119 results on '"Mikelis, Constantinos M."'
Search Results
2. Angiopoietin-2-induced lymphatic endothelial cell migration drives lymphangiogenesis via the β1 integrin-RhoA-formin axis
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Akwii, Racheal Grace, Sajib, Md. Sanaullah, Zahra, Fatema Tuz, Tullar, Paul, Zabet-Moghaddam, Masoud, Zheng, Yi, Silvio Gutkind, J., Doci, Colleen L., and Mikelis, Constantinos M.
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- 2022
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3. Chlorinated benzothiadiazines inhibit angiogenesis through suppression of VEGFR2 phosphorylation
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Huwaimel, Bader I., Jonnalagadda, Sravan, Jonnalagadda, Shirisha, Zahra, Fatema T., Nocentini, Alessio, Supuran, Claudiu T., Mikelis, Constantinos M., and Trippier, Paul C.
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- 2022
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4. Targeting the Angiopoietin/Tie Pathway: Prospects for Treatment of Retinal and Respiratory Disorders
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Akwii, Racheal Grace and Mikelis, Constantinos M.
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- 2021
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5. Design, synthesis and structure-activity relationship study of novel urea compounds as FGFR1 inhibitors to treat metastatic triple-negative breast cancer
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Ashraf-Uz-Zaman, Md, Shahi, Sadisna, Akwii, Racheal, Sajib, Md Sanaullah, Farshbaf, Mohammad Jodeiri, Kallem, Raja Reddy, Putnam, William, Wang, Wei, Zhang, Ruiwen, Alvina, Karina, Trippier, Paul C., Mikelis, Constantinos M., and German, Nadezhda A.
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- 2021
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6. Pleiotrophin selectively binds to vascular endothelial growth factor receptor 2 and inhibits or stimulates cell migration depending on ανβ3 integrin expression
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Lamprou, Margarita, Kastana, Pinelopi, Kofina, Fani, Tzoupis, Ηaralampos, Barmpoutsi, Spyridoula, Sajib, Md Sanaullah, Koutsioumpa, Marina, Poimenidi, Evangelia, Zompra, Aikaterini A., Tassopoulos, Dimitrios, Choleva, Effrosyni, Tselios, Theodore, Mikelis, Constantinos M., and Papadimitriou, Evangelia
- Published
- 2020
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- View/download PDF
7. Advanced bioinformatic analysis and pathway prediction of NSCLC cells upon cisplatin resistance
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Hossian, A K M Nawshad, Zahra, Fatema Tuz, Poudel, Sagun, Abshire, Camille F., Polk, Paula, Garai, Jone, Zabaleta, Jovanny, Mikelis, Constantinos M., and Mattheolabakis, George
- Published
- 2021
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8. Stable Dual miR-143 and miR-506 Upregulation Inhibits Proliferation and Cell Cycle Progression.
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Shrestha, Archana, Lahooti, Behnaz, Hossian, A. K. M. Nawshad, Madadi, Mahboubeh, Mikelis, Constantinos M., and Mattheolabakis, George
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INHIBITION of cellular proliferation ,CELL cycle ,CELL morphology ,CARCINOGENESIS ,CELL motility - Abstract
The mainstays of lung cancer pathogenesis are cell cycle progression dysregulation, impaired apoptosis, and unregulated cell proliferation. While individual microRNA (miR) targeting or delivering is a promising approach that has been extensively studied, combination of miR targeting can enhance therapeutic efficacy and overcome limitations present in individual miR regulations. We previously reported on the use of a miR-143 and miR-506 combination via transient transfections against lung cancer. In this study, we evaluated the effect of miR-143 and miR-506 under stable deregulations in A549 lung cancer cells. We used lentiviral transductions to either up- or downregulate the two miRs individually or in combination. The cells were sorted and analyzed for miR deregulation via qPCR. We determined the miR deregulations' effects on the cell cycle, cell proliferation, cancer cell morphology, and cell motility. Compared to the individual miR deregulations, the combined miR upregulation demonstrated a miR-expression-dependent G2 cell cycle arrest and a significant increase in the cell doubling time, whereas the miR-143/506 dual downregulation demonstrated increased cellular motility. Furthermore, the individual miR-143 and miR-506 up- and downregulations exhibited cellular responses lacking an apparent miR-expression-dependent response in the respective analyses. Our work here indicates that, unlike the individual miR upregulations, the combinatorial miR treatment remained advantageous, even under prolonged miR upregulation. Finally, our findings demonstrate potential advantages of miR combinations vs. individual miR treatments. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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9. Anti-angiogenic effects of VEGF stimulation on endothelium deficient in phosphoinositide recycling
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Stratman, Amber N., Farrelly, Olivia M., Mikelis, Constantinos M., Miller, Mayumi F., Wang, Zhiyong, Pham, Van N., Davis, Andrew E., Burns, Margaret C., Pezoa, Sofia A., Castranova, Daniel, Yano, Joseph J., Kilts, Tina M., Davis, George E., Gutkind, J. Silvio, and Weinstein, Brant M.
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- 2020
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10. Mechanisms of angiogenesis in microbe-regulated inflammatory and neoplastic conditions
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Sajib, Sanaullah, Zahra, Fatema Tuz, Lionakis, Michail S., German, Nadezhda A., and Mikelis, Constantinos M.
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- 2018
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11. Endothelial RhoA GTPase is essential for in vitro endothelial functions but dispensable for physiological in vivo angiogenesis
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Zahra, Fatema Tuz, Sajib, Md Sanaullah, Ichiyama, Yusuke, Akwii, Racheal Grace, Tullar, Paul E., Cobos, Christopher, Minchew, Shelby A., Doçi, Colleen L., Zheng, Yi, Kubota, Yoshiaki, Gutkind, J. Silvio, and Mikelis, Constantinos M.
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- 2019
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12. Genetic deletion or tyrosine phosphatase inhibition of PTPRZ1 activates c‐Met to up‐regulate angiogenesis and lung adenocarcinoma growth.
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Kastana, Pinelopi, Ntenekou, Despoina, Mourkogianni, Eleni, Enake, Michaela‐Karina, Xanthopoulos, Athanasios, Choleva, Effrosyni, Marazioti, Antonia, Nikou, Sophia, Akwii, Racheal G., Papadaki, Eleni, Gramage, Esther, Herradón, Gonzalo, Stathopoulos, Georgios T., Mikelis, Constantinos M., and Papadimitriou, Evangelia
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PROTEIN-tyrosine phosphatase ,CELL migration inhibition ,CRIZOTINIB ,VASCULAR endothelial growth factors ,VASCULAR endothelial cells ,MET receptor ,VASCULAR endothelial growth factor receptors - Abstract
Protein tyrosine phosphatase receptor zeta 1 (PTPRZ1) is a transmembrane tyrosine phosphatase (TP) expressed in endothelial cells and required for stimulation of cell migration by vascular endothelial growth factor A165 (VEGFA165) and pleiotrophin (PTN). It is also over or under‐expressed in various tumor types. In this study, we used genetically engineered Ptprz1−/− and Ptprz1+/+ mice to study mechanistic aspects of PTPRZ1 involvement in angiogenesis and investigate its role in lung adenocarcinoma (LUAD) growth. Ptprz1−/− lung microvascular endothelial cells (LMVEC) have increased angiogenic features compared with Ptprz1+/+ LMVEC, in line with the increased lung angiogenesis and the enhanced chemically induced LUAD growth in Ptprz1−/− compared with Ptprz1+/+ mice. In LUAD cells isolated from the lungs of urethane‐treated mice, PTPRZ1 TP inhibition also enhanced proliferation and migration. Expression of beta 3 (β3) integrin is decreased in Ptprz1−/− LMVEC, linked to enhanced VEGF receptor 2 (VEGFR2), c‐Met tyrosine kinase (TK) and Akt kinase activities. However, only c‐Met and Akt seem responsible for the enhanced endothelial cell activation in vitro and LUAD growth and angiogenesis in vivo in Ptprz1−/− mice. A selective PTPRZ1 TP inhibitor, VEGFA165 and PTN also activate c‐Met and Akt in a PTPRZ1‐dependent manner in endothelial cells, and their stimulatory effects are abolished by the c‐Met TK inhibitor (TKI) crizotinib. Altogether, our data suggest that low PTPRZ1 expression is linked to worse LUAD prognosis and response to c‐Met TKIs and uncover for the first time the role of PTPRZ1 in mediating c‐Met activation by VEGFA and PTN. [ABSTRACT FROM AUTHOR]
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- 2023
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13. Multipronged activity of combinatorial miR-143 and miR-506 inhibits Lung Cancer cell cycle progression and angiogenesis in vitro
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Hossian, A. K. M. Nawshad, Sajib, Md. Sanaullah, Tullar, Paul E., Mikelis, Constantinos M., and Mattheolabakis, George
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- 2018
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14. CDP-diacylglycerol synthetase-controlled phosphoinositide availability limits VEGFA signaling and vascular morphogenesis
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Pan, Weijun, Pham, Van N., Stratman, Amber N., Castranova, Daniel, Kamei, Makoto, Kidd, Kameha R., Lo, Brigid D., Shaw, Kenna M., Torres-Vazquez, Jesus, Mikelis, Constantinos M., Gutkind, J. Silvio, Davis, George E., and Weinstein, Brant M.
