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Losson, Hélène. "Combinaisons de nouveaux inhibiteurs de désacétylase d’histones 6 avec des inhibiteurs de tyrosine kinase pour le traitement de la leucémie myéloïde chronique." Thesis, Université de Lorraine, 2020. http://www.theses.fr/2020LORR0003.
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Les patients atteints de leucémie myéloïde chronique (LMC) breakpoint cluster region-Abelson (BCR-ABL)+ sont traités avec des inhibiteurs de tyrosine kinase (ITK), comme l’imatinib, cependant certains développent des résistances et des effets secondaires sévères. Des traitements combinés à base d’inhibiteurs d’histone désacétylase (HDAC)6 (HDAC6i), pouvant potentiellement réduire l’expression de BCR-ABL, apparaît être une approche intéressante pour prévenir l’apparition de résistances aux ITK. De plus, l’implication d’HDAC6 dans les voies de dégradation des protéines rend son inhibition couplée à celle du protéasome susceptible de sensibiliser les cellules aux ITK. Notre hypothèse est que la combinaison ITK-HDAC6i pourrait être efficace pour le traitement de la LMC. Dans un premier temps, les effets anti-cancéreux d’un HDAC6i identifié dans notre laboratoire, le composé 7b, à celui de référence, la tubacine, en combinaison avec l’imatinib ont été comparés. La combinaison imatinib-7b a généré des effets anti- cancéreux plus importants que la combinaison imatinib-tubacine et a provoqué une mort synergique apoptotique dépendante des caspases dans les cellules K-562 et réduit la proportion de cellules souches leucémiques alors qu’elle n’a eu qu’un effet modéré sur des cellules saines. Enfin, la combinaison a diminué plus fortement la capacité de formation de colonies et la masse tumorale des cellules de LMC respectivement en milieu semi-solide et dans des poissons zèbres xénogreffés, par rapport aux composés seuls. D’un point de vue mécanistique, la combinaison induit l’ubiquitination et la dégradation de BCR-ABL, et la dérégulation de protéines de ses voies de signalisation impliquées dans la prolifération et la survie cellulaire. La protéine HDAC6 possédant deux sites catalytiques, nos résultats tendent à montrer que le composé 7b cible le deuxième. Dans un second temps, une étude a été initiée sur un nouvel HDAC6i de type hydroxamate, le MAKV-15, qui diminue l’expression de BCR-ABL, et qui en pré-traitement avec le bortezomib, sensibilise les cellules à l’imatinib, entrainant une augmentation de la mort apoptotique dépendante des caspases dans les cellules sensibles et résistantes à l’imatinib. Enfin, nos résultats suggèrent que l’inhibition d’HDAC6 potentialise l’effet de l’imatinib, pourrait prévenir l’apparition de résistances et que de telles combinaisons pourraient représenter une approche thérapeutique prometteuse pour les patients atteints de LMCBreakpoint cluster region-Abelson (BCR-ABL)+ chronic myeloid leukemia (CML) patients receive tyrosine kinase inhibitors (TKIs) such as imatinib as the first-line treatment; however, some patients develop resistances and severe adverse effects. Combination treatments, especially with histone deacetylase (HDAC)6 inhibitors (HDAC6i), appear as an attractive option to prevent TKI resistances considering the capacity of HDAC6i to downregulate BCR-ABL. Moreover, HDAC6 is implicated in protein degradation pathways, so that its inhibition combined with that of the proteasome could sensitize cells to TKIs. Thus, we hypothesized that HDAC6i combined to TKIs could be effective for CML treatment. In the first part, we compared the anti-CML effects of a HDAC6i identified in our laboratory, compound 7b, to the reference HDAC6i tubacin, in combination with imatinib. Results showed that the imatinib-7b combination generated stronger anti- CML effects than imatinib-tubacin. Especially, the imatinib-7b combination elicited a potent synergistic caspase- dependent apoptotic cell death and drastically reduced the proportion of cancer stem cells in K562 CML cells, whereas it only moderately impacted various healthy cell models. Ultimately, the imatinib-7b combination decreased more potently the colony forming capacities and tumor mass formation of CML cells in a semisolid methylcellulose medium and in xenografted zebrafishes, respectively, compared to each compound alone. Mechanistically, the combination induced BCR-ABL ubiquitination and downregulation leading to a dysregulation of multiple key proteins of its downstream pathways involved in CML proliferation and survival. Results tend to demonstrate that 7b could target the second site. In the second part, we initiated a study of a novel hydroxamate-based HDAC6i, MAKV-15, and preliminary results demonstrated it triggered BCR-ABL downregulation. Accordingly, in pre-treatment with bortezomib it sensitizes CML cells to imatinib leading to enhanced caspase-dependent apoptotic death in imatinib-sensitive and imatinib-resistant CML cells. Considering that HDAC6 is reported to possess two functional catalytic sites, we finally attempted to determine which catalytic site is targeted by these HDAC6i. Taken together, our results suggest that HDAC6i potentiate the effect of imatinib and could overcome TKI resistance in CML cells and therefore such combination may represent a promising therapeutic approach for CML patients
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Shor, Audrey Cathryn. "Src kinase inhibitors for the treatment of sarcomas : cellular and molecular mechanisms of action." [Tampa, Fla] : University of South Florida, 2007. http://purl.fcla.edu/usf/dc/et/SFE0001906.
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Zhang, Wen. "Identification of novel pyruvate dehydrogenase kinase 1 (PDK1) inhibitors for anticancer therapeutics." Thesis, University of Macau, 2018. http://umaclib3.umac.mo/record=b3953604.
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Alsfouk, Aisha. "Synthesis and biological evaluation of selective inhibitory kappa B kinase-alpha (IKKa) inhibitors for the treatment of prostate and pancreatic cancer." Thesis, University of Strathclyde, 2018. http://digitool.lib.strath.ac.uk:80/R/?func=dbin-jump-full&object_id=29272.
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D'Cunha, Ronilda Raymond. "Treatment strategies to reverse efflux transporter-mediated resistance to Tyrosine kinase inhibitors." Diss., University of Iowa, 2018. https://ir.uiowa.edu/etd/6563.
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Multidrug resistance (MDR), a phenomenon in which tumors that were initially sensitive, recur and start showing resistance not only to the initial chemotherapeutic agent but also to various anticancer drugs that are structurally and functionally different from the initial drug, constitutes one of the main reasons for the failure of chemotherapy. An important mechanism of MDR is the enhanced cellular efflux of anticancer agents due to an overexpression of ATP-binding cassette (ABC) transporters (i.e. efflux transporters), especially P-glycoprotein (Pgp), Multidrug Resistance-associated Protein 1 (MRP1) and Breast Cancer Resistance Protein (BCRP), in cancer cells. In order to reverse this resistance, there has been a lot of emphasis on the development of Pgp, MRP1 and BCRP inhibitors. Although this search has been ongoing for three decades, there are still no clinically available efflux transporter modulators.
Tyrosine kinase inhibitors (TKIs) are a novel, rapidly growing class of anticancer agents that have a target-based mechanism of action, and their use transformed cancer chemotherapy due to higher specificity and enhanced safety profiles compared to conventional chemotherapeutic agents. Despite their tremendous success in treating various types of tumors, patients develop resistance to TKIs over time. Most of the FDA- approved TKIs are substrates of Pgp and/or BCRP, and as a result, these efflux transporters are also an important cause of conferred resistance against TKIs in cancer cells. Additionally, none of the 31 approved TKIs have an indication for use in brain tumors and interestingly, this may also due to the presence of Pgp and BCRP at the blood-brain barrier (BBB) and in the tumor cells, which prevent the TKI from crossing the BBB and reaching its target tumor site. Since Pgp- and BCRP- mediated TKI efflux has been shown to be involved in TKI resistance, the inhibition of these transporters could represent a potential TKI resistance reversal strategy.
Over the last three decades, a large number of Pgp and/or BCRP inhibitors have been identified, but none of them have successfully made it to the clinic. It was observed that most drugs identified as inhibitors were either unable to achieve Pgp and BCRP inhibitory concentrations in-vivo without imparting severe toxicity, or did not possess adequate bioavailability and tissue distribution profiles in order to reach the tumor site. From these identified candidate inhibitors, after much thought and consideration, we chose to investigate TKIs and methylated flavones as modulators of efflux transporter-mediated TKI resistance.
The overall goal of this project was to investigate the promising chemosensitizing potential of TKIs and methylated flavones in efflux transporter-mediated TKI resistance, both in-vitro and in-vivo. To identify potent efflux transporter inhibitor TKIs, we evaluated the effect of various TKIs on the accumulation of afatinib, the model TKI substrate, in Pgp- and BCRP- overexpressing cell lines. Afatinib was chosen as the model TKI substrate for our study because it undergoes very minimal metabolism in several species. Afatinib is a substrate of both Pgp and BCRP, but is not a substrate of uptake transporters. Therefore, it was anticipated that an in-vivo efflux transporter-mediated interaction with afatinib would most likely not be confounded or masked by other factors influencing its disposition. From the in-vitro cell uptake studies, we found that nilotinib is a potent inhibitor of both Pgp and BCRP, and it reversed Pgp- and BCRP- mediated afatinib efflux. Subsequently, an in-vivo study was carried out in mice to investigate the interaction between afatinib and nilotinib; and also the impact of nilotinib on the pharmacokinetics and tissue distribution of afatinib. Afatinib exposure in the plasma and in most tissues, namely liver, lung, kidney, heart, muscle, fat, and skin, was found to be significantly increased when nilotinib was coadministered with afatinib. Further, the nilotinib concentrations in most mice tissues was above that needed for Pgp and BCRP inhibition. These results showed that nilotinib could be a potent chemosensitizing agent for Pgp- and BCRP- mediated TKI resistance. Additionally, a significant increase in afatinib brain exposure was observed in the mice which were administered afatinib in combination with nilotinib. This is an interesting and important finding that could potentially be very useful in the treatment of primary and metastasized brain tumors. We also developed a physiologically based pharmacokinetic model of afatinib to characterize its tissue disposition in mice organs, and this model was then scaled up to humans. The developed model accurately predicted afatinib plasma exposure in healthy volunteers and patients with solid malignant tumors, renal impairment, and hepatic impairment.
To investigate the chemosensitizing potential of methylated flavones in efflux transporter-mediated TKI resistance, the Bcrp1 inhibitory effect of 5,7-DMF and its effect on sorafenib accumulation was evaluated in-vitro. 5,7- DMF was found to be a potent inhibitor of Bcrp1 and consequently, its impact on the pharmacokinetics and tissue distribution of sorafenib was evaluated in mice. Results showed that co-administration with 5,7-DMF led to significantly greater sorafenib exposure in plasma and in most tissues collected. This indicated that 5,7-DMF may represent a promising chemosensitizing agent for Bcrp1-mediated TKI resistance due to its low toxicity and potent Bcrp1 inhibition.
Our results may have important clinical implications as TKIs are currently the most widely used anticancer agents. 5,7-DMF may show great potential in reversing MDR in tumors expressing BCRP. On the other hand, TKI-TKI combination therapy, especially with nilotinib as the perpetrator, is an attractive strategy to combat both Pgp- and BCRP-mediated TKI resistance. Additionally, since nilotinib has a wide volume of distribution and can reach various tissues at concentrations sufficient enough to inhibit Pgp and BCRP; it could potentially be used as a chemosensitizer in the treatment of numerous types of cancers. Furthermore, its chemosensitizing potential could particularly be useful in the treatment of primary and metastatic brain tumors. Further studies are warranted to assess the chemosensitizing effect of nilotinib in tumor xenograft models.
