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Your search keyword '"Kinase activity"' showing total 3 results
3 results on '"Kinase activity"'

1. Distinct effects of leukocyte and cardiac phosphoinositide 3-kinase γ activity in pressure overload-induced cardiac failure

Author

Alessia Perino, Daniela Carnevale, Ornella Azzolino, Giuseppe Lembo, Irene Franco, Riccardo Ragona, Pierluigi Carullo, Giuseppe Cifelli, Cinzia Perrino, Emilio Hirsch, Alessandra Ghigo, Federico Damilano, Katrin Schaefer, Damilano, F, Franco, I, Perrino, Cinzia, Schaefer, K, Azzolino, O, Carnevale, D, Cifelli, G, Carullo, P, Ragona, R, Ghigo, A, Perino, A, Lembo, G, and Hirsch, E.

Subjects
Cardiac function curve, medicine.medical_specialty, heart failure, Inflammation, 030204 cardiovascular system & hematology, Mice, 03 medical and health sciences, pi3kγ, 0302 clinical medicine, Fibrosis, Physiology (medical), Internal medicine, Leukocytes, Animals, Class Ib Phosphatidylinositol 3-Kinase, Medicine, pi3k gamma, Gene Knock-In Techniques, Kinase activity, 030304 developmental biology, Pressure overload, 0303 health sciences, Phosphoinositide 3-kinase, Ventricular Remodeling, biology, business.industry, Myocardium, fibrosis, Heart, medicine.disease, pi3k?, inflammation, signal transduction, Mice, Inbred C57BL, inflammation signal transduction heart failure, Endocrinology, Heart failure, Immunology, cardiovascular system, biology.protein, Signal transduction, medicine.symptom, Cardiology and Cardiovascular Medicine, business
Abstract

Background— Signaling from phosphoinositide 3-kinase γ (PI3Kγ) is crucial for leukocyte recruitment and inflammation but also contributes to cardiac maladaptive remodeling. To better understand the translational potential of these findings, this study investigates the role of PI3Kγ activity in pressure overload–induced heart failure, addressing the distinct contributions of bone marrow–derived and cardiac cells. Methods and Results— After transverse aortic constriction, mice knock-in for a catalytically inactive PI3Kγ (PI3Kγ KD) showed reduced fibrosis and normalized cardiac function up to 16 weeks. Accordingly, treatment with a selective PI3Kγ inhibitor prevented transverse aortic constriction–induced fibrosis. To define the cell types involved in this protection, bone marrow chimeras, lacking kinase activity in the immune system or the heart, were studied after transverse aortic constriction. Bone marrow–derived cells from PI3Kγ KD mice were not recruited to wild-type hearts, thus preventing fibrosis and preserving diastolic function. After prolonged pressure overload, chimeras with PI3Kγ KD bone marrow–derived cells showed slower development of left ventricular dilation and higher fractional shortening than controls. Conversely, in the presence of a wild-type immune system, KD hearts displayed bone marrow–derived cell infiltration and fibrosis at early stages but reduced left ventricular dilation and preserved contractile function at later time points. Conclusions— Together, these data demonstrate that, in response to transverse aortic constriction, PI3Kγ contributes to maladaptive remodeling at multiple levels by modulating both cardiac and immune cell functions.

Published
2011

2. Identification of a Novel MAN1A1-ROS1 Fusion Gene Through mRNA-based Screening for Tyrosine Kinase Gene Aberrations in a Patient with Leiomyosarcoma.

Author

Yoshiyuki Suehara, Shinji Kohsaka, Takuo Hayashi, Keisuke Akaike, Aiko Kurisaki-Arakawa, Shingo Sato, Eisuke Kobayashi, Sho Mizuno, Toshihide Ueno, Takeshi Morii, Tomotake Okuma, Taisei Kurihara, Nobuhiko Hasegawa, Kei Sano, Keita Sasa, Taketo Okubo, Youngji Kim, Hiroyuki Mano, Tsuyoshi Saito, and Suehara, Yoshiyuki