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- 2012
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15. Chlorotoxin and Lung Cancer: A Targeting Perspective for Drug Delivery.
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Shrestha, Archana, Lahooti, Behnaz, Mikelis, Constantinos M., and Mattheolabakis, George
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LUNG cancer ,DRUG carriers ,SCORPION venom ,TOXINS ,PEPTIDES ,PEPTIDE antibiotics ,CANCER research ,GLIOBLASTOMA multiforme - Abstract
In the generational evolution of nano-based drug delivery carriers, active targeting has been a major milestone for improved and selective drug accumulation in tissues and cell types beyond the existing passive targeting capabilities. Among the various active targeting moieties, chlorotoxin, a peptide extracted from scorpions, demonstrated promising tumor cell accumulation and selection. With lung cancer being among the leading diagnoses of cancer-related deaths in both men and women, novel therapeutic methodologies utilizing nanotechnology for drug delivery emerged. Given chlorotoxin's promising biological activity, we explore its potential against lung cancer and its utilization for active targeting against this cancer's tumor cells. Our analysis indicates that despite the extensive chlorotoxin's research against glioblastoma, lung cancer research with the molecule has been limited, despite some promising early results. [ABSTRACT FROM AUTHOR]
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- 2022
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16. ORAL CANCER METASTASIS IS INFLUENCED BY GΑ12 SIGNALING: OP189.
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Abraham, Mannil Thomas, Gan, Chai Phei, Patel, Vyomesh, Mikelis, Constantinos M., Zain, Rosnah binti, Molinolo, Alfredo A., Abdul Rahman, Zainal Ariff, Gutkind, Silvio J., and Cheong, Sok Ching
- Published
- 2015
17. Protein tyrosine phosphatase receptor-ξ1 deletion triggers defective heart morphogenesis in mice and zebrafish.
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Katraki-Pavlou, Stamatiki, Kastana, Pinelopi, Bousis, Dimitris, Ntenekou, Despoina, Varela, Aimilia, Davos, Constantinos H., Nikou, Sophia, Papadaki, Eleni, Tsigkas, Grigorios, Athanasiadis, Emmanouil, Herradon, Gonzalo, Mikelis, Constantinos M., Beis, Dimitris, and Papadimitriou, Evangelia
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PROTEIN-tyrosine phosphatase ,PHOSPHOPROTEIN phosphatases ,FETAL heart ,BRACHYDANIO ,EMBRYONIC stem cells ,DOBUTAMINE ,CONTRACTILE proteins - Abstract
Protein tyrosine phosphatase receptor-ξ1 (PTPRZ1) is a transmembrane tyrosine phosphatase receptor highly expressed in embryonic stem cells. In the present work, gene expression analyses of Ptprz1-/- and Ptprz1þ/þ mice endothelial cells and hearts pointed to an unidentified role of PTPRZ1 in heart development through the regulation of heart-specific transcription factor genes. Echocardiography analysis in mice identified that both systolic and diastolic functions are affected in Ptprz1-/-compared with Ptprz1þ/þ hearts, based on a dilated left ventricular (LV) cavity, decreased ejection fraction and fraction shortening, and increased angiogenesis in Ptprz1-/- hearts, with no signs of cardiac hypertrophy. A zebrafish ptprz1-/- knockout was also generated and exhibited misregulated expression of developmental cardiac markers, bradycardia, and defective heart morphogenesis characterized by enlarged ventricles and defected contractility. A selective PTPRZ1 tyrosine phosphatase inhibitor affected zebrafish heart development and function in a way like what is observed in the ptprz1-/- zebrafish. The same inhibitor had no effect in the function of the adult zebrafish heart, suggesting that PTPRZ1 is not important for the adult heart function, in line with data from the human cell atlas showing very low to negligible PTPRZ1 expression in the adult human heart. However, in line with the animal models, Ptprz1 was expressed in many different cell types in the human fetal heart, such as valvar, fibroblast-like, cardiomyocytes, and endothelial cells. Collectively, these data suggest that PTPRZ1 regulates cardiac morphogenesis in a way that subsequently affects heart function and warrant further studies for the involvement of PTPRZ1 in idiopathic congenital cardiac pathologies. [ABSTRACT FROM AUTHOR]
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- 2022
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18. MiRNAs as Anti-Angiogenic Adjuvant Therapy in Cancer: Synopsis and Potential.
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Lahooti, Behnaz, Poudel, Sagun, Mikelis, Constantinos M., and Mattheolabakis, George
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NEOVASCULARIZATION inhibitors ,CANCER treatment ,DRUG target ,MICRORNA ,REGULATOR genes - Abstract
Angiogenesis is a key mechanism for tumor growth and metastasis and has been a therapeutic target for anti-cancer treatments. Intensive vascular growth is concomitant with the rapidly proliferating tumor cell population and tumor outgrowth. Current angiogenesis inhibitors targeting either one or a few pro-angiogenic factors or a range of downstream signaling molecules provide clinical benefit, but not without significant side effects. miRNAs are important post-transcriptional regulators of gene expression, and their dysregulation has been associated with tumor progression, metastasis, resistance, and the promotion of tumor-induced angiogenesis. In this mini-review, we provide a brief overview of the current anti-angiogenic approaches, their molecular targets, and side effects, as well as discuss existing literature on the role of miRNAs in angiogenesis. As we highlight specific miRNAs, based on their activity on endothelial or cancer cells, we discuss their potential for anti-angiogenic targeting in cancer as adjuvant therapy and the importance of angiogenesis being evaluated in such combinatorial approaches. [ABSTRACT FROM AUTHOR]
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- 2021
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19. Pleiotrophin selectively binds to vascular endothelial growth factor receptor 2 and inhibits or stimulates cell migration depending on ανβ3 integrin expression.
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Lamprou, Margarita, Kastana, Pinelopi, Kofina, Fani, Tzoupis, Ηaralampos, Barmpoutsi, Spyridoula, Sajib, Md Sanaullah, Koutsioumpa, Marina, Poimenidi, Evangelia, Zompra, Aikaterini A., Tassopoulos, Dimitrios, Choleva, Effrosyni, Tselios, Theodore, Mikelis, Constantinos M., and Papadimitriou, Evangelia
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VASCULAR endothelial growth factor receptors ,CELL migration ,CELLULAR control mechanisms ,PROTHROMBIN - Abstract
Pleiotrophin (PTN) has a moderate stimulatory effect on endothelial cell migration through α
ν β3 integrin, while it decreases the stimulatory effect of vascular endothelial growth factor A (VEGFA) and inhibits cell migration in the absence of αν β3 through unknown mechanism(s). In the present work, by using a multitude of experimental approaches, we show that PTN binds to VEGF receptor type 2 (VEGFR2) with a KD of 11.6 nM. Molecular dynamics approach suggests that PTN binds to the same VEGFR2 region with VEGFA through its N-terminal domain. PTN inhibits phosphorylation of VEGFR2 at Tyr1175 and still stimulates endothelial cell migration in the presence of a selective VEGFR2 tyrosine kinase inhibitor. VEGFR2 downregulation by siRNA or an anti-VEGFR2 antibody that binds to the ligand-binding VEGFR2 domain also induce endothelial cell migration, which is abolished by a function-blocking antibody against αν β3 or the peptide PTN112−136 that binds αν β3 and inhibits PTN binding. In cells that do not express αν β3 , PTN decreases both VEGFR2 Tyr1175 phosphorylation and cell migration in a VEGFR2-dependent manner. Collectively, our data identify VEGFR2 as a novel PTN receptor involved in the regulation of cell migration by PTN and contribute to the elucidation of the mechanism of activation of endothelial cell migration through the interplay between VEGFR2 and αν β3 . [ABSTRACT FROM AUTHOR]- Published
- 2020
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20. An iPSC-Derived Neuron Model of CLN3 Disease Facilitates Small Molecule Phenotypic Screening.
