-
Cell Death & Disease Aug 2019Extracorporeal photochemotherapy (ECP) is employed for the management of cutaneous T cell lymphoma (CTCL). ECP involves the extracorporeal exposure of white blood cells...
Extracorporeal photochemotherapy (ECP) is employed for the management of cutaneous T cell lymphoma (CTCL). ECP involves the extracorporeal exposure of white blood cells (WBCs) to a photosensitizer, 8-methoxypsoralen (8-MOP), in the context of ultraviolet A (UVA) radiation, followed by WBC reinfusion. Historically, the therapeutic activity of ECP has been attributed to selective cytotoxicity on circulating CTCL cells. However, only a fraction of WBCs is exposed to ECP, and 8-MOP is inactive in the absence of UVA light, implying that other mechanisms underlie the anticancer effects of ECP. Recently, ECP has been shown to enable the physiological differentiation of monocytes into dendritic cells (DCs) that efficiently cross-present tumor-associated antigens (TAAs) to CD8 T lymphocytes to initiate cognate immunity. However, the source of TAAs and immunostimulatory signals for such DCs remains to be elucidated. Here, we demonstrate that 8-MOP plus UVA light reduces melanoma cell viability along with the emission of ICD-associated danger signals including calreticulin (CALR) exposure on the cell surface and secretion of ATP, high mobility group box 1 (HMGB1) and type I interferon (IFN). Consistently, melanoma cells succumbing to 8-MOP plus UVA irradiation are efficiently engulfed by monocytes, ultimately leading to cross-priming of CD8 T cells against cancer. Moreover, malignant cells killed by 8-MOP plus UVA irradiation in vitro vaccinate syngeneic immunocompetent mice against living cancer cells of the same type, and such a protection is lost when cancer cells are depleted of calreticulin or HMGB1, as well as in the presence of an ATP-degrading enzyme or antibodies blocking type I IFN receptors. ECP induces bona fide ICD, hence simultaneously providing monocytes with abundant amounts of TAAs and immunostimulatory signals that are sufficient to initiate cognate anticancer immunity.
Topics: Adenosine Triphosphate; Animals; Antigens, Neoplasm; Apoptosis; CD8-Positive T-Lymphocytes; Cell Differentiation; Cell Line, Tumor; Cell Survival; Dendritic Cells; HMGB1 Protein; Humans; Immunogenic Cell Death; Leukocytes; Lymphoma, T-Cell, Cutaneous; Methoxsalen; Mice; Monocytes; Photopheresis; Photosensitizing Agents; Receptor, Interferon alpha-beta; Ultraviolet Rays
PubMed: 31371700
DOI: 10.1038/s41419-019-1819-3 -
Blood Advances Jul 2019The investigation of extracorporeal photopheresis (ECP) plus standard of care (SoC) (SoC+ECP) in chronic graft-versus-host disease (cGVHD) within prospective, randomized... (Randomized Controlled Trial)
Randomized Controlled Trial
The investigation of extracorporeal photopheresis (ECP) plus standard of care (SoC) (SoC+ECP) in chronic graft-versus-host disease (cGVHD) within prospective, randomized clinical studies is limited, despite its frequent clinical use. This phase 1/pilot study was the first randomized, prospective study to investigate ECP use as first-line therapy in cGVHD, based on the 2015 National Institutes of Health (NIH) consensus criteria for diagnosis and response assessment. Adult patients with new-onset (≤3 years of hematopoietic stem cell transplantation) moderate or severe cGVHD were randomized 1:1 to 26 weeks of SoC+ECP vs SoC (corticosteroids and cyclosporine A/tacrolimus) between 2011 and 2015. The primary endpoint was overall response rate (ORR), defined as complete or partial response, at week 28 in the intention-to-treat population (ITT). Other outcomes included quality of life (QoL) measures and safety. Sixty patients were randomized; ITT included 53 patients (SoC+ECP: 29; SoC: 24). Week 28 ORR was 74.1% (SoC+ECP) and 60.9% (SoC). Investigator-assessed ORR was 56.0% (SoC+ECP) and 66.7% (SoC). Patients treated with SoC experienced a decline in QoL over the 28-week study period; QoL remained unchanged in SoC+ECP patients. Most frequent treatment-emergent adverse events (TEAEs) in SoC+ECP patients were hypertension (31.0%), cough (20.7%), dyspnea (17.2%), and fatigue (17.2%). Seventeen patients (SoC+ECP: 8; SoC: 9) experienced 35 serious adverse events (SAEs). No TEAEs or SAEs were considered related to the ECP instrument or methoxsalen. The encouraging short-term results of this study could inform the design of subsequent studies. This trial was registered at www.clinicaltrials.gov as #NCT01380535.