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Padi, Sathish K. R., Libia A. Luevano, Ningfei An, Ritu Pandey, Neha Singh, Jin H. Song, Jon C. Aster, Xue-Zhong Yu, Shikhar Mehrotra, and Andrew S. Kraft. "Targeting the PIM protein kinases for the treatment of a T-cell acute lymphoblastic leukemia subset." IMPACT JOURNALS LLC, 2017. http://hdl.handle.net/10150/624055.
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New approaches are needed for the treatment of patients with T-cell acute lymphoblastic leukemia (T-ALL) who fail to achieve remission with chemotherapy. Analysis of the effects of pan-PIM protein kinase inhibitors on human T-ALL cell lines demonstrated that the sensitive cell lines expressed higher PIM1 protein kinase levels, whereas T-ALL cell lines with NOTCH mutations tended to have lower levels of PIM1 kinase and were insensitive to these inhibitors. NOTCH-mutant cells selected for resistance to gamma secretase inhibitors developed elevated PIM1 kinase levels and increased sensitivity to PIM inhibitors. Gene profiling using a publically available T-ALL dataset demonstrated overexpression of PIM1 in the majority of early T-cell precursor (ETP)-ALLs and a small subset of non-ETP ALL. While the PIM inhibitors blocked growth, they also stimulated ERK and STAT5 phosphorylation, demonstrating that activation of additional signaling pathways occurs with PIM inhibitor treatment. To block these pathways, Ponatinib, a broadly active tyrosine kinase inhibitor (TKI) used to treat chronic myelogenous leukemia, was added to this PIM-inhibitor regimen. The combination of Ponatinib with a PIM inhibitor resulted in synergistic T-ALL growth inhibition and marked apoptotic cell death. Treatment of mice engrafted with human T-ALL with these two agents significantly decreased the tumor burden and improved the survival of treated mice. This dual therapy has the potential to be developed as a novel approach to treat T-ALL with high PIM expression.
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McIntyre, Neil A. "Synthesis of ring-constrained thiazolylpyrimidines : inhibitors of cyclin-dependent kinases." Thesis, University of St Andrews, 2006. http://hdl.handle.net/10023/353.
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One current approach in the treatment of cancer is the inhibition of cyclin dependent kinase (CDK) enzymes with small molecules. Here the discovery and development of 2-anilino-4-(thiazol-5-yl)pyrimidine CDK inhibitors is described, including details of the design and successful synthesis of novel ring-constrained thiazolylpyrimidines. The structure-activity relationship (SAR) trends exhibited by this constrained thiazolylpyrimidine family of CDK inhibitors are presented and compared with those from an unconstrained series of analogues. One significant finding from this aspect of the project was that ring-constrained thiazolylpyrimidines in general inhibit CDK2-cyclin E with greater potency than the corresponding unconstrained forms. Furthermore, an X-ray crystal structure of 2-methyl-N-[3-nitrophenyl]-4,5-dihydrothiazolo[4,5-h]quinazolin-8-amine, a representative from the constrained thiazolylpyrimidine series, in complex with CDK2-cyclin A is reported; confirming the binding mode within the CDK2 ATP binding pocket. A further assessment of SARs through the synthesis of control compounds and an extended study into the synthesis of N-substituted derivatives is described. The identification of CDK inhibitors that possess a strong selectivity profile across the CDK family is important. For example, the identification of highly CDK4-selective inhibitors should enable researchers to study the biological role of this important enzyme and to enable a block of cell division in the G1 phase. Here synthetic attempts to prepare a potentially CDK4 selective inhibitor compound, namely 5-methyl-N8-[4-(piperazin-1-yl)phenyl]thiazolo[4,5-h]quinazoline-2,8-diamine, are described. This approach was inspired by SAR data published on a structurally related inhibitor, 8-cyclopentyl-5-methyl-2-[4-(piperazin-1-yl)phenylamino]pyrido[2,3-d]pyrimidin-7(8H)-one.
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Haaning, Kelsey L. "Deletion of the phosphoinositide-3-kinase RhoGAP domain to assess inhibition of Staphylococcus aureus infection." Virtual Press, 2008. http://liblink.bsu.edu/uhtbin/catkey/1398713.
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It is important to understand the mechanism of endocytic invasion into the host cell by Staphylococcus aureus. Activation of phosphoinositide-3-kinase (PI3K) is essential to S. aureus invasion. In a normal cell, the p85 subunit of PI3K is bound at the Rho GTPase activating protein (RhoGAP) domain to small guanosine triphosphate binding proteins (GTPases), which are attached to the cell membrane by a prenyl group. This association anchors PI3K near the cellular membrane. PI3K must be anchored near the membrane in order to phosphorylate its substrate. The hypothesis for this project is that deletion of the binding domain between PI3K and small GTPases will block endocytic bacterial invasion by sequestering PI3K in the cytosol. To investigate this hypothesis, the RhoGAP binding domain of PI3K p85 was mutated using site-directed mutagenesis and S. aureus invasion was reduced by up to 86% (p<0.05), which shows that this domain is important to bacterial invasion.Department of Biology
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Deininger, Michael Werner Nikolaus. "STI571, a novel tyrosine kinase inhibitor : pre-clinical evaluation and application to identify downstream targets of BCR-ABL." Thesis, Imperial College London, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.325912.
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Trousil, Sebastian. "Choline kinase inhibition as a treatment strategy of cancers with deregulated lipid metabolism." Thesis, Imperial College London, 2014. http://hdl.handle.net/10044/1/25146.
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Aberrant choline metabolism is a characteristic shared by many human cancers. It is predominantly caused by elevated expression of choline kinase alpha, which catalyses the phosphorylation of choline to phosphocholine, an essential precursor of membrane lipids. In this thesis, a novel choline kinase inhibitor has been developed and its therapeutic potential evaluated. Furthermore the probe was used to elaborate choline kinase biology. A lead compound, ICL-CCIC-0019 (IC50 of 0.27 ± 0.06 μM), was identified through a focused library screen. ICL-CCIC-0019 was competitive with choline and non-competitive with ATP. In a selectivity screen of 131 human kinases, ICL-CCIC-0019 inhibited only 5 kinases more than 20% at a concentration of 10 μM (< 35% in all 131 kinases). ICL- CCIC-0019 potently inhibited cell growth in a panel of 60 cancer cell lines (NCI-60 screen) with a median GI50 of 1.12 μM (range: 0.00389-16.2 μM). Importantly, proliferation of normal cells was only minimally affected (MCF-10A, ST-T1b and CCD-18Co; GI50 30-120 μM). In HCT116 cells, ICL-CCIC-0019 potently inhibited the formation of phosphocholine (EC50 0.67 ± 0.28 μM), which consequently decreased the formation of phosphatidylcholine. The compound arrested cells in the G1 phase of the cell cycle, and induced endoplasmic reticulum stress and apoptosis. A single injection of ICL-CCIC-0019 at 10 mg/kg decreased tumour uptake of the choline kinase specific PET tracer [18F]fluoromethyl-[1,2-2H4]-choline at 24 hours (AUC0-60-23%). Treatment of HCT116 colon cancer cell xenograft bearing mice with 5 mg/kg ICL-CCIC-0019 i.p. resulted in strong tumour growth inhibition. Human breast cancer cell lines oncogenically transformed by HER2 exhibit increased levels of phosphocholine and are therefore more likely respond to CHKA inhibition. To identify such patients more readily, a novel, non-invasive, PET-imaging-based HER2- targeting diagnostic tool, [18F]GE-226, was developed. [18F]GE-226 (KD = 76 pM) uptake was 11 to 67-fold higher in 10 HER2 positive versus negative cell lines in vitro. Tumour uptake correlated with HER2 expression in 5 different tumour models (r2 = 0.78), and a fluorophore-labelled tracer analogue co-localised with HER2 expression. Tracer uptake was not influenced by short-term or continuous treatment with trastuzumab, but reflected HER2 degradation by short-term HSP90 inhibition. Taken together, these data further validate CHKA as a drug target and warrant the further development of ICL-CCIC-0019, potentially in the setting of HER2 positive cancers.
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Ashcroft, Jonathan William. "Assessment of the potential of a narrow spectrum kinase inhibitor in the treatment of influenza infection." Thesis, Imperial College London, 2014. http://hdl.handle.net/10044/1/44083.
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Severe outcome following influenza infection has been linked to the over induction of the host innate immune response resulting in a 'cytokine storm'. In certain patient populations, a therapeutic approach to help control the innate immune response may be of benefit. However concerns have been expressed that suppression of the host immune response might also lead to an increase in virus replication and directly enhance viral induced pathology. Using cultures of primary well-differentiated human airway epithelium (HAEs), the ability of a small molecule, narrow spectrum kinase inhibitor, developed by RespiVert (RV1088) to inhibit the extent of influenza virus induced expression of an array of human cytokines including IL6, IL8, IP10, and RANTES was investigated. The virus-induced response following infection with influenza virus was found to be diminished at the mRNA and protein level in the presence of the drug. Importantly, drug treatment did not adversely increase viral replication. In contrast, treatment with a steroid did not suppress the cytokine/chemokine response and resulted in increased viral titres. RV1088 inhibited the viral induction of transcription from the interferon promoter acting at or below the level of MAVS, preventing nuclear translocation of both IRF-3 and NFkB. Used alone, RV1088 inhibited cytokine production by all currently circulating subtypes and lineages of influenza A and B virus. Used in combination with currently licenced antivirals, the virus titre released from HAE cells was suppressed even further than for either drug alone, suggesting a synergistic antiviral effect. Finally, a novel murine model of influenza infection using nebulized virus to infect the mouse airways was developed. Drug also administered through the nebulised route suppressed the interferon response in the mouse lung and did not result in increased viral lung titre or weight loss. Administered intranasally with or without Relenza to mice infected with pH1N1 2009 virus, RV1088 suppressed interferon levels in mouse lung and reduced weight loss and mortality. This, or similar molecules, may represent a new generation of compounds suitable for the treatment of respiratory virus infections.
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Stillman, Anthony D. "Targeting Sphingosine Kinase 2 as a Treatment for Cholangiocarcinoma." VCU Scholars Compass, 2019. https://scholarscompass.vcu.edu/etd/6067.
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Cholangiocarcinoma (CCA) has a high mortality rate and its occurrence is rising. This increase prompts the need for improved CCA treatments. Studies have suggested that CCA is highly reliant on the sphingosine-1-phosphate-receptor-2 (S1PR2) and sphingosine kinase 2 (SphK2). Recently, a competitive SphK2 inhibitor, ABC294640, has been approved for clinical trial. ABC294640 has the potential to treat CCA, which is support by a phase I clinical study that was able to temporarily treat a patient suffering from metastasized CCA with ABC294640. To determine the viability of ABC294640 as a treatment for CCA, this study focused on determining the effects of ABC294640 on rat CCA cell lines. We found that ABC294640 inhibited the growth and migration of CCA and CAFs cells. The growth and count of 3-D organotypic co-culture of CCA and CAFs, which forms the “duct-like” structures, were reduced by ABC294640. The potential of inhibiting SphK2 as a treatment for CCA is supported by our finding of increased expression of S1PR2 and SphK2 in CCA patient liver samples. In conclusion, ABC294640 represents a potential therapeutic agent for CCA.
