A major obstacle in the implementation of adoptive cell therapy (ACT) for solid tumors is CD8+ T cell quantity and functional quality. In order to address this issue, the ACT field has directed considerable effort toward the generation of less-differentiated memory T cells (Tm), which demonstrate superior effector function and engraftment over effector T cells. An obstacle in using Tm for ACT is their requirement for in vivo activation before full effector function can be acquired. We sought to determine if a rhabdovirus expressing a defined tumor antigen (i.e. a rhabdoviral oncolytic vaccine) could activate adoptively-transferred Tm in vivo and eliminate established tumors. We used ex vivo cultured DUC18 TCR-transgenic Tm combined with a rhabdoviral oncolytic vaccine to target established CMS5 fibrosarcomas in both balb/c and NRG mice, and we compared the efficacy of the combination treatment versus monotherapies. Our data demonstrate that the rhabdoviral oncolytic vaccine was capable of expanding adoptively-transferred Tm in order to eliminate established tumors. Furthermore, synergy between ACT and oncolytic vaccination was required for optimal therapeutic outcome. Interestingly, we observed a population of endogenous, tumor-primed lymphocytes which appeared to be required for complete tumor elimination and subsequent memory formation. This was in contrast to the current consensus in the ACT field which is that endogenous lymphocytes are detrimental to therapeutic outcome, thus necessitating lymphodepletion prior to the commencement of therapy. Our data suggest that endogenous lymphocytes may be a beneficial cell population which is overlooked by current approaches to ACT. / Thesis / Master of Science (MSc) / Current approaches to the T cell therapy of cancer are hindered by poor cell quality. It is simple to grow higher quality T cells, but it is difficult to grow very large numbers of them. Furthermore, higher quality T cells need a signal in order to “switch on” before they can start killing cancer cells. Here, we use a cancer-targeting virus as a signal for these cells to activate, grow to very large numbers in the patient, and destroy their tumor. Our vaccine also switches on other immune cells in the patient, which help guarantee the destruction of the tumor. The significance of this work is that it will improve T cell therapy for cancer by opening the possibility of using higher-quality T cells which are much better at killing cancer than the currently used type of T cells.

No description available.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Martina Sýkorová Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Study on impact of selected tyrosine kinase inhibitors on multidrug resistance mediated by ABC drug efflux transporters Tyrosine kinases are an important class of enzymes controlling cell proliferation, carcinogenesis, apoptosis and cell differentiation. Deregulation of these enzymes can transform normal cell into a cancerous one. Blocking their function by tyrosine kinase inhibitors (TKi) is considered a promising treatment for various types of cancer. ATP-binding cassette (ABC) transporters form a family of transmembrane proteins that can transport a wide variety of substrates across biological membranes via ATP-dependent drug efflux pumps. They modulate drug pharmacokinetics, but on the other hand, lead to therapy failure due to overexpression in cancer cells. In our previous study, we evaluated inhibition properties of two selected TKi (alectinib, brivanib) in MDCKII cell lines (parent one and those transduced with human ABCB1, ABCC1 and ABCG2). Alectinib significantly inhibited ABCB1, ABCG2 but not ABCC1 transporter. Brivanib showed triple inhibition of all studied transporters. In the present work, we...

<p>Malaria is a health, social and economic burden in Nigeria and consistently ranks amongst the four most common causes of childhood deaths. Treatment of malaria is usually started at home / care is only sought from the health facility when the treatment is ineffective (McCombie, 1996). Patent medicine vendors (PMVs) have been identified as a widely patronized source for drugs used in the home treatment of malaria (Breiger et al, 2001 / Goodman, et al, 2007 / Salako et al, 2001). Inadequate or poor knowledge and practices in the use of anti-malaria drugs (AMDs) increases morbidity and mortality, undermines therapeutic efficacy, and promotes the emergence and spread of drugresistant malaria. Aim: The aim of the study was to describe and quantify the knowledge and self-reported practices of PMVs in the use of antimalarials, particularly artemisinin-based combination therapies (ACTs), in a poor urban community in Lagos state, Nigeria.</p>