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Kinarivala, Nihar, Morsy, Ahmed, Patel, Ronak, Carmona, Angelica V., Sajib, Md. Sanaullah, Raut, Snehal, Mikelis, Constantinos M., Al-Ahmad, Abraham, and Trippier, Paul C.
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- 2020
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21. Nanoparticle Delivery and Tumor Vascular Normalization: The Chicken or The Egg?
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Mattheolabakis, George and Mikelis, Constantinos M.
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EGGS ,TUMORS ,CANCER cells ,BLOOD vessels ,NEOVASCULARIZATION - Abstract
Tumor-induced angiogenesis has been a significant focus of anti-cancer therapies for several decades. The immature and "leaky" tumor vasculature leads to significant cancer cell intravasation, increasing the metastatic potential, while the disoriented and hypo-perfused tumor vessels hamper the anti-tumor efficacy of immune cells and prevent the efficient diffusion of chemotherapeutic drugs. Therefore, tumor vascular normalization has emerged as a new treatment goal, aiming to provide a mature tumor vasculature, with higher perfusion, decreased cancer cell extravasation, and higher efficacy for anti-cancer therapies. Here we propose an overview of the nanodelivery approaches that target tumor vasculature, aiming to achieve vascular normalization. At the same time, abnormal vascular architecture and leaky tumor vessels have been the cornerstone for nanodelivery approaches through the enhanced permeability and retention (EPR) effect. Vascular normalization presents new opportunities and requirements for efficient nanoparticle delivery against the tumor cells and overall improved anti-cancer therapies. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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22. Epidermal loss of Gαq confers a migratory and differentiation defect in keratinocytes.
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Doçi, Colleen L., Mikelis, Constantinos M., Callejas-Valera, Juan Luis, Hansen, Karina K., Molinolo, Alfredo A., Inoue, Asuka, Offermanns, Stefan, and Gutkind, J. Silvio
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CELL differentiation , *KERATINOCYTES , *MEMBRANE proteins , *G protein coupled receptors , *CELLULAR signal transduction , *GENETIC overexpression - Abstract
G-protein coupled receptors (GPCRs), which activate heterotrimeric G proteins, are an essential class of transmembrane receptors that are responsible for a myriad of signaling events in normal and pathologic conditions. Two members of the G protein family, Gαq and Gα11, activate one of the main GPCR pathways and function as oncogenes by integrating mitogen-stimulated signaling cascades that are active under malignant conditions. Recently, it has been shown that targeted deletion of Gα11 and Gαq from endothelial cells impairs the Rho-mediated formation of focal adherens junctions, suggesting that Gα11/q signaling may also play a significant role in cytoskeletal-mediated cellular responses in epithelial cells. Indeed, combined deletion of Gα11 and Gαq confers a significant migratory defect in keratinocytes that delays cutaneous wound healing in an in vivo setting. This delay can be attributed to a defect during the reepithelialization phase due to significantly attenuated migratory capacity of Gαq-null keratinocytes under combined Gα11 deficiency. In fact, cells lacking Gα11/q demonstrate a severely reduced ability to respond to mitogenic and migratory signals in the microenvironment, leading to inappropriate and premature terminal differentiation. These results suggest that Gα11/q signaling pathways may be critical for integrating mitogenic signals and cytoskeletal function to achieve normal physiological responses. Emergence of a malignant phenotype may therefore arise from both under- and overexpression of Gα11/q signaling, implicating its upstream regulation as a potential therapeutic target in a host of pathologic conditions. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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23. CARD9-Dependent Neutrophil Recruitment Protects against Fungal Invasion of the Central Nervous System.
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Drummond, Rebecca A., Collar, Amanda L., Swamydas, Muthulekha, Rodriguez, Carlos A., Lim, Jean K., Mendez, Laura M., Fink, Danielle L., Hsu, Amy P., Zhai, Bing, Karauzum, Hatice, Mikelis, Constantinos M., Rose, Stacey R., Ferre, Elise M. N., Yockey, Lynne, Lemberg, Kimberly, Kuehn, Hye Sun, Rosenzweig, Sergio D., Lin, Xin, Chittiboina, Prashant, and Datta, Sandip K.
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NEUTROPHILS ,GRANULOCYTES ,PHAGOCYTES ,FUNGI ,CRYPTOGAMS - Abstract
Candida is the most common human fungal pathogen and causes systemic infections that require neutrophils for effective host defense. Humans deficient in the C-type lectin pathway adaptor protein CARD9 develop spontaneous fungal disease that targets the central nervous system (CNS). However, how CARD9 promotes protective antifungal immunity in the CNS remains unclear. Here, we show that a patient with CARD9 deficiency had impaired neutrophil accumulation and induction of neutrophil-recruiting CXC chemokines in the cerebrospinal fluid despite uncontrolled CNS Candida infection. We phenocopied the human susceptibility in Card9
-/- mice, which develop uncontrolled brain candidiasis with diminished neutrophil accumulation. The induction of neutrophil-recruiting CXC chemokines is significantly impaired in infected Card9-/- brains, from both myeloid and resident glial cellular sources, whereas cell-intrinsic neutrophil chemotaxis is Card9-independent. Taken together, our data highlight the critical role of CARD9-dependent neutrophil trafficking into the CNS and provide novel insight into the CNS fungal susceptibility of CARD9-deficient humans. [ABSTRACT FROM AUTHOR]- Published
- 2015
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24. Genetic Identification of SEMA3F as an Antilymphangiogenic Metastasis Suppressor Gene in Head and Neck Squamous Carcinoma.
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Doçi, Colleen L., Mikelis, Constantinos M., Lionakis, Michail S., Molinolo, Alfredo A., and Gutkind, J. Silvio
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SQUAMOUS cell carcinoma , *TUMOR suppressor proteins , *HEAD & neck cancer , *METASTASIS , *BIOMARKERS - Abstract
Head and neck squamous cell carcinomas (HNSCC) often metastasize to locoregional lymph nodes, and lymph node involvement represents one of the most important prognostic factors of poor clinical outcome. HNSCCs are remarkably lymphangiogenic and represent a clear example of a cancer that utilizes the lymphatic vasculature for malignant dissemination; however, the molecular mechanisms underlying lymphangio-genesis in HNSCC is still poorly understood. Of interest, we found that an axon guidance molecule, Semaphorin 3F (SEMA3F), is among the top 1% underexpressed genes in HNSCC, and that genomic loss of SEMA3F correlates with increased metastasis and decreased survival. SEMA3F acts on its coreceptors, plexins and neuropilins, among which neuro-pilin-2 (NRP2) is highly expressed in lymphatic endothelial cells (LEC) but not in oral epithelium and most HNSCCs. We show that recombinant SEMA3F promotes LEC collapse and potently inhibits lymphangiogenesis in vivo. By reconstituting all possible plexin and neuropilin combinations, we found that SEMA3F acts through multiple receptors, but predominantly requires NRP2 to signal in LECs. Using orthotopic HNSCC metastasis mouse models, we provide direct evidence that SEMA3F re-expression diminishes lymphangiogenesis and lymph node metastasis. Furthermore, analysis of a large tissue collection revealed that SEMA3F is progressively lost during HNSCC progression, concomitant with increased tumor lymphangiogenesis. SEMA3F is localized to 3p21, an early and frequently deleted locus in HNSCC and many other prevalent human malignancies. Thus, SEMA3F may represent an anti-lymphangiogenic metastasis suppressor gene widely lost during cancer progression, hence serving as a prognostic biomarker and an attractive target for therapeutic intervention to halt metastasis. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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25. SDF-1/CXCL12 induces directional cell migration and spontaneous metastasis via a CXCR4/Gai/mTORC1 axis.