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Tawati, Salha M. "Design, synthesis and biological evaluation of sphingosine kinase inhibitors for the treatment of prostate cancer." Thesis, University of Strathclyde, 2018. http://digitool.lib.strath.ac.uk:80/R/?func=dbin-jump-full&object_id=30298.
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Sphingosine is phosphorylated via the action of the enzymes sphingosine kinase 1 (SK1) and sphingosine kinase 2 (SK2) to produce the bioactive signalling molecule sphingosine 1-phosphate (S1P). S1P drives cancer cell proliferation and migration whilst also promoting cell survival. Many studies have demonstrated that SK is a promising target for the treatment of cancer and the development of novel isoform-selective SK inhibitors to treat human cancers is of major interest. To date, inhibitors for this enzyme have either been selective for SK1 or non-selective for both isoforms. However, most SK inhibitors have only weak potency. This project involves the design and synthesis of small molecule inhibitors of SK as potential anti-cancer compounds. In this drug discovery project, a series of potent and selective inhibitors of SK (SK1 or SK2 or SK1/SK2) were developed based on the structure of PF-543, a known potent SK1 selective inhibitor. Analogues of PF-543 were prepared that were potent selective inhibitors of SK1 over SK2 and nM potent SK2 inhibitors with selectivity over SK1. These compounds represent some of the first nM potent SK2 inhibitors with selectivity over SK1. Indeed, the studies identified a structural determinant in the catalytic site of SK1 and SK2 that confers selectivity, with the heel and toe regions of the so-called J-channel in either enzyme providing a means toward selectivity. Exemplars from the series were shown to have potent cellular activity but poor in vitro microsomal stability. Effective target engagement and selectivity for SK1 in prostate cancer cell lines (LNCaP and LNCaP-AI) and proliferating human pulmonary artery smooth muscle cells (hPSMAC) were also established. A variety of biological assays associated with SK inhibition were used to evaluate their ability to induce cancer cell death, which was shown to involve a caspase-3/7-independent mechanism. Our SK1 and SK1/SK2 inhibitors, but not SK2 inhibitors, also reduced expression of dihydroceramide desaturase 1 (Des1) in a dose dependent manner, causing growth arrest and caspase-independent cell death. This project highlighted the importance for combining SK1 with Des1 inhibition in terms of endowing compounds with cytotoxicity against cancer cells.
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Chakraborty, Kanishka, John B. Bossaer, R. Patel, and K. Krishnan. "Successful Treatment of Nilotinib-Induced Pleural Effusion with Prednisone." Digital Commons @ East Tennessee State University, 2012. https://dc.etsu.edu/etsu-works/2318.
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Chronic myeloid leukemia is characterized by a unique reciprocal translocation between chromosomes 9 and 22 resulting in deregulated tyrosine kinase activity. Tyrosine kinase inhibitors, such as imatinib, dasatinib, and nilotinib have revolutionized treatment of Chronic myeloid leukemia. However, tyrosine kinase inhibitors? use has presented new challenges in managing both acute and chronic toxicities, particularly ?off-target? toxicities like pleural effusion. Pleural effusions are seen less often with imatinib and very rarely with nilotinib. A 66-year-old male presented to emergency department with complaints of mild chest pain and dyspnea of 3 days duration with progressive worsening, including dyspnea at rest. Patient was currently taking nilotinib after failing imatinib for chronic myeloid leukemia. Nilotinib was put on hold. After exclusion of cardiac and pulmonary etiologies patient was treated for community acquired pneumonia with minimal improvement. Despite the very low incidence of pleural effusion with nilotinib (<1%), he was started on 20?mg of prednisone PO for 3 days. Patient had a dramatic improvement within 48?h after beginning prednisone. This treatment approach suggests that pleural effusions associated with nilotinib can be successfully treated in the same way as pleural effusions associated with dasatinib.
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Nakano, Kenji. "Risk factors of pneumothorax in advanced and/or metastatic soft tissue sarcoma patients during pazopanib treatment: a single-institute analysis." Kyoto University, 2018. http://hdl.handle.net/2433/232072.
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Mazanetz, Michael Philip. "Approaches towards the design and synthesis of selective kinase inhibitors for the treatment of neurodegenerative diseases." Thesis, University of Nottingham, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.508226.
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Sykora, Vladimir. "Computational approaches in the development of cyclin-dependent kinase 2 inhibitors for the treatment of cancer." Thesis, University of Newcastle Upon Tyne, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.506664.
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Farina, Anne Kata. "Regulation of cadherin-11 by GSK3 inhibition and TGFbeta1 treatment in cancer cells." Connect to Electronic Thesis (CONTENTdm), 2008. http://worldcat.org/oclc/457179663/viewonline.
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Schneider, Melanie. "Integrative chemoinformatics to guide drug design : application to re-design a clinical protein kinase inhibitor." Thesis, Montpellier, 2019. http://www.theses.fr/2019MONTT057.
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Malgré des années de recherche et de développement intensifs, le cancer reste l’une des principales causes de décès dans le monde. La chimiothérapie est le traitement le plus couramment utilisé contre le cancer, car la chirurgie et la radiothérapie ne sont souvent pas efficaces pour traiter le cancer à tous les endroits où il se propage. Cependant, la pharmacorésistance des cellules cancéreuses aux agents chimiothérapeutiques et / ou la réduction de l’efficacité d’un médicament est la principale cause d’échec de la chimiothérapie. Les médicaments sont développés pour se lier efficacement à une cible thérapeutique donnée, appelée cible primaire. Malheureusement, les traitements médicamenteux peuvent souffrir de la liaison à une cible secondaire qui perturbe l’activité du médicament et / ou a un impact sur son métabolisme. L’objectif principal du projet de thèse était de développer uneapproche chémo-informatique intégrative pour optimiser la conception de médicaments en étudiant non seulement la cible primaire, mais également les effets secondaires putatifs au niveau atomique afin de calculer des modes de liaison plus précis et d’obtenir de meilleures estimations d’affinité.Le projet de conception de médicament présenté vise un inhibiteur amélioré du mutant de la sérine/thréonine kinase BRAFV600E avec perte simultanée de la liaison à la cible secondaire, le récepteur PXR. L’accent est mis sur l’étude à la fois de la protéine kinase BRAF et du récepteur nucléaire PXR, impliqué dans la régulation du métabolisme xénobiotique. Un outil d’apprentissage automatique est d’abord développé sur le récepteur nucléaire bien étudié ERα en raison de grandes quantités de données expérimentales, puis généré de manière similaire pour BRAFV600E. Malgré son importance reconnue dans le métabolisme des médicaments, nous manquons toujours d’informations structurelles et de mesures d’affinité suffisantes pour développer l’apprentissage automatique sur PXR. Aussi, une approche alternative reposant sur la dynamique moléculaire associée à la méthode «Molecular Mechanics Poisson-Boltzmann Surface Area» est utilisée afin d’obtenir une estimation précise des affinités des ligands. Enfin, divers outils informatiques sont utilisés pour concevoir de nouveaux dérivés du médicament initial, métabolisé trop rapidement chez de nombreux patients, entraînant une résistance et une rechute du cancer. Les propriétés des nouveaux composés empêchent l’activation des enzymes métabolisantes qui dégradent le médicament initial. Ceci devrait fournir un nouveau médicament candidat aux propriétés pharmacocinétiques bien meilleures et à une efficacité accrue.Cette thèse comprend un pipeline complet de conception de médicaments et présente une stratégie intégrée comprenant la modélisation, la conception et la synthèse in silico, le criblage virtuel, les prédictions d’affinité, les tests in vitro et la cristallographie de rayon X. L’accent est mis principalement sur la partie informatique qui comprend des approches complémentaires du point de vue du médicament et des protéinesDespite years of intensive research and development, cancer remains one of the leading causes of death worldwide. Chemotherapy is the most commonly used treatment for cancer, as surgery and radiation therapy are often not effective in treating cancer at every location where it spreads. However,drug resistance of cancer cells to chemotherapeutic agents and/or reduction in effectiveness of a drug is the leading cause of failure of chemotherapy. Drugs are developed to bind efficiently to a given therapeutic target, called the primary target. Unfortunately, drug treatments can suffer from bindingto a secondary target that perturbs drug activity and/or impacts its metabolism. The main aim of the PhD project was to develop an integrative chemoinformatics approach to optimize drug design by studying not only the primary target but also putative secondary effects at the atomic level in order to compute more accurate binding modes and to derive better affinity estimates.The presented drug design project aims for an improved inhibitor of the serine/threonine kinase mutant BRAFV600E with simultaneous loss of binding to the secondary target PXR. Focus is on the study of both, protein kinase BRAF and nuclear receptor PXR, which is involved in regulation of xenobiotic metabolism. A machine learning tool is first developed on the well studied nuclear receptor ERα due to large amounts of experimental data, and subsequently similarly generated for BRAFV600E. Despite its recognized importance in drug metabolism, we are still lacking sufficient structural information and affinity measurements to develop machine learning models for PXR. So, an alternative approach that relies on molecular dynamics combined with the Molecular Mechanics Poisson-Boltzmann Surface Area method is employed in order to obtain a precise estimation of ligand affinities. Finally, diverse computational tools are applied to design new derivatives of the initial drug, which is too rapidly metabolized in many patients resulting in resistance and cancer relapse. The properties of the new compounds prevent activation of metabolizing enzymes that are degrading the original drug. This is expected to provide a new drug-candidate with much better pharmacokintics properties and enhanced efficacy.This thesis comprises a complete drug design pipeline and presents an integrated strategy that includes modeling, in silico design and synthesis, virtual screening, affinity predictions, in vitro tests and X-ray crystallography. The main focus is on the computational part that comprises complementary approaches from the drug’s and from the proteins’ point of view
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Choi, Ho-ying, and 蔡可盈. "Review of clinical benefits and cost effectiveness of epidermal growthfactor receptor-tyrosine kinase inhibitor (EGFR-TKI) as first linetreatment for patients with advanced non-small cell lung cancer(NSCLC)." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46935320.
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Lilly, Scott Matthew. "Protein Kinase A Alterations Following Chronic Flurazepam Treatment: Implications for Inhibitory and Excitatory Synaptic Plasticity in Rat Hippocampal CA1." Connect to full-text via OhioLINK ETD Center, 2006. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=mco1145293063.
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Thesis (Ph.D.)--Medical University of Ohio, 2006."In partial fulfillment of the requirements for the degree of Doctor of Philosophy in Medical Sciences." Major advisor: Elizabeth I. Tietz. Includes abstract. Document formatted into pages: iv, 234 p. Title from title page of PDF document. Bibliography: pages 86-95,126-135,167-174,190-232.
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Yacoub, Jeanine. "Synthesis of Agents for the Treatment and Analysis of Tropical Diseases." Scholar Commons, 2015. http://scholarcommons.usf.edu/etd/6441.
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Toxoplasmosis is an opportunistic disease caused by the protozoan parasite Toxoplasma gondii. The parasite is usually staved off by a healthy immune system and remains dormant in the body. In immunocompromised patients, the parasite can become active and spread throughout the body causing symptoms such as encephalitis, cognitive disorders, seizures, and death. Combination drug therapy is the usual treatment for toxoplasmosis; however, patients suffer from problems of intolerance, allergic reactions, and cytotoxicity. In an effort to identify new drug targets for toxoplasmosis, a series of compounds have been synthesized that can be used as tools to probe the unique pathways used by T. gondii to survive in the human host. One class of these compounds is pyridinyl imidazoles, which have been shown to be active against T. gondii MAP kinases. To set up a protein pull down assay, a biotinylated linker was synthesized. We have also synthesized a compound that’s being used to study the pathways involved in the most active and proliferative form of T. gondii.
