P6494Activation of focal adhesion kinase is involved in pathogenesis of aortic dissection in mice

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
R Majima ◽  
H Aoki ◽  
Y Hashimoto ◽  
M Hayashi ◽  
S Ohno-Urabe ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Western Blot ◽  
Focal Adhesion ◽  
Transcriptome Analysis ◽  
Aortic Wall ◽  
Treated Group ◽  
Immunofluorescence Staining ◽  
Actin Dynamics ◽  
Aortic Tissue ◽  
Immunochemical Staining

Abstract Background Aortic dissection (AD) is a fatal disease where the media of the aorta suddenly fail. Currently, Molecular pathogenesis of AD is unknown. Recently, we discovered that the activity of MRTF-A, a mechanosensitive transcriptional regulator, promotes AD development. The activity of MRTF-A is regulated by mechanical stress to cells, which is transduced through focal adhesion and actin dynamics. However, it is currently unknown whether the mechanotransduction mechanism is involved in AD pathogenesis. Purpose We investigated the role of focal adhesion kinase (FAK), a signaling molecule that transduces mechanostress from focal adhesion to actin dynamics, in AD pathogenesis. Methods We created a mouse model of AD with a continuous infusion of beta-aminopropionitrile (150 mg/kg/day), a collagen crosslink inhibitor, and angiotensin II (1,000 ng/kg/min) (BAPN + AngII) by an osmotic pump. This model caused about 60% death in all mice due to AD rupture within 2 weeks. In this model, we examined the severity and mortality rate of aortic dissection after 2 weeks in mice administered with PND-1186, an orally available FAK inhibitor, and in those treated with vehicle (n=20 for each group). We performed immunochemical staining, immunofluorescence staining and Western blot for activated (phosphorylated) FAK (pFAK) to evaluate the activation status of FAK in the aortic tissue. We also performed transcriptome analysis of the aortic tissue in with and without PND-1186 with BAPN + AngII stimulation before AD development. Results Immunochemical staining revealed that FAK was inactive in normal mouse aorta, but was strongly activated in the aortic walls after AD development. Immunofluorescence staining showed that FAK was activated mainly in smooth muscle cells after AD development. Western blot analysis also revealed that FAK was activated in 3 days after BAPN + AngII infusion before AD development, followed by transient reduction at day 7, and re-activation after AD at day 14. Significantly, administration of PND-1186 resulted in a significant reduction in the severity of AD in the aortic arch (1.96±0.41 mm in vehicle group, 0.66±0.29 mm in PND group, P<0.05). In addition, survival rate improved from 36.4% to 80.0% by administration of PND-1186 (P<0.01). In immunofluorescence staining, the PND-1186 treated group showed weaker staining of pFAK. Transcriptome analysis showed that genes for hematopoiesis and immune system were suppressed in PND-1186 treated group. Conclusions These findings proved that FAK plays a central role in the pathogenesis of AD probably by transmitting pathological stress to the aortic wall to cause tissue destruction. We propose that FAK is a potential therapeutic target for limiting the fatal destruction of the aortic wall of AD.

Abstract 18391: Accumulation of Adiponectin in Association With Up-regulated T-cadherin in the Aortic Wall of Acute and Chronic Aortic Dissection

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Yoshiki Watanabe ◽  
Shigeru Miyagawa ◽  
Satsuki Fukushima ◽  
Norikazu Maeda ◽  
Atsuhiro Saito ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Western Blot ◽  
Blood Concentration ◽  
Aortic Wall ◽  
Acute Type ◽  
Type B ◽  
Rt Pcr ◽  
Multiple Organs ◽  
Aortic Media

