48 research outputs found
Therapeutic Angiogenesis: Foundations and Practical Application
Angiogenesis as therapeutic target has emerged since early works by Judah Folkman, yet his βholy grailβ was inhibiting vascular growth to block tumor nutrition. However, in modern biomedicine, βtherapeutic angiogenesisβ became a large field focusing on stimulation of blood vessel growth for ischemia relief to reduce its detrimental effects in the tissues. In this review, we introduce basic principles of tissue vascularization in response to ischemia exploited in this field. An overview of recent status in therapeutic angiogenesis is given with introduction to emerging technologies, including gene therapy, genetic modification of cells ex vivo and tissue engineering
Oligonucleotide Microarrays Identified Potential Regulatory Genes Related to Early Outward Arterial Remodeling Induced by Tissue Plasminogen Activator
Constrictive vascular remodeling limiting blood flow, as well as compensatory outward remodeling, has been observed in many cardiovascular diseases; however, the underlying mechanisms regulating the remodeling response of the vessels remain unclear. Plasminogen activators (PA) are involved in many of the processes of vascular remodeling. We have shown previously that increased levels of tissue-type PA (tPA) contributes to outward vascular remodeling. To elucidate the mechanisms involved in the induction of outward remodeling we characterized changes in the expression profiles of 8799 genes in injured rat carotid arteries 1 and 4 days after recombinant tPA treatment compared to vehicle. Periadventitial tPA significantly increased lumen size and vessel area, encompassed by the external elastic lamina, at both one and 4 days after treatment. Among 41 differentially expressed known genes 1 day after tPA application, five genes were involved in gene transcription, five genes were related to the regulation of vascular tone [for example, thromboxane A2 receptor (D32080) or non-selective-type endothelin receptor (S65355)], and eight genes were identified as participating in vascular innervation [for example, calpain (D14478) or neural cell adhesion molecule L1 (X59149)]. Four days after injury in tPA-treated arteries, four genes, regulating vascular tone, were differentially expressed. Thus, tPA promotes outward arterial remodeling after injury, at least in part, by regulating expression of genes in the vessel wall related to function of the nervous system and vascular tone
Urokinase-type Plasminogen Activator (uPA) Promotes Angiogenesis by Attenuating Proline-rich Homeodomain Protein (PRH) Transcription Factor Activity and De-repressing Vascular Endothelial Growth Factor (VEGF) Receptor Expression
Urokinase-type plasminogen activator (uPA) regulates angiogenesis and vascular permeability through proteolytic degradation of extracellular matrix and intracellular signaling initiated upon its binding to uPAR/CD87 and other cell surface receptors. Here, we describe an additional mechanism by which uPA regulates angiogenesis. Ex vivo VEGF-induced vascular sprouting from Matrigel-embedded aortic rings isolated from uPA knock-out (uPA(β/β)) mice was impaired compared with vessels emanating from wild-type mice. Endothelial cells isolated from uPA(β/β) mice show less proliferation and migration in response to VEGF than their wild type counterparts or uPA(β/β) endothelial cells in which expression of wild type uPA had been restored. We reported previously that uPA is transported from cell surface receptors to nuclei through a mechanism that requires its kringle domain. Intranuclear uPA modulates gene transcription by binding to a subset of transcription factors. Here we report that wild type single-chain uPA, but not uPA variants incapable of nuclear transport, increases the expression of cell surface VEGF receptor 1 (VEGFR1) and VEGF receptor 2 (VEGFR2) by translocating to the nuclei of ECs. Intranuclear single-chain uPA binds directly to and interferes with the function of the transcription factor hematopoietically expressed homeodomain protein or proline-rich homeodomain protein (HHEX/PRH), which thereby lose their physiologic capacity to repress the activity of vehgr1 and vegfr2 gene promoters. These studies identify uPA-dependent de-repression of vegfr1 and vegfr2 gene transcription through binding to HHEX/PRH as a novel mechanism by which uPA mediates the pro-angiogenic effects of VEGF and identifies a potential new target for control of pathologic angiogenesis
UKβRussia Researcher Links Workshop: extracellular vesicles β mechanisms of biogenesis and roles in disease pathogenesis, M.V. Lomonosov Moscow State University, Moscow, Russia, 1β5 March 2015
The UKβRussia extracellular vesicles (EVs) workshop was held at the Medical Center of the M.V. Lomonosov Moscow State University, Moscow, Russia, with 56 attendees from UK and Russian universities and research institutes. The program consisted of 6 research sessions and was focused on studies of EVs isolated from in vitro model systems or biological fluids, including blood and urine. The multidisciplinary program included presentations on mechanisms of EV biogenesis, the role of EVs in disease pathogenesis, the diagnostic value of EVs, including their quantitation and cargo load, as well as the clinical use of EVs in regenerative medicine. Methodological challenges imposed by the nanoscale size of EVs as well as targeted delivery approaches for therapeutics were considered in a separate session on technologies. The main aim of the workshop was to overview challenges confronting EV researchers and to facilitate knowledge exchange between researchers with different backgrounds and skills. Given the lack of definitive EV nomenclature, specific terms (exosomes or microvesicles) were only applied in the meeting report to studies that carried out full EV characterization, including differential ultracentrifugation isolation approaches, comprehensive protein marker characterization, and single vesicle analysis (electron microscopy and nanoparticle analysis), to ascertain EV size and morphology following the International Society for Extracellular Vesicles standardization recommendations (1,2). In studies where characterization was not conclusive, the term EV is used
ΠΠΏΠΈΠΊΠ°ΡΠ΄ΠΈΠ°Π»ΡΠ½Π°Ρ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΡ ΠΏΠ»Π°ΡΡΠΎΠ² ΠΈΠ· ΠΌΠ΅Π·Π΅Π½Ρ ΠΈΠΌΠ°Π»ΡΠ½ΡΡ ΡΡΡΠΎΠΌΠ°Π»ΡΠ½ΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΆΠΈΡΠΎΠ²ΠΎΠΉ ΠΊΠ»Π΅ΡΡΠ°ΡΠΊΠΈ ΡΠΏΠΎΡΠΎΠ±ΡΡΠ²ΡΠ΅Ρ Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠΈ ΡΠΏΠΈΠΊΠ°ΡΠ΄Π° ΠΈ ΡΡΠΈΠΌΡΠ»ΠΈΡΡΠ΅Ρ Π°Π½Π³ΠΈΠΎΠ³Π΅Π½Π΅Π· ΠΏΡΠΈ ΠΈΠ½ΡΠ°ΡΠΊΡΠ΅ ΠΌΠΈΠΎΠΊΠ°ΡΠ΄Π° (ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΠΎΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅)
Aim: to evaluate the impact of tissue-engineered structures (TES) transplantation based on mesenchymal stromal cell (MSC) sheets in myocardial infarction on the activation of the epicardial cell pool and vascularization of the damaged zone.Materials and methods. Mesenchymal stromal cells were obtained from samples of subcutaneous fat of Wistar rats and C57Bl/6 mice. Tissue engineering structures were obtained by culturing cell sheets on thermosensitive plates (Nunc Dishes with UpCell Surface). Transplantation of TESs was performed after myocardial infarction modeling in rats by ligation of the anterior descending coronary artery. Transplant cells and damaged zones were assessed using immunofluorescent staining of myocardial cryosections. The impact of MSC secretion products on the migration activity of epicardial cells in vitro was evaluated using the explant culture method.Results. MSCs in TESs after transplantation remain viable and induce activation of the epicardial cell pool and local increase of the damaged zone vascularization. The in vitro experiments showed that the conditioned environment of MSCs stimulates the migratory activity of epicardial cells and initiates the formation of activated Wt1/POD1 precursor cells.Conclusion. TES transplantation on the basis of MSC sheets seems to be a promising approach for effective delivery of viable cells into myocardium to activate the epicardial cellular niche and reparative angiogenesis.