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
Human induced pluripotent stem cell line was generated from commercially available primary human prostate fibroblasts HPrF derived from a fetus, aged 18-24 weeks of gestation. The fibroblast cell line was reprogrammed with Yamanaka factors (OCT4, SOX2, c-MYC, KLF4) using CytoTuneTM-iPS 2.0 Sendai Reprogramming Kit. Pluripotency of the derived transgene-free iPS cell line was confirmed both in vitro by detecting the expression of factors of pluripotency on a single-cell level, and in vivo using teratoma formation assay. This iPS cell line will be a useful tool for studying both normal prostate development and prostate cancer disease.
- MeSH
- fibroblasty * cytologie metabolismus MeSH
- indukované pluripotentní kmenové buňky * cytologie metabolismus MeSH
- lidé MeSH
- plod * cytologie embryologie MeSH
- přeprogramování buněk MeSH
- prostata * cytologie embryologie MeSH
- techniky buněčného přeprogramování * MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The stability of in vitro cell cultures is an important issue for any clinical, bio-industrial, or pharmacological use. Embryonic stem cells are pluripotent; consequently, they possess the ability to differentiate into all three germ layers and are inherently prone to respond to differentiation stimuli. However, long-term culture inevitably yields clones that are best adapted to the culture conditions, passaging regimes, or differentiation sensitivity. This cellular plasticity is a major obstacle in the development of bio-industrial or clinical-grade cultures. At present, the quality control of cell cultures is limited by the lack of reliable (epi)genetic or molecular markers or by the focus on a particular type of instability such as karyotype abnormalities or adverse phenotypic traits. Therefore, there is an ongoing need for robust, feasible, and sensitive methods of determining or confirming cell status and for revealing potential divergences from the optimal state. We modeled both intrinsic and extrinsic changes in human embryonic stem cell (hESC) states using different experimental strategies and addressed the changes in cell status by intact cell mass spectrometry fingerprinting. The analysis of spectral fingerprints by methods routinely used in analytical chemistry clearly distinguished the morphologically and biochemically similar populations of hESCs and provided a biomarker-independent tool for the quality control of cell culture. Stem Cells Translational Medicine 2018;7:109-114.
- MeSH
- biologické markery analýza MeSH
- buněčné kultury MeSH
- fenotyp MeSH
- kultivované buňky MeSH
- lidé MeSH
- lidské embryonální kmenové buňky fyziologie MeSH
- plasticita buňky fyziologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A human induced pluripotent stem cell line was generated from cancer-associated fibroblasts of a 68-years old patient with diagnosed prostate adenocarcinoma (PCa). The fibroblast cell line was reprogrammed with Epi5TM Episomal iPSC Reprogramming Kit. Pluripotency of the derived transgene-free iPS cell line was confirmed both in vitro by detecting expression of factors of pluripotency on a single-cell level, and also in vivo using teratoma formation assay. This new iPS cell line may be used for differentiation into different prostate-specific cell types in differentiation studies.
- MeSH
- fibroblasty asociované s nádorem metabolismus MeSH
- fibroblasty metabolismus MeSH
- indukované pluripotentní kmenové buňky metabolismus MeSH
- lidé MeSH
- nádory prostaty genetika MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
Chromosomal instability evoked by abnormalities in centrosome numbers has been traditionally considered as a hallmark of aberrant, typically cancerous or senescent cells. We have reported previously that pristine human embryonic stem cells (hESC) suffer from high frequency of supernumerary centrosomes and hence may be prone to undergo abnormal mitotic divisions. We have also unraveled that this phenomenon of multicentrosomal mitoses vanishes with prolonged time in culture and with initiation of differentiation, and it is strongly affected by the culture substratum. In this study, we report for the first time that Cripto-1 protein (teratocarcinoma-derived growth factor 1, epidermal growth factor-Cripto/FRL-1/Cryptic) produced by hESC represents a factor capable of inducing formation of supernumerary centrosomes in cultured hESC. Elimination of Cripto-1 signaling on the other hand restores the normal number of centrosomes in hESC. Linking the secretory phenotype of hESC to the centrosomal metabolism may help to develop better strategies for propagation of stable and safe bioindustrial and clinical grade cultures of hESC. From a broader point of view, it may lead to unravelling Cripto-1 as a micro-environmental factor contributing to adverse cell behaviors in vivo.
- MeSH
- buněčná diferenciace genetika MeSH
- centrozom * MeSH
- GPI-vázané proteiny antagonisté a inhibitory genetika MeSH
- lidé MeSH
- lidské embryonální kmenové buňky cytologie metabolismus MeSH
- mezibuněčné signální peptidy a proteiny genetika MeSH
- mitóza genetika MeSH
- nádorové proteiny antagonisté a inhibitory genetika MeSH
- signální transdukce genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Opioidní analgetika jsou standardem léčby střední a silné bolesti nádorového i nenádorového původu. Léčbu však mohou komplikovat dobře známé vedlejší účinky opioidů, zahrnující nauzeu, zvracení, pruritus, obstipaci, sedaci, retenci moče nebo výskyt tolerance a závislosti. Použití smíšeného opioidního agonisty-antagonisty nalbufinu v léčbě bolesti může přispět k redukci těchto nežádoucích projevů léčby. Jako monoterapie je s úspěchem používán pro léčbu středně silné až silné pooperační bolesti. Kombinace nalbufinu s čistými opioidními agonisty přináší silný analgetický účinek s poklesem výskytu vedlejších účinků léčby.
Opiate analgesics represent the golden standard of treatment in cases of intermediate to severe pain of both malignant and non-malignant origin. However, this treatment may be complicated by the well-known side effects of opioids, including nausea/vomiting, pruritus, constipation, sedation, urinary retention, the development of tolerance and dependence. Use of the mixed opioid agonist-antagonist nalbuphine to treat pain can reduce these side effects. In monotherapy, it can satisfactorily control moderate to severe post-operative pain. It can be used concurrently with opioid agonists, yielding good analgesia while simultaneously decreasing the incidence of opioid side effects. This paper provides information about the pharmacological profile and clinical experience with nalbuphine in different indications. .
- MeSH
- bolest farmakoterapie MeSH
- hodnocení léčiv MeSH
- klinické zkoušky jako téma MeSH
- lidé MeSH
- nalbufin * aplikace a dávkování farmakokinetika kontraindikace terapeutické užití MeSH
- opioidní analgetika * aplikace a dávkování farmakokinetika kontraindikace škodlivé účinky terapeutické užití MeSH
- Check Tag
- lidé MeSH
