scholarly journals Regorafenib Regulates AD Pathology, Neuroinflammation, and Dendritic Spinogenesis in Cells and a Mouse Model of AD

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1655
Author(s):  
Kyung-Min Han ◽  
Ri Jin Kang ◽  
Hyongjun Jeon ◽  
Hyun-ju Lee ◽  
Ji-Soo Lee ◽  
...  
Keyword(s):  
Mouse Model ◽  
Dendritic Spine ◽  
Kinase Inhibitor ◽  
Glycogen Synthase ◽  
Regulatory Function ◽  
Spine Density ◽  
App Processing ◽  
Beneficial Effects

The oral multi-target kinase inhibitor regorafenib, which targets the oncogenic receptor tyrosine kinase (RTK), is an effective therapeutic for patients with advanced gastrointestinal stromal tumors or metastatic colorectal cancer. However, whether regorafenib treatment has beneficial effects on neuroinflammation and Alzheimer’s disease (AD) pathology has not been carefully addressed. Here, we report the regulatory function of regorafenib in neuroinflammatory responses and AD-related pathology in vitro and in vivo. Regorafenib affected AKT signaling to attenuate lipopolysaccharide (LPS)-mediated expression of proinflammatory cytokines in BV2 microglial cells and primary cultured microglia and astrocytes. In addition, regorafenib suppressed LPS-induced neuroinflammatory responses in LPS-injected wild-type mice. In 5x FAD mice (a mouse model of AD), regorafenib ameliorated AD pathology, as evidenced by increased dendritic spine density and decreased Aβ plaque levels, by modulating APP processing and APP processing-associated proteins. Furthermore, regorafenib-injected 5x FAD mice displayed significantly reduced tau phosphorylation at T212 and S214 (AT100) due to the downregulation of glycogen synthase kinase-3 beta (GSK3β) activity. Taken together, our results indicate that regorafenib has beneficial effects on neuroinflammation, AD pathology, and dendritic spine formation in vitro and in vivo.

CHIR258, a Novel Multi-Targeted Tyrosine Kinase Inhibitor, for the Treatment of t(4;14) Multiple Myeloma.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 641-641 ◽  
Author(s):  
Suzanne Trudel ◽  
Zhi Hua Li ◽  
Ellen Wei ◽  
Marion Wiesmann ◽  
Katherine Rendahl ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Mouse Model ◽  
Tyrosine Kinase ◽  
Cell Lines ◽  
Small Molecule ◽  
Kinase Inhibitor ◽  
Wild Type ◽  
Myeloma Cells

