model for experimental analysis of meningeal lymphatic development and opens up new avenues for probing meningeal lymphatic function in health and disease.


Asunto(s)
Linfangiogénesis , Vasos Linfáticos/fisiología , Meninges/fisiología , Microscopía Confocal , Imagen Óptica , Animales , Animales Modificados Genéticamente , Linfangiogénesis/efectos de los fármacos , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/inmunología , Meninges/inmunología , Infiltración Neutrófila , Neutrófilos/inmunología , Factor C de Crecimiento Endotelial Vascular/farmacología , Pez Cebra/genética
20.
J Musculoskelet Neuronal Interact ; 20(4): 526-534, 2020 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-33265080

RESUMEN

OBJECTIVE: The lymphatic system plays an important role in joint diseases. This study aimed to evaluate the effects of ginsenoside Rg1 on lymphatic drainage and accumulation of inflammatory products in the joints. METHODS: Two-month-old transgenic mice that overexpress tumor necrosis factor alpha (TNF-α; TNF-Tg) were used as the animal models. Ginsenoside Rg1 was administered for 12 weeks and the lymphatic drainage in the mice was evaluated using near infrared-indocyanine green (NIR-ICG) lymphatic imaging system. The clinical symptoms of arthritis were evaluated weekly. The ankle and knee joints were harvested for hematoxylin-eosin (HE), alcian blue/orange G (ABOG), and tartrate-resistant acid phosphatase (TRAP) staining, and the foot dorsal skin was used for whole-mount immuno-staining. Simultaneously, the serum levels of IL-6 and TNF-α were detected using enzyme-linked immunosorbent assay (ELISA). RESULTS: Ginsenoside Rg1 significantly improved the lymphatic drainage function, reduced synovial inflammation and bone erosion, decreased serum IL-6 and TNF-α concentration, and increased smooth muscle coverage on the collecting lymphatic vessels in the foot skin of the TNF-Tg mice. Furthermore, ginsenoside Rg1 treatment for 12 weeks did not cause any damage to the liver and kidney tissues. CONCLUSION: Ginsenoside Rg1 improves lymphatic drainage and joint inflammation in TNF-Tg mice. Therefore, ginsenoside Rg1 has the potential to be a candidate drug for the treatment of inflammatory arthritis.


Asunto(s)
Artritis Experimental/patología , Ginsenósidos/farmacología , Vasos Linfáticos/efectos de los fármacos , Animales , Artritis Psoriásica/patología , Artritis Reumatoide/patología , Ratones , Ratones Transgénicos , Espondilitis Anquilosante/patología
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