Combination therapy has proven to be a popular treatment strategy for tighter diabetes control. Since the preliminary evidence is suggestive of complementary actions of American (AG) and Korean Red Ginseng (KRG) in improving glycemia, this project was designed to investigate the therapeutic potential of AG and KRG in combination. Following a randomized, double-blind, placebo-controlled parallel design in a population with diabetes at two centres, the combined use of AG and KRG for 12 weeks was safe, but did not significantly affect glycemic control, blood lipids or blood pressure. However, there was a trend toward lower glycated hemoglobin by 0.7% (p=0.1) and office systolic blood pressure by 5 mm Hg (p=0.052) compared to placebo. These findings encourage further investigation of the mechanism and roles of AG, KRG and their effective components. They also highlight limitations in ginseng research and the need to impose strict regulations to facilitate its standardization.

Combination therapy has proven to be a popular treatment strategy for tighter diabetes control. Since the preliminary evidence is suggestive of complementary actions of American (AG) and Korean Red Ginseng (KRG) in improving glycemia, this project was designed to investigate the therapeutic potential of AG and KRG in combination. Following a randomized, double-blind, placebo-controlled parallel design in a population with diabetes at two centres, the combined use of AG and KRG for 12 weeks was safe, but did not significantly affect glycemic control, blood lipids or blood pressure. However, there was a trend toward lower glycated hemoglobin by 0.7% (p=0.1) and office systolic blood pressure by 5 mm Hg (p=0.052) compared to placebo. These findings encourage further investigation of the mechanism and roles of AG, KRG and their effective components. They also highlight limitations in ginseng research and the need to impose strict regulations to facilitate its standardization.

<p>Malaria is a health, social and economic burden in Nigeria and consistently ranks amongst the four most common causes of childhood deaths. Treatment of malaria is usually started at home / care is only sought from the health facility when the treatment is ineffective (McCombie, 1996). Patent medicine vendors (PMVs) have been identified as a widely patronized source for drugs used in the home treatment of malaria (Breiger et al, 2001 / Goodman, et al, 2007 / Salako et al, 2001). Inadequate or poor knowledge and practices in the use of anti-malaria drugs (AMDs) increases morbidity and mortality, undermines therapeutic efficacy, and promotes the emergence and spread of drugresistant malaria. Aim: The aim of the study was to describe and quantify the knowledge and self-reported practices of PMVs in the use of antimalarials, particularly artemisinin-based combination therapies (ACTs), in a poor urban community in Lagos state, Nigeria.</p>