Introduction: Adiponectin (APN) is a major adipokine, which has been reported to accumulate into the damaged tissues in multiple organs. However, the role of APN and its receptor, T-cadherin (T-cad), in the pathology of the human aortic wall (AW) is poorly understood. In this study, we examined the distributions of APN and T-cad in the dissected AW and measured the serum APN concentration in patients with aortic dissection (AD). Methods: Diseased AW tissues were collected from patients with acute or chronic AD at the time of surgery, and a healthy aorta was used as a control (n = 6 per group). Blood was serially sampled to measure the APN concentrations in non-surgically-treated patients with acute type B AD (n = 10). Results: Immunohistochemically, normal aortic walls weakly expressed APN or T-cad on the intimal surface, whereas the AWs of acute AD patients showed marked expression of both APN and T-cad on the surface of the dissected aortic media. In chronic AD, APN and T-cad were diffusely expressed on the medial layers of the thickened AW (Fig. A-C). Western blot analysis showed that APN expression in the AW was significantly greater in acute and chronic AD than in the normal aorta (151 ± 2% and 220 ± 27% vs. normal, respectively) (Fig. D). RT-PCR revealed no expression of APN mRNA in any AW, and stronger expression of T-cad mRNA in acute and chronic AD than in the normal aorta (108 ± 9% and 194 ± 101% vs. normal, respectively). The blood concentration of APN in type B AD patients decreased by 13.3 ± 3.8% in 24-78 hours, and by 22.7 ± 7.2% in over 78 hours, as compared to those monitored within 24 hours after the onset, indicating that APN was supplied to the affected aortic wall from the blood. Conclusions: APN markedly accumulated into the dissected aortic media in acute and chronic AD, and T-cad was upregulated in the corresponding area. In contrast, the blood concentration of APN reciprocally decreased after AD onset, indicating a possibility of APN contribution to protect a diseased aortic wall of AD.

scholarly journals Increased Risk of Aortic Dissection with Perlecan Deficiency

2021 ◽  
Vol 23 (1) ◽  
pp. 315
Author(s):  
Risa Nonaka ◽  
Takafumi Iesaki ◽  
Aurelien Kerever ◽  
Eri Arikawa-Hirasawa
Keyword(s):  
Aortic Dissection ◽  
High Frequency ◽  
Aortic Wall ◽  
Elastic Fibers ◽  
Aortic Tissue ◽  
Increased Risk ◽  
Fibrillin 1 ◽  
Membrane Type ◽  
Deficient Mice ◽  
Elastic Lamina

Perlecan (HSPG2), a basement membrane-type heparan sulfate proteoglycan, has been implicated in the development of aortic tissue. However, its role in the development and maintenance of the aortic wall remains unknown. Perlecan-deficient mice (Hspg2−/−-Tg: Perl KO) have been found to show a high frequency (15–35%) of aortic dissection (AD). Herein, an analysis of the aortic wall of Perl KO mice revealed that perlecan deficiency caused thinner and partially torn elastic lamina. Compared to the control aortic tissue, perlecan-deficient aortic tissue showed a significant decrease in desmosine content and an increase in soluble tropoelastin levels, implying the presence of immature elastic fibers in Perl KO mice. Furthermore, the reduced expression of the smooth muscle cell contractile proteins actin and myosin in perlecan-deficient aortic tissue may explain the risk of AD. This study showed that a deficiency in perlecan, which is localized along the elastic lamina and at the interface between elastin and fibrillin-1, increased the risk of AD, largely due to the immaturity of extracellular matrix in the aortic tissue. Overall, we proposed a new model of AD that considers the deficiency of extracellular molecule perlecan as a risk factor.

Abstract 105: Association of Viscoelastic Material Properties and Extracellular Matrix Remodeling in Human Abdominal Aortic Aneurysmal Tissue

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Doran S Mix ◽  
Sandra A Toth ◽  
Ibrahima Bah ◽  
Michael C Stoner ◽  
Bruce I Goldman ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Western Blot ◽  
Viscoelastic Material ◽  
Material Properties ◽  
Aortic Wall ◽  
Cyclic Strain ◽  
Wall Stress ◽  
Aortic Tissue ◽  
Matrix Remodeling ◽  
Abdominal Aortic