Abstract The t(4;14) translocation that occurs uniquely in a subset (15%) of multiple myeloma (MM) patients results in the ectopic expression of the receptor tyrosine kinase, Fibroblast Growth Factor Receptor3 (FGFR3). Wild-type FGFR3 induces proliferative signals in myeloma cells and appears to be weakly transforming in a hematopoeitic mouse model. The subsequent acquisition of FGFR3 activating mutations in some MM is associated with disease progression and is strongly transforming in several experimental models. The clinical impact of t(4;14) translocations has been demonstrated in several retrospective studies each reporting a marked reduction in overall survival. We have previously shown that inhibition of activated FGFR3 causes morphologic differentiation followed by apoptosis of FGFR3 expressing MM cell lines, validating activated FGFR3 as a therapeutic target in t(4;14) MM and encouraging the clinical development of FGFR3 inhibitors for the treatment of these poor-prognosis patients. CHIR258 is a small molecule kinase inhibitor that targets Class III–V RTKs and inhibits FGFR3 with an IC50 of 5 nM in an in vitro kinase assay. Potent anti-tumor and anti-angiogenic activity has been demonstrated in vitro and in vivo. We employed the IL-6 dependent cell line, B9 that has been engineered to express wild-type FGFR3 or active mutants of FGFR3 (Y373C, K650E, G384D and 807C), to screen CHIR258 for activity against FGFR3. CHIR258 differentially inhibited FGF-mediated growth of B9 expressing wild-type and mutant receptors found in MM, with an IC50 of 25 nM and 80 nM respectively as determined by MTT proliferation assay. Growth of these cells could be rescued by IL-6 demonstrating selectivity of CHIR258 for FGFR3. We then confirmed the activity of CHIR258 against FGFR3 expressing myeloma cells. CHIR258 inhibited the viability of FGFR3 expressing KMS11 (Y373C), KMS18 (G384D) and OPM-2 (K650E) cell lines with an IC50 of 100 nM, 250 nM and 80 nM, respectively. Importantly, inhibition with CHIR258 was still observed in the presence of IL-6, a potent growth factors for MM cells. U266 cells, which lack FGFR3 expression, displayed minimal growth inhibition demonstrating that at effective concentrations, CHIR258 exhibits minimal nonspecific cytotoxicity on MM cells. Further characterization of this finding demonstrated that inhibition of cell growth corresponded to G0/G1 cell cycle arrest and dose-dependent inhibition of downstream ERK phosphorylation. In responsive cell lines, CHIR258 induced apoptosis via caspase 3. In vitro combination analysis of CHIR258 and dexamethasone applied simultaneously to KMS11 cells indicated a synergistic interaction. In vivo studies demonstrated that CHIR258 induced tumor regression and inhibited growth of FGFR3 tumors in a plasmacytoma xenograft mouse model. Finally, CHIR258 produced cytotoxic responses in 4/5 primary myeloma samples derived from patients harboring a t(4;14) translocation. These data indicate that the small molecule inhibitor, CHIR258 potently inhibits FGFR3 and has activity against human MM cells setting the stage for a Phase I clinical trial of this compound in t(4;14) myeloma.

scholarly journals Electrospun PCL/PLA Scaffolds Are More Suitable Carriers of Placental Mesenchymal Stromal Cells Than Collagen/Elastin Scaffolds and Prevent Wound Contraction in a Mouse Model of Wound Healing

2020 ◽  
Vol 8 ◽  
Author(s):  
Eva Vonbrunn ◽  
Marc Mueller ◽  
Melanie Pichlsberger ◽  
Monika Sundl ◽  
Alexander Helmer ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mouse Model ◽  
Stromal Cells ◽  
Wound Care ◽  
Skin Wound ◽  
Beneficial Effects ◽  
New Therapeutic Approaches

Mesenchymal stem/stromal cells (MSCs) exert beneficial effects during wound healing, and cell-seeded scaffolds are a promising method of application. Here, we compared the suitability of a clinically used collagen/elastin scaffold (Matriderm) with an electrospun Poly(ε-caprolactone)/poly(l-lactide) (PCL/PLA) scaffold as carriers for human amnion-derived MSCs (hAMSCs). We created an epidermal-like PCL/PLA scaffold and evaluated its microstructural, mechanical, and functional properties. Sequential spinning of different PCL/PLA concentrations resulted in a wide-meshed layer designed for cell-seeding and a dense-meshed layer for apical protection. The Matriderm and PCL/PLA scaffolds then were seeded with hAMSCs, with or without Matrigel coating. The quantity and quality of the adherent cells were evaluated in vitro. The results showed that hAMSCs adhered to and infiltrated both scaffold types but on day 3, more cells were observed on PCL/PLA than on Matriderm. Apoptosis and proliferation rates were similar for all carriers except the coated Matriderm, where apoptotic cells were significantly enhanced. On day 8, the number of cells decreased on all carrier types except the coated Matriderm, which had consistently low cell numbers. Uncoated Matriderm had the highest percentage of proliferative cells and lowest apoptosis rate of all carrier types. Each carrier also was topically applied to skin wound sites in a mouse model and analyzed in vivo over 14 days via optical imaging and histological methods, which showed detectable hAMSCs on all carrier types on day 8. On day 14, all wounds exhibited newly formed epidermis, and all carriers were well-integrated into the underlying dermis and showing signs of degradation. However, only wounds treated with uncoated PCL/PLA maintained a round appearance with minimal contraction. Overall, the results support a 3-day in vitro culture of scaffolds with hAMSCs before wound application. The PCL/PLA scaffold showed higher cell adherence than Matriderm, and the effect of the Matrigel coating was negligible, as all carrier types maintained sufficient numbers of transplanted cells in the wound area. The anti-contractive effects of the PCL/PLA scaffold offer potential new therapeutic approaches to wound care.