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Dillenburg-Pilla, Patricia, Patel, Vyomesh, Mikelis, Constantinos M., Zárate-Bladés, Carlos Rodrigo, Doçi, Colleen L., Amornphimoltham, Panomwat, Zhiyong Wang, Martin, Daniel, Leelahavanichkul, Kantima, Dorsam, Robert T., Masedunskas, Andrius, Weigert, Roberto, Molinolo, Alfredo A., and Gutkind, J. Silvio
- Subjects
CELL migration ,METASTASIS ,CHEMOKINE receptors ,CHEMOKINES ,EXPERIMENTAL biology - Abstract
Translocation of bacteria and their products across the intestinal barrier is common in patients with liver disease, and there is evidence that experimental liver fibrosis depends on bacterial translocation. The purpose of our study was to investigate liver fibrosis in conventional and germ-free (GF) C57BL/6 mice. Chronic liver injury was induced by administration of thioacetamide (TAA) in the drinking water for 21 wk or by repeated intraperitoneal injections of carbon tetrachloride (CCl
4 ). Increased liver fibrosis was observed in GF mice compared with conventional mice. Hepatocytes showed more toxin-induced oxi-dative stress and cell death. This was accompanied by increased activation of hepatic stellate cells, but hepatic mediators of inflammation were not significantly different. Similarly, a genetic model using Myd88/Trif-deficient mice, which lack downstream innate immunity signaling, had more severe fibrosis than wild-type mice. Isolated Myd88/Trif-deficient hepatocytes were more susceptible to toxin-induced cell death in culture. In conclusion, the commensal microbiota prevents fibrosis upon chronic liver injury in mice. This is the first study describing a beneficial role of the commensal microbiota in maintaining liver homeostasis and preventing liver fibrosis. [ABSTRACT FROM AUTHOR]- Published
- 2015
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26. Endothelial‐specific nanodelivery of Rho kinase inhibitors: Targeting tumor angiogenesis and metastasis.
- Author
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Lahooti, Behnaz, Akwii, Racheal, Mattheolabakis, George, and Mikelis, Constantinos M.
- Published
- 2022
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27. PDZ-RhoGEF and LARG Are Essential for Embryonic Development and Provide a Link between Thrombin and LPA Receptors and Rho Activation.
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Mikelis, Constantinos M., Palmby, Todd R., Simaan, May, Wenling Li, Szabo, Roman, Lyons, Ruth, Martin, Daniel, Yagi, Hiroshi, Fukuhara, Shigetomo, Chikumi, Hiroki, Galisteo, Rebeca, Mukouyama, Yoh-suke, Bugge, Thomas H., and Gutkind, J. Silvio
- Subjects
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G protein coupled receptors , *THROMBIN , *GUANOSINE triphosphatase , *BIOCHEMISTRY , *EMBRYOLOGY - Abstract
G protein-coupled receptors (GPCRs) linked to both members of the Gα12 family of heterotrimeric G proteins α subunits, Gα12 and Gα13, regulate the activation of Rho GTPases, thereby contributing to many key biological processes. Multiple Rho GEFs have been proposed to link Gα12/13 GPCRs to Rho activation, including PDZ-RhoGEF (PRG), leukemia-associated Rho GEF (LARG), p115-RhoGEF (p115), lymphoid blast crisis (Lbc), and Dbl. PRG, LARG, and p115 share the presence of a regulator of G protein signaling homology (RGS) domain. There is limited information on the biological roles of this RGS-containing family of RhoGEFs in vivo. p115-deficient mice are viable with some defects in the immune system and gastrointestinal motor dysfunctions, whereas in an initial study we showed that mice deficient for Larg are viable and resistant to salt-induced hypertension. Here, we generated knock-out mice for Prg and observed that these mice do not display any overt phenotype. However, deficiency in Prg and Larg leads to complex developmental defects and early embryonic lethality. Signaling from Gα11/q-linked GPCRs to Rho was not impaired in mouse embryonic fibroblasts defective in all three RGS-containing RhoGEFs. However, a combined lack of Prg, Larg, and p115 expression abolished signaling through Gα12/13 to Rho and thrombin-induced cell proliferation, directional migration, and nuclear signaling through JNK and p38. These findings provide evidence of an essential role for the RGS-containing RhoGEF family in signaling to Rho by Gα12/13-coupled GPCRs, which may likely play a critical role during embryonic development. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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28. Role of bFGF in Acquired Resistance upon Anti-VEGF Therapy in Cancer.
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Zahra, Fatema Tuz, Sajib, Md. Sanaullah, Mikelis, Constantinos M., and Ribatti, Domenico
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VASCULAR endothelial growth factor antagonists ,NEOVASCULARIZATION inhibitors ,FIBROBLASTS ,GROWTH factors ,NEOVASCULARIZATION ,CANCER ,MOLECULAR biology ,TUMORS ,DRUG resistance in cancer cells ,MEDICAL research - Abstract
Simple Summary: Anti-angiogenic therapies targeting the vascular endothelial growth factor (VEGF) signaling are established in the arsenal of cancer treatments. Despite the expectations, their benefits are temporary in cancer patients, partly due to the compensatory function of other angiogenic growth factors. This review focuses on the role of basic fibroblast growth factor (bFGF), one of the highly implicated players in the emergence of resistance to anti-angiogenic approaches. Here, we summarize data from various tumor types where bFGF is upregulated after anti-angiogenic treatment, the molecular mechanisms involved, and we highlight the current status and future perspectives of multi-target anti-angiogenic drugs for cancer. Anti-angiogenic approaches targeting the vascular endothelial growth factor (VEGF) signaling pathway have been a significant research focus during the past decades and are well established in clinical practice. Despite the expectations, their benefit is ephemeral in several diseases, including specific cancers. One of the most prominent side effects of the current, VEGF-based, anti-angiogenic treatments remains the development of resistance, mostly due to the upregulation and compensatory mechanisms of other growth factors, with the basic fibroblast growth factor (bFGF) being at the top of the list. Over the past decade, several anti-angiogenic approaches targeting simultaneously different growth factors and their signaling pathways have been developed and some have reached the clinical practice. In the present review, we summarize the knowledge regarding resistance mechanisms upon anti-angiogenic treatment, mainly focusing on bFGF. We discuss its role in acquired resistance upon prolonged anti-angiogenic treatment in different tumor settings, outline the reported resistance mechanisms leading to bFGF upregulation, and summarize the efforts and outcome of combined anti-angiogenic approaches to date. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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29. Glycolysis is integral to histamine-induced endothelial hyperpermeability.
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Ziogas, Athanasios, Sajib, Md Sanaullah, Lim, Jong-Hyung, Alves, Tiago C., Das, Anupam, Witt, Anke, Hagag, Eman, Androulaki, Nikolais, Grossklaus, Sylvia, Gerlach, Michael, Noll, Thomas, Grinenko, Tatyana, Mirtschink, Peter, Hajishengallis, George, Chavakis, Triantafyllos, Mikelis, Constantinos M., and Sprott, David
- Published
- 2021
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30. Assessing the Current State of Lung Cancer Chemoprevention: A Comprehensive Overview.
- Author
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Ashraf-Uz-Zaman, Md, Bhalerao, Aditya, Mikelis, Constantinos M., Cucullo, Luca, and German, Nadezhda A.
- Subjects
ANTINEOPLASTIC agents ,HUMAN microbiota ,CHEMOPREVENTION ,CLINICAL trials ,HEALTH status indicators ,LUNG tumors ,MOLECULAR structure ,RACE ,SEX distribution ,NEOPLASTIC cell transformation - Abstract
Chemoprevention of lung cancer is thought to significantly reduce the risk of acquiring these conditions in the subpopulation of patients with underlying health issues, such as chronic obstructive pulmonary disorder and smoking-associated lung problems. Many strategies have been tested in the previous decades, with very few translating to successful clinical trials in specific subpopulations of patients. In this review, we analyze these strategies, as well as new approaches that have emerged throughout the last few years, including synthetic lethality concept and microbiome-induced regulation of lung carcinogenesis. Overall, the continuous effort in the area of lung chemoprevention is required to develop practical therapeutical approaches. Given the inconsistency of results obtained in clinical trials targeting lung cancer chemoprevention in various subgroups of patients that differ in the underlying health condition, race, and gender, we believe that individualized approaches will have more promise than generalized treatments. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
31. Low Dose of Penfluridol Inhibits VEGF-Induced Angiogenesis.
- Author
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Srivastava, Suyash, Zahra, Fatema Tuz, Gupta, Nehal, Tullar, Paul E., Srivastava, Sanjay K., and Mikelis, Constantinos M.