Topics: Adult; Aged; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Male; Methoxsalen; Middle Aged; Photopheresis; Quality of Life; Severity of Illness Index; Treatment Outcome; Young Adult
PubMed: 31332045
DOI: 10.1182/bloodadvances.2019000145 -
Microbiology and Immunology Oct 2019Bovine mastitis is the most common disease in dairy cattle. Bacterial infections are the main cause of mastitis. Lipopolysaccharide (LPS), a major structural component...
Bovine mastitis is the most common disease in dairy cattle. Bacterial infections are the main cause of mastitis. Lipopolysaccharide (LPS), a major structural component of the cell wall of Escherichia coli, is a good inducer used to replicate inflammation models. 8-Methoxypsoralen (8-MOP), a formerly considered photosensitizing agent, has been used in immunotherapy. This study investigated the protective effects of 8-MOP on LPS-induced inflammatory injury in bovine mammary epithelial cells (BMECs). LPS treatment (50 μg/mL for 12 hr) caused a decrease in cell viability, morphological damage, and cell apoptosis. Pretreatment with 8-MOP at concentrations of 25 and 50 μg/ml significantly attenuated LPS-induced inflammation in BMECs. qRT-PCR analysis revealed that the messenger RNA expression of inflammatory cytokines and chemokine (interleukin-1β [IL-1β], IL-6, tumor necrosis factor-α, and IL-8) was suppressed by 8-MOP in LPS-stimulated BMECs. Western blot analysis showed that 8-MOP could also reduce the protein levels of cyclooxygenase-2 and promote the translocation of high-mobility group box 1 from the nucleus to cytoplasm. Furthermore, the anti-inflammatory property of 8-MOP was mediated by inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells activation and STAT1 phosphorylation. Taken together, 8-MOP could protect cells from inflammatory injury induced by LPS, and may be a potential agent against bovine mastitis.
Topics: Animals; Cattle; Epithelial Cells; Female; Inflammation; Janus Kinases; Lipopolysaccharides; Mammary Glands, Animal; Mastitis, Bovine; Methoxsalen; NF-kappa B; STAT Transcription Factors; Signal Transduction
PubMed: 31313848
DOI: 10.1111/1348-0421.12730 -
Bioorganic Chemistry Aug 2019Seventy-one 7-oxycoumarins, 66 synthesized and 5 commercially sourced, were tested for their ability to inhibit growth in murine PAM212 keratinocytes. Forty-nine...
Seventy-one 7-oxycoumarins, 66 synthesized and 5 commercially sourced, were tested for their ability to inhibit growth in murine PAM212 keratinocytes. Forty-nine compounds from the library demonstrated light-induced lethality. None was toxic in the absence of UVA light. Structure-activity correlations indicate that the ability of the compounds to inhibit cell growth was dependent not only on their physiochemical characteristics, but also on their ability to absorb UVA light. Relative lipophilicity was an important factor as was electron density in the pyrone ring. Coumarins with electron withdrawing moieties - cyano and fluoro at C - were considerably less active while those with bromines or iodine at that location displayed enhanced activity. Coumarins that were found to inhibit keratinocyte growth were also tested for photo-induced DNA plasmid nicking. A concentration-dependent alteration in migration on neutral gels caused by nicking was observed.
Topics: Animals; Cell Proliferation; Cells, Cultured; Coumarins; Dose-Response Relationship, Drug; Keratinocytes; Mice; Molecular Structure; Photochemical Processes; Photosensitizing Agents; Structure-Activity Relationship
PubMed: 31170642
DOI: 10.1016/j.bioorg.2019.103014 -
International Journal of Nanomedicine 20198-methoxypsoralen (8-MOP) is one of the most commonly utilized drugs in psoralen-ultraviolet A therapy for treatment of vitiligo. However, poor skin retention and...
BACKGROUND
8-methoxypsoralen (8-MOP) is one of the most commonly utilized drugs in psoralen-ultraviolet A therapy for treatment of vitiligo. However, poor skin retention and systemic side effects limit the clinical application of 8-MOP.
METHODS
Microemulsions (MEs) and chitosan derivative-coated 8-MOP MEs were developed and compared for dermal delivery of 8-MOP. Ex vivo skin retention/permeation study was performed to select the ME formulation with the highest retention:permeation ratio. Four different chitosan-coated MEs were prepared and compared with the ME formulation for their ability to distribute 8-MOP in the skin.
