- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Monocyte-derived dendritic cell (DC)-based vaccines loaded with tumor self-antigens represent a novel approach in anticancer therapy. We evaluated DC-based anticancer immunotherapy (ITx) in an academic Phase I/II clinical trial for children, adolescent, and young adults with progressive, recurrent, or primarily metastatic high-risk tumors. The primary endpoint was safety of intradermal administration of manufactured DCs. Here, we focused on relapsing high-risk sarcoma subgroup representing a major diagnosis in DC clinical trial. As a part of peripheral blood immunomonitoring, we evaluated quantitative association between basic cell-based immune parameters. Furthermore, we describe the pattern of these parameters and their time-dependent variations during the DC vaccination in the peripheral blood immunograms. The peripheral blood immunograms revealed distinct patterns in particular patients in the study group. As a functional testing, we evaluated immune response of patient T-cells to the tumor antigens presented by DCs in the autoMLR proliferation assay. This analysis was performed with T-cells obtained prior to DC ITx initiation and with T-cells collected after the fifth dose of DCs, demonstrating that the anticancer DC-based vaccine stimulates a preexisting immune response against self-tumor antigens. Finally, we present clinical and immunological findings in a Ewing's sarcoma patient with an interesting clinical course. Prior to DC therapy, we observed prevailing CD8+ T-cell stimulation and low immunosuppressive monocytic myeloid-derived suppressor cells (M-MDSC) and regulatory T-cells (Tregs). This patient was subsequently treated with 19 doses of DCs and experienced substantial regression of metastatic lesions after second disease relapse and was further rechallenged with DCs. In this patient, functional ex vivo testing of autologous T-cell activation by manufactured DC medicinal product during the course of DC ITx revealed that personalized anticancer DC-based vaccine stimulates a preexisting immune response against self-tumor antigens and that the T-cell reactivity persisted for the period without DC treatment and was further boosted by DC rechallenge. Trial Registration Number: EudraCT 2014-003388-39.
- Publikační typ
- časopisecké články MeSH
Despite efforts to develop novel treatment strategies, refractory and relapsing sarcoma, and high-risk neuroblastoma continue to have poor prognoses and limited overall survival. Monocyte-derived dendritic cell (DC)-based anti-cancer immunotherapy represents a promising treatment modality in these neoplasias. A DC-based anti-cancer vaccine was evaluated for safety in an academic phase-I/II clinical trial for children, adolescents, and young adults with progressive, recurrent, or primarily metastatic high-risk tumors, mainly sarcomas and neuroblastomas. The DC vaccine was loaded with self-tumor antigens obtained from patient tumor tissue. DC vaccine quality was assessed in terms of DC yield, viability, immunophenotype, production of IL-12 and IL-10, and stimulation of allogenic donor T-cells and autologous T-cells in allo-MLR and auto-MLR, respectively. Here, we show that the outcome of the manufacture of DC-based vaccine is highly variable in terms of both DC yield and DC immunostimulatory properties. In 30% of cases, manufacturing resulted in a product that failed to meet medicinal product specifications and therefore was not released for administration to a patient. Focusing on the isolation of monocytes and the pharmacotherapy preceding monocyte harvest, we show that isolation of monocytes by elutriation is not superior to adherence on plastic in terms of DC yield, viability, or immunostimulatory capacity. Trial patients having undergone monocyte-interfering pharmacotherapy prior to monocyte harvest was associated with an impaired DC-based immunotherapy product outcome. Certain combinations of anti-cancer treatment resulted in a similar pattern of inadequate DC parameters, namely, a combination of temozolomide with irinotecan was associated with DCs showing poor maturation and decreased immunostimulatory features, and a combination of pazopanib, topotecan, and MTD-based cyclophosphamide was associated with poor monocyte differentiation and decreased DC immunostimulatory parameters. Searching for a surrogate marker predicting an adverse outcome of DC manufacture in the peripheral blood complete blood count prior to monocyte harvest, we observed an association between an increased number of immature granulocytes in peripheral blood and decreased potency of the DC-based product as quantified by allo-MLR. We conclude that the DC-manufacturing yield and the immunostimulatory quality of anti-cancer DC-based vaccines generated from the monocytes of patients were not influenced by the monocyte isolation modality but were detrimentally affected by the specific combination of anti-cancer agents used prior to monocyte harvest.