Das Harnblasenkarzinom (BCa) stellt in Deutschland die vierthäufigste Tumorneuerkrankung und die zehnthäufigste krebsbedingte Todesursache bei Männern dar. Nichtmuskelinvasive BCa werden organerhaltend aus der Blasenwand entfernt und zur Rezidiv- und Progressionsprophylaxe mittels intravesikaler Chemo- oder Immuntherapien behandelt. Trotz dieser adjuvanten Therapien, die mit starken Nebenwirkungen verbunden sein können, ist nur eine bedingte Minimierung des Rezidivrisikos möglich. Besonders im fortgeschrittenen Stadium weisen Harnblasenkarzinome eine schlechte Prognose auf. Obwohl das BCa eine chemosensitive Erkrankung darstellt, wird das Ansprechen auf lokale oder systemische Chemotherapien häufig durch auftretende Resistenzmechanismen limitiert. Daher stehen sowohl die Verbesserung konventioneller Chemotherapien als auch die Suche nach neuartigen Behandlungsstrategien im Fokus der experimentellen BCa-Forschung. Die Apoptose, eine Form des programmierten Zelltodes, ist ein essenzieller, streng regulierter biologischer Prozess, welcher der Aufrechterhaltung der Gewebshomöostase und der gezielten, entzündungsfreien Eliminierung geschädigter Zellen dient. Fehlregulationen in den Apoptosesignalwegen stellen ein zentrales Ereignis in der Tumorgenese dar und tragen außerdem zur Entstehung von Chemo- und Radiotherapieresistenzen bei. Eine wichtige Rolle in der Apoptoseregulation spielen die Mitglieder der BCL2- und der Inhibitor of Apoptosis Protein (IAP)-Familien, deren wichtigste antiapoptotische Vertreter BCL2, BCL-XL, XIAP und Survivin häufig in Tumoren, einschließlich des BCa, überexprimiert sind. Unter Verwendung von small interfering RNAs (siRNAs), synthetischen Nukleinsäurekonstrukten zur selektiven Geninhibition, wurde im Rahmen der Arbeit in vitro und in vivo untersucht, ob die Hemmung der Apoptoseinhibitoren BCL2, BCL-XL, XIAP und Survivin – allein und in Kombination mit Chemotherapie – eine Therapieoption zur Behandlung des BCa darstellen könnte. Da zur Tumorentstehung und -progression eine Vielzahl von genetischen Veränderungen beitragen, erscheint der Angriff eines einzelnen Zielgens unzureichend für eine effektive Tumortherapie. Aufgrund dessen wurde untersucht, ob durch simultane Reduktion der ausgewählten Apoptoseinhibitoren in BCa-Zellen stärkere wachstumsinhibitorische Effekte erzielt werden können. In der vorliegenden Arbeit wurde gezeigt, dass insbesondere die siRNA-vermittelte Hemmung von BCL-XL und Survivin in den BCa-Zelllinien EJ28 und J82 antiproliferative Effekte hervorruft und diese Tumorzellen gegenüber einer nachgeschalteten Chemotherapie mit Mitomycin C oder Cisplatin sensitiviert. Hingegen bewirkte sowohl die transiente als auch die stabile RNAi-induzierte Hemmung von BCL2 und XIAP in den untersuchten BCa-Monolayerzellkulturen, möglicherweise infolge kontinuierlicher Versorgung der Tumorzellen mit Sauerstoff und Nährstoffen, keine Reduktion des Tumorwachstums. Eine gegenüber den Einzelbehandlungen deutliche Verstärkung der antitumoralen und insbesondere der chemosensitivierenden Effekte in den BCa-Zelllinien wurde durch simultane Hemmung von BCL-XL und Survivin erzielt. Beispielsweise stieg der Anteil apoptotischer Zellen von 64 % nach Survivin-siRNA+Cisplatin-Behandlung auf 94 % nach gleichzeitiger BCL-XL+Survivin-Inhibition in Kombination mit Cisplatin. Folglich stellt die simultane Inhibition von BCL-XL und Survivin in Kombination mit Chemotherapeutika eine äußert viel versprechende BCa-Therapieoption dar. Tierexperimentelle Studien belegen die wachstumsinhibitorische Wirkung der Survivin-Reduktion und der kombinierten BCL-XL-siRNA+Chemotherapie-Behandlung, so wurde das Tumorendvolumen im Vergleich zur Kontrollbehandlung um 43 % bzw. um 48 % reduziert.

In this study, we have explored the combination of a novel Purine Nucleoside Phosphorylase mediated Gene-Directed Enzyme Prodrug Therapy (PNP-GDEPT) with chemotherapeutics, Taxotere and/or Carboplatin to target prostate and ovarian cancer (PC & OC). PNP converts the prodrug (Fludarabine-phosphate) to a toxic purine, 2-fluoroadenine (2FA) that inhibits RNA/DNA synthesis. Taxotere is active against late stage PC whilst carboplatin is first line therapy for OC. Neither modality is adequately effective. We expect that a combination will target heterogeneity via cytotoxicity to diverse cancer cell populations leading to effective synergies, which may improve efficacy and quality of life. For PC, Synergy between Ad-PNP-GDEPT and Taxotere were assessed in vitro and in vivo. Cell killing effects of combination led to significant synergistic killing of human PC-3 & murine RM1 PC cells accompanied by enhanced apoptosis. A lower individual dose (by up to 8 fold) led to enhanced efficacy. In vivo, the combination regimen given at the suboptimal doses led to reduction in local tumour (PC-3 & RM1) growth in nude and in C57BL/6 mice, respectively. A significant reduction in lung RM1 colony numbers indicated enhanced systemic efficacy. Combination treated mice also displayed significantly improved survival (25 days vs 15 days for control mice). Importantly, the condition of combination treated mice (e.g. weight loss) was better than those given individual treatments. The possible involvement of the immune system in this enhanced effect is under investigation. For OC, three-way synergy between Ad-PNP-GDEPT, Taxotere and carboplatin was effectively demonstrated in SKOV-3 and OVCAR-3 cells. This was significantly greater than bimodal or individual treatments. A 10-50 fold dose reduction of individual treatments was effective when combined, accompanied by enhanced apoptosis. Western-blotting analyses revealed a shift in the expression of anti-apoptotic and proapoptotic proteins upon treatment with various combinations. This is the first demonstration of synergy between these modalities.