RESULTS
Among various ME formulations developed, a formulation containing 2.9% ethyl oleate, 17.2% Cromophor EL35, 8.6% ethanol and 71.3% water showed the highest ex vivo skin retention:permeation ratio (1.98). Of four chitosan-coated MEs prepared, carboxymethyl chitosan-coated MEs (CC-MEs) and hydroxypropyl chitosan-coated MEs (HC-MEs) showed higher ex vivo skin retention:permeation ratio (1.46 and 1.84). and were selected for in vivo pharmacokinetic study. AUC (0-12 h) for 8-MOP MEs (4578.56 h·ng·mL) was higher than HC-MEs (3422.47 h·ng·mL), CC-MEs (2808.51 h·ng·mL) and tincture (1500.16 h·ng·mL). Also, AUCplasma (0-12 h) for MEs (39.35±13.90 h·ng·mL) was significantly lower than HC-MEs (66.32 h·ng·mL), CC-MEs (59.70 h·ng·mL) and tincture (73.02 h·ng·mL).
CONCLUSION
These combined results suggested that the MEs developed could be a promising and safe alternative for targeted skin delivery of 8-MOP.
Topics: Administration, Cutaneous; Animals; Chitosan; Drug Delivery Systems; Emulsions; Humans; Male; Methoxsalen; Microdialysis; Permeability; Rats, Sprague-Dawley; Skin; Skin Absorption; Swine
PubMed: 31015760
DOI: 10.2147/IJN.S191940 -
International Journal of Molecular... Mar 2019This study evaluated whether bergapten and methoxsalen could prevent diabetes-induced osteoporosis and its underlying mechanism. For 10 weeks, bergapten or methoxsalen...
This study evaluated whether bergapten and methoxsalen could prevent diabetes-induced osteoporosis and its underlying mechanism. For 10 weeks, bergapten or methoxsalen (0.02%, /) was applied to diabetic mice that were provided with a high-fat diet and streptozotocin. Bone mineral density (BMD) and microarchitecture quality were significantly reduced in the diabetic control group; however, both bergapten and methoxsalen reversed serum osteocalcin, bone-alkaline phosphatase and femur BMD. These coumarin derivatives significantly increased bone volume density and trabecular number, whereas they decreased the structure model index of femur tissue in diabetic mice. Conversely, tartrate-resistant acid phosphatase 5 (TRAP) staining revealed that these derivatives reduced osteoclast numbers and formation in diabetic bone tissue. Additionally, both bergapten and methoxsalen tended to downregulate the expression of osteoclast-related genes such as receptor activator of nuclear factor kappa-B ligand (), nuclear of activated T-cells, cytoplasmic 1 () and in diabetic femurs, with and expression showing significant reductions. Our data suggest that both bergapten and methoxsalen prevent diabetic osteoporosis by suppressing bone resorption.
Topics: 5-Methoxypsoralen; Alkaline Phosphatase; Animals; Bone Density; Diabetes Mellitus, Experimental; Diet, High-Fat; Disease Models, Animal; Gene Expression Regulation; Humans; Male; Methoxsalen; Mice; NFATC Transcription Factors; Osteocalcin; Osteogenesis; Osteoporosis; RANK Ligand; Streptozocin; Tartrate-Resistant Acid Phosphatase
PubMed: 30875838
DOI: 10.3390/ijms20061298 -
PloS One 2019Extracorporeal photopheresis (ECP) is an autologous immunomodulatory cell therapy that consists of the ex vivo collection of mononuclear cells (MNCs), which are...
Extracorporeal photopheresis (ECP) is an autologous immunomodulatory cell therapy that consists of the ex vivo collection of mononuclear cells (MNCs), which are irradiated with UVA in the presence of the photosensitizing agent 8-methoxypsoralen (8-MOP) to induce cell apoptosis. This photoactivated cell preparation is then reinfused into the patient. While the clinical benefits of ECP are well-demonstrated, no study has yet characterized the influence of variations in the composition of the cell preparation on the efficacy of ECP in vitro. Here, we describe a standardized methodology for the in vitro assessment of ECP that uses the human lymphoma T-cell line and mimics the clinical procedure. By quantifying cell apoptosis, inhibition of cell proliferation, and 8-MOP consumption, we used this approach to characterize the specific influence of key variables on the cellular response to ECP. We found that (i) increases in hematocrit and plasma concentrations attenuated the cellular response to ECP; (ii) plasma concentration was the only variable tested that influenced 8-MOP consumption; and (iii) the loss of efficacy due to variations in the concentration of certain blood components could be counteracted by modulating the UVA dose. This methodology may enable evaluation of other leukapheresis preparation protocols and better determination of the optimal working parameters for ECP.
Topics: Apoptosis; Cell Culture Techniques; Cell Proliferation; Feasibility Studies; Humans; Jurkat Cells; Lymphoma, T-Cell; Methoxsalen; Photopheresis; Photosensitizing Agents; Treatment Outcome; Ultraviolet Rays
PubMed: 30822323
DOI: 10.1371/journal.pone.0212835 -
Bioorganic & Medicinal Chemistry Letters Feb 2019Linear furocoumarins, also known as psoralens, are clinically useful photo-activated pharmaceuticals employed to address hyperproliferative skin diseases. Seven diverse...
