Emerging evidence shows the crucial role of inflammation (particularly NF‐κB pathway) in the development and progression of myelofibrosis (MF), becoming a promising therapeutic target. Furthermore, ...tailoring treatment with currently available JAK inhibitors (such as ruxolitinib or fedratinib) does not modify the natural history of the disease and has important limitations, including cytopenias. Since recent studies have highlighted the role of miR‐146a, a negative regulator of the NF‐κB pathway, in the pathogenesis of MF; here we used miR‐146a−/− (KO) mice, a MF‐like model lacking driver mutations, to investigate whether pharmacological inhibition of JAK/STAT and/or NF‐κB pathways may reverse the myelofibrotic phenotype of these mice. Specifically, we tested the JAK1/2 inhibitor, ruxolitinib; the NF‐κB inhibitor via IKKα/β, BMS‐345541; both inhibitors in combination; or a dual inhibitor of both pathways (JAK2/IRAK1), pacritinib. Although all treatments decreased spleen size and partially recovered its architecture, only NF‐κB inhibition, either using BMS‐345541 (alone or in combination) or pacritinib, resulted in a reduction of extramedullary hematopoiesis, bone marrow (BM) fibrosis and osteosclerosis, along with an attenuation of the exacerbated inflammatory state (via IL‐1β and TNFα). However, although dual inhibitor improved anemia and reversed thrombocytopenia, the combined therapy worsened anemia by inducing BM hypoplasia. Both therapeutic options reduced NF‐κB and JAK/STAT signaling in a context of JAK2V617F‐driven clonal hematopoiesis. Additionally, combined treatment reduced both COL1A1 and IL‐6 production in an in vitro model mimicking JAK2‐driven fibrosis. In conclusion, NF‐κB inhibition reduces, in vitro and in vivo, disease burden and BM fibrosis, which could provide benefits in myelofibrosis patients.
We used miR‐146a−/− (KO) mice, a MF‐like model lacking driver mutations, to investigate whether pharmacological inhibition of JAK/STAT and/or NF‐κB pathways may reverse the myelofibrotic phenotype of these mice. Specifically, we tested the JAK1/2 inhibitor, ruxolitinib; the NF‐κB inhibitor via IKKα/β, BMS‐345541; both inhibitors in combination; or a dual inhibitor of both pathways (JAK2/IRAK1), pacritinib. Although all treatments decreased spleen size and partially recovered its architecture, only NF‐κB inhibition, either using BMS‐345541 (alone or in combination) or pacritinib, resulted in a reduction of extramedullary hematopoiesis, bone marrow (BM) fibrosis and osteosclerosis, along with an attenuation of the exacerbated inflammatory state (via IL‐1β and TNFα). However, whereas dual inhibitor improved anemia and reversed thrombocytopenia, the combined therapy worsened anemia by inducing BM hypoplasia. Additionally, combined treatment reduced both COL1A1 and IL‐6 production in an in vitro model mimicking JAK2‐driven fibrosis. In summary, NF‐κB inhibition reduces, in vitro and in vivo, disease burden, and BM fibrosis, which could provide benefits in myelofibrosis patients
Abstract
New detector approaches in Positron Emission Tomography
imaging will play an important role in reducing costs, lowering
administered radiation doses, and improving overall
performance. ...PETALO employs liquid xenon as the active scintillating
medium and UV-sensitive silicon photomultipliers for scintillation
readout. The scintillation time in liquid xenon is fast enough to
register time-of-flight information for each detected coincidence,
and sufficient scintillation is produced with low enough
fluctuations to obtain good energy resolution. The present
simulation study examines a full-body-sized PETALO detector and
evaluates its potential performance in PET image reconstruction.
We prove a negative result for the approximation of functions defined on compact subsets of Rd (where d≥2) using feedforward neural networks with one hidden layer and arbitrary continuous activation ...function. In a nutshell, this result claims the existence of target functions that are as difficult to approximate using these neural networks as one may want. We also demonstrate an analogous result (for general d∈N) for neural networks with an arbitrary number of hidden layers, for activation functions that are either rational functions or continuous splines with finitely many pieces.