- Subjects
METASTATIC breast cancer ,NEOVASCULARIZATION ,VASCULAR endothelial growth factors ,ENDOTHELIAL cells ,CELL migration ,BLOOD-brain barrier - Abstract
Metastasis is considered a major burden in cancer, being responsible for more than 90% of cancer-related deaths. Tumor angiogenesis is one of the main processes that lead to tumor metastasis. Penfluridol is a classic and commonly used antipsychotic drug, which has a great ability to cross the blood–brain barrier. Recent studies have revealed that penfluridol has significant anti-cancer activity in diverse tumors, such as metastatic breast cancer and glioblastoma. Here, we aim to identify the effect of low doses of penfluridol on tumor microenvironment and compare it with its effect on tumor cells. Although low concentration of penfluridol was not toxic for endothelial cells, it blocked angiogenesis in vitro and in vivo. In vitro, penfluridol inhibited VEGF-induced primary endothelial cell migration and tube formation, and in vivo, it blocked VEGF- and FGF-induced angiogenesis in the matrigel plug assay. VEGF-induced VEGFR2 phosphorylation and the downstream p38 and ERK signaling pathways were not affected in endothelial cells, although VEGF-induced Src and Akt activation were abrogated by penfluridol treatment. When cancer cells were treated with the same low concentration of penfluridol, basal Src activation levels were mildly impaired, thus impacting their cell migration and wound healing efficiency. The potential of cancer-induced paracrine effect on endothelial cells was explored, although that did not seem to be a player for angiogenesis. Overall, our data demonstrates that low penfluridol levels, similar to the ones clinically used for anti-psychotic conditions, suppress angiogenic efficiency in the tumor microenvironment. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
32. Role of Angiopoietin-2 in Vascular Physiology and Pathophysiology.
- Author
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Akwii, Racheal G., Sajib, Md S., Zahra, Fatema T., and Mikelis, Constantinos M.
- Subjects
ANGIOPOIETIN-2 ,VASCULAR endothelial growth factors ,GROWTH factors ,ANGIOPOIETINS ,ENDOTHELIAL cells ,MOLECULAR pathology - Abstract
Angiopoietins 1–4 (Ang1–4) represent an important family of growth factors, whose activities are mediated through the tyrosine kinase receptors, Tie1 and Tie2. The best characterized are angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2). Ang1 is a potent angiogenic growth factor signaling through Tie2, whereas Ang2 was initially identified as a vascular disruptive agent with antagonistic activity through the same receptor. Recent data demonstrates that Ang2 has context-dependent agonist activities. Ang2 plays important roles in physiological processes and the deregulation of its expression is characteristic of several diseases. In this review, we summarize the activity of Ang2 on blood and lymphatic endothelial cells, its significance in human physiology and disease, and provide a current view of the molecular signaling pathways regulated by Ang2 in endothelial cells. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
33. DNA Repair Response Adaptors: Novel Targets for Vasoproliferative Retinopathy?
- Author
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Sajib, Md Sanaullah and Mikelis, Constantinos M.
- Published
- 2019
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- View/download PDF
34. GM-CSF therapy in human caspase recruitment domain–containing protein 9 deficiency.
- Author
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Drummond, Rebecca A., Zahra, Fatema Tuz, Natarajan, Mukil, Swamydas, Muthulekha, Hsu, Amy P., Wheat, L. Joseph, Gavino, Christina, Vinh, Donald C., Holland, Steven M., Mikelis, Constantinos M., and Lionakis, Michail S.
- Published
- 2018
- Full Text
- View/download PDF
35. RhoA and ROCK mediate histamine-induced vascular leakage and anaphylactic shock.
- Author
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Mikelis, Constantinos M., Simaan, May, Ando, Koji, Fukuhara, Shigetomo, Sakurai, Atsuko, Amornphimoltham, Panomwat, Masedunskas, Andrius, Weigert, Roberto, Chavakis, Triantafyllos, Adams, Ralf H., Offermanns, Stefan, Mochizuki, Naoki, Zheng, Yi, and Gutkind, J. Silvio
- Published
- 2015
- Full Text
- View/download PDF
36. Delayed Exercise-induced Upregulation of Angiogenic Proteins and Recovery of Motor Function after Photothrombotic Stroke in Mice.
- Author
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Al Shoyaib, Abdullah, Alamri, Faisal F., Biggers, Abbie, Karamyan, Serob T., Arumugam, Thiruma V., Ahsan, Fakhrul, Mikelis, Constantinos M., Al-Hilal, Taslim A., and Karamyan, Vardan T.
- Subjects
- *
VASCULAR endothelial growth factors , *MOLECULAR motor proteins , *ISCHEMIC stroke , *CEREBRAL cortex , *MICE - Abstract
• Dose-dependent effect of delayed physiotherapy on post-stroke recovery was studied. • Overnight but not 5 h voluntary wheel running promoted post-stroke motor recovery. • Recovery of motor function was accompanied by induction of angiogenic proteins. • Induction of angiogenic proteins was observed in peri-infract region and infarct core. • Infarct core maybe involved in mechanisms governing post-stroke functional recovery. Treatments promoting post-stroke functional recovery continue to be an unmet therapeutic problem with physical rehabilitation being the most reproduced intervention in preclinical and clinical studies. Unfortunately, physiotherapy is typically effective at high intensity and early after stroke – requirements that are hardly attainable by stroke survivors. The aim of this study was to directly evaluate and compare the dose-dependent effect of delayed physical rehabilitation (daily 5 h or overnight voluntary wheel running; initiated on post-stroke day 7 and continuing through day 21) on recovery of motor function in the mouse photothrombotic model of ischemic stroke and correlate it with angiogenic potential of the brain. Our observations indicate that overnight but not 5 h access to running wheels facilitates recovery of motor function in mice in grid-walking test. Western blotting and immunofluorescence microscopy experiments evaluating the expression of angiogenesis-associated proteins VEGFR2, doppel and PDGFRβ in the peri-infarct and corresponding contralateral motor cortices indicate substantial upregulation of these proteins (≥2-fold) in the infarct core and surrounding cerebral cortex in the overnight running mice on post-stroke day 21. These findings indicate that there is a dose-dependent relationship between the extent of voluntary exercise, motor recovery and expression of angiogenesis-associated proteins in this expert-recommended mouse ischemic stroke model. Notably, our observations also point out to enhanced angiogenesis and presence of pericytes within the infarct core region during the chronic phase of stroke, suggesting a potential contribution of this tissue area in the mechanisms governing post-stroke functional recovery. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
37. Analogs of penfluridol as chemotherapeutic agents with reduced central nervous system activity.
- Author
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Ashraf-Uz-Zaman, Md, Sajib, Md Sanaullah, Cucullo, Luca, Mikelis, Constantinos M., and German, Nadezhda A.
- Subjects
- *
PENFLURIDOL , *CANCER chemotherapy , *CENTRAL nervous system diseases , *LUNG cancer treatment , *G protein coupled receptors , *ANTIPSYCHOTIC agents - Abstract
Graphical abstract Highlights • Anticancer doses of penfluridol inhibit a majority of G-protein coupled receptors. • Analysis of antipsychotic pharmacophore allowed to design less toxic analogs. • Confirmed in vitro cytotoxicity of analogs, in vivo permeability of the BBB. • Clinical chemistry analysis confirmed optimized toxicity profile. Abstract Several recent reports have highlighted the feasibility of the use of penfluridol, a well-known antipsychotic agent, as a chemotherapeutic agent. In vivo experiments have confirmed the cytotoxic activity of penfluridol in triple-negative breast cancer model, lung cancer model, and further studies have been proposed to assess its anticancer activity and viability for the treatment of glioblastomas. However, penfluridol anticancer activity was observed at a dosage significantly higher than that administered in antipsychotic therapy, thus raising the concern for the potential onset of CNS side effects in patients undergoing intensive pharmacological treatment. In this study, we evaluate the potential CNS toxicity of penfluridol side by side with a set of analogs. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