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Carpenter, Kent James. "Inhibition of PIM and AXL Kinases As Potential Treatments for a Variety of Hematological Malignancies and Solid Tumors." BYU ScholarsArchive, 2014. https://scholarsarchive.byu.edu/etd/3842.
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This thesis is divided into three chapters. In each case, the goal is to achieve inhibition of a growth kinase (PIM or AXL) and subsequent arrest of cell growth and induction of apoptosis (in vitro cell culture models) or decrease in tumor volume (in vivo xenograft studies). Chapter one and chapter two discuss inhibition of proviral integration site for Moloneymurine leukemia virus (PIM) kinases. The three PIM kinases, PIM-1, PIM-2, and PIM-3, are a subfamily of serine/threonine kinases that are known to be involved in signaling pathways as downstream effectors of signal transducer and activator of transcription-5 (STAT5) signaling and inhibitors of apoptosis. PIM kinases are implicated in a large percentage of hematological malignancies and solid tumors. Because they have been shown to correlate with disease progression and poor prognosis in many of these conditions, PIM kinase inhibitors are being developed and investigated for therapeutic use. The aim of this study in chapter one was to evaluate the role of PIM 1, 2 and 3 in urothelial carcinomas, using second generation Pan-PIM kinase inhibitor TP-3654. Retrospective immunohistochemical analysis of bladder cancer specimens found that PIM 1, 2, and 3 was expressed in a significant number of cases. PIM-1 was expressed in 4 bladder cancer cell lines and TP-3654 treatment was able to inhibit BAD phosphorylation to induce apoptosis. The second aim of this study was to investigate the effects of TP-3654 on the interaction of c-MYC with PIM kinase family members. The data indicate that PIM-1 only interacts with c-MYC in the acute myeloid leukemia (AML) and multiple myeloma (MM) cell lines studied, and that PIM-1 siRNA knockdown or treatment with TP-3654 is able to decrease this interaction. The third chapter discusses inhibition of the receptor tyrosine kinase Axl. Pancreatic cancer is a highly lethal disease characterized by malignant cells that rapidly disseminate from the primary tumor to form local and distant metastases. Axl is overexpressed in over 50% of pancreatic cancers and expression of Axl in these cancers is highly associated with a poor prognostic outcome for patients. Small molecule inhibitors of AXL are currently under investigation, as AXL is associated with cell migration mediated by epithelial-mesenchymal transition (EMT). The aim of this study was to investigate the effects of a small molecule inhibitor of AXL, TP-0903, on pancreatic cancer cell lines. Consistent with the known function of Axl, TP-0903 inhibited Gas6-induced migration and invasion of pancreatic cancer cells invitro and potently induced apoptosis. Additionally, we found that inhibition of AXL decreased expression of EMT marker genes and induced mesenchymal pancreatic cancer cell lines to take on an epithelial phenotype. TP-0903 also significantly inhibited the growth of pancreatic cancer cell lines grown in xenograft tumor mouse model and taken together, the results suggest Axl is a potential therapeutic target in pancreatic cancer and TP-0903 as a potential therapeutic agent.
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Johnson, Neil. "Investigation into the therapeutic potential of novel cyclin dependent kinase (CDK) inhibitors in the treatment of antiestrogen sensitive and resistant breast cancer." Thesis, University of Newcastle Upon Tyne, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.427304.
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Mohedas, Agustin Humberto. "Development of BMP type I receptor kinase inhibitors for the treatment of fibrodysplasia ossificans progressiva and the study of the BMP signaling pathway." Thesis, Massachusetts Institute of Technology, 2014. http://hdl.handle.net/1721.1/90173.
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Thesis: Ph. D., Harvard-MIT Program in Health Sciences and Technology, 2014.164
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Includes bibliographical references (pages 121-129).
The BMP signaling pathway is essential for embryonic development and the maintenance of tissue homeostasis. Dysregulated BMP signaling, both loss and gain-of-function, has been demonstrated in the pathogenesis of diseases including cancer, atherosclerosis, anemia and particularly hereditary disorders such as pulmonary arterial hypertension, hereditary hemorrhagic telangiectasia, and fibrodysplasia ossificans progressiva (FOP). FOP is a rare and disabling condition caused by a highly recurrent mutation in the ACVR1 gene encoding the BMP type I receptor activin-like kinase 2 (ALK2), characterized by the progressive heterotopic ossification (HO) of skeletal muscle and connective tissue leading to widespread joint immobilization, with significant morbidity and premature mortality. There are currently no effective treatments for FOP. The goal of this thesis is to develop and characterize highly selective BMP type I receptor inhibitors targeting ALK2 for the treatment of FOP. Despite the high degree of structural homology between all the BMP and TGF-[beta] type I receptors, I hypothesized that potent and selective inhibitors targeting a single BMP type I receptor, ALK2, could be developed based on a previously identified pyrazolo[1,5-a]pyrimidine core scaffold. I screened a library of pyrazolo[1,5-a]pyrimidine derivatives in a high throughout sensitive radiometric assay of BMP and TGF-[beta] type I receptor kinase activities. I identified a derivative with a unique chemical moiety (5-quinoline) that demonstrated high selectivity for ALK2, but with lower potency than the parent molecule. We synthesized a new 5-quinoline derivative with increased potency and selectivity for ALK2 over the other BMP type I receptors and greatly improved selectivity against the TGF--[beta] type I receptors. I used this highly selective compound to examine ALK2-mediated BMP signaling in vitro and demonstrated in vivo efficacy in two mouse models of HO. In a complementary approach, we generated a library of novel BMP type I receptor inhibitors based on the 2-aminopyridine core scaffold. I developed a structure activity relationship to determine the key structural elements responsible for potency and selectivity. We identified a several novel derivative compounds with improved potency and selectivity for ALK2 over the parent. We successfully used this set of derivatives to address a specific question in FOP biology, of whether ATP-competitive kinase inhibitors exert differential activity against wild-type or diverse FOP-causing ALK2 mutants. Finally, in our SAR of pyrazolopyrimidine compounds, we identified a highly potent inhibitor of both BMP and TGF-[beta] type I receptor activity. I characterized the ability of this compound to inhibit ligand-induced BMP and TGF-[beta] signaling in a variety of cell culture models, as well as inhibit the activity of individual type I receptors. We then used this compound to examine the contribution of individual BMP and TGF-[beta] receptors to signal transduction. We used the broad activity of this inhibitor to limit signaling of all endogenous BMP and TGF-[beta] type I receptors in cells, while reconstituting the activity of specific type I receptors using engineered, inhibitor-resistant mutant receptor kinases which we developed by modifying gatekeeper residues critical for interactions with inhibitor. These mutant receptor kinases demonstrated preserved basal and ligand-mediated signaling functions which were unaffected by inhibitor. These results demonstrate proof-of-principle of a system for examining the function of individual receptors of this pathway in isolation. The work presented in this thesis advances the development of novel BMP type I receptor kinase inhibitors of high selectivity and potency which could serve as important tools for the study of BMP signaling and as therapies for diseases of excessive BMP signaling such as FOP. Development of highly potent and selective inhibitors of ALK2 offers the hope of rational disease modifying therapy for the treatment of FOP.
by Agustin Humberto Mohedas.
Ph. D.
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Tohme, Rita. "DIRECT PP2A ACTIVATION FOR THE TREATMENT OF KRAS- AND EGFR-DRIVEN LUNG ADENOCARCINOMA." Case Western Reserve University School of Graduate Studies / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case1526073628472942.
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Ghelli, Luserna di Rorà Andrea <1987>. "The Inhibition of Chk1/Chk2 and Wee-1 Kinases as a Promising Therapy for the Treatment of Adult Acute Lymphoblastic Leukemia." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7529/.
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Due to inadequate treatments, the survival rate of adult patients with acute lymphoblastic leukemia (ALL) is still very poor. Thus there is a need to improve the efficacy of conventional therapy. In this study we evaluated the effectiveness of checkpoint kinase inhibitors (Chk-i) in single agent and in combination with different compounds conventionally used for the treatment of B-/T-ALL. We showed that Chk1 and Chk2 kinases are highly expressed and hyper-activated in tumor samples in comparison to normal tissue. On these bases we speculate that the inhibition of these kinases could mine the genetic stability and enhance cell death in ALL cells. We firstly evaluate the efficacy in single agent of the Chk1/Chk2 (PF-0477736 and LY2606368) and of the Wee1 (MK-1775) inhibitors on different cell lines and on primary cells isolated from adult B-ALL patients. We demonstrated that the inhibition of Chk1/Chk2 kinases reduces of the cell viability, activates the apoptosis and modify the expression of different elements of the G2/M checkpoint. To assess the chemo-sensitizer activity of different checkpoint kinase inhibitors, several combination studies were performed. To this purpose, LY2606368 and MK-1775 were combined with different tyrosine kinase inhibitors (imatinb, dasatinib and bosutinib) and with the purine nucleoside analogue, clofarabine. The efficacy of the combinations was not only evaluated in term of reduction of the cell viability but also in term of induction of apoptosis and induction of DNA damages. The results found were then confirmed on primary cells of B-ALL patients. Finally different class of checkpoint kinase inhibitors were combined together in order to evaluate their interaction. In our opinion the preclinical data presented in this study are the basis for a future evaluation of this class of compound in clinical trials in the treatment of adult ALL patients.
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Xiao, Zhiguang [Verfasser], Bernhard [Akademischer Betreuer] Küster, and Axel [Akademischer Betreuer] Ullrich. "Combinatorial treatment of lung cancer monolayer cells and their spheroids with tyrosine kinase inhibitors and Salinomycin / Zhiguang Xiao. Gutachter: Bernhard Küster ; Axel Ullrich. Betreuer: Bernhard Küster." München : Universitätsbibliothek der TU München, 2015. http://d-nb.info/1069127760/34.
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Hähnel, Tom, Christoph Baldow, Joëlle Guilhot, François Guilhot, Susanne Saussele, Satu Mustjoki, Stefanie Jilg, et al. "Model-based inference and classification of immunological control mechanisms from TKI cessation and dose reduction in CML patients." American Association for Cancer Research (AACR), 2020. https://tud.qucosa.de/id/qucosa%3A74320.
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Recent clinical findings in chronic myeloid leukemia (CML) patients suggest that the risk of molecular recurrence after stopping tyrosine kinase inhibitor (TKI) treatment substantially depends on an individual’s leukemia-specific immune response. However, it is still not possible to prospectively identify patients that will remain in treatment-free remission (TFR). Here, we used an ordinary differential equation (ODE) model for CML, which explicitly includes an anti-leukemic immunological effect and applied it to 21 CML patients for whom BCR-ABL1/ABL1 time courses had been quantified before and after TKI cessation. Immunological control was conceptually necessary to explain TFR as observed in about half of the patients. Fitting the model simulations to data, we identified patient-specific parameters and classified patients into three different groups
according to their predicted immune system configuration ('immunological landscapes”). While one class of patients required complete CML eradication to achieve TFR, other patients were able to control residual leukemia levels after treatment cessation. Among them were a third class of patients, that maintained TFR only if an optimal balance between leukemia abundance and immunological activation was achieved before treatment cessation. Model simulations further suggested that changes in the BCR-ABL1 dynamics resulting from TKI dose reduction convey information about the patient-specific immune system and allow prediction of outcome after treatment cessation. This inference of individual immunological configurations based on treatment alterations can also be applied to other cancer types in which the endogenous immune system supports maintenance therapy, long-term disease control or even cure.