Inhibiting the IGF-1 Receptor Tyrosine Kinase with Picropodophillin: An In Vitro and In Vivo Study in the 5T33MM Mouse Model.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 640-640
Author(s):  
Karin Vanderkerken ◽  
Eline Menu ◽  
Thomas Stromberg ◽  
Hendrik De Raeve ◽  
Kewal Asosingh ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Mouse Model ◽  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Microvessel Density ◽  
Kinase Inhibitor ◽  
High Dose

Abstract Multiple myeloma (MM) represents a B-cell malignancy, characterized by monoclonal proliferation of plasma cells in the bone marrow (BM) and is associated with osteolysis and angiogenesis. Insulin-like growth factor-1 (IGF-1), produced by the BM stromal cells, has been described as an important factor in the survival, proliferation and migration of MM cells. The latter process is involved in the homing of the MM cells to the BM. IGF-1 also induces VEGF secretion by the MM cells, thus stimulating angiogenesis in the BM. As IGF-1 is a pleiotropic factor in MM, therapeutic strategies targeting the IGF-1R may be effective as anti-tumor treatments. In this work we investigated the effect of an IGF-1 receptor tyrosine kinase inhibitor (picropodophyllin or PPP1) in the murine, syngeneic 5T33MM model of multiple myeloma. This mouse model is representative for the human disease and can combine in vitro and in vivo studies. We first investigated the effects of PPP on the MM cells in vitro. We and others have previously demonstrated that IGF-1 induced ERK activation, involved in VEGF secretion and proliferation. When the 5T33MM cells were preincubated with 1microM PPP, Western blot analysis demonstrated the blocking of this activation. Furthermore, when the 5T33MM cells were preincubated with PPP for 30 min, IGF-1 induced VEGF secretion and proliferation of the 5T33MM cells were completely blocked. Next, we used the tyrosine kinase inhibitor PPP in vivo. 5T33MM cells were injected intravenously in C57BLKaLwRij mice and the development of the disease was monitored by measuring the serum paraprotein concentration. Mice were either treated with a low (17mM, IP, twice a day) or a high dose of PPP (50mM, IP, twice a day) or with the vehicle (DMSO/oil 9/1) from the day of injection with 5T33MM onward. At week 3, vehicle controls showed signs of morbidity and were sacrificed. The presence of tumor was measured by assessing serum paraprotein concentrations and determining the proportion of idiotype positive cells in the BM by flow cytometry. Angiogenesis was assessed by measuring the microvessel density on CD31 stained paraffin sections. The tumor burden in the bone marrow in the PPP treated mice was 77% lower than in vehicle treated animals (p< 0,0001) and the serum paraprotein concentration was 90% lower (p< 0,0001). The microvessel density in the BM of the PPP treated group was reduced by 60% (p< 0,02). In a separate survival experiment the mice were either treated with the vehicle or with the high dose (50mM) of PPP, from the time of tumor injection. Kaplan-Meier analysis demonstrated a significant increase in survival after treatment with PPP when compared with vehicle (28 vs. 18 days, p<0,001). These data demonstrate that the IGF-1RTK inhibitor PPP possesses strong anti-tumor activity, as demonstrated both in vitro and in vivo in a syngeneic model of multiple myeloma, and may therefore be an effective therapeutic candidate for MM treatment.

scholarly journals AZD2014, A Dual mTOR Inhibitor, Attenuates Cardiac Hypertrophy in Vitro and in Vivo