The phase III trial GEM05MENOS65 randomized 390 patients 65 years old or younger with newly diagnosed symptomatic multiple myeloma (MM) to receive induction with thalidomide/dexamethasone, ...bortezomib/thalidomide/dexamethasone and Vincristine, BCNU, melphalan, cyclophosphamide, prednisone/vincristine, BCNU, doxorubicin, dexamethasone bortezomib (VBMCP/VBAD/B) followed by autologous stem cell transplantation (ASCT) with MEL-200. After ASCT, a second randomization was performed to compare thalidomide/bortezomib (TV), thalidomide (T) and alfa-2b interferon (alfa2-IFN). Maintenance treatment consisted of TV (thalidomide 100 mg daily plus one cycle of intravenous bortezomib at 1.3 mg/m
on days 1, 4, 8 and 11 every 3 months) versus T (100 mg daily) versus alfa2-IFN (3 MU three times per week) for up to 3 years. A total of 271 patients were randomized (TV: 91; T: 88; alfa2-IFN: 92). The complete response (CR) rate with maintenance was improved by 21% with TV, 11% with T and 17% with alfa2-IFN (P, not significant). After a median follow-up of 58.6 months, the progression-free survival (PFS) was significantly longer with TV compared with T and alfa2-IFN (50.6 vs 40.3 vs 32.5 months, P=0.03). Overall survival was not significantly different among the three arms. Grade 2-3 peripheral neuropathy was observed in 48.8%, 34.4% and 1% of patients treated with TV, T and alfa2-IFN, respectively. In conclusion, bortezomib and thalidomide maintenance resulted in a significantly longer PFS when compared with thalidomide or alfa2-IFN. (no. EUDRA 2005-001110-41).
Lurbinectedin (PM01183) has synergistic antitumor activity when combined with doxorubicin in mice with xenografted tumors. This phase I trial determined the recommended dose (RD) of doxorubicin ...(bolus) and PM01183 (1-h intravenous infusion) on day 1 every 3 weeks (q3wk), and obtained preliminary evidence of antitumor activity for this combination in small-cell lung cancer (SCLC).
Patients with advanced solid tumors received doxorubicin and PM01183 following a standard dose escalation design and expansion at the RD. Twenty-seven patients had relapsed SCLC: 12 with sensitive disease (platinum-free interval ≥90 days) and 15 with resistant disease (platinum-free interval <90 days).
Doxorubicin 50 mg/m2 and PM01183 4.0 mg flat dose was the RD. In relapsed SCLC, treatment tolerance at the RD was manageable. Transient and reversible myelosuppression (including neutropenia, thrombocytopenia, and febrile neutropenia) was the main toxicity, managed with dose adjustment and colony-stimulating factors. Fatigue (79%), nausea/vomiting (58%), decreased appetite (53%), mucositis (53%), alopecia (42%), diarrhea/constipation (42%), and asymptomatic creatinine (68%) and transaminase increases (alanine aminotransferase 42%; aspartate aminotransferase 32%) were common, and mostly mild or moderate. Complete (n = 2, 8%) and partial response (n = 13, 50%) occurred in relapsed SCLC, mostly at the RD. Response rates at second line were 91.7% in sensitive disease median progression-free survival (PFS)=5.8 months and 33.3% in resistant disease (median PFS = 3.5 months). At third line, response rate was 20.0% (median PFS = 1.2 months), all in resistant disease.
Doxorubicin 50 mg/m2 and PM01183 4.0 mg flat dose on day 1 q3wk has shown remarkable activity, mainly in second line, with manageable tolerance in relapsed SCLC, leading to further evaluation of this combination within an ongoing phase III trial.
PM00104 binds guanines at DNA minor grooves, impacting DNA replication and transcription. A phase I study was undertaken to investigate safety, dose-limiting toxicities (DLTs), recommended phase II ...dose (RP2D), pharmacokinetics (PKs) and preliminary antitumour activity of PM00104 as a 1- or 3-h infusion three-weekly.
Patients with advanced solid tumours received PM00104 in a dose escalation trial, as guided by toxicity and PK data.
A total of 47 patients were treated; 27 patients on the 1-h schedule (0.23-3.6 mg m(-2)) and 20 patients on the 3-h schedule (1.8-3.5 mg m(-2)). Dose-limiting toxicities comprised reversible nausea, vomiting, fatigue, elevated transaminases and thrombocytopenia, establishing the 1-h schedule RP2D at 3.0 mg m(-2). With the 3-h schedule, DLTs of reversible hypotension and neutropenia established the RP2D at 2.8 mg m(-2). Common PM00104-related adverse events at the RP2D comprised grade 1-2 nausea, fatigue and myelosuppression. In both schedules, PKs increased linearly, but doses over the 1-h schedule RP2D resulted in higher than proportional increases in exposure. A patient with advanced urothelial carcinoma had RECIST shrinkage by 49%, and three patients had RECIST stable disease ≥6 months.
PM00104 is well tolerated, with preliminary evidence of antitumour activity observed. The 1-h 3-weekly schedule is being assessed in phase II clinical trials.