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Horn, Matthias. "Optimierung der Therapie von chronischer myeloischer Leukämie mit Hilfe eines dynamischen Modells normaler und leukämischer Stammzellorganisation." Doctoral thesis, Universitätsbibliothek Leipzig, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-154574.
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Unter Verwendung eines mathematischen Hämatopoese-Modells werden verschiedene Fragen adressiert, die im Zusammenhang mit einer möglichen Optimierung der gegenwärtigen Therapie chronischer myeloischer Leukämie (CML) stehen. Es handelt sich um ein agentenbasiertes Modell, das heißt, jede Zelle wird als einzelnes Objekt repräsentiert und gemäß festgelegter Regeln im Computer simuliert. Es werden proliferative von ruhenden Stammzellen unterschieden, wobei sich der Proliferationszustand reversibel ändern kann. Das Modell basiert auf der Annahme, dass sich normale und maligne Stammzellen in einem Wettbewerb um gemeinsame Ressourcen befinden, wobei der CML-Klon einen kompetitiven Vorteil besitzt.
Es ist ungeklärt, ob Tyrosinkinaseinhibitoren wie Imatinib (IM) in der Lage sind, die Erkrankung zu heilen. Es gibt Evidenz, dass residuale leukämische Stammzellen im Knochenmark persistieren, welche in einem Ruhezustand (G0-Phase des Zellzyklus) von IM nicht eradiziert werden können. Proliferativ aktive Zellen sind der IM-Wirkung hingegen ausgesetzt. Das Modell sagt voraus, unter welchen Bedingungen eine Kombinationsstrategie von IM mit stammzellaktivierenden Substanzen Synergieeffekte hervorbringen könnte.
Ein verwandtes Problem ist die Frage, in welchen Fällen nach Reduktion der Tumorlast auf ein mittels hochsensitiver Messmethoden undetektierbares Niveau ein Therapieabbruch gerechtfertigt ist. Basierend auf dem dynamischen Modell wird in dieser Arbeit ein Prädiktor vorgeschlagen, der vorhersagt, ob ein Patient nach Abbruch der Therapie einen molekularen Rückfall zu erwarten hat. Zusätzlich wird approximativ ein modellunabhängiger Prädiktor angegeben, der die Vorhersage nur auf Basis klinisch messbarer Größen gestattet.
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Frémond, Marie-Louise. "Clinical and molecular characterisation of the type I interferonopathies and approaches to therapy Efficacy of the Janus kinase 1/2 inhibitor ruxolitinib in the treatment of vasculopathy associated with TMEM173-activating mutations in three children Blockade of TANK-binding kinase 1/IKKε mutant stimulator of interferon genes (STING)-mediated inflammatory responses in human peripheral blood mononuclear cells." Thesis, Sorbonne Paris Cité, 2018. http://www.theses.fr/2018USPCB098.
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Le concept d'interféronopathie de type I émerge en 2011 et fait référence à un ensemble de pathologies Mendéliennes caractérisées par une hyperactivation des interférons (IFN) de type I. Tous les gènes associés au syndrome d'Aicardi-Goutières (SAG), la première interféronopathie de type I décrite, sont impliqués dans la détection ou le métabolisme des acides nucléiques. Les autres protéines mutées associées aux interféronopathies de type I modifient toutes la voie de signalisation des acides nucléiques, de manière directe, indirecte ou encore non définie. Les IFN de type I se fixent à un récepteur unique et activent la Janus kinase 1 (JAK1) et la tyrosine kinase 2 conduisant à l'expression de gènes stimulés par les IFN (IFN-stimulated genes, ISGs) via la phosphorylation des facteurs de transcription STAT1 et STAT2. Notre équipe a développé des outils diagnostiques des interféronopathies de type I, comprenant la signature IFN, analyse combinée de l'expression de 6 ISGs, et, plus récemment, une méthode de détection de l'IFN alpha à l'aide de la technologie «single molecule array». Les mutations monogéniques associées jusqu'à présent aux interféronopathies de type I causent des phénotypes variables. Leurs points communs sont une morbidité et une mortalité importantes, notamment en raison de leur réponse faible aux immunomodulateurs classiques. Les mutations activatrices de TMEM173 codant pour STING (Stimulator of IFN genes) sont responsables d'une inflammation sévère, connue sous le nom de STING-associated vasculopathy with onset in infancy (SAVI), et caractérisée par une vascularite cutanée et une atteinte interstitielle pulmonaire conduisant à une insuffisance respiratoire terminale. STING, une protéine du réticulum endoplasmique (RE), agit comme un adaptateur cytosolique de la détection de l'ADN, permettant la synthèse d'IFN de type I via la phosphorylation d'IRF3. Une cohorte internationale de 20 patients SAVI est décrite dans cette thèse soulignant l'hétérogénéité clinique de cette maladie. Nous avons également étudié le lien entre des mutations hétérozygotes de COPA et une activation de la voie des IFN de type I. COPA code pour la sous-unité alpha du complexe du coatomère I, impliqué dans le transport rétrograde entre le RE et le Golgi. Les mutations hétérozygotes de COPA sont à l'origine d'un phénotype proche du SAVI et entraînent une hausse du stress du RE et une réponse immunitaire de type Th17. Cependant, la physiopathologie de cette maladie reste peu connue. Nous avons étudié un groupe de 8 patients qui illustre l'hétérogénéité phénotypique de cette affection nouvellement décrite. Nous avons observé des similitudes entre l'histologie pulmonaire du syndrome COPA et du SAVI, ainsi qu'une signature IFN, des taux élevés d'IFN alpha dans le sérum et une phosphorylation de STAT1 dans les lymphocytes des patients. Dans un modèle cellulaire, la coexpression de COPA muté et de STING sauvage entraîne la phosphorylation d'IRF3 et à une induction d'ISGs, suggérant que les mutations de COPA conduisent à une activation dépendante de STING de la voie des IFN de type I. Nous avons mené avec succès le premier essai clinique d'un inhibiteur de JAK1, le ruxolitinib, dans le contexte du SAVI. L'amélioration clinique remarquable a été confirmée in vitro et ex vivo. La gravité de la maladie nous a également poussé à chercher des alternatives thérapeutiques pour contrôler la voie des IFN de type I. Nous avons montré que l'inhibition d'IKK bloquait efficacement la production et la signalisation des IFN de type I dans les cellules de patients STING in vitro. Devant les résultats prometteurs de l'inhibition de JAK1 dans le SAVI, nous avons ensuite testé le ruxolitinib dans le cadre d'autres interféronopathies de type I monogéniques (COPA, TREX1) mais aussi chez une enfant ayant une dermatomyosite sévère, une maladie pour laquelle le rôle pathogénique de l'IFN de type I a été suggéréThe term 'type I interferonopathies', first coined in 2011, refers to a set of Mendelian disorders associated with constitutive up-regulation of type I interferon (IFN) signalling. All of the genes associated with Aicardi-Goutières syndrome (AGS), the first Mendelian type I interferonopathy described, have been implicated in either the processing or sensing of nucleic acids. Beyond AGS, the other mutated proteins associated with type I interferonopathies have a direct, indirect, or currently undefined action on nucleic acid signalling. Type I IFNs drive the expression of IFN-stimulated genes (ISGs) through the engagement of a common receptor and the subsequent activation of Janus kinase 1 (JAK1) and tyrosine kinase 2, and phosphorylation of STAT1 and STAT2. Our team has developed diagnostic tools to identify type I interferonopathies, comprising a so-called IFN signature, involving the assessment of mRNA expression of 6 ISGs and, more recently, a high sensitivity assay of IFN alpha protein using single molecule array technology. Monogenic mutations so far recognised as type I interferonopathies are associated with a wide spectrum of phenotype. The hallmark of these diseases is their significant morbidity and mortality, associated with an apparent absence of response to conventional immunosuppressive therapies. Activating mutations in TMEM173, encoding stimulator of IFN genes (STING), cause a severe inflammatory condition referred to as STING-associated vasculopathy with onset in infancy (SAVI), characterised by skin vasculopathy and interstitial lung disease leading to end-stage respiratory failure. The endoplasmic reticulum (ER) protein STING is a central component of DNA sensing that induces type I IFNs through phosphorylation of IRF3. An international cohort of 20 STING patients is reported in this thesis, emphasising the clinical heterogeneity of this condition. We also investigated the link between heterozygous mutations in COPA and type I IFN signalling. COPA encodes the alpha subunit of the 7 member coatomer complex I, involved in retrograde transport from the golgi to the ER. Heterozygous mutations in COPA cause a phenotype showing some overlap with SAVI, and are associated with increased ER stress and priming of a Th17 response. However, the precise pathophysiology of this disease is so far undefined. We have studied a group of 8 patients illustrating the phenotypic variability of this emerging disease. We observed commonalities in the lung pathology in COPA and SAVI, as well as an IFN signature, raised levels of IFN alpha in the serum and phosphorylation of STAT1 in patient T cells. In a cellular model, phosphorylation of IRF3 and increased ISG expression were observed in cells co-transfected with wild type STING and mutant COPA plasmids, suggesting that mutations in COPA lead to constitutive activation of type IFN signalling through STING. We reported, for the first time, the successful use of a JAK1 inhibitor, ruxolitinib, in the context of SAVI. We observed a marked clinical effect, which was mirrored by the results of in vitro and ex vivo experiments. Because of the severity of SAVI, we also aimed to evaluate alternative therapeutic approaches to block type I IFN signalling and showed that IKK inhibition efficiently abrogated in vitro constitutive activation of type I IFN production and signalling in cells from STING patients. Considering the promising results of JAK1 blockade in SAVI, we then trialled ruxolitinib in other monogenic type I interferonopathies (TREX1, COPA) and in a child with severe dermatomyositis, a disease where type I IFN has been suggested to play a key pathogenic role
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Van, Sebille Ysabella. "Characterisation and Treatment of Pan-Human Epidermal Receptor Tyrosine Kinase Inhibitor-Induced Gastrointestinal Toxicity." Thesis, 2017. http://hdl.handle.net/2440/122124.