2021 ◽  
Author(s):  
Byung-Hyun Cha ◽  
Minjin Jung ◽  
Angela S. Kim ◽  
Victoria C. Lepak ◽  
Brett A. Colson ◽  
...  
Keyword(s):  
Mouse Model ◽  
Cardiac Hypertrophy ◽  
Mammalian Target Of Rapamycin ◽  
Underlying Mechanism ◽  
Regulatory Function ◽  
Cardiomyocyte Hypertrophy ◽  
Therapeutic Effects ◽  
Cardiac Morbidity

Abstract Cardiac hypertrophy is one of the most common genetic heart disorders and considered a risk factor for cardiac morbidity and mortality. The mammalian target of rapamycin (mTOR) pathway plays a key regulatory function in cardiovascular physiology and pathology in hypertrophy. AZD2014 is a small-molecule ATP competitive mTOR inhibitor working on both mTORC1 and mTORC2 complexes. Little is known about the therapeutic effects of AZD2014 in cardiac hypertrophy and its underlying mechanism. Here, AZD2014 is examined in in vitro model of phenylephrine (PE)-induced human cardiomyocyte hypertrophy and a myosin-binding protein-C (Mybpc3)-targeted knockout (KO) mouse model of cardiac hypertrophy. Our results demonstrate that cardiomyocytes treated with AZD2014 retain the normal phenotype and AZD2014 attenuates cardiac hypertrophy in the Mybpc3-KO mouse model through inhibition of dual mTORC1 and mTORC2, which in turn results in the down-regulation of the Akt/mTOR signaling pathway.

scholarly journals Toxicity and Anti-inflammatory Activities of an Extract of the Eleutherine bulbosa Rhizome on Collagen Antibody-induced Arthritis in a Mouse Model

2018 ◽  
Vol 13 (7) ◽  
pp. 1934578X1801300
Author(s):  
Pham Thi Bich Hanh ◽  
Do Thi Thao ◽  
Nguyen Thi Nga ◽  
Ngo Thi Phuong ◽  
Le Ngoc Hung ◽  
...  
Keyword(s):  
Mouse Model ◽  
Ethanol Extract ◽  
Serum Levels ◽  
Negative Control ◽  
High Dose ◽  
Necrosis Factor Alpha ◽  
Beneficial Effects ◽  
Anti Inflammatory

As a continuation of our interest in the anti-inflammatory activities of Vietnamese plants, we searched for novel anti-inflammatory agents in Eleutherine bulbosa and evaluated the anti-inflammatory effects of an ethanol extract of the rhizome of E. bulbosa (EBE) on lipopolysaccharide-stimulated RAW 264.7 macrophages in vitro and in a collagen antibody-induced arthritic (CAIA) mouse model in vivo. Treatment of the CAIA mice with EBE decreased the incidence of arthritis, especially at a dose of 1000 mg/kg body weight. A significant ( P<0.05) decrease in the arthritis score was seen after high-dose EBE treatment between days 10 and 14 in comparison with the negative control. The serum levels of the inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, and IL-10 in the mice were measured using commercial ELISA kits. The results suggest that an ethanol extract of the E. bulbosa rhizome has beneficial effects on inflammatory cytokine regulation in an experimental CAIA model.

scholarly journals Assessing the therapeutic potential of APPs⍺ in a tau transgenic mouse model of Alzheimers disease

2020 ◽  
Author(s):  
Charlotte Bold ◽  
Danny Baltissen ◽  
Susann Ludewig ◽  
Roman Spilger ◽  
Karl Rohr ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Mouse Model ◽  
Therapeutic Potential ◽  
Large Body ◽  
Transgenic Mouse Model ◽  
Spine Density ◽  
General Applicability ◽  
Plaque Deposition

A large body of evidence indicates a neuroprotective and neurotrophic function for APPs⍺ not only in vitro, but also when expressed by AAV vectors in vivo such as in APP/PS1 transgenic AD model mice with Aβ-induced pathology. Previously, we could show that APPs⍺ rescued deficits of APP/PS1 in synaptic plasticity and spine density and also reduced plaque deposition. Thus, it is crucial to test a more general applicability of APPs⍺ as a treatment for AD and to assess whether APPs⍺ is also beneficial in mice with tau-induced pathology.

scholarly journals Benzydamine, a CDK2 Kinase Inhibitor, Suppresses The Growth of Esophageal Squamous Cell Carcinoma in Vitro and in Vivo.