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ: ΠΎΡΠ΅Π½ΠΈΡΡ Π²Π»ΠΈΡΠ½ΠΈΠ΅ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΠΈ ΡΠΊΠ°Π½Π΅ΠΈΠ½ΠΆΠ΅Π½Π΅ΡΠ½ΡΡ
ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΉ (Π’ΠΠ) Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ ΠΏΠ»Π°ΡΡΠΎΠ² ΠΌΠ΅Π·Π΅Π½Ρ
ΠΈΠΌΠ°Π»ΡΠ½ΡΡ
ΡΡΡΠΎΠΌΠ°Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ (ΠΠ‘Π) ΠΏΡΠΈ ΠΈΠ½ΡΠ°ΡΠΊΡΠ΅ ΠΌΠΈΠΎΠΊΠ°ΡΠ΄Π° Π½Π° Π°ΠΊΡΠΈΠ²Π°ΡΠΈΡ ΡΠΏΠΈΠΊΠ°ΡΠ΄ΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΏΡΠ»Π° ΠΊΠ»Π΅ΡΠΎΠΊ ΠΈ Π²Π°ΡΠΊΡΠ»ΡΡΠΈΠ·Π°ΡΠΈΡ Π·ΠΎΠ½Ρ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π΅Π½ΠΈΡ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠ‘Π ΠΏΠΎΠ»ΡΡΠΈΠ»ΠΈ ΠΈΠ· ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² ΠΏΠΎΠ΄ΠΊΠΎΠΆΠ½ΠΎΠΉ ΠΆΠΈΡΠΎΠ²ΠΎΠΉ ΠΊΠ»Π΅ΡΡΠ°ΡΠΊΠΈ ΠΊΡΡΡ Π»ΠΈΠ½ΠΈΠΈ Wistar ΠΈ ΠΌΡΡΠ΅ΠΉ Π»ΠΈΠ½ΠΈΠΈ C57Bl/6. Π’ΠΠ ΠΏΠΎΠ»ΡΡΠΈΠ»ΠΈ ΠΏΡΡΠ΅ΠΌ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΏΠ»Π°ΡΡΠΎΠ² ΠΊΠ»Π΅ΡΠΎΠΊ Π½Π° ΡΠ°ΡΠΊΠ°Ρ
Ρ ΡΠ΅ΡΠΌΠΎΡΡΠ²ΡΡΠ²ΠΈΡΠ΅Π»ΡΠ½ΡΠΌ ΠΏΠΎΠΊΡΡΡΠΈΠ΅ΠΌ (Nunc Dishes with UpCell Surface). Π’ΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΡ Π’ΠΠ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΏΠΎΡΠ»Π΅ ΠΌΠΎΠ΄Π΅Π»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΈΠ½ΡΠ°ΡΠΊΡΠ° ΠΌΠΈΠΎΠΊΠ°ΡΠ΄Π° Ρ ΠΊΡΡΡΡ ΠΏΡΡΠ΅ΠΌ ΠΏΠ΅ΡΠ΅Π²ΡΠ·ΠΊΠΈ ΠΏΠ΅ΡΠ΅Π΄Π½Π΅ΠΉ Π½ΠΈΡΡ
ΠΎΠ΄ΡΡΠ΅ΠΉ ΠΊΠΎΡΠΎΠ½Π°ΡΠ½ΠΎΠΉ Π°ΡΡΠ΅ΡΠΈΠΈ. ΠΡΠ΅Π½ΠΊΡ ΡΠΎΡΡΠΎΡΠ½ΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠ° ΠΈ Π·ΠΎΠ½Ρ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π΅Π½ΠΈΡ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΈΠΌΠΌΡΠ½ΠΎΡΠ»ΡΠΎΡΠ΅ΡΡΠ΅Π½ΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΡΠ°ΡΠΈΠ²Π°Π½ΠΈΡ ΠΊΡΠΈΠΎΡΡΠ΅Π·ΠΎΠ² ΠΌΠΈΠΎΠΊΠ°ΡΠ΄Π°. ΠΠ»Ρ ΠΎΡΠ΅Π½ΠΊΠΈ Π²Π»ΠΈΡΠ½ΠΈΡ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΡΠ΅ΠΊΡΠ΅ΡΠΈΠΈ ΠΠ‘Π Π½Π° ΠΌΠΈΠ³ΡΠ°ΡΠΈΠΎΠ½Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠΏΠΈΠΊΠ°ΡΠ΄Π° in vitro ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ ΡΠΊΡΠΏΠ»Π°Π½ΡΠ½ΠΎΠΉ ΠΊΡΠ»ΡΡΡΡΡ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΠ‘Π Π² ΡΠΎΡΡΠ°Π²Π΅ Π’ΠΠ ΠΏΠΎΡΠ»Π΅ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΠΈ ΡΠΎΡ
ΡΠ°Π½ΡΡΡ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΡ ΠΈ Π²ΡΠ·ΡΠ²Π°ΡΡ Π°ΠΊΡΠΈΠ²Π°ΡΠΈΡ ΡΠΏΠΈΠΊΠ°ΡΠ΄ΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΏΡΠ»Π° ΠΊΠ»Π΅ΡΠΎΠΊ ΠΈ Π»ΠΎΠΊΠ°Π»ΡΠ½ΠΎΠ΅ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΠ΅ Π²Π°ΡΠΊΡΠ»ΡΡΠΈΠ·Π°ΡΠΈΠΈ Π·ΠΎΠ½Ρ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π΅Π½ΠΈΡ. ΠΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΡ in vitro ΠΏΠΎΠΊΠ°Π·Π°Π»ΠΈ, ΡΡΠΎ ΠΊΠΎΠ½Π΄ΠΈΡΠΈΠΎΠ½ΠΈΡΠΎΠ²Π°Π½Π½Π°Ρ ΡΡΠ΅Π΄Π° ΠΠ‘Π ΠΎΠΊΠ°Π·ΡΠ²Π°Π΅Ρ ΡΡΠΈΠΌΡΠ»ΠΈΡΡΡΡΠ΅Π΅ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ Π½Π° ΠΌΠΈΠ³ΡΠ°ΡΠΈΠΎΠ½Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠΏΠΈΠΊΠ°ΡΠ΄Π° ΠΈ Π²ΡΠ·ΡΠ²Π°Π΅Ρ ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠ΅ Π°ΠΊΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Wt1/POD1 ΠΊΠ»Π΅ΡΠΎΠΊ-ΠΏΡΠ΅Π΄ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΈΡ.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. Π’ΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΡ Π’ΠΠ Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ ΠΏΠ»Π°ΡΡΠΎΠ² ΠΠ‘Π ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅ΡΡΡ ΡΠ°ΡΠΈΠΎΠ½Π°Π»ΡΠ½ΡΠΌ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄ΠΎΠΌ Π΄Π»Ρ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΠΉ Π΄ΠΎΡΡΠ°Π²ΠΊΠΈ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π² ΠΌΠΈΠΎΠΊΠ°ΡΠ΄ Ρ ΡΠ΅Π»ΡΡ Π°ΠΊΡΠΈΠ²ΠΈΡΡΡΡΠ΅Π³ΠΎ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΡ Π½Π° ΡΠΏΠΈΠΊΠ°ΡΠ΄ΠΈΠ°Π»ΡΠ½ΡΡ ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ Π½ΠΈΡΡ ΠΈ ΡΠ΅ΠΏΠ°ΡΠ°ΡΠΈΠ²Π½ΡΠΉ Π°Π½Π³ΠΈΠΎΠ³Π΅Π½Π΅Π·