38. Decreased Lymphangiogenesis and Lymph Node Metastasis by mTOR Inhibition in Head and Neck Cancer.
- Author
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Patel, Vyomesh, Marsh, Christina A., Dorsam, Robert T., Mikelis, Constantinos M., Masedunskas, Andrius, Amornphimoltham, Panomwat, Nathan, Cherie Ann, Singh, Bhuvanesh, Weigert, Roberto, Molinolo, Alfredo A., and Gutkind, J. Silvio
- Subjects
- *
SQUAMOUS cell carcinoma , *LYMPH node cancer , *LYMPHATIC cancer , *LYMPHATIC metastasis , *RAPAMYCIN - Abstract
Despite our improved understanding of cancer, the 5-year survival rate for head and neck squamous cell carcinomas (HNSCC) patients remains relatively unchanged at 50% for the past three decades. HNSCCs often metastasize to locoregional lymph nodes, and lymph node involvement represents one of the most important prognostic factors of poor clinical outcome. Among the multiple dysregulated molecular mechanismin HNSCCs, emerging basic, preclinical, and clinical findings support the importance of the mTOR signaling route in HNSCC progression. Indeed, we observed here that the activation of mTOR is a widespread event in clinical specimens of HNSCCs invading locoregional lymph nodes. We developed an orthotopic model of HNSCC consisting of the implantation of HNSCC cells into the tongues of immunocompromised mice. These orthotopic tumors spontaneously metastasize to the cervical lymph nodes, where the presence of HNSCC cells can be revealed by histologic and immunohistochemical evaluation. Both primary and metastatic experimental HNSCC lesions exhibited elevated mTOR activity. The ability tomonitor and quantitate lymph node invasion in this model system enabled us to explore whether the blockade of mTOR could impact HNSCC metastasis. We found that inhibition of mTOR with rapamycin and the rapalog RAD001 diminished lymphangiogenesis in the primary tumors and prevented the dissemination of HNSCC cancer cells to the cervical lymph nodes, thereby prolonging animal survival. These findings may provide a rationale for the future clinical evaluation of mTOR inhibitors, including rapamycin and its analogues, as part of a molecular-targeted metastasis preventive strategy for the treatment of patients with HNSCC. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
39. The Role of Soluble CD163 (sCD163) in Human Physiology and Pathophysiology.
- Author
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Plevriti A, Lamprou M, Mourkogianni E, Skoulas N, Giannakopoulou M, Sajib MS, Wang Z, Mattheolabakis G, Chatzigeorgiou A, Marazioti A, and Mikelis CM
- Subjects
- Humans, Inflammation metabolism, Inflammation blood, Macrophages metabolism, Solubility, Biomarkers metabolism, Biomarkers blood, Animals, Receptors, Cell Surface metabolism, Antigens, Differentiation, Myelomonocytic metabolism, Antigens, Differentiation, Myelomonocytic blood, Antigens, CD metabolism, Antigens, CD blood
- Abstract
Soluble CD163 (sCD163) is a circulating inflammatory mediator, indicative of acute and chronic, systemic and non-systemic inflammatory conditions. It is the cleavage outcome, consisting of almost the entire extracellular domain, of the CD163, a receptor expressed in monocytic lineages. Its expression is proportional to the abundance of CD163
+ macrophages. Various mechanisms trigger the shedding of the CD163 receptor or the accumulation of CD163-expressing macrophages, inducing the sCD163 concentration in the circulation and bodily fluids. The activities of sCD163 range from hemoglobin (Hb) scavenging, macrophage marker, decoy receptor for cytokines, participation in immune defense mechanisms, and paracrine effects in various tissues, including the endothelium. It is an established marker of macrophage activation and thus participates in many diseases, including chronic inflammatory conditions, such as atherosclerosis, asthma, and rheumatoid arthritis; acute inflammatory conditions, such as sepsis, hepatitis, and malaria; insulin resistance; diabetes; and tumors. The sCD163 levels have been correlated with the severity, stage of the disease, and clinical outcome for many of these conditions. This review article summarizes the expression and role of sCD163 and its precursor protein, CD163, outlines the sCD163 generation mechanisms, the biological activities, and the known underlying molecular mechanisms, with an emphasis on its impact on the endothelium and its contribution in the pathophysiology of human diseases.- Published
- 2024
- Full Text
- View/download PDF
40. Tumor-induced endothelial RhoA activation mediates tumor cell transendothelial migration and metastasis.
- Author
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Sajib MS, Zahra FT, Lamprou M, Akwii RG, Park JH, Osorio M, Tullar P, Doci CL, Zhang C, Huveneers S, Van Buul JD, Wang MH, Markiewski MM, Srivastava SK, Zheng Y, Gutkind JS, Hu J, Bickel U, Maeda DY, Zebala JA, Lionakis MS, Trasti S, and Mikelis CM
- Abstract
The endothelial barrier plays an active role in transendothelial tumor cell migration during metastasis, however, the endothelial regulatory elements of this step remain obscure. Here we show that endothelial RhoA activation is a determining factor during this process. Breast tumor cell-induced endothelial RhoA activation is the combined outcome of paracrine IL-8-dependent and cell-to-cell contact β
1 integrin-mediated mechanisms, with elements of this pathway correlating with clinical data. Endothelial-specific RhoA blockade or in vivo deficiency inhibited the transendothelial migration and metastatic potential of human breast tumor and three murine syngeneic tumor cell lines, similar to the pharmacological blockade of the downstream RhoA pathway. These findings highlight endothelial RhoA as a potent, universal target in the tumor microenvironment for anti-metastatic treatment of solid tumors.- Published
- 2024
- Full Text
- View/download PDF
41. Comprehensive analysis of lipid nanoparticle formulation and preparation for RNA delivery.
- Author
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Haque MA, Shrestha A, Mikelis CM, and Mattheolabakis G
- Abstract
Nucleic acid-based therapeutics are a common approach that is increasingly popular for a wide spectrum of diseases. Lipid nanoparticles (LNPs) are promising delivery carriers that provide RNA stability, with strong transfection efficiency, favorable and tailorable pharmacokinetics, limited toxicity, and established translatability. In this review article, we describe the lipid-based delivery systems, focusing on lipid nanoparticles, the need of their use, provide a comprehensive analysis of each component, and highlight the advantages and disadvantages of the existing manufacturing processes. We further summarize the ongoing and completed clinical trials utilizing LNPs, indicating important aspects/questions worth of investigation, and analyze the future perspectives of this significant and promising therapeutic approach., Competing Interests: The authors declare no competing interest., (© 2024 The Authors.)
- Published
- 2024
- Full Text
- View/download PDF
42. Mechanistic Aspects of Biphenyl Urea-Based Analogues in Triple-Negative Breast Cancer Cell Lines.
- Author
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Bandy R, Shahi S, Quagraine N, Shahbazi Nia S, Howlader MSI, Srivenugopal K, Stephan C, Das H, Mikelis CM, and German NA
- Abstract
Triple-negative breast cancer (TNBC) poses significant challenges due to its aggressive nature and limited treatment options. In this study, we investigated the impact of urea-based compounds on TNBC cells to uncover their mechanisms of action and therapeutic potential. Notably, polypharmacology urea analogues were found to work via p53-related pathways, and their cytotoxic effects were amplified by the modulation of oxidative phosphorylation pathways in the mitochondria of cancer cells. Specifically, compound 1 demonstrated an uncoupling effect on adenosine triphosphate (ATP) synthesis, leading to a time- and concentration-dependent shift toward glycolysis-based ATP production in MDA-MB-231 cells. At the same time, no significant changes in ATP synthesis were observed in noncancerous MCF10A cells. Moreover, the unique combination of mitochondrial- and p53-related effects leads to a higher cytotoxicity of urea analogues in cancer cells. Notably, the majority of tested clinical agents, but sorafenib, showed significantly higher toxicity in MCF10A cells. To test our hypothesis of sensitizing cancer cells to the treatment via modulation of mitochondrial health, we explored the combinatorial effects of urea-based analogues with established chemotherapeutic agents commonly used in TNBC treatment. Synergistic effects were evident in most tested combinations in TNBC cell lines, while noncancerous MCF10A cells exhibited higher resistance to these combination treatments. The combination of compound 1 with SN38 displayed nearly 60-fold selectivity toward TNBC cells over MCF10A cells. Encouragingly, combinations involving compound 1 restored the sensitivity of TNBC cells to cisplatin. In conclusion, our study provides valuable insights into the mechanisms of action of urea-based compounds in TNBC cells. The observed induction of mitochondrial membrane depolarization, inhibition of superoxide dismutase activity, disruption of ATP synthesis, and cell-line-specific responses contribute to their cytotoxic effects. Additionally, we demonstrated the synergistic potential of compound 1 to enhance the efficacy of existing TNBC treatments. However, the therapeutic potential and underlying molecular mechanisms of urea-based analogues in TNBC cell lines require further exploration., Competing Interests: The authors declare no competing financial interest., (© 2023 American Chemical Society.)