- Publikační typ
- časopisecké články MeSH
Cíle: Za akademické klinické studie se označují takové klinické studie, na jejichž plánování a realizaci se nepodílí žádný komerční subjekt. Akademické klinické studie často řeší relevantní klinické otázky. Oblast jejich zájmu není ovlivněna potřebami farma‑ ceutických společností, ale potřebami pacientů. Výsledky takového výzkumu jsou považovány za nezávislé a často hodnoceny jako klinicky významné a pro praxi potřebné. Předmětem tohoto výzkumu byla analýza mezinárodních registrů klinických studií s cílem zmapovat situaci akademických intervenčních klinických studií (AIKS) v České republice (ČR). Analyzovány byly také zdroje financování akademického klinického výzkumu. Metody: Pomocí klíčových slov „Non‑commercial“, „Interventional studies“ a „Czech Republic“ byly systematicky prohledá‑ ny čtyři různé mezinárodní registry klinických studií: ClinicalTrials.gov, The European Union Clinical Trials Register, BioMed Central International Standard Randomised Controlled Trial Number Registry (ISCTRN) a Australia and New Zealand Clinical Trials Registry (ANZCTR), za období od 1. 1. 2004 do 31. 12. 2017. Výsledky byly kontrolovány a vzájemně porovnány dvěma nezávislými recenzenty. Nalezené AIKS byly rozděleny do kategorií dle intervence, fáze hodnocení, dle terapeutické ob‑ lasti, designu, národního či mezinárodního provedení a typu financování. Extrahovaná data byla podrobena deskriptivní statistické analýze. Výsledky: V registru ClinicalTrials.gov bylo nalezeno 239 studií, v registru The European Union Clinical Trials Register 235, v registru ISCTRN 52 a v ANZCTR 26 studií. Celkově bylo zaznamenáno 552 nálezů, z nichž duplicit bylo objeveno 40, celkem 67 studií mělo komerčního zadavatele, i když byly v registrech vedené jako akademické, a celkem 42 studií bylo z další analýzy vyloučeno, protože nesplňovaly daná kritéria (začátek studie byl mimo vyhledávané období nebo se jednalo o neintervenční studii). Za období 1. 1. 2004 – 31. 12. 2017 bylo ve čtyřech výše zmíněných registrech evidováno 403 unikátních AIKS probí‑ hajících v ČR. Nejčastěji měly prováděné AIKS charakter klinického hodnocení léčivého přípravku a nejvíc jich bylo zaměřeno na oblasti onkologie a kardiologie. Nejčastější zdroj financování AIKS představovaly neziskové organizace, a to mezinárodní a národní grantové agentury. Na financování AIKS se dále podíleli nadace, vládní organizace, samotné univerzity, fakultní nemocnice a další. Závěr: Vyhledávání přesných údajů o klinických studiích je časově náročný proces, neboť i přes snahu národní a mezinárodní vědecké komunity jsou registry neúplné a neobsahují transparentní data. V současnosti v ČR probíhá jen několik mezinárodních randomizovaných klinických studií, které jsou iniciovány a koordinovány českými výzkumníky. Vzhledem k zásadnímu dopadu AIKS, které jsou zdrojem nezávislých a objektivních výsledků přímo uplatnitelných v klinické praxi, vyvstává reálná potřeba pod‑ pory provádění mezinárodních AIKS iniciovaných z ČR, a to zejména ze strany státu, grantových agentur i samotných výzkumných institucí a zdravotnických zařízení. Nezastupitelnou roli v této oblasti hraje velká národní výzkumná infrastruktura CZECRIN, která nabízí výzkumníkům kompletní servis při provádění AIKS.