Combined Transfer of Human VEGF165 and HGF Genes Renders Potent Angiogenic Effect in Ischemic Skeletal Muscle
Increased interest in development of combined gene therapy emerges from results of recent clinical trials that indicate good safety yet unexpected low efficacy of βsingle-geneβ administration. Multiple studies showed that vascular endothelial growth factor 165 aminoacid form (VEGF165) and hepatocyte growth factor (HGF) can be used for induction of angiogenesis in ischemic myocardium and skeletal muscle. Gene transfer system composed of a novel cytomegalovirus-based (CMV) plasmid vector and codon-optimized human VEGF165 and HGF genes combined with intramuscular low-voltage electroporation was developed and tested in vitro and in vivo. Studies in HEK293T cell culture, murine skeletal muscle explants and ELISA of tissue homogenates showed efficacy of constructed plasmids. Functional activity of angiogenic proteins secreted by HEK293T after transfection by induction of tube formation in human umbilical vein endothelial cell (HUVEC) culture. HUVEC cells were used for in vitro experiments to assay the putative signaling pathways to be responsible for combined administration effect one of which could be the ERK1/2 pathway. In vivo tests of VEGF165 and HGF genes co-transfer were conceived in mouse model of hind limb ischemia. Intramuscular administration of plasmid encoding either VEGF165 or HGF gene resulted in increased perfusion compared to empty vector administration. Mice injected with a mixture of two plasmids (VEGF165+HGF) showed significant increase in perfusion compared to single plasmid injection. These findings were supported by increased CD31+ capillary and SMA+ vessel density in animals that received combined VEGF165 and HGF gene therapy compared to single gene therapy. Results of the study suggest that co-transfer of VEGF and HGF genes renders a robust angiogenic effect in ischemic skeletal muscle and may present interest as a potential therapeutic combination for treatment of ischemic disorders
Adipose-Derived Stem Cells Stimulate Regeneration of Peripheral Nerves: BDNF Secreted by These Cells Promotes Nerve Healing and Axon Growth De Novo
Transplantation of adipose-derived mesenchymal stem cells (ASCs) induces tissue regeneration by accelerating the growth of blood vessels and nerve. However, mechanisms by which they accelerate the growth of nerve fibers are only partially understood. We used transplantation of ASCs with subcutaneous matrigel implants (well-known in vivo model of angiogenesis) and model of mice limb reinnervation to check the influence of ASC on nerve growth. Here we show that ASCs stimulate the regeneration of nerves in innervated mice's limbs and induce axon growth in subcutaneous matrigel implants. To investigate the mechanism of this action we analyzed different properties of these cells and showed that they express numerous genes of neurotrophins and extracellular matrix proteins required for the nerve growth and myelination. Induction of neural differentiation of ASCs enhances production of brain-derived neurotrophic factor (BDNF) as well as ability of these cells to induce nerve fiber growth. BDNF neutralizing antibodies abrogated the stimulatory effects of ASCs on the growth of nerve sprouts. These data suggest that ASCs induce nerve repair and growth via BDNF production. This stimulatory effect can be further enhanced by culturing the cells in neural differentiation medium prior to transplantation