- Published
- 2023
- Full Text
- View/download PDF
43. Murine Dermal Lymphatic Endothelial Cell Isolation.
- Author
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Akwii RG, Lamprou M, Georgomanoli M, Plevriti A, Marazioti A, Mouzaki A, Mattheolabakis G, and Mikelis CM
- Subjects
- Mice, Animals, Mice, Transgenic, Dissection, Extracellular Fluid, Endothelial Cells, Lymphatic Vessels
- Abstract
The lymphatic system participates in the regulation of immune surveillance, lipid absorption, and tissue fluid balance. The isolation of murine lymphatic endothelial cells is an important process for lymphatic research, as it allows the performance of in vitro and biochemical experiments on the isolated cells. Moreover, the development of Cre-lox technology has enabled the tissue-specific deficiency of genes that cannot be globally targeted, leading to the precise determination of their role in the studied tissues. The dissection of the role of certain genes in lymphatic physiology and pathophysiology requires the use of lymphatic-specific promoters, and thus, the experimental verification of the expression levels of the targeted genes. Methods for efficient isolation of lymphatic endothelial cells from wild-type or transgenic mice enable the use of ex vivo and in vitro assays to study the mechanisms regulating the lymphatic functions and the identification of the expression levels of the studied proteins. We have developed, standardized and present a protocol for the efficient isolation of murine dermal lymphatic endothelial cells (DLECs) via magnetic bead purification based on LYVE-1 expression. The protocol outlined aims to equip researchers with a tool to further understand and elucidate important players of lymphatic endothelial cell functions, especially in facilities where fluorescence-activated cell sorting equipment is not available.
- Published
- 2023
- Full Text
- View/download PDF
44. Endothelial-Specific Targeting of RhoA Signaling via CD31 Antibody-Conjugated Nanoparticles.
- Author
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Lahooti B, Akwii RG, Patel D, ShahbaziNia S, Lamprou M, Madadi M, Abbruscato TJ, Astrinidis A, Bickel U, Al-Ahmad A, German NA, Mattheolabakis G, and Mikelis CM
- Subjects
- Humans, HEK293 Cells, Liposomes, rho-Associated Kinases metabolism, rhoA GTP-Binding Protein metabolism, Endothelial Cells metabolism, Vascular Endothelial Growth Factor A metabolism
- Abstract
Existing vascular endothelial growth factor-oriented antiangiogenic approaches are known for their high potency. However, significant side effects associated with their use drive the need for novel antiangiogenic strategies. The small GTPase RhoA is an established regulator of actin cytoskeletal dynamics. Previous studies have highlighted the impact of endothelial RhoA pathway on angiogenesis. Rho-associate kinase (ROCK), a direct RhoA effector, is potently inhibited by Fasudil, a clinically relevant ROCK inhibitor. Here, we aimed to target the RhoA signaling in endothelial cells by generating Fasudil-encapsulated CD31-targeting liposomes as a potential antiangiogenic therapy. The liposomes presented desirable characteristics, preferential binding to CD31-expressing HEK293T cells and to endothelial cells, inhibited stress fiber formation and cytoskeletal-related morphometric parameters, and inhibited in vitro angiogenic functions. Overall, this work shows that the nanodelivery-mediated endothelial targeting of RhoA signaling can offer a promising strategy for angiogenesis inhibition in vascular-related diseases. SIGNIFICANCE STATEMENT: Systemic administration of antiangiogenic therapeutics induces side effects to non-targeted tissues. This study, among others, has shown the impact of the RhoA signaling in the endothelial cells and their angiogenic functions. Here, to minimize potential toxicity, this study generated CD31-targeting liposomes with encapsulated Fasudil, a clinically relevant Rho kinase inhibitor, and successfully targeted endothelial cells. In this proof-of-principle study, the efficient Fasudil delivery, its impact on the endothelial signaling, morphometric alterations, and angiogenic functions verify the benefits of site-targeted antiangiogenic therapy., (Copyright © 2023 by The American Society for Pharmacology and Experimental Therapeutics.)
- Published
- 2023
- Full Text
- View/download PDF
45. Human Dectin-1 deficiency impairs macrophage-mediated defense against phaeohyphomycosis.
- Author
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Drummond RA, Desai JV, Hsu AP, Oikonomou V, Vinh DC, Acklin JA, Abers MS, Walkiewicz MA, Anzick SL, Swamydas M, Vautier S, Natarajan M, Oler AJ, Yamanaka D, Mayer-Barber KD, Iwakura Y, Bianchi D, Driscoll B, Hauck K, Kline A, Viall NS, Zerbe CS, Ferré EM, Schmitt MM, DiMaggio T, Pittaluga S, Butman JA, Zelazny AM, Shea YR, Arias CA, Ashbaugh C, Mahmood M, Temesgen Z, Theofiles AG, Nigo M, Moudgal V, Bloch KC, Kelly SG, Whitworth MS, Rao G, Whitener CJ, Mafi N, Gea-Banacloche J, Kenyon LC, Miller WR, Boggian K, Gilbert A, Sincock M, Freeman AF, Bennett JE, Hasbun R, Mikelis CM, Kwon-Chung KJ, Belkaid Y, Brown GD, Lim JK, Kuhns DB, Holland SM, and Lionakis MS
- Subjects
- Animals, Humans, Male, Mice, CARD Signaling Adaptor Proteins genetics, Lectins, C-Type genetics, Macrophages metabolism, Tumor Necrosis Factor-alpha genetics, beta-Glucans, Phaeohyphomycosis microbiology
- Abstract
Subcutaneous phaeohyphomycosis typically affects immunocompetent individuals following traumatic inoculation. Severe or disseminated infection can occur in CARD9 deficiency or after transplantation, but the mechanisms protecting against phaeohyphomycosis remain unclear. We evaluated a patient with progressive, refractory Corynespora cassiicola phaeohyphomycosis and found that he carried biallelic deleterious mutations in CLEC7A encoding the CARD9-coupled, β-glucan-binding receptor, Dectin-1. The patient's PBMCs failed to produce TNF-α and IL-1β in response to β-glucan and/or C. cassiicola. To confirm the cellular and molecular requirements for immunity against C. cassiicola, we developed a mouse model of this infection. Mouse macrophages required Dectin-1 and CARD9 for IL-1β and TNF-α production, which enhanced fungal killing in an interdependent manner. Deficiency of either Dectin-1 or CARD9 was associated with more severe fungal disease, recapitulating the human observation. Because these data implicated impaired Dectin-1 responses in susceptibility to phaeohyphomycosis, we evaluated 17 additional unrelated patients with severe forms of the infection. We found that 12 out of 17 carried deleterious CLEC7A mutations associated with an altered Dectin-1 extracellular C-terminal domain and impaired Dectin-1-dependent cytokine production. Thus, we show that Dectin-1 and CARD9 promote protective TNF-α- and IL-1β-mediated macrophage defense against C. cassiicola. More broadly, we demonstrate that human Dectin-1 deficiency may contribute to susceptibility to severe phaeohyphomycosis by certain dematiaceous fungi.
- Published
- 2022
- Full Text
- View/download PDF
46. Evaluation of Urea-Based Inhibitors of the Dopamine Transporter Using the Experimental Autoimmune Encephalomyelitis Model of Multiple Sclerosis.
- Author
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Ashraf-Uz-Zaman M, Ji G, Tidwell D, Yin L, Thakolwiboon S, Pan J, Junell R, Griffin Z, Shahi S, Barthels D, Sajib MS, Trippier PC, Mikelis CM, Das H, Avila M, Neugebauer V, and German NA
- Subjects
- Animals, Dopamine Plasma Membrane Transport Proteins, Neuroinflammatory Diseases, Urea, Encephalomyelitis, Autoimmune, Experimental drug therapy, Multiple Sclerosis drug therapy
- Abstract
The dopaminergic system is involved in the regulation of immune responses in various homeostatic and disease conditions. For conditions such as Parkinson's disease and multiple sclerosis (MS), pharmacological modulation of dopamine (DA) system activity is thought to have therapeutic relevance, providing the basis for using dopaminergic agents as a treatment of relevant states. In particular, it was proposed that restoration of DA levels may inhibit neuroinflammation. We have recently reported a new class of dopamine transporter (DAT) inhibitors with high selectivity to the DAT over other G-protein coupled receptors tested. Here, we continue their evaluation as monoamine transporter inhibitors. Furthermore, we show that the urea-like DAT inhibitor (compound 5 ) has statistically significant anti-inflammatory effects and attenuates motor deficits and pain behaviors in the experimental autoimmune encephalomyelitis model mimicking clinical signs of MS. To the best of our knowledge, this is the first study reporting the beneficial effects of DAT inhibitor-based treatment in animals with induced autoimmune encephalomyelitis, and the observed results provide additional support to the model of DA-related neuroinflammation.