2021 ◽  
Author(s):  
Yubing Zhou ◽  
Xinyu He ◽  
Yanan Jiang ◽  
Zitong Wang ◽  
Yin Yu ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Esophageal Squamous Cell Carcinoma ◽  
Mouse Model ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Kinase Inhibitor ◽  
Xenograft Mouse Model ◽  
Potential Target

Abstract Background: Esophageal squamous cell carcinoma (ESCC) is among one of the leading causes of cancer death worldwide owing to late detection and low survival rate. The clinical outcome of ESCC remains dismal. To date, the disease lacks available targeted therapies. Recently, drugs approved by the Food and Drug Administration have been reported to have potential as cancer chemoprevention agents. Methods: Benzydamine, available as a hydrochloride salt, a locally acting non-steroidal anti-inflammatory drug, was screened out among FDA-approved drugs owing to its effective cytotoxic effect on KYSE450 cells, which remained unexplored. Mass spectrometry, kinase prediction and Swiss Target Prediction were used to verify the potential target(s) of benzydamine. Patient-derived oesophageal xenograft mouse model were used to investigate the effect of benzydamine on tumor growth in vivo.Results: We found that benzydamine inhibited anchorage-dependent and -independent growth of ESCC cells. Kyoto Encyclopedia of Genes and Genomes pathway enrichment revealed that benzydamine attenuated five signaling pathways, including the DNA replication pathway. We further found that benzydamine could bind to CDK2 in its ATP-binding site. Inhibition of the activity of CDK2 suppressed the growth of ESCC cells and led to a G1/S cell cycle arrest. Additionally, knocking-down CDK2 decreased the sensitivity of ESCC cells to benzydamine hydrochloride. Notably, benzydamine suppressed tumour growth in a patient-derived oesophageal xenograft mouse model of ESCC in vivo.Conclusions: We have identified CDK2 as a potential target of benzydamine for the treatment and prevention of ESCC. Benzydamine suppressed the growth of ESCC cells by inhibiting the activity of CDK2.

scholarly journals Fasudil Attenuates Adriamycin-Induced Cardiac Damage by Modifying Oxidative Stress and Apoptosis Mediated Cellular Signaling

2021 ◽  
Author(s):  
Yi Yan ◽  
Chengyu Xiang ◽  
Dingguo Zhang
Keyword(s):  
Mouse Model ◽  
Kinase Inhibitor ◽  
Rho Kinase ◽  
Control Group ◽  
H9c2 Cell ◽  
Protective Mechanism ◽  
High Dose ◽  
Cardiac Damage

Abstract PURPOSETo explore the protective mechanism of fasudil,a Rho kinase inhibitor, on acute cardiac injury induced by adriamycin(ADR).METHODSIn vitro investigations on H9C2 cell line, as well as an in vivo study in a mouse model of ADR-induced acute cardiomyopathy, were performed. In vitro, H9C2 cells were treated with fasudil for 30mins then incubated with ADR for 24 hours. Cells were collected for immunohistochemistry and western blot study, respectively. In vivo, C57BL6 mice were randomly divided into the following four groups: ①ADR group;②low-dose fasudil ( ADR+L);③high-dose fasudil ( ADR+H); and ④control group(CON). Animals were injected i.p 20 mg/kg ADR once in group①~③. And animals were injected i.p fasudil (2 or 10 mg/kg/day ) daily once for six times in group ②and ③,respectively. Blood samples and heart tissues were collected for assays.RESULTSIn vitro, fasudil treatment ameliorated ADR-induced immunofluorescence reaction of 8-OHdG, decreased the expression of TUNEL cells and protein of Bax、Caspase-3 and p53,and increased the expression of protein of Bcl-2 and SIRT 1. In the mouse model, administration of fasudil significantly ameliorated ADR-induced cardiac damage, suppressed cell apoptosis and senescence, ameliorated redox imbalance and DNA damage.CONCLUSIONFasudil has the protective effect on adriamycin induced acute cardiotoxicity, which partially attributed to its antioxidant, anti-senescence, and anti-apoptotic effects of inhibiting the RhoA/Rho kinase signaling pathway.