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The format of my thesis is as follows: two literature reviews, three research chapters, a general discussion, references and appendices. During my candidature, I made a significant effort to publish my research findings. Each chapter is presented in its original publication format, with the exception of spelling changes to keep consistent English spelling, and the references which are listed in the final chapter. This may result in slight repetition between chapters arising from the same study. My thesis has two distinct themes relating to the pathobiology of gastrointestinal toxicity second to targeted cancer treatment. The first theme aims to characterise gastrointestinal toxicity induced by dacomitinib, a pan-human epidermal receptor (HER) tyrosine kinase inhibitor (TKI) associated with high levels of diarrhoea. This theme gives rise to the first four chapters. Chapter 1 was an invited review introducing dacomitinib, highlighting the increasing incidence of gastrointestinal toxicity seen with second generation small molecule HER-TKIs and incorporating basic information required for understanding of subsequent chapters. Chapter 2 is a detailed literature review proposing chloride secretion as a mechanistic hypothesis for development of HER TKI-induced diarrhoea. Chapter 3 is my first original research chapter, which characterised dacomitinib-induced gastrointestinal toxicity in a novel rat model. Chapter 4, my second original research chapter, advances on this characterisation, and introduces crofelemer, a chloride channel blocker as a therapeutic intervention for the prevention of dacomitinib-induced diarrhoea. During my candidature, I had the opportunity to work with Assistant Professor Tom Carney from the Institute of Molecular and Cell Biology (IMCB), Biopolis, Singapore. This collaboration comprises Chapter 5, the second theme of this thesis. This chapter attempted to address the need for an innovative model for the study of gastrointestinal toxicity, investigating transgenic zebrafish reporter lines as a novel model for the study of chemotherapy and targeted therapy-induced gastrointestinal toxicity. Parts of this thesis were funded by Pfizer Pharmaceuticals under an unrestricted investigator-initiated grant (Reference #WI175212). The report I generated for this grant, which partly formed the basis of the publications presented in Chapters 3 and 4, is included as an appendix in this thesis.Thesis (Ph.D.) -- University of Adelaide, Adelaide Medical School, 2017
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Yen, Nai-Jung, and 顏乃容. "Polo-like kinase 1 modulated by HDAC inhibitors treatment in gastric cancer cells." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/36137596198636985692.
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碩士國立陽明大學
醫學生物技術暨檢驗學系暨研究所
96
Histone deacetylase inhibitors (HDIs), Sodium butyrate (NaB) and Tritrostatin A (TSA), are promising chemotherapeutic candidates. Although they could block proliferation of cancer cells and induce their apoptosis, it is known that some HDIs treatment also brings about unwanted cytotoxicity. Therefore, it is our interests to find a kinase target could have combinatory effect for HDI treatment. Only limited numbers of studies have examined the HDI modulated kinases, which were aberrantly expressed in cancer cells resulting in increasing cell survival, progression and malignancies. We used a restriction analysis of gene expression (RAGE) screening tool which had previously established and could generate comprehensive kinase profiles by using degenerate PCR primers targeting the conserved kinase motifs. Based on the PTK profiling results of this technique, we further found that both TSA and NaB treatments in NUGC cells generated surprisingly similar modulation effect on protein kinase expression. According to subsequent real-time PCR validated results, we demonstrated several kinases were up regulated in TSA or NaB treated NUGC cells, like Fyn, Tnk1, PdgfrB, and a few kinases down regulated such as ErbB2 and Plk1. Based on the validation results and additional evidence, we selected Plk1 (polo-like kinase 1) for further studies. Plk1 is a serine/threonine kinase that functions in cell cycle regulation and spindle assembly during mitosis. It is found to be over-expressed in many human cancer types and has become a new target for cancer therapeutics. However, little is known about Plk1 regulation by HDI treatment. From our observation on HDI modulation of Plk1 expression, it is possible that Plk1 could play critical roles in HDI modulated cancer cell apoptosis. We further analyzed the combinatory effects of the Plk1 inhibitor and HDI to enhance gastric cancer cell death by reducing HDI dosage and toxicity with the addition of Plk1 inhibitor. We demonstrated that Plk1 seems to be important in HDI mediated cell death and could have important applications in enhancing the clinical treatment strategy of TSA in human cancers. The use of RAGE kinase expression profiling method is beneficial in discovering important oncogenic kinases in cancer malignant progression. With the discovery of noval kinase gene targets, new chemotherapeutic strategies could be formulated by taking advantages of many new kinase inhibitors developed.
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Yeung, David Tak On. "Prognostic markers associated with tyrosine kinase inhibitor treatment response and maintenance of treatment free remission in chronic myeloid leukaemia." Thesis, 2016. http://hdl.handle.net/2440/119800.
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Treatment outcomes in Chronic Phase Chronic Myeloid Leukaemia (CP-CML) have dramatically improved with the introduction of highly active tyrosine kinase inhibitors (TKIs). However, treatment responses are highly heterogeneous. The aim of this thesis is to identify prognostic markers that may help individualise treatment and optimise outcome by stratifying patients into risk groups. Selective treatment intensification is important - more potent treatment may be associated with increased toxicity, and universal adoption of the most potent treatment does not optimally balance risk versus benefit, nor is such a strategy cost effective. This thesis will summarise factors with prognostic significance in CP-CML. Such predictive factors include the metric called the “halving time”, which measures the velocity of BCR-ABL1 decline with initial TKI treatment. This correlates well with future treatment response and risk of disease progression. Conversely, the treatment resistance can be measured by the speed at which the BCR-ABL1 rises, in a similar metric called the “doubling time”. A patient’s Killer Immunoglobulin-like Receptor (KIR) genotype is also correlated with survival as well as molecular outcomes. Whilst the biological basis for this interaction is poorly understand, it is believed to be underpinned by the innate immune system’s role in tumour surveillance and suppression. In the setting of disease resistance mediated by BCR-ABL1 kinase domain mutations, demonstrating an increased number of low level mutants via the use of ultra sensitive mutation detection techniques may help prognosticate patients with a history of the T315I mutation. Early molecular response (EMR, BCR-ABL1 ≤10% at 3 months after starting treatment) is currently acknowledged as one of the strongest prognostic markers, and salvage strategies targeting patients failing to achieve time dependent molecular targets may be an optimal point of intervention. The TIDEL-II study examined such a strategy, firstly by imatinib dose escalation, followed by switching to nilotinib, in patients who fail to achieve time dependent molecular responses. This study also examined the effectiveness of increasing imatinib dose in patients with serum trough levels <1000 ng/mL, a threshold thought to be necessary to achieve cytogenetic response. Although survival and overall molecular response in TIDEL-II is excellent, this strategy was found to be of only marginal benefit in those who failed to achieve EMR. The subsequent study, Pinnacle, aims to study the combination of pegylated interferon and nilotinib in a similar context. Enrolment in this study is ongoing. The same prognostic marker (EMR) may identify patients unlikely to achieve deep molecular responses (DMR), a milestone associated with excellent long term event free survival. It is also commonly stipulated as a pre-requisite to participation in treatment cessation studies. Consistent performance of BCR-ABL1 qRT-PCR assays of a sufficient quality to determine this may be a challenge for some laboratories. To avoid false negatives which may lead to inappropriate reassurance and cessation attempts, we have developed an improved protocol that reliably achieves detection sensitivity required to classify patients in DMR. This assay may also be used as a basis for further enhancing BCR-ABL1 qRT-PCR sensitivity, aimed at identifying patients with an ultra-low level of residual disease. Such patients are hypothesised to have negligible risk of molecular recurrence upon treatment cessation, versus patients who have residual disease just below the current limit of detection. Such ultra-sensitive BCR-ABL1 qRT-PCR may, in this manner, contribute to further elucidation of the prognostic markers of successful treatment cessation. The thesis will conclude with prospects in prognostic markers in CML research and clinical management.Thesis (Ph.D.) (Research by Publication) -- University of Adelaide, School of Medicine, 2016.
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"Src kinase inhibitors for the treatment of sarcomas: Cellular and molecular mechanisms of action." UNIVERSITY OF SOUTH FLORIDA, 2007. http://pqdtopen.proquest.com/#viewpdf?dispub=3260112.
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Tsai, Yu-Chieh, and 蔡育傑. "Utilizing New-Generation EGFR Tyrosine Kinase Inhibitor as Radiosensitizer in the Treatment of Urinary Bladder Cancer." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/01747075121732497583.
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博士國立臺灣大學
臨床醫學研究所
103
Bladder cancer is the ninth most common cancer in the world and in Taiwanese male population. For many patients with localized muscle-invasive bladder cancer, radical cystectomy is not a feasible treatment, and considerable interest was focused on the optimal use of radiotherapy in ”bladder preservation” protocol. However, the long-term survival of patients receiving radiation-based therapy in muscle invasive bladder cancer is about 10% inferior to patients receiving standard radical cystectomy. Traditionally chemotherapeutic agents are used as radiosensitizer but they have many well-known toxicities. Therefore, there is a strong need to find agents enhancing the radiation effect in urinary bladder cancer treatment while not increasing the toxicities. A reasonable way to enhance the outcome of radiotherapy is by concomitantly using agents that inhibit radiation-activated signaling pathways. Epidermal growth factor receptor (EGFR) is the most important target. Cetuximab, an anti-EGFR antibody, has shown clinical benefit in head and neck cancer when combined with radiotherapy. In bladder cancer, gefitinib, an EGFR tyrosine kinase inhibitor (TKI), has moderate in vitro and marginal in vivo radiosensitizing activities. Therefore the topic deserves further investigation. In pilot study, I tested the radiosensitizing activities of erlotinib (EGFR inhibitor), trastuzumab (HER2 inhibitor) and lapatinib (EGFR/HER2 inhibitor) in bladder cancer cells. None of them showed good potential. Instead, afatinib, a new-generation EGFR inhibitor with activity against both EGFR and HER2, is more promising. In murine bladder cancer model, I demonstrated for the first time the in vitro and in vivo radiosensitizing activity of afatinib, an EGFR/HER2 dual inhibitor. The animal study was performed in immunocompetent mice and mimic human physiologic status. Afatinib likely mediates its effect on bladder cancer cells by suppressing radiation-activated EGFR and HER2 signals and thereby causing enhanced DNA damage and cell apoptosis. Based on the findings I hypothesized that in bladder cancer cells, the concomitant inhibition of EGFR and HER2 tyrosine kinase activity by afatinib has greater radiosensitizing activity than the inhibition of EGFR tyrosine kinase activity alone by erlotinib To confirm the hypothesis, in human bladder cancer model the radiosensitizing effects of different generations of clinically useful EGFR TKIs were compared for the first time. I showed the inadequacy of EGFR inhibition alone and the advantage of concomitant blockade of radiation-activated EGFR and HER2 signaling to inhibit the in vitro and in vivo growth of bladder cancer cells. The radiosensitizing effect of an EGFR inhibitor was much higher in HER2 knocked-down than wild-type cells, therefore HER2 may play a synergistic role with EGFR in determining radiosensitivity. I also showed evidence to support that receptor heterodimerization plays an important role in the radiosensitizing effect of afatinib. In Prospect I mentioned how to continue current project and apply the data to clinical use. I hope that the results of this study can help to meet the need of enhancing the radiation effect in urinary bladder cancer treatment while not increasing the toxicities.
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Bansal, Ruchi. "An inhibitor of the mitotic kinase, MPS1, is selective towards pancreatic cancer cells." Thesis, 2014. http://hdl.handle.net/1805/6184.
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Indiana University-Purdue University Indianapolis (IUPUI).The abysmal five year pancreatic cancer survival rate of less than 6% highlights the need for new treatments for this deadly malignancy. Cytotoxic drugs normally target rapidly dividing cancer cells but unfortunately often target stem cells resulting in toxicity. This warrants the development of compounds that selectively target tumor cells. An inhibitor of the mitotic kinase, MPS1, which has been shown to be more selective towards cancer cells than non-tumorigenic cells, shows promise but its effects on stem cells has not been investigated. MPS1 is an essential component of the Spindle Assembly Checkpoint and is proposed to be up-regulated in cancer cells to maintain chromosomal segregation errors within survivable limits. Inhibition of MPS1 kinase causes cancer cell death accompanied by massive aneuploidy. Our studies demonstrate that human adipose stem cells (ASCs) and can tolerate higher levels of a small molecule MPS1 inhibitor than pancreatic cancer cells. In contrast to PANC-1 cancer cells, ASCs and telomerase-immortalized pancreatic ductal epithelial cells did not exhibit elevated chromosome mis-segregation after treatment with the MPS1 inhibitor for 72hrs. In contrast, PANC-1 pancreatic cancer cells exhibited a large increase in chromosomal mis-segregation under similar conditions. Furthermore, growth of ASCs was minimally affected post treatment whereas PANC-1 cells were severely growth impaired suggesting a favorable therapeutic index. Our studies, demonstrate that MPS1 inhibition is selective towards pancreatic cancer cells and that stem cells are less affected in vitro. These data suggest MPS1 inhibition should be further investigated as a new treatment approach in pancreatic cancer.