Objectives: Predicting rupture of abdominal aortic aneurysm (AAA) requires knowledge of both the rate of extracellular matrix (ECM) degradation and the pulsatile stress on the aortic tissue. The activity of matrix metallopeptidase 9 (MMP9) and its inhibitor, TIMP1, are associated with alterations in aortic ECM but it is unknown if these changes effect the dynamic viscoelastic properties. We hypothesize that increased levels of MMP9 within AAA tissue will be associated with a greater dynamic modulus (E*), as a surrogate of increased aortic wall stress. Methods: Human aneurysmal aortic tissue was obtained at the time of open AAA repair (n=11) and age-matched non-aneurysmal cadavers (n=10). Uniaxial viscoelastic material properties were measured in the circumferential orientation under physiologic preload (110 mmHg) and cyclic strain (± 5%@1Hz). Quantitative histologic and immunohistochemistry were preformed using Fiji imaging software. Aortic MMP9 and TIMP1 content and activity were quantified using western blot and zymography. Results: E* was greater (1862±464 vs 1362±405 kPa, p=0.02) in the AAA tissue as compared to non-aneurysmal tissue. AAA tissue contained less elastin (6.7±6.7 vs 23.4±8.7%, p=0.01) and a greater collagen/elastin ratio (19.9±20.6 vs 2.3±2.5%, p=0.05). Immunohistochemistry revealed 200% greater MMP9 content in the AAA tissue (Figure A & B, 0.61 vs 0.03%, p=0.03). Increased MMP9 content was confirmed using a western blot (0.43 vs 0.06 AU, p<0.01). No difference in relative MMP9 activity (4307 vs 2324 AU, p=0.25) or level of TIMP1 (0.03 vs 0.02, p=0.6) were observed. There was a positive linear correlation (Figure C, r 2 =0.47) between E* and MMP9 as determined by quantitative immunohistochemistry. Conclusions: Our data suggests a positive relationship between E* and MMP9 content. Increased tissue stiffness may trigger MMP9 production resulting in a positive-feedback loop, progressively increasing aortic wall stress and rupture risk.

Identification and Expression Analysis of CD73 Inhibitors in Cervical Cancer

2020 ◽  
Vol 16 ◽  
Author(s):  
Jamshed Iqbal ◽  
Ayesha Basharat ◽  
Sehrish Bano ◽  
Syed Mobasher Ali Abid ◽  
Julie Pelletier ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Western Blot ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Apoptosis ◽  
Immunofluorescence Staining ◽  
Cell Cycle Analysis ◽  
Hela Cell Line ◽  
Cell Line Hela

Aims: The present study was conducted to examine the inhibitory effects of synthesized sulfonylhydrazones on the expression of CD73 (ecto-5′-NT). Background: CD73 (ecto-5′-NT) represents the most significant class of ecto-nucleotidases which are mainly responsible for dephosphorylation of adenosine monophosphate to adenosine. Inhibition of CD73 played an important role in the treatment of cancer, autoimmune disorders, precancerous syndromes, and some other diseases associated with CD73 activity. Objective: Keeping in view the significance of CD73 inhibitor in the treatment of cervical cancer, a series of sulfonylhydrazones (3a-3i) derivatives synthesized from 3-formylchromones were evaluated. Methods: All sulfonylhydrazones (3a-3i) were evaluated for their inhibitory activity towards CD73 (ecto-5′-NT) by the malachite green assay and their cytotoxic effect was investigated on HeLa cell line using MTT assay. Secondly, most potent compound was selected for cell apoptosis, immunofluorescence staining and cell cycle analysis. After that, CD73 mRNA and protein expression were analyzed by real-time PCR and Western blot. Results: Among all compounds, 3h, 3e, 3b, and 3c were found the most active against rat-ecto-5′-NT (CD73) enzyme with IC50 (µM) values of 0.70 ± 0.06 µM, 0.87 ± 0.05 µM, 0.39 ± 0.02 µM and 0.33 ± 0.03 µM, respectively. These derivatives were further evaluated for their cytotoxic potential against cancer cell line (HeLa). Compound 3h and 3c showed the cytotoxicity at IC50 value of 30.20 ± 3.11 µM and 86.02 ± 7.11 µM, respectively. Furthermore, compound 3h was selected for cell apoptosis, immunofluorescence staining and cell cycle analysis which showed promising apoptotic effect in HeLa cells. Additionally, compound 3h was further investigated for its effect on expression of CD73 using qRT-PCR and western blot. Conclusion: Among all synthesized compounds (3a-3i), Compound 3h (E)-N'-((6-ethyl-4-oxo-4H-chromen-3-yl) methylene)-4-methylbenzenesulfonohydrazide was identified as most potent compound. Additional expression studies conducted on HeLa cell line proved that this compound successfully decreased the expression level of CD73 and thus inhibiting the growth and proliferation of cancer cells.