Subjects
*PROTEIN-tyrosine kinases, *GENE fusion, *SCHWANNOMAS, *SMOOTH muscle tumors, *CHIMERIC proteins, *LEIOMYOSARCOMA, *ANAPLASTIC lymphoma kinase, *SARCOMA, *PROTEIN-tyrosine kinase inhibitors, *PROTEINS, *REVERSE transcriptase polymerase chain reaction, *SEQUENCE analysis, *ANTHROPOMETRY, *PROTEIN kinase inhibitors, *RNA, *ANTINEOPLASTIC agents, *SOFT tissue tumors, *GLYCOSIDASES, *GENES, *GENE expression profiling, *CELLS, *POLYMERASE chain reaction, *MICE, *ANIMALS, *PHARMACODYNAMICS
Abstract

Background: Soft tissue sarcomas are a heterogeneous group of rare malignant tumors. Advanced soft tissue sarcomas have a poor prognosis, and effective systemic therapies have not been established. Tyrosine kinases are increasingly being used as therapeutic targets for a variety of cancers and soft tissue sarcomas. Although complex karyotype sarcomas typically tend to carry more potentially actionable genetic alterations than do translocation-associated sarcomas (fusion gene sarcomas), based on our database review, we found that leiomyosarcoma and malignant peripheral nerve sheath tumors have lower frequencies of potential targets than other nontranslocation soft tissue sarcomas. We theorized that both leiomyosarcoma and malignant peripheral nerve sheath tumors might be included in any unique translocations. Furthermore, if tyrosine kinase imbalances, especially fusion genes, occur in patients with leiomyosarcomas and malignant peripheral nerve sheath tumors, tyrosine kinase inhibitors might be a drug development target for this sarcoma. In this study, we used a tyrosine kinase screening system that could detect an imbalance in mRNA between 5'- and 3'-sides in tyrosine kinase genes to identify potential novel therapeutic tyrosine kinase targets for soft tissue sarcomas.Questions/purposes: (1) Are there novel therapeutic tyrosine kinase targets in tumors from patients with soft tissue sarcomas that are detectable using mRNA screening focusing on imbalance expressions between the 5' and 3' end of the kinase domain? (2) Can potential targets be verified by RNA sequencing and reverse transcription PCR (RT-PCR)? (3) Will potential fusion gene(s) transform cells in in vitro assays? (4) Will tumors in mice that have an identified fusion gene respond to treatment with a therapeutic drug directed at that target?Methods: We used mRNA screening to look for novel tyrosine kinase targets that might be of therapeutic potential. Using functional assays, we verified whether the identified fusion genes would be good therapeutic candidates for soft tissue sarcomas. Additionally, using in vivo assays, we assessed whether suppressing the fusion's kinase activity has therapeutic potential. Study eligibility was based on a patient having high-grade spindle cell and nontranslocation sarcomas, including leiomyosarcoma, malignant peripheral nerve sheath tumor, and high-grade myxofibrosarcoma. Between 2015 and 2019, of the 172 patients with soft tissue sarcomas treated with surgical resection at Juntendo University Hospital, 72 patients had high-grade nontranslocation sarcomas. The analysis was primarily for leiomyosarcoma and malignant peripheral nerve sheath tumors, and there was a limitation of analysis size (reagent limitations) totaling 24 samples at the start of the study. We collected additional samples from a sample bank at the Tokyo Medical and Dental University to increase the number of sarcomas to study. Therefore, in this study, a total of 15 leiomyosarcoma samples, five malignant peripheral nerve sheath tumors samples, and four high-grade myxofibrosarcoma samples were collected to achieve the sample size of 24 patients. To identify tyrosine kinase fusion genes, we designed a NanoString-based assay (NanoString Technologies Inc, Seattle, WA, USA) to query the expression balances regarding transcripts of 90 tyrosine kinases at two points: the 5' end of the kinase domain and within the kinase domain or 3' end of the kinase domain. The tumor's RNA was hybridized to the NanoString probes and analyzed for the expression ratios of outliers from the 3' to 5' end of the kinase domain. Presumed novel fusion events in these positive tumors that were defined by NanoString-based assays were confirmed tyrosine kinase fusion genes by RNA sequencing and confirmatory RT-PCR. Functional analyses consisting of in vitro and in vivo assays were also performed to elucidate whether the identified tyrosine kinase gene fusions were associated with oncogenic abilities and drug responses.Results: We identified aberrant expression ratios regarding the 3' to 5' end of the kinase domain ratios in ROS1 transcripts in a leiomyosarcoma in a 90-year-old woman. A novel MAN1A1-ROS1 fusion gene was identified from her thigh tumor through RNA sequencing, which was confirmed with real-time PCR. In functional assays, MAN1A1-ROS1 rearrangement revealed strong transforming potential in 3T3 cells. Moreover, in an in vivo assay, crizotinib, a ROS1 inhibitor, markedly inhibited the growth of MAN1A1-ROS1 rearrangement-induced transformed cells in a dose-dependent manner.Conclusion: We conducted tyrosine kinase screening to identify new therapeutic targets in soft tissue sarcomas. We found a novel MAN1A1-ROS1 fusion gene that may be a therapeutic target in patients with leiomyosarcoma. This study demonstrates that the mRNA screening system may aid in the development of useful therapeutic options for soft tissue sarcomas.Clinical Relevance: If novel tyrosine fusions such as MAN1A1-ROS1 fusion can be found in sarcomas from other patients, they could offer avenues for new molecular target therapies for sarcomas that currently do not have effective chemotherapeutic options. Therefore, the establishment of a screening system that includes both genomic and transcript analyses in the clinical setting is needed to verify our discoveries and take the developmental process of treatment to the next step. [ABSTRACT FROM AUTHOR]

