Dynamic Survival Risk Prognostic Model and Genomic Landscape for Atypical Teratoid/Rhabdoid Tumors: A Population-Based, Real-World Study

Background: An atypical teratoid/rhabdoid tumor (AT/RT) is an uncommon and aggressive pediatric central nervous system neoplasm. However, a universal clinical consensus or reliable prognostic evaluation system for this malignancy is lacking. Our study aimed to develop a risk model based on comprehensive clinical data to assist in clinical decision-making. Methods: We conducted a retrospective study by examining data from the Surveillance, Epidemiology, and End Results (SEER) repository, spanning 2000 to 2019. The external validation cohort was sourced from the Children’s Hospital Affiliated to Chongqing Medical University, China. To discern independent factors affecting overall survival (OS) and cancer-specific survival (CSS), we applied Least Absolute Shrinkage and Selection Operator (LASSO) and Random Forest (RF) regression analyses. Based on these factors, we structured nomogram survival predictions and initiated a dynamic online risk-evaluation system. To contrast survival outcomes among diverse treatments, we used propensity score matching (PSM) methodology. Molecular data with the most common mutations in AT/RT were extracted from the Catalogue of Somatic Mutations in Cancer (COSMIC) database. Results: The annual incidence of AT/RT showed an increasing trend (APC, 2.86%; 95% CI:0.75–5.01). Our prognostic study included 316 SEER database participants and 27 external validation patients. The entire group had a median OS of 18 months (range 11.5 to 24 months) and median CSS of 21 months (range 11.7 to 29.2). Evaluations involving C-statistics, DCA, and ROC analysis underscored the distinctive capabilities of our prediction model. An analysis via PSM highlighted that individuals undergoing triple therapy (integrating surgery, radiotherapy, and chemotherapy) had discernibly enhanced OS and CSS. The most common mutations of AT/RT identified in the COSMIC database were SMARCB1, BRAF, SMARCA4, NF2, and NRAS. Conclusions: In this study, we devised a predictive model that effectively gauges the prognosis of AT/RT and briefly analyzed its genomic features, which might offer a valuable tool to address existing clinical challenges

Real-world effectiveness of an intranasal spray A8G6 antibody cocktail in the post-exposure prophylaxis of COVID-19

Abstract Previously, we identified an antibody combination A8G6 that showed promising efficacy in COVID-19 animal models and favorable safety profile in preclinical models as well as in a first-in-human trial. To evaluate the real-word efficacy of A8G6 neutralizing antibody nasal spray in post-exposure prophylaxis of COVID-19, an open-label, non-randomized, two-arm, blank-controlled, investigator-initiated trial was conducted in Chongqing, China (the register number: ChiCTR2200066416). High-risk healthy participants (18–65 years) within 72 h after close contact to COVID-19 patients were recruited and received a three-dose (1.4 mg/dose) A8G6 treatment daily or no treatment (blank control) for 7 consecutive days. SARS-CoV-2 infection occurred in 151/340 (44.4%) subjects in the blank control group and 12/173 (6.9%) subjects in the A8G6 treatment group. The prevention efficacy of the A8G6 treatment within 72 h exposure was calculated to be 84.4% (95% CI: 74.4–90.4%). Moreover, compared to the blank-control group, the time from the SARS-CoV-2 negative to the positive COVID-19 conversion was significantly longer in the AG86 treatment group (mean time: 3.4 days vs 2.6 days, p = 0.019). In the secondary end-point analysis, the A8G6 nasal treatment had no effects on the viral load at baseline SARS-CoV-2 RT-PCR positivity and the time of the negative COVID-19 conversion. Finally, except for 5 participants (3.1%) with general adverse effects, we did not observe any severe adverse effects related to the A8G6 treatment. In this study, the intranasal spray AG86 antibody cocktail showed potent efficacy for prevention of SARS-CoV-2 infection in close contacts of COVID-19 patients

MED16 Promotes Tumour Progression and Tamoxifen Sensitivity by Modulating Autophagy through the mTOR Signalling Pathway in ER-Positive Breast Cancer

Recent studies have shown that the mediator complex (MED) plays a vital role in tumorigenesis and development, but the role of MED16 (mediator complex subunit 16) in breast cancer (BC) is not clear. Increasing evidence has shown that the mTOR pathway is important for tumour progression and therapy. In this study, we demonstrated that the mTOR signalling pathway is regulated by the expression level of MED16 in ER+ breast cancer. With the analysis of bioinformatics data and clinical specimens, we revealed an elevated expression of MED16 in luminal subtype tumours. We found that MED16 knockdown significantly inhibited cell proliferation and promoted G1 phase cell cycle arrest in ER+ BC cell lines. Downregulation of MED16 markedly reduced the sensitivity of ER+ BC cells to tamoxifen and increased the stemness and autophagy of ER+ BC cells. Bioinformatic analysis of similar genes to MED16 were mainly enriched in autophagy, endocrine therapy and mTOR signalling pathways, and the inhibition of mTOR-mediated autophagy restored sensitivity to tamoxifen by MED16 downregulation in ER+ BC cells. These results suggest an important role of MED16 in the regulation of tamoxifen sensitivity in ER+ BC cells, crosstalk between the mTOR signalling pathway-induced autophagy, and together, with the exploration of tamoxifen resistance, may indicate a new therapy option for endocrine therapy-resistant patients