scholarly journals Aggrecan: a new biomarker for acute type A aortic dissection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Karl C. König ◽  
Harald Lahm ◽  
Martina Dreßen ◽  
Stefanie A. Doppler ◽  
Stefan Eichhorn ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Troponin T ◽  
Type A ◽  
Aortic Aneurysms ◽  
Aortic Tissue ◽  
Acute Type ◽  
Type A Aortic Dissection ◽  
Potential Biomarker ◽  
Aortic Pathology ◽  
Creatine Kinase Mb

AbstractAcute type A aortic dissection (ATAAD) constitutes a life-threatening aortic pathology with significant morbidity and mortality. Without surgical intervention the usual mortality rate averages between 1 and 2% per hour. Thus, an early diagnosis of ATAAD is of pivotal importance to direct the affected patients to the appropriate treatment. Preceding tests to find an appropriate biomarker showed among others an increased aggrecan (ACAN) mRNA expression in aortic tissue of ATAAD patients. As a consequence, we investigated whether ACAN is a potential biomarker for diagnosing ATAAD. Mean ACAN protein concentration showed a significantly higher plasma concentration in ATAAD patients (38.59 ng/mL, n = 33) compared to plasma of patients with thoracic aortic aneurysms (4.45 ng/mL, n = 13), patients with myocardial infarction (11.77 ng/mL, n = 18) and healthy volunteers (8.05 ng/mL, n = 12). Cardiac enzymes like creatine kinase MB and cardiac troponin T showed no correlation with ACAN levels in ATAAD patients. Receiver-operator characteristics (ROC) curve analysis for ATAAD patients versus control subjects an optimum discrimination limit of ACAN plasma levels at 14.3 ng/mL with a corresponding sensitivity of 97% and specificity of 81%. According to our findings ACAN is a reliable potential biomarker in plasma samples to detect ATAAD with high sensitivity and specificity.

scholarly journals Mimicking the Mechanical Properties of Aortic Tissue with Pattern-Embedded 3D Printing for a Realistic Phantom

Materials ◽  
2020 ◽  
Vol 13 (21) ◽  
pp. 5042
Author(s):  
Jaeyoung Kwon ◽  
Junhyeok Ock ◽  
Namkug Kim
Keyword(s):  
Mechanical Properties ◽  
Tensile Strength ◽  
3D Printing ◽  
Mechanical Characteristics ◽  
Aortic Wall ◽  
Tensile Tests ◽  
Human Aorta ◽  
Aortic Tissue ◽  
Medical Field ◽  
Base Materials

3D printing technology has been extensively applied in the medical field, but the ability to replicate tissues that experience significant loads and undergo substantial deformation, such as the aorta, remains elusive. Therefore, this study proposed a method to imitate the mechanical characteristics of the aortic wall by 3D printing embedded patterns and combining two materials with different physical properties. First, we determined the mechanical properties of the selected base materials (Agilus and Dragonskin 30) and pattern materials (VeroCyan and TPU 95A) and performed tensile testing. Three patterns were designed and embedded in printed Agilus–VeroCyan and Dragonskin 30–TPU 95A specimens. Tensile tests were then performed on the printed specimens, and the stress-strain curves were evaluated. The samples with one of the two tested orthotropic patterns exceeded the tensile strength and strain properties of a human aorta. Specifically, a tensile strength of 2.15 ± 0.15 MPa and strain at breaking of 3.18 ± 0.05 mm/mm were measured in the study; the human aorta is considered to have tensile strength and strain at breaking of 2.0–3.0 MPa and 2.0–2.3 mm/mm, respectively. These findings indicate the potential for developing more representative aortic phantoms based on the approach in this study.

scholarly journals In Vivo Role of Focal Adhesion Kinase in Regulating Pancreatic  -Cell Mass and Function Through Insulin Signaling, Actin Dynamics, and Granule Trafficking