Persistence of minimal residual disease (MRD) after treatment for myeloma predicts inferior outcomes, but within MRD-positive patients there is great heterogeneity with both early and very late ...relapses. Among different MRD techniques, flow cytometry provides additional information about antigen expression on tumor cells, which could potentially contribute to stratify MRD-positive patients. We investigated the prognostic value of those antigens required to monitor MRD in 1265 newly diagnosed patients enrolled in the GEM2000, GEM2005MENOS65, GEM2005MAS65 and GEM2010MAS65 protocols. Overall, CD19
, CD27
, CD38
, CD45
, CD81
, CD117
and CD138
expression predicted inferior outcomes. Through principal component analysis, we found that simultaneous CD38
CD81
CD117
expression emerged as the most powerful combination with independent prognostic value for progression-free survival (HR:1.69; P=0.002). This unique phenotypic profile retained prognostic value among MRD-positive patients. We then used next-generation flow to determine antigen stability throughout the course of the disease, and found that the expression of antigens required to monitor MRD is mostly stable from diagnosis to MRD stages, except for CD81 whose expression progressively increased from baseline to chemoresistant tumor cells (14 vs 28%). Altogether, we showed that the phenotypic profile of tumor cells provides additional prognostic information, and could be used to further predict risk of relapse among MRD-positive patients.
Abstract
Study question
To compare embryo development and clinical outcomes between two commercial heavy oils using sibling donor oocytes collected.
Summary answer
Our study suggests that both ...commercial heavy oils achieve similar embryo development and clinical outcomes rates.
What is known already
Current tendencies in IVF laboratories, such as extending the embryo culture uninterruptedly until day 6/7 or the use of dry time-lapse incubators, have enhanced the importance to use good quality oils supporting human embryo culture in vitro. The coating of the culture dishes with oil is highly important to maintain the ideal conditions that embryos need for an optimal development. Specifically, oil plays an essential role in maintaining a stable temperature, provides a barrier against external agents and contributes preventing the media evaporation, and thus, to the maintenance of an optimal pH and osmolality for the correct embryo development.
Study design, size, duration
This is a single-centre prospective study performed between February and November 2022 that included 180 donors and 213 recipients. Donors were randomized using a computer-generated randomization list. Each case was processed and cultured with a commercial single medium coated with a layer of the commercial heavy oil assigned, a mineral oil (A) or a paraffin oil (B).
Participants/materials, setting, methods
Oocytes were injected by ICSI and then cultured in 16-well dishes (EmbryoSlide+®, EmbryoScope+™,Vitrolife) prepared with each heavy oil (1700µl oil/dish). These were cultured in a time-lapse incubator (EmbryoScope+™,Vitrolife) at 37.29 ± 0.05 °C in an atmosphere of 6.5% CO2 and 5% O2. These parameters were controlled periodically (T+Button,BrightSentinel and G100,Geotech). Laboratory conditions, such as temperature, humidity and volatile organic compounds levels were monitored continuously (Octax Log&Guard™,Vitrolife) during the study period, and pH was measured in a weekly basis.
Main results and the role of chance
A total of 2554 MII oocytes were injected by ICSI (oil A, n = 1304 and oil B, n = 1250). The proportion of fertilized oocytes was identical between the two oils (A:80.00% vs B:80.53%), as well as, abnormal fertilized oocyte rate (A:6.85% vs B:5.62%) and oocyte degeneration rate post-ICSI (A:6.14% vs B:6.00%). The mean number of embryos that reached the blastocyst stage and the proportion of blastocysts suitable for clinical use (transferred or cryopreserved) was almost the same independently of the oil used (A:69.84% vs B:67.15% and A:62.39% vs B:60.71%, respectively). Statistical data analysis was performed without referring to statistical significance (p > 0,05).
205 patients had an embryo transfer on day 5/6 with either fresh or cryopreserved blastocysts cultured coated with a layer of oil A (n = 101) or B (n = 104), with a mean number of 1.42 ± 0,55 and 1.33 ± 0,53 blastocysts transferred/patient in each group, respectively. No differences were found in terms of clinical pregnancy (A:69.30% vs B:67.31%) or implantation rates (A:62.37% vs B:61.53%) between both groups. Miscarriage rates were similar between group A (11.88%) and group B (12.62%).
The pH average value during the study was 7.26 ± 0.06. The mean values of the room temperature, humidity and VOCs were stable at 21.7 ± 0.4 °C, 66.7 ± 6.9% and 0.098 ± 0.01ppm, respectively.
Limitations, reasons for caution
Although heavy oils are the most competent in keeping optimal culture conditions over time, there are several culture oils with different features available in IVF market. Thus, further studies should be performed comparing among them. Future research is also needed to compare peroxidation rates of our culture oils studied.
Wider implications of the findings
The present study suggests that both commercial heavy oils used in a continuous approach may provide similar in vitro fertilization rates regarding fertilization, blastocysts suitable for clinical use or clinical pregnancy. Heavy oil features, laboratory conditions and the culture environment should be properly validated independently on each IVF center.
Trial registration number
not applicable
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