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Yang, I.-Chi, and 楊毅奇. "Study on the Potential Usage of Phosphatidylinositol 3-kinase/ Akt Pathway Inhibitor for the Treatment of Oral Cancer." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/79517331164098657092.
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碩士國立臺灣大學
口腔生物科學研究所
92
Oral cancer is the fourth leading cause of cancer-related deaths in male population in Taiwan. Despite recent advances in radiotherapy and chemotherapy, the survival of patients with oral cancer has not improved significantly. Continued investigation of new chemotherapeutic agents is thus needed. Recent studies have shown that feeding rotenone inhibited carcinogen-induced mouse oral carcinogenesis. However, the mechanisms by which it inhibits oral carcinogenesis are not well understood. We examined the effects of rotenone on proliferation, cell cycle and apoptosis of oral cancer cell lines SAS using MTT assay, flow cytometry analysis, TUNEL assay, DNA fragmentation assay and Western blotting of cleaved PARP. Rotenone significantly inhibited the proliferation of SAS oral cancer cell lines in a dose-dependent manner with a 50% inhibition concentration (IC50) of rotenone about 2.35 �嵱. However, the IC50 of normal oral mucosa fibroblasts (OMF) was 44.55�嵱. It was almost 22 times higher than the IC50 of SAS. DNA flow cytometric analysis showed that rotenone treatment induced a G2/M arrest. Rotenone treatment also caused significant apoptosis of SAS cells as evidenced by Hoechst 33258 staining, TUNEL labeling, DNA fragmentation and cleavage of PARP. These results indicate that the inhibitory effects of rotenone on oral carcinogenesis may be related to the G2/M phase arrest and induction of apoptosis. Furthermore, nuclear p53 protein and its downstream targets, p21CIP1/WAF1 and Bax, could be induced in SAS (p53-wild type) cells after treatment with rotenone. Rotenone could induce the activation of caspase 8 and caspase 9 in SAS cells, which was different from the previous studies found in other cell types.
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Ross, David Morrall. "Minimal residual disease in chronic myeloid leukaemia after imatinib treatment." 2010. http://hdl.handle.net/2440/60064.
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Around 50% of chronic myeloid leukaemia (CML) patients who remain on imatinib treatment for more than 5 years will achieve a complete molecular response (CMR), defined by undetectable BCR-ABL mRNA in a sensitive reverse transcriptase real-time quantitative PCR (RQ-PCR) assay. Given the increasing importance of CMR on imatinib therapy the primary aim of this study was to improve the accuracy and sensitivity of MRD detection to allow a more accurate estimation of relapse risk when therapy is withdrawn. Firstly, we investigated ways of improving the sensitivity of RT-PCR methods for the detection of BCR-ABL mRNA. Secondly, we investigated the use of the patient-specific BCR-ABL gene for the detection of MRD. Thirdly, we conducted a multi-centre clinical trial of imatinib withdrawal in selected CML patients in a stable CMR. This clinical trial provided patient samples that could be used to test our optimized MRD assays, and provided clinical data on the risk and patterns of relapse after withdrawal of imatinib therapy. The trial is ongoing, but an interim analysis of the study data was performed. In 22 patients the estimated probability of molecular relapse after imatinib withdrawal was 54%, and 60% of relapses occurred within the first 4 months. The average detection limit of BCR-ABL mRNA by RQ-PCR is estimated at around 4.5 log below the level of BCR-ABL prior to commencing treatment. The number of leukaemic cells at diagnosis is around 10¹ ², so the number of residual leukaemic cells in CMR might vary from zero to over a million. We hypothesized that the amount of residual leukaemia in CMR is variable between patients, and that this heterogeneity is a determinant of the risk of relapse when treatment is withdrawn. We developed more sensitive methods for the detection of BCR-ABL and tested these methods in samples from our study patients. We showed that random pentadecamer (15-mer) primers improved the efficiency of reverse transcriptase PCR (RT-PCR), and resulted in a lower detection limit of BCR-ABL mRNA. We also developed a novel nested RT-PCR method using real-time PCR for the second round of the reaction, and this resulted in a lower detection limit of BCR-ABL in patient samples. The utility of this nested RT-PCR method was limited by a false positive rate of 2-3% in the HeLa cell line that we used as our negative control. Consequently, we examined the detection of the patient-specific genomic BCR-ABL sequence as an alternative to RT-PCR. Breakpoints in BCR and ABL1 in CML patients are widely dispersed over 3 kb and 150 kb, respectively. Therefore, the BCR-ABL genomic sequence is essentially unique to each patient. We sequenced the genomic breakpoints of 43 CML patients. We showed that the distribution of breakpoints in BCR and ABL1 was non-random, but we were unable to identify any genomic feature that determined the specific location of individual breakpoints. We developed a novel BCR-ABL DNA Q-PCR method for 12 of the study patients, and in 11 of the patients BCR-ABL DNA was detected when the patient was in a CMR, confirming that this method was more sensitive than RQ-PCR. Contrary to our hypothesis, the detection of BCR-ABL DNA was not predictive of relapse. In most patients who relapsed there was a significant increase in BCR-ABL DNA prior to mRNA relapse. Two patients had stable levels of BCR-ABL DNA measurable on multiple occasions, but remained in remission after 6 months and 15 months, respectively. We have shown that a stable CMR after the withdrawal of imatinib therapy does not necessarily indicate the eradication of leukaemia.Thesis (Ph.D.) -- University of Adelaide, School of Medicine, 2010
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Ross, David Morrall. "Minimal residual disease in chronic myeloid leukaemia after imatinib treatment." Thesis, 2010. http://hdl.handle.net/2440/60064.
Full textAbstract:
Around 50% of chronic myeloid leukaemia (CML) patients who remain on imatinib treatment for more than 5 years will achieve a complete molecular response (CMR), defined by undetectable BCR-ABL mRNA in a sensitive reverse transcriptase real-time quantitative PCR (RQ-PCR) assay. Given the increasing importance of CMR on imatinib therapy the primary aim of this study was to improve the accuracy and sensitivity of MRD detection to allow a more accurate estimation of relapse risk when therapy is withdrawn. Firstly, we investigated ways of improving the sensitivity of RT-PCR methods for the detection of BCR-ABL mRNA. Secondly, we investigated the use of the patient-specific BCR-ABL gene for the detection of MRD. Thirdly, we conducted a multi-centre clinical trial of imatinib withdrawal in selected CML patients in a stable CMR. This clinical trial provided patient samples that could be used to test our optimized MRD assays, and provided clinical data on the risk and patterns of relapse after withdrawal of imatinib therapy. The trial is ongoing, but an interim analysis of the study data was performed. In 22 patients the estimated probability of molecular relapse after imatinib withdrawal was 54%, and 60% of relapses occurred within the first 4 months.
The average detection limit of BCR-ABL mRNA by RQ-PCR is estimated at around 4.5 log below the level of BCR-ABL prior to commencing treatment. The number of leukaemic cells at diagnosis is around 10¹ ², so the number of residual leukaemic cells in CMR might vary from zero to over a million. We hypothesized that the amount of residual leukaemia in CMR is variable between patients, and that this heterogeneity is a determinant of the risk of relapse when treatment is withdrawn. We developed more sensitive methods for the detection of BCR-ABL and tested these methods in samples from our study patients.
We showed that random pentadecamer (15-mer) primers improved the efficiency of reverse transcriptase PCR (RT-PCR), and resulted in a lower detection limit of BCR-ABL mRNA. We also developed a novel nested RT-PCR method using real-time PCR for the second round of the reaction, and this resulted in a lower detection limit of BCR-ABL in patient samples. The utility of this nested RT-PCR method was limited by a false positive rate of 2-3% in the HeLa cell line that we used as our negative control. Consequently, we examined the detection of the patient-specific genomic BCR-ABL sequence as an alternative to RT-PCR.
Breakpoints in BCR and ABL1 in CML patients are widely dispersed over 3 kb and 150 kb, respectively. Therefore, the BCR-ABL genomic sequence is essentially unique to each patient. We sequenced the genomic breakpoints of 43 CML patients. We showed that the distribution of breakpoints in BCR and ABL1 was non-random, but we were unable to identify any genomic feature that determined the specific location of individual breakpoints. We developed a novel BCR-ABL DNA Q-PCR method for 12 of the study patients, and in 11 of the patients BCR-ABL DNA was detected when the patient was in a CMR, confirming that this method was more sensitive than RQ-PCR.
Contrary to our hypothesis, the detection of BCR-ABL DNA was not predictive of relapse. In most patients who relapsed there was a significant increase in BCR-ABL DNA prior to mRNA relapse. Two patients had stable levels of BCR-ABL DNA measurable on multiple occasions, but remained in remission after 6 months and 15 months, respectively. We have shown that a stable CMR after the withdrawal of imatinib therapy does not necessarily indicate the eradication of leukaemia.Thesis (Ph.D.) -- University of Adelaide, School of Medicine, 2010
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Tang, En-kuei, and 湯恩魁. "Usefulness of delE746-A750 and L858R Mutation-Specific Antibodies of EGFR for Predicting Treatment Outcome of Tyrosine Kinase Inhibitors." Thesis, 2012. http://ndltd.ncl.edu.tw/handle/95277095378599958170.
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碩士國立中山大學
生物科學系研究所
100
Efficacy of tyrosine kinase inhibitor (TKI) therapy depends on epidermal growth factor receptor (EGFR) mutation status in patients with non-small cell lung cancer (NSCLC). There has been an increasing interest in studying mutation-specific rabbit monoclonal antibodies of delE746-A750 mutation in exon 19 and L858R point mutation in exon 21 for detecting EGFR mutants. These two mutations account for approximately 90% of all EGFR mutations. We evaluated the two mutation-specific monoclonal antibodies for the detection of EGFR mutations by immunohistochemistry (IHC) and the relationship with treatment outcome and survival. Twenty-five patients (58.1%) harbored EGFR mutations. These mutations include delE746-A750 mutation for seven patients, L858R point mutation for in eighteen patients. IHC showed, for the delE746-A750 and L858R mutations, sensitivity (57.1% and 66.7%), specificity (97.3% and 100%), positive predictive value (80.0% and 100%), and negative predictive value (94.7% and 80.6%). Analysis for progression-free survival was not correlated to IHC staining, but the overall survival was correlated to IHC staining. These mutation-specific antibodies for delE746-A750 and L858R mutations have high positive predictive value and specificity for predefined EGFR mutations and may be suitable for screening for these predefined mutations. However, negative IHC results required further mutation analyses before excluding EGFR TKI therapy.
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"New Approaches For The Treatment Of Erectile Dysfunction In Conditions Of Low Nitric Oxide Formation Or Bioavailability: Investigation Of Rho-kinase Inhibitors And Soluble Guanylate Cyclase-targeted Therapies." Tulane University, 2014.