- Published
- 2022
- Full Text
- View/download PDF
47. Protein tyrosine phosphatase receptor-ζ1 deletion triggers defective heart morphogenesis in mice and zebrafish.
- Author
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Katraki-Pavlou S, Kastana P, Bousis D, Ntenekou D, Varela A, Davos CH, Nikou S, Papadaki E, Tsigkas G, Athanasiadis E, Herradon G, Mikelis CM, Beis D, and Papadimitriou E
- Subjects
- Animals, Gene Deletion, Mice, Receptor-Like Protein Tyrosine Phosphatases, Class 5 metabolism, Zebrafish, Zebrafish Proteins metabolism, Heart embryology, Myocardium metabolism, Organogenesis, Receptor-Like Protein Tyrosine Phosphatases, Class 5 genetics, Zebrafish Proteins genetics
- Abstract
Protein tyrosine phosphatase receptor- ζ 1 (PTPRZ1) is a transmembrane tyrosine phosphatase receptor highly expressed in embryonic stem cells. In the present work, gene expression analyses of Ptprz1
-/- and Ptprz1+/+ mice endothelial cells and hearts pointed to an unidentified role of PTPRZ1 in heart development through the regulation of heart-specific transcription factor genes. Echocardiography analysis in mice identified that both systolic and diastolic functions are affected in Ptprz1-/- compared with Ptprz1+/+ hearts, based on a dilated left ventricular (LV) cavity, decreased ejection fraction and fraction shortening, and increased angiogenesis in Ptprz1-/- hearts, with no signs of cardiac hypertrophy. A zebrafish ptprz1-/- knockout was also generated and exhibited misregulated expression of developmental cardiac markers, bradycardia, and defective heart morphogenesis characterized by enlarged ventricles and defected contractility. A selective PTPRZ1 tyrosine phosphatase inhibitor affected zebrafish heart development and function in a way like what is observed in the ptprz1-/- zebrafish. The same inhibitor had no effect in the function of the adult zebrafish heart, suggesting that PTPRZ1 is not important for the adult heart function, in line with data from the human cell atlas showing very low to negligible PTPRZ1 expression in the adult human heart. However, in line with the animal models, Ptprz1 was expressed in many different cell types in the human fetal heart, such as valvar, fibroblast-like, cardiomyocytes, and endothelial cells. Collectively, these data suggest that PTPRZ1 regulates cardiac morphogenesis in a way that subsequently affects heart function and warrant further studies for the involvement of PTPRZ1 in idiopathic congenital cardiac pathologies. NEW & NOTEWORTHY Protein tyrosine phosphatase receptor ζ 1 (PTPRZ1) is expressed in fetal but not adult heart and seems to affect heart development. In both mouse and zebrafish animal models, loss of PTPRZ1 results in dilated left ventricle cavity, decreased ejection fraction, and fraction shortening, with no signs of cardiac hypertrophy. PTPRZ1 also seems to be involved in atrioventricular canal specification, outflow tract morphogenesis, and heart angiogenesis. These results suggest that PTPRZ1 plays a role in heart development and support the hypothesis that it may be involved in congenital cardiac pathologies.- Published
- 2022
- Full Text
- View/download PDF
48. In Vivo Ear Sponge Lymphangiogenesis Assay.
- Author
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Akwii RG, Sajib MS, Zahra FT, Madala HR, Srivenugopal KS, and Mikelis CM
- Subjects
- Angiopoietin-2 genetics, Animals, Ear surgery, Humans, Immunity immunology, Immunity physiology, Lymphangiogenesis genetics, Lymphangiogenesis immunology, Lymphatic Vessels immunology, Mice, Signal Transduction genetics, Vascular Remodeling genetics, Vascular Remodeling immunology, Vascular Remodeling physiology, Wound Healing genetics, Wound Healing physiology, Biological Assay methods, Ear physiopathology, Lymphangiogenesis physiology, Lymphatic Vessels physiology
- Abstract
Lymphangiogenesis, the formation of lymphatic vessels from preexisting ones, is an important process in wound-healing physiology. Deregulation of lymphangiogenesis and lymphatic vascular remodeling have been implicated in a range of inflammatory conditions, such as lymphedema, lymphadenopathy, tumor growth, and cancer metastasis. Any attempt in understanding various parameters of the lymphangiogenic process and developing desirable therapeutic targets requires recapitulating these conditions in in vivo models. One pitfall with some experimental models is the absence of immune response, an important regulatory factor for lymphangiogenesis. We overcome this issue by using immune competent mice. In this chapter, by using Angiopoietin-2 (Ang2), a protein that belongs to the Ang/Tie signaling pathway, we describe the ear sponge assay with important adaptations, highlighting a reproducible and quantitative tool for assessment of in vivo lymphangiogenesis.
- Published
- 2021
- Full Text
- View/download PDF
49. Investigating Epidermal Interactions Through an In Vivo Cutaneous Wound-Healing Assay.
- Author
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Zemkewicz JL, Akwii RG, Mikelis CM, and Doçi CL
- Subjects
- Animals, Disease Models, Animal, Epidermis growth & development, Mice, Biological Assay methods, Cellular Microenvironment physiology, Skin growth & development, Wound Healing physiology
- Abstract
Cutaneous wound healing is an intricate and multifaceted process. Despite these complexities, the distinct phases of wound healing provide a unique opportunity to evaluate the roles of different targets in these coordinated responses. This protocol details an in vivo wound healing assay to study the intersection of cellular, molecular, and systemic effector pathways. The role of certain proteins in the wound healing process can be efficiently explored in vivo through the generation of tissue-specific deficient mice. This approach, although optimized for use with animal models displaying epithelial deficiencies, can be used for other tissue-specific deficiencies, and utilizes simple and cost-effective methods, allowing investigators to precisely devise their experimental design. The coordination of immunological, epithelial, vascular, and microenvironmental factors in wound healing makes this technique a valuable tool for investigators across fields.
- Published
- 2021
- Full Text
- View/download PDF
50. Identification of Rho GEF and RhoA Activation by Pull-Down Assays.
- Author
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Sajib MS, Zahra FT, Akwii RG, and Mikelis CM
- Subjects
- GTPase-Activating Proteins isolation & purification, Guanosine Diphosphate genetics, Guanosine Triphosphate genetics, Humans, Protein Binding genetics, Rho Guanine Nucleotide Exchange Factors genetics, Rho Guanine Nucleotide Exchange Factors isolation & purification, rhoA GTP-Binding Protein isolation & purification, GTPase-Activating Proteins genetics, Molecular Biology methods, rhoA GTP-Binding Protein genetics
- Abstract
The small GTPase RhoA participates in actin and microtubule machinery, cell migration and invasion, gene expression, vesicular trafficking and cell cycle, and its dysregulation is a determining factor in many pathological conditions. Similar to other Rho GTPases, RhoA is a key component of the wound-healing process, regulating the activity of different participating cell types. RhoA gets activated upon binding to guanine nucleotide exchange factors (GEFs), which catalyze the exchange of guanosine diphosphate (GDP) for guanosine triphosphate (GTP). GTPase-activating proteins (GAPs) mediate the exchange of GTP to GDP, inactivating RhoA, whereas guanine nucleotide dissociation inhibitors (GDIs) preserve the inactive pool of RhoA proteins in the cytosol. RhoA and Rho GEF activation is detected by protein pull-down assays, which use chimeric proteins with Rhotekin and G17A mutant RhoA as "bait" to pull down active RhoA and RhoA GEFs, respectively. In this chapter, we describe an optimized protocol for performing RhoA and GEF pull-down assays.
- Published
- 2021
- Full Text
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