Published
2021
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3. Ponatinib Combined With Rapamycin Causes Regression of Murine Venous Malformation

Author

Elisa Boscolo, Xian Li, Adam Lane, Lars Brichta, Timothy D. Le Cras, Jillian Goines, Yuqi Cai, and Patricia A. Pastura

Subjects
0301 basic medicine, MAPK/ERK pathway, Male, Vascular Malformations, Drug Evaluation, Preclinical, Apoptosis, Receptor tyrosine kinase, chemistry.chemical_compound, Mice, 0302 clinical medicine, Translational Sciences, Medicine, Proto-Oncogene Proteins c-abl, Cells, Cultured, ABL, biology, Chemotaxis, Ponatinib, Imidazoles, Receptor, TIE-2, endothelial cells, Pyridazines, 030220 oncology & carcinogenesis, embryonic structures, cardiovascular system, ComputingMethodologies_DOCUMENTANDTEXTPROCESSING, Heterografts, Drug Therapy, Combination, Cardiology and Cardiovascular Medicine, Cell Division, Signal Transduction, Combination therapy, MAP Kinase Signaling System, Mutation, Missense, Mice, Nude, 03 medical and health sciences, In vivo, Human Umbilical Vein Endothelial Cells, Animals, Humans, Protein kinase B, Protein Kinase Inhibitors, umbilical vein, Sirolimus, Cell growth, business.industry, Phospholipase C gamma, 030104 developmental biology, cell proliferation, chemistry, Cancer research, biology.protein, mutation, business, Proto-Oncogene Proteins c-akt
Abstract

Supplemental Digital Content is available in the text., Objective— Venous malformations (VMs) arise from developmental defects of the vasculature and are characterized by massively enlarged and tortuous venous channels. VMs grow commensurately leading to deformity, obstruction of vital structures, bleeding, and pain. Most VMs are associated with the activating mutation L914F in the endothelial cell (EC) tyrosine kinase receptor TIE2. Therapeutic options for VM are limited and ineffective while therapy with the mammalian target of rapamycin inhibitor rapamycin shows moderate efficacy. Here, we investigated novel therapeutic targets promoting VM regression. Approach and Results— We performed an unbiased screen of Food and Drug Administration–approved drugs in human umbilical vein ECs expressing the TIE2-L914F mutation (HUVEC-TIE2-L914F). Three ABL (Abelson) kinase inhibitors prevented cell proliferation of HUVEC-TIE2-L914F. Moreover, c-ABL, common target of these inhibitors, was highly phosphorylated in HUVEC-TIE2-L914F and VM patient–derived ECs with activating TIE2 mutations. Knockdown of c-ABL/ARG in HUVEC-TIE2-L914F reduced cell proliferation and vascularity of murine VM. Combination treatment with the ABL kinase inhibitor ponatinib and rapamycin caused VM regression in a xenograft model based on injection of HUVEC-TIE2-L914F. A reduced dose of this drug combination was effective in this VM murine model with minimal side effects. The drug combination was antiproliferative, enhanced cell apoptosis and vascular channel regression both in vivo and in a 3-dimensional fibrin gel assay. Conclusions— This is the first report of a combination therapy with ponatinib and rapamycin promoting regression of VM. Mechanistically, the drug combination enhanced AKT inhibition compared with single drug treatment and reduced PLCγ (phospholipase C) and ERK (extracellular signal-regulated kinase) activity.

Published
2019

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