scholarly journals AZD2014, a dual mTOR inhibitor, attenuates cardiac hypertrophy in vitro and in vivo

2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Byung-Hyun Cha ◽  
Minjin Jung ◽  
Angela S. Kim ◽  
Victoria C. Lepak ◽  
Brett A. Colson ◽  
...  
Keyword(s):  
Mouse Model ◽  
Cardiac Hypertrophy ◽  
Mtor Inhibitor ◽  
Underlying Mechanism ◽  
Regulatory Function ◽  
Cardiomyocyte Hypertrophy ◽  
Therapeutic Effects ◽  
Cardiac Morbidity

AbstractCardiac hypertrophy is one of the most common genetic heart disorders and considered a risk factor for cardiac morbidity and mortality. The mammalian target of rapamycin (mTOR) pathway plays a key regulatory function in cardiovascular physiology and pathology in hypertrophy. AZD2014 is a small-molecule ATP competitive mTOR inhibitor working on both mTORC1 and mTORC2 complexes. Little is known about the therapeutic effects of AZD2014 in cardiac hypertrophy and its underlying mechanism. Here, AZD2014 is examined in in vitro model of phenylephrine (PE)-induced human cardiomyocyte hypertrophy and a myosin-binding protein-C (Mybpc3)-targeted knockout (KO) mouse model of cardiac hypertrophy. Our results demonstrate that cardiomyocytes treated with AZD2014 retain the normal phenotype and AZD2014 attenuates cardiac hypertrophy in the Mybpc3-KO mouse model through inhibition of dual mTORC1 and mTORC2, which in turn results in the down-regulation of the Akt/mTOR signaling pathway.

scholarly journals Testing Fmr1KO Phenotypes in Response to GSK3 Inhibitors: SB216763 versus AFC03127

2021 ◽  
Vol 14 ◽  
Author(s):  
Pamela R. Westmark ◽  
Beatrice Garrone ◽  
Rosella Ombrato ◽  
Claudio Milanese ◽  
Francesco Paolo Di Giorgio ◽  
...  
Keyword(s):  
Metabotropic Glutamate Receptor ◽  
Glycogen Synthase ◽  
Fragile X ◽  
Drug Dose ◽  
Amyloid Beta Protein ◽  
Spine Density ◽  
Spine Length ◽  
Gsk3 Inhibitor

Glycogen synthase kinase 3 (GSK3) is a proline-directed serine-threonine kinase that is associated with several neurological disorders, including Alzheimer’s disease and fragile X syndrome (FXS). We tested the efficacy of a novel GSK3 inhibitor AFC03127, which was developed by Angelini Pharma, in comparison to the metabotropic glutamate receptor 5 inhibitor 2-Methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP) and the GSK3 inhibitor SB216763 in in vivo and in vitro assays in Fmr1KO mice, a mouse model useful for the study of FXS. The in vivo assay tested susceptibility to audiogenic-induced seizures (AGS) whereas the in vitro assays assessed biomarker expression and dendritic spine length and density in cultured primary neurons as a function of drug dose. MPEP and SB216763 attenuated AGS in Fmr1KO mice, whereas AFC03127 did not. MPEP and AFC03127 significantly reduced dendritic expression of amyloid-beta protein precursor (APP). All drugs rescued spine length and the ratio of mature dendritic spines. Spine density was not statistically different between vehicle and GSK3 inhibitor-treated cells. The drugs were tested over a wide concentration range in the in vitro assays to determine dose responses. A bell-shaped dose response decrease in APP expression was observed in response to AFC03127, which was more effective than SB216763. These findings confirm previous studies demonstrating differential effects of various GSK3 inhibitors on AGS propensity in Fmr1KO mice and confirm APP as a downstream biomarker that is responsive to GSK3 activity.

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