Linear furocoumarins, also known as psoralens, are clinically useful photo-activated pharmaceuticals employed to address hyperproliferative skin diseases. Seven diverse cytotoxic pharmacophores have been synthetically attached to 8-methoxypsoralen via a 5-amino functionality. The resulting unique set of compounds was evaluated for dark and light toxicity against PAM212 keratinocytes in culture.
Topics: Cell Survival; Cells, Cultured; Darkness; Humans; Keratinocytes; Light; Methoxsalen; Photosensitizing Agents; Skin Diseases
PubMed: 30638875
DOI: 10.1016/j.bmcl.2018.12.048 -
Molecules (Basel, Switzerland) Oct 2018Eight alkaloids (⁻) were isolated from , and their herbicide activities were evaluated through in vitro, semivivo, and in vivo assays. The most relevant results were...
Eight alkaloids (⁻) were isolated from , and their herbicide activities were evaluated through in vitro, semivivo, and in vivo assays. The most relevant results were observed for Compounds and ⁻ at 150 μM, which decreased dry biomass by 20% and 23%, respectively. These are significant results since they presented similar values with the positive control, commercial herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Based on the performed assays, Compound (graveoline) is classified as an electron-transport inhibitor during the light phase of photosynthesis, as well as a plant-growth regulator. On the other hand, Compounds ⁻ inhibited electron and energy transfers, and are also plant-growth inhibitors. These phytotoxic behaviors based on acridone and quinolone alkaloids may serve as a valuable tool in the further development of a new class of herbicides since natural products represent an interesting alternative to replace commercial herbicides, potentially due their low toxicity.
Topics: Alkaloids; Biomass; Diuron; Electron Transport; Herbicides; Methoxsalen; Photosynthesis; Ruta
PubMed: 30347671
DOI: 10.3390/molecules23102693 -
Beijing Da Xue Xue Bao. Yi Xue Ban =... Oct 2018To establish a high performance liquid chromatography (HPLC) method for the determination of 8-methoxypsoralen (8-MOP) in mouse plasma and apply it to a pharmacokinetic...
OBJECTIVE
To establish a high performance liquid chromatography (HPLC) method for the determination of 8-methoxypsoralen (8-MOP) in mouse plasma and apply it to a pharmacokinetic study of 8-MOP.
METHODS
8-MOP was separated on a Waters Symmetry18 column (250 mm × 4.6 mm, 5 μm) and determined by HPLC using isocratic elution, and 5-methoxypsoralen was used as internal standard. The mobile phase consisted of methanol-water (55:45, V/V) at a flow rate of 1.0 mL/min. The excitation and emission wavelength of fluorescence detector were set at 334 nm and 484 nm respectively, and the internal standard method was used for quantitative analysis. In the study, 60 healthy ICR male mice were randomly divided into twelve groups. The mice in control group were administered intragastrically with 1% Tween 80, and the mice in the other eleven groups were administered intragastrically with 8-MOP (40 mg/kg). Plasma concentrations of 8-MOP in the mice at different time points after treatment were determined by HPLC. Pharmacokinetic parameters were calculated by DAS 2.0 software.
RESULTS
The calibration curve of 8-MOP was linear with a correlation coefficient of 0.999 3 over the concentration range of 0.05 to 10 mg/L, and the limit of detection was 0.015 mg/L. The average recoveries of 8? MOP at three different concentrations (0.10, 0.50, 2.5 mg/L) were from 92.5% to 100.6%. The intra-day precision of 8-MOP was from 3.3% to 8.2%, while the inter-day precision was from 3.4% to 6.7% at three spiked concentration levels. The extraction recoveries of 8-MOP were from 90.9% to 92.0%, and the plasma samples could be stored at -80°C for 15 days at least at three spiked concentration levels. 8-MOP could be detected in mouse plasma 5 min after intragastrical administration to the mice (1.4 mg/L). The concentration of 8-MOP in the mouse plasma reached a maximum 2 h after administration, and 8-MOP could still be detected 24 h after administration (1.1 mg/L). t was (39.21±3.65) h, C was (2.31±0.02) mg/L, t was (2.00±0.00) h, and AUC0-t was (33.34±1.19) (h×mg)/L.
CONCLUSION
The proposed method is accurate and simple,suitable for pharmacokinetics of 8-MOP in mice.
Topics: Animals; Calibration; Chromatography, High Pressure Liquid; Male; Methoxsalen; Mice; Mice, Inbred ICR; Photosensitizing Agents; Plasma; Random Allocation
PubMed: 30337737
DOI: No ID Found