Objetivos: Avaliar a eficácia anti-isquêmica e efeito metabólico da trimetazidina em pacientes com angina refratária e diabete melito (DM) tipo 2, em uso de pelo menos duas medicações de efeito hemodinâmico, sem condições de revascularização. Métodos: Ensaio clínico randomizado, cruzado, duplo-cego, conduzido em 10 pacientes com DM tipo 2 e angina estável, em tratamento com pelo menos 2 anti-anginosos clássicos. Pacientes foram randomizados para receber trimetazidina (20 mg 3 vezes ao dia) ou placebo, por períodos de 6 semanas. Avaliação clínica, laboratorial, ergométrica e monitorização de pressão arterial (MAPA) e Holter de 24 horas foram realizadas no início do estudo e ao término de 6 semanas de cada intervenção. Resultados: Os pacientes em tratamento com trimetazidina apresentaram melhora significativa na classe funcional da angina (p<0,05), com diminuição significativa no número de episódios de crise anginosa por semana e na dose de nitrato sub-lingual utilizada (p<0,05). O tempo de início da isquemia no teste ergométrico foi maior após o uso da trimetazidina (229 ± 126 s no basal, 276 ± 101 s após o placebo e 348 ± 145 s após a trimetazidina; p<0,05). Não houve diferença nos níveis de pressão arterial, freqüência cardíaca e duplo-produto nas 24h, avaliadas concomitantemente com Holter e MAPA, ou no controle glicêmico e lipídico entre as intervenções. Conclusões: O uso da trimetazidina mostrou-se eficaz como tratamento coadjuvante da angina estável de difícil manejo em pacientes com DM tipo 2 em uso de múltiplos anti-anginosos, sem alteração de variáveis hemodinâmicas avaliadas nas 24 h. Estes dados ampliam o espectro de emprego da trimetazidina no manejo de pacientes sintomáticos e com limitadas opções de tratamento. / Aims: To evaluate the anti-ischemic and metabolic effect of trimetazidine in patients with refractory angina and type 2 diabetes (DM2) not eligible for revascularization and using at least two hemodynamic agents. Methods: Randomized, double-blind, crossover clinical trial conducted in 10 patients. Patients were randomized to receive trimetazidine (20 mg 3 times a day) or placebo for 6-week periods. Clinical and exercise evaluations, in addition to 24-h ambulatory blood pressure and Holter monitoring were carried out at baseline and at the end of each 6-week intervention period. Results: The patients receiving trimetazidine presented significant improvement in terms of angina functional class (p<0.05), with decrease in the number of weekly angina episodes at rest and in the sublingual nitrate dose (p<0.05). Time to 1 mm ST-segment depression was increased after trimetazidine use (229.2 ± 126.1 s at baseline, 276.3 ± 100.9 s after placebo and 347.5 ± 144.6 s after trimetazidine; p<0.001). No differences were observed between treatments group in terms of 24h-blood pressure, heart rate and ratepressure product as evaluated concomitantly with ambulatory blood pressure and Holter monitoring, or in terms of glycemic and lipid profile. Conclusions: The combined use of trimetazidine and haemodynamic agents was efficient to treat stable refractory angina in DM2 patients, significantly improving clinical and exercise parameters. The present data support the use of trimetazidine in the management of symptomatic patients with limited treatment options.