Diabetes ◽  
2012 ◽  
Vol 61 (7) ◽  
pp. 1708-1718 ◽  
Author(s):  
E. P. Cai ◽  
M. Casimir ◽  
S. A. Schroer ◽  
C. T. Luk ◽  
S. Y. Shi ◽  
...  
Keyword(s):  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Insulin Signaling ◽  
Cell Mass ◽  
Actin Dynamics ◽  
Pancreatic Cell ◽  
And Function

scholarly journals Visualization and Analysis of Gene Expression in Stanford Type A Aortic Dissection Tissue Section by Spatial Transcriptomics

2021 ◽  
Vol 12 ◽  
Author(s):  
Yan-Hong Li ◽  
Ying Cao ◽  
Fen Liu ◽  
Qian Zhao ◽  
Dilare Adi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Aortic Dissection ◽  
Cell Structure ◽  
Type A ◽  
Cell Types ◽  
Biological Tissues ◽  
Human Aorta ◽  
Aortic Tissue ◽  
Specific Gene ◽  
Type A Aortic Dissection

Background: Spatial transcriptomics enables gene expression events to be pinpointed to a specific location in biological tissues. We developed a molecular approach for low-cell and high-fiber Stanford type A aortic dissection and preliminarily explored and visualized the heterogeneity of ascending aortic types and mapping cell-type-specific gene expression to specific anatomical domains.Methods: We collected aortic samples from 15 patients with Stanford type A aortic dissection and a case of ascending aorta was randomly selected followed by 10x Genomics and spatial transcriptomics sequencing. In data processing of normalization, component analysis and dimensionality reduction analysis, different algorithms were compared to establish the pipeline suitable for human aortic tissue.Results: We identified 19,879 genes based on the count level of gene expression at different locations and they were divided into seven groups based on gene expression trends. Major cell that the population may contain are indicated, and we can find different main distribution of different cell types, among which the tearing sites were mainly macrophages and stem cells. The gene expression of these different locations and the cell types they may contain are correlated and discussed in terms of their involvement in immunity, regulation of oxygen homeostasis, regulation of cell structure and basic function.Conclusion: This approach provides a spatially resolved transcriptome− and tissue-wide perspective of the adult human aorta and will allow the application of human fibrous aortic tissues without any effect on genes in different layers with low RNA expression levels. Our findings will pave the way toward both a better understanding of Stanford type A aortic dissection pathogenesis and heterogeneity and the implementation of more effective personalized therapeutic approaches.

scholarly journals Rab40/Cullin5 complex regulates EPLIN and actin cytoskeleton dynamics during cell migration and invasion

2021 ◽  
Author(s):  
Erik S Linklater ◽  
Emily Duncan ◽  
Ke Jun Han ◽  
Algirdas Kaupinis ◽  
Mindaugas Valius ◽  
...  
Keyword(s):  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Leading Edge ◽  
Stress Fibers ◽  
Actin Dynamics ◽  
Migration And Invasion ◽  
Matrix Remodeling ◽  
Cell Migration And Invasion

Rab40b is a SOCS box containing protein that regulates the secretion of MMPs to facilitate extracellular matrix remodeling during cell migration. Here we show that Rab40b interacts with Cullin5 via the Rab40b SOCS domain. We demonstrate that loss of Rab40b/Cullin5 binding decreases cell motility and invasive potential, and show that defective cell migration and invasion stem from alteration to the actin cytoskeleton, leading to decreased invadopodia formation, decreased actin dynamics at the leading edge, and an increase in stress fibers. We also show that these stress fibers anchor at less dynamic, more stable focal adhesions. Mechanistically, changes in the cytoskeleton and focal adhesion dynamics are mediated in part by EPLIN, which we demonstrate to be a binding partner of Rab40b and a target for Rab40b/Cullin5 dependent localized ubiquitylation and degradation. Thus, we propose a model where the Rab40b/Cullin5 dependent ubiquitylation regulates EPLIN localization to promote cell migration and invasion by altering focal adhesion and cytoskeletal dynamics.

scholarly journals Histopathological evaluation of aortic dissection: a comparison of congenital versus acquired aortic wall weakness

2018 ◽  
Vol 27 (2) ◽  
pp. 277-283 ◽  
Author(s):  
Hiroaki Osada ◽  
Masahisa Kyogoku ◽  
Tekehiko Matsuo ◽  
Naoki Kanemitsu
Keyword(s):  
Aortic Dissection ◽  
Aortic Wall ◽  
Histopathological Evaluation
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