Aims: Investigator-initiated (“academic”) clinical trials (IICTs) are designed and carried out without a participation of any com‑ mercial subject. IICTs adopt a patient-oriented approach and attempt to answer relevant questions from the clinical practice that may not be attractive for the pharmaceutical companies. The results obtained by IICTs are considered independent and clinically significant. The purpose of this study was to analyse data from the clinical trial public registers to describe a current status of IICTs and their financial support in the Czech Republic (CR). Methods: The search was performed in four international clinical trials databases – ClinicalTrials.gov, The European Union Clinical Trials Register, BioMed Central International Standard Randomised Controlled Trial Number Registry (ISCTRN) and Australia and New Zealand Clinical Trials Registry (ANZCTR). Entries were filtered by date from 01/01/2004 until 31/12/2017. The key words “Czech Republic”, “Interventional studies“, and “Non-commercial” were used. The results were separately checked and compared by two independent reviewers. Identified IICTs were categorized by intervention, phase, therapeutic area, design characteristics, and type of funding. The descriptive statistics were used for the analysis of the data. Results: 239 trials were found in ClinicalTrials.gov register, 235 in The European Union Clinical Trials Register, 52 in ISCTRN, and 26 trials in ANZCTR. In total, 552 trials were identified, from these 40 were recognized as duplicates, and 67 were found having a commercial sponsor, although marked as non-commercial trials. Other 42 were also discarded because they did not meet the inclusion criteria (the start of the study was beyond the time period or they were only observational). To sum up, the search identified 403 unique IITCs conducting in the Czech Republic. Most trials investigated the use of medicinal products and were focused on cancer or cardiovascular diseases. The most common sources of the Czech IITCs’ funding were non-profit organi‑ zations, including international and national grant agencies, endowment funds and foundations, governmental organizations, universities, university hospitals, or others. Conclusion: Despite efforts of the national and international scientific community, collecting data about clinical trials from the public registers is a time-consuming process, as the registers are incomplete and do not contain transparent information. Cur‑ rently, only a few multinational randomized IICTs are coordinated by national investigators from the Czech Republic. Considering the importance of IITCs providing more independent and objective results directly applicable in the clinical practice, there is a need of increasing of the support incentives of the government, grant agencies, research institutions, and healthcare providers. The large research infrastructure CZECRIN plays an indispensable role and offers researchers the complete services in the imple‑ mentation of IICTs.
- MeSH
- finanční podpora výzkumu jako téma MeSH
- financování organizované MeSH
- financování vládou MeSH
- klinická studie jako téma * ekonomika MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
Growing evidence suggests that microRNAs are involved in the development and progression of colorectal cancer (CRC). In the present study, deregulation and functioning of tumor-suppressive miR-215-5p was evaluated in CRC. In total, 448 tumor tissues and 325 paired adjacent healthy tissues collected from Czech and Spain cohorts of CRC patients have been used for miR-215-5p expression analyses. A series of in vitro experiments have been performed using transient transfection of miR-215-5p mimics into four CRC cell lines to identify specific cellular processes affected by miR-215-5p. Further, the effects of miR-215-5p on tumor growth were evaluated in vivo using NSG mice and stable cell line overexpressing miR-215-5p. Target mRNAs of miR-215-5p were tested using luciferase assay and western blot analyses. We found that miR-215-5p is significantly downregulated in tumor tissues compared with non-tumor adjacent tissues and its decreased levels correlate with the presence of lymph node metastases, tumor stage, and shorter overall survival in CRC patients. Overexpression of miR-215-5p significantly reduced proliferation, clonogenicity, and migration of CRC cells, lead to cell cycle arrest in G2/M phase and p53-dependent induction of apoptosis. The ability of miR-215-5p to inhibit tumor growth was confirmed in vivo. Finally, we confirmed epiregulin and HOXB9 to be the direct targets of miR-215-5p. As epiregulin is EGFR ligand and HOXB9 is its transcriptional inducer, we suggest that the main molecular link between miR-215-5p and CRC cells phenotypes presents the EGFR signaling pathway, which is one of the canonical pathogenic pathways in CRC.