Characterization of pTAK1-peptide-specific RaMoAbs.

a<p>Frequency indicates numbers of clones showing the identical amino acid sequences.</p>b<p>The data are presented as average of at least two experiments.</p

A Novel Rabbit Immunospot Array Assay on a Chip Allows for the Rapid Generation of Rabbit Monoclonal Antibodies with High Affinity

<div><p>Antigen-specific rabbit monoclonal antibodies (RaMoAbs) are useful due to their high specificity and high affinity, and the establishment of a comprehensive and rapid RaMoAb generation system has been highly anticipated. Here, we present a novel system using immunospot array assay on a chip (ISAAC) technology in which we detect and retrieve antigen-specific antibody-secreting cells from the peripheral blood lymphocytes of antigen-immunized rabbits and produce antigen-specific RaMoAbs with 10^–12 M affinity within a time period of only 7 days. We have used this system to efficiently generate RaMoAbs that are specific to a phosphorylated signal-transducing molecule. Our system provides a new method for the comprehensive and rapid production of RaMoAbs, which may contribute to laboratory research and clinical applications.</p> </div

Relationship between rabbit-ISAAC immunospots and affinities.

<p>(A) Representative ISAAC immunospots of secreted HEL-binding RaMoAbs with high (<i>K</i>D: 10⁻¹² M) and low (<i>K</i>D: 10⁻⁹ M) affinity. (B) A decay curve of fluorescence intensities in individual ISAAC immunospots. The fluorescence intensity of individual ISAAC immunospots (<i>y</i>-axis) is plotted against the distance (<i>x</i>-axis). <i>I</i>_1/2 is the value at which the immunospot fluorescence intensity reaches 50%, and <i>D</i>_1/2 indicates the distance at which <i>I</i>_1/2 is achieved. The <i>D</i>_1/2 value is shown in each individual plot. (C) Relationship between the <i>D</i>_1/2 of individual immunospots (<i>x</i>-axis) and <i>K</i>D (<i>y</i>-axis). The dotted line indicates <i>D</i>_1/2 = 60. The bar indicates the average <i>K</i>D for antibodies with <i>D</i>_1/2<60 and those with <i>D</i>_1/2>60. The <i>p</i>-value was determined using Student’s t-test. (D) Frequency of antibodies with the indicated order of <i>K</i>D values for antibodies with <i>D</i>_1/2<60 and <i>D</i>_1/2>60. The colored pie segment indicates the frequency of antibodies with the indicated order of <i>K</i>D. The number in the center of the pie chart denotes the number of antibodies analyzed. The <i>p</i>-value was determined using Fisher’s test.</p

Western blot analysis of pTAK1-peptide-specific RaMoAbs.

<p>(A) Examination of the specificity of pTAK1-peptide-specific RaMoAbs. Whole-cell lysates obtained from HeLa cells that were transfected with plasmids containing FLAG-tagged wild type (WT) TAK1 or a phosphorylation site-substituted mutant (T187A) together with HA-tagged TAB1 were separated by SDS-PAGE and immunoblotted with Ra_pTAK01, 04, 05, 06, 14, 19, 21, and 23 antibodies, a commercial antibody specific to phosphorylated TAK1, a TAK1-specific antibody or a TAB1-specific antibody. (B) TNF-α-induced phosphorylation of endogenous TAK1 at Thr187. Whole cell lysates from HeLa cells that had been stimulated with 20 ng ml⁻¹ TNF-α for the indicated time periods were separated by SDS-PAGE and immunoblotted with the Ra_pTAK23 antibody, a commercial pTAK1-pep-specific antibody, or a TAK1-specific antibody.</p

Efficient screening for phosphorylated peptide-specific RaMoAbs with rabbit-ISAAC.

<p>(A) Non-blocking procedure using the TAK1-peptide. Antibodies recognizing the non-phosphorylated peptide (a) and the phosphorylated peptide (b) can bind to biotinylated pTAK1-peptide, and a signal can be detected. (B) Blocking procedure using the TAK1-peptide. Microwell array chips were pre-treated with TAK1-peptide before the addition of biotinylated pTAK1-peptide. a) The TAK1-peptide binds to antibodies that recognize the non-phosphorylated site of the pTAK1-peptide. As a result, the biotinylated pTAK1-peptide cannot bind to the antibodies, and no signal is detected. b) The TAK1-peptide does not bind to antibodies that recognize the phosphorylated site of the pTAK1-peptide. As a result, the biotinylated pTAK1-peptide binds to antibodies, and a signal can be detected. (C) Acquisition efficiency of phosphorylated peptide-specific antibodies in the non-blocking and blocking procedures. The colored pie segment indicates the frequency of RaMoAbs that are specific to a phosphorylated peptide (red) and non-phosphorylated peptide (blue) in the non-blocking procedure (left) and the blocking procedure (right). The number in the center of the pie chart denotes the number of antibodies analyzed. The <i>p</i>-value was determined using Fisher’s test.</p