Find full textAbstract:
Nitric oxide (NO) is the principal mediator of erectile function. NO is released from the nerves and endothelium of small arteries in the penis and diffuses into surrounding smooth muscle to vasodilate through activation of soluble guanylate cyclase (sGC). Erectile dysfunction (ED) occurs in 50% of men between the ages of 40 and 70. It is likely that the pathology of ED results from impairment of NO formation or bioavailability in penile tissue. Iatrogenic nerve damage occurring during prostatectomy can attenuate neurotransmission and release of vasodilators from cavernosal nerves. Oxidative stress from chronic conditions such as diabetes and cardiovascular disease generates reactive oxygen species that can oxidize NO and decrease the molecule's bioavailability. The "gold standard" treatment for ED involves use of oral PDE-5 inhibitors that rely on an intact NO-signaling mechanism for efficacy. Although these therapies are easy to use, they are not effective in many patients suffering from ED associated with pathological conditions of decreased NO bioavailability. Rho-kinase inhibitors, sGC stimulators and sGC activators offer three new interventions that may demonstrate efficacy in treating ED associated with low NO bioavailability. Our results suggest that erectile responses to Rho-kinase inhibitors are not modulated by muscarinic receptor blockade, soluble guanylate cyclase inhibition or cavernosal nerve injury in the rat and that Rho-kinase inhibitors are additive and do not potentiate the endogenous NO-mediated erectile response. Our results with BAY 41-8543 show that this sGC stimulator has significant erectile activity and can potentiate erectile responses to low levels of exogenous and endogenously released NO. These results suggest that BAY 41-8543 would be useful in the treatment of ED occurring following nerve damage from prostatectomy. The sGC activator BAY 60-2770 has very potent erectile activity that is enhanced significantly in conditions of oxidative stress when erectile responses to endogenous NO or sGC stimulators are severely diminished. In oxidizing conditions erectile activity of sGC activators may be enhanced further with concomitant PDE-5 inhibitor therapy, providing evidence that sGC activators may be used alone and in combination with existing treatments to improve erectile function in patients who are non-responsive to standard therapeutic options for ED.acase@tulane.edu
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Wen-ChangTzeng and 曾文璋. "Identification of metastasis related phosphotyrosine proteins in response to tyrosine kinase inhibitor treatment in human lung cancer cells using label-free quantitative analysis." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/53742724473299540703.
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Chen-FangWei and 魏辰芳. "Evidence Based Research on Tyrosine Kinase Inhibitors Use for Non-Small Cell Lung Cancer Treatment: Reimbursement Policy Assessment and Comparative Effectiveness Analysis." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/35396671925915139644.
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山村, 和生, and Kazuo Yamamura. "Combination Treatment of Human Pancreatic Cancer Xenograft Models with the Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Erlotinib and Oncolytic Herpes Simplex Virus HF10." Thesis, 2014. http://hdl.handle.net/2237/20368.
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"Toward an Improved Chronic Myelogenous Leukemia Treatment: Blocking the Stem Cell Factor–Mediated Innate Resistance With Anti–c-Kit Synthetic-Antibody Inhibitors." Thesis, 2015. http://hdl.handle.net/10388/ETD-2015-03-1990.
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Chronic Myelogenous Leukemia (CML) is a blood cancer that arises when hematopoietic cells acquire an abnormal protein known as BCR-ABL. Current therapies for CML include drugs that inhibit BCR-ABL. However, these drugs only suppress the disease and do not cure it. One reason is that BCR-ABL drugs fail to kill the primitive population of CML cells, referred to as leukemia stem cells (LSCs), which are responsible for initiating and propagating CML. Since LSCs are not killed, the cancer is not cured and many affected patients eventually relapse. Recent studies suggest that LSCs are protected from current therapies by the bone marrow micro-environment where they reside. There, cytokine signaling molecules are present, which mediate processes that protect LSCs from BCR-ABL drugs. The stem cell factor (SCF) is one of these signaling molecules. It activates the receptor c-Kit located on the surface of LSCs, and this activation in turn allows proliferating LSCs to resist BCR-ABL drugs, even without prior exposure to these drugs, i.e., innate resistance is observed.
In this thesis, the mechanism of this innate resistance is investigated, so that a suitable treatment strategy can be developed. To this end, a co-agent approach based on synthetic antibodies (sABs) is proposed to inhibit the receptor c-Kit, with the goal of disrupting its activation by the ligand SCF. This disruption should in turn block the SCF-mediated innate resistance, thus potentially restoring BCR-ABL drug apoptotic activity. The method for this disruption involves targeting the c-Kit structural susceptibility. Specifically, the sABs are designed via antibody phage display technology to target the D1–D2–D3 domains representing the SCF binding sites, hence preventing downstream pathway activation. The hypothesis is that, by blocking the SCF-mediated innate resistance, a suitable combination of such an sAB co-agent and a BCR-ABL drug should be conducive to suppressing LSCs, thereby providing a potential means to improve CML treatment.
In addition, to assess the performance of the proposed treatment strategy, a set of in vitro tests is conducted, focusing on performance behaviors such as cell binding, cell death, and the progenitor inhibition. The experimental results support the hypothesis that the proposed combinatorial strategy is indeed a promising approach to mitigate the innate resistance, thus restoring BCR-ABL drug apoptotic activity.
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Tonsing-Carter, Eva Y. "Modulation of the Mdm2 signaling axis sensitizes triple-negative breast cancer cells to carboplatin." Thesis, 2014. http://hdl.handle.net/1805/6306.
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Triple-negative breast cancers (TNBCs) are highly refractive to current treatment strategies, and new multi-targeted treatments need to be elucidated. Combination therapy that includes targeting the murine double minute 2 (Mdm2) signaling axis offers a promising approach. Protein-protein interaction inhibitors such as Nutlin-3a block the binding of key signaling molecules such as p53, p73α, and E2F1 to the hydrophobic pocket of Mdm2 and can lead to activation of cell-death signaling pathways. Since clinical trials for TNBC are evaluating the DNA damaging agent carboplatin, the objective of this thesis was to evaluate the therapeutic potential and mechanism of action of combination carboplatin and Nutlin-3a to treat TNBC. In TNBC cell lines with a mutant p53 background, we determined if modulation of Mdm2 function in the context of carboplatin-mediated DNA damage resulted in a synergistic inhibition of cell growth. Several ratios of carboplatin:Nutlin-3a were strongly synergistic in increasing cell death, with combination indices of 0.5 and lower. Mechanistic studies indicated that drug sensitivity and Mdm2 expression were dependent on p73. Mdm2 localized to a larger degree in the chromatin fraction isolated from cells treated with the combination treatment consistent with observations by others that Mdm2 binds to the Mre11/Rad50/Nbs1 complex, inhibits the DNA damage response, and increases drug sensitivity. In vivo efficacy experiments were conducted in the TMD231 orthotopic mammary fat pad model in NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. For assessment of baseline tumor burden and randomization, fluorescent imaging of E2-Crimson expressing TMD231 cells was performed. Following Nutlin-3a and carboplatin combination treatment, there was a statistically significant reduction in primary tumor volume as well as lung metastases with significantly increased probability of survival compared to Vehicle and single drug treatments (p<0.001). While there was a decrease in bone-marrow cellularity, this did not lead to bone-marrow aplasia, and body weights recovered to normal levels within 7 days post-treatment. The present studies demonstrate the promise of Mdm2 as a therapeutic target in combination with conventional therapy, increase our understanding of how to potentiate DNA damage in cancers, and may lead to new clinical therapies for triple-negative primary and metastatic breast cancer.
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Chen, Yi-Wen, and 陳依玟. "Ⅰ. Proteomics study of oxidative stress, Src kinase inhibition and quercetin in H9C2 cardiomyocytes: a cell model of heart ischemia reperfusion injury and treatment Ⅱ. The application of proteomics for disease biomarker discovery and mechanism study." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/81922667673395367359.
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博士國立清華大學
生物資訊與結構生物研究所
103
PART І. Proteomics study of oxidative stress, Src kinase inhibition (PP1) and Quercetin in H9C2 cardiomyocytes: a cell model of heart ischemia reperfusion injury and treatment. Oxidative stress production of myocardial ischemia/reperfusion injury leads to protein phosphorylation in regulating gene expression, metabolism, cell adhesion and survival. In this thesis, we used hydrogen peroxide treatment of H9C2 rat cardiomyocytes as a model of oxidative stress in heart ischemia reperfusion injury. A proteomics approach using anti-phosphotyrosine affinity purification and LC-MS/MS was then used to identify the stress-induced protein phosphorylation. We showed that oxidative stress induces a robust tyrosine phosphorylation of multiple proteins in this cell type. Most of identified tyrosine phosphorylated proteins were relative to cell-cell junctions, the actin cytoskeleton and cell adhesion. This suggested that oxidative stress may have a profound effect on intercellular connections and the cytoskeleton to affect cell adhesion, morphology and survival. After stress-induced phosphotyrosine proteins were analyzed by STRING, Src kinase was shown to be a major upstream regulator of these events. Furthermore, immunofluorescence studies, fluorescent activated cell sorting and cell-based assays were used to demonstrate H2O2-induced modifications of cell adhesion structures and cytoskeleton, de-adhesion and apoptosis, which were reversed by treatment with the Src kinase inhibitor PP1 or quercetin. Moreover, quercetin likely blocked the H2O2-induced inflammatory response through STAT3 modulation, which also contributed in preventing ischemia/reperfusion injury in cardiomyocytes. These findings provide the critical role of Src kinase in oxidative stress-induced phosphorylation and cell damage in cardiomyocytes and suggested that targeting Src kinase or quercetin may be an effective strategy for preventing ischemia reperfusion injury in the heart. PART П. The application of Proteomics for disease biomarker discovery Cancer and diabetic are high incidence and mortality in worldwide; however, early detection, surgical resection and postoperative therapy can lead to survival improvement for cancer or diabetes. In recent study, body fluids of patient were used to screen markers, such as plasma, urine and cerebrospinal fluid. Here, plasma of critical limb ischemia (CLI), type 1 diabetic (T1DM), transition carcinoma cancer and uterine leiomyoma were collected and analyzed by 2D-DIGE and MALDI-TOF. Then, particular protein markers were found in specific diseases, such as dual adapter for phosphotyrosine and 3-phosphotyrosine and 3-phosphoinositide (DAPP1) in CLI, hemopexin in T1DM, selenocysteine-specific elongation factor in TCC and vitamin D-binding protein in uterine leiomyoma. Nevertheless, most identified plasma proteins are related to inflammatory responses and blood coagulation. Therefore, a cell-based platform was established to screen protein markers relating to gemcitabine (GEM)-induced drug resistance pancreatic cells and tumorigenic breast cells. In GEM-induced drug resistant pancreatic cells, ribonucleoside-diphosphate reductase large subunit significantly overexpressed and tumor suppressor protein p53 may interplay with GEM-induced pancreatic cell resistance. In addition, in breast cancer cells, the level of calcium-binding mitochondrial carrier protein SCaMC-1 in tumorigenetic breast cancer cells or breast cancer patients’ plasma was higher than that of normal cell or health donors’ plasma. These data demonstrate that plasma proteomics provides a lot of common proteins between various diseases, but a cell based strategy provides a good platform for specific protein markers discovery in particular disease and afterwards these protein markers are potential for disease screening.