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Drug addiction is commonly associated with depression and comorbid patients also suffer from higher cravings and increased relapse rate. To address this issue preclinically we combined the olfactory bulbectomy (OBX) model of depression and intravenous methamphetamine self-administration procedure in rats to assess differences in relapse-like behavior. Male Sprague-Dawley rats were divided randomly into two groups; in one group the bilateral olfactory bulbectomy (OBX) was performed while the other group was sham operated. After recovery, intracardiac catheter was implanted. Intravenous self-administration procedure was conducted in operant boxes using nose-poke operandi (Coulbourn Instruments, Inc., USA) under fixed ratio 1 schedule of reinforcement. Methamphetamine was available at dose 0.08 mg/kg/infusion. After stable methamphetamine intake was maintained, a period of forced abstinence was initiated and rats were kept in their home-cages for 14 days. Finally, one reinstatement session was conducted in operant boxes with no drug delivery. In the reinstatement session the mean of 138.4 active nose-pokes was performed by the OBX group, while the sham group displayed 41 responses, i.e. 140 % and 48 % of basal nose-poking during maintenance phase in OBX and sham operated group respectively. OBX group also showed significantly more passive nose-pokes indicating hyperactive behavioral traits in bulbectomized rats. However, the % of active operandum preference was equal in both groups. Olfactory bulbectomy model significantly increased reinstatement of methamphetamine seeking behavior. This paradigm can be used to evaluate potential drugs that are able to suppress the drug-seeking behavior.
- MeSH
- autoaplikace MeSH
- bulbus olfactorius patofyziologie chirurgie MeSH
- chování při shánění drogy fyziologie MeSH
- depresivní poruchy komplikace patofyziologie MeSH
- intravenózní podání MeSH
- methamfetamin aplikace a dávkování MeSH
- modely nemocí na zvířatech MeSH
- náhodné rozdělení MeSH
- operantní podmiňování účinky léků fyziologie MeSH
- poruchy spojené s užíváním amfetaminu komplikace patofyziologie MeSH
- potkani Sprague-Dawley MeSH
- recidiva MeSH
- stimulanty centrálního nervového systému aplikace a dávkování MeSH
- zaváděcí katétry MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
MiRNAs are important regulators of gene expression and changes in their levels are linked with various pathological states, including solid tumors. MiR-215 has been identified as a tumor suppressor in colorectal cancer (CRC). Following our previous in vitro and in vivo experiments, the aim of this project was to study the possibility of increasing the levels of miR-215 in tumor cells by systemic administration of miRNA mimics in liposomal delivery system in vivo. By subcutaneous xenotransplantation of human cancer cells to NSG mice, CRC model was established. The treatment (miR-215 mimics in liposomes [20 and 40 microg/mouse], control oligonucleotide in liposomes, or saline) was administered repeatedly by i.v. injection via tail-vein. Animals were sacrificed, tumor were dissected and measured by a caliper. Expression of miR-215 in tumors, lungs and liver was quantified by RT-PCR. There was no significant differences in tumor volume and miR-215 expression between all three treatment groups. Therefore, the decrease in tumor volume was not achieved. By comparing the levels of miR-215 in lungs, liver and tumors after the treatment, we suggest that the liposomes are accumulated in the lungs and do not concentrate sufficiently in the tumor site to exert significant tumor-suppressive effect.
- MeSH
- biomimetické materiály aplikace a dávkování metabolismus MeSH
- HCT116 buňky MeSH
- kolorektální nádory farmakoterapie metabolismus patologie MeSH
- lidé MeSH
- liposomy MeSH
- mikro RNA aplikace a dávkování biosyntéza MeSH
- modely nemocí na zvířatech * MeSH
- myši MeSH
- systémy cílené aplikace léků metody MeSH
- xenogenní modely - testy antitumorózní aktivity metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
