Blimp1 And Nr4a3 Transcription Factors Reciprocally Regulate Antitumor Car T-Cell Stemness And Exhaustion

Chimeric antigen receptor (CAR) T-cells have not induced meaningful clinical responses in solid tumor indications. Loss of T-cell stemness, poor expansion capacity and exhaustion during prolonged tumor antigen exposure are major causes of CAR T-cell therapeutic resistance. scRNA-sequencing analysis of CAR T-cells from a first-in-human trial in metastatic prostate cancer identified two distinct and independently validated cell states associated with antitumor potency or lack of efficacy. Low levels of the PRDM1 gene encoding the BLIMP1 transcription factor defined highly potent TCF7+CD8+ CAR T-cells, while enrichment of TIM3+CD8+ T-cells with elevated PRDM1 expression predicted poor outcome. PRDM1 single knockout promoted TCF7-dependent CAR T-cell stemness and proliferation resulting in marginally enhanced leukemia control. However, in the setting of PRDM1 deficiency, a negative epigenetic feedback program of NFAT-driven T-cell dysfunction characterized by compensatory upregulation of NR4A3 and multiple other genes encoding exhaustion-related transcription factors hampered effector function in solid tumors. PRDM1 and NR4A3 combined ablation skewed CAR T-cell phenotypes away from TIM3+CD8+ and toward TCF7+CD8+ to counter exhaustion of tumor-infiltrating CAR T-cells and improve in vivo antitumor responses, effects that were not achieved with BLIMP1 or NR4A3 single disruption alone. These data reveal a novel molecular targeting strategy to enrich stem-like CAR T-cells resistant to exhaustion and underscore dual inhibition of PRDM1/NR4A3 expression or activity as a promising approach to advance adoptive cell immuno-oncotherapy

The influence of game genre on Internet gaming disorder

Although Internet gaming disorder (IGD) has been investigated in detail, minimal research has been conducted regarding the influence of different game genres on IGD. The aim of this study is to compare the characteristics of members of game genre-specific groups with IGD and to identify factors associated with IGD status in each group in a large sample of adults. Methods Internet games were categorized into four genres: real-time strategy games, massive multiplayer online role-playing games (MMORPG), sport games, and first-person shooter (FPS) games. Participants (n = 2,923) who usually played one of these games completed an anonymous online survey that collected sociodemographic, game usage pattern, and psychopathological assessment data. Results MMORPG and FPS game players more frequently met the criteria for IGD than participants in the other two groups. Differences between the IGD-suspected gamers within the genre-specific groups were observed for a few items, such as average game-playing time and the subscales of the behavioral activation system; however, the factors that contributed to the development of IGD within each game genre-specific group were found to be considerably different. Discussion and conclusions The findings of this study suggest that IGD is a stable psychiatric diagnosis encompassing users of a broad range of game genres. In addition, the development of strategies for the prevention of and early intervention on individuals at high risk for developing IGD may require consideration of the distinct characteristics identified as effective predictors of IGD in users of each game genre

Directed evolution of CRISPR-Cas9 to increase its specificity

The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed S. pyogenes Cas9 (SpCas9) variants that display higher specificities than the wild-type SpCas9 protein are available, these attenuated Cas9 variants are often poorly efficient in human cells. Here, we develop a directed evolution approach in E. coli to obtain Sniper-Cas9, which shows high specificities without killing on-target activities in human cells. Unlike other engineered Cas9 variants, Sniper-Cas9 shows WT-level on-target activities with extended or truncated sgRNAs with further reduced off-target activities and works well in a preassembled ribonucleoprotein (RNP) format to allow DNA-free genome editing.

BLIMP1 and NR4A3 Transcription Factors Reciprocally Regulate Antitumor Car T-Cell Stemness and Exhaustion

Chimeric antigen receptor (CAR) T-cells have not induced meaningful clinical responses in solid tumor indications. Loss of T-cell stemness, poor expansion capacity and exhaustion during prolonged tumor antigen exposure are major causes of CAR T-cell therapeutic resistance. scRNA-sequencing analysis of CAR T-cells from a first-in-human trial in metastatic prostate cancer identified two distinct and independently validated cell states associated with antitumor potency or lack of efficacy. Low levels of the PRDM1 gene encoding the BLIMP1 transcription factor defined highly potent TCF7+CD8+ CAR T-cells, while enrichment of TIM3+CD8+ T-cells with elevated PRDM1 expression predicted poor outcome. PRDM1 single knockout promoted TCF7-dependent CAR T-cell stemness and proliferation resulting in marginally enhanced leukemia control. However, in the setting of PRDM1 deficiency, a negative epigenetic feedback program of NFAT-driven T-cell dysfunction characterized by compensatory upregulation of NR4A3 and multiple other genes encoding exhaustion-related transcription factors hampered effector function in solid tumors. PRDM1 and NR4A3 combined ablation skewed CAR T-cell phenotypes away from TIM3+CD8+ and toward TCF7+CD8+ to counter exhaustion of tumor-infiltrating CAR T-cells and improve in vivo antitumor responses, effects that were not achieved with BLIMP1 or NR4A3 single disruption alone. These data reveal a novel molecular targeting strategy to enrich stem-like CAR T-cells resistant to exhaustion and underscore dual inhibition of PRDM1/NR4A3 expression or activity as a promising approach to advance adoptive cell immuno-oncotherapy

Directed evolution of CRISPR-Cas9 to increase its specificity

The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed S. pyogenes Cas9 (SpCas9) variants that display higher specificities than the wild-type SpCas9 protein are available, these attenuated Cas9 variants are often poorly efficient in human cells. Here, we develop a directed evolution approach in E. coli to obtain Sniper-Cas9, which shows high specificities without killing on-target activities in human cells. Unlike other engineered Cas9 variants, Sniper-Cas9 shows WT-level on-target activities with extended or truncated sgRNAs with further reduced off-target activities and works well in a preassembled ribonucleoprotein (RNP) format to allow DNA-free genome editing. © 2018, The Author(s

Risk factors for postoperative delirium in elderly patients after spinal fusion surgery

Background Postoperative delirium (POD) has an incidence rate of 9% to 41%. It is directly linked to decreasing cognitive function, increasing length of hospitalization and cost, as well as other complications and mortality. We aimed to assess the risk factors for POD among elderly patients by analyzing data from those who underwent spinal surgery. Methods This study included 446 patients aged 65 years or older who underwent spinal surgery at our institution between March 2013 and May 2018. Data were collected retrospectively from the patients’ electronic medical records, and logistic regression was used to identify the risk factors associated with POD. The diagnosis of POD was based on the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, and was made through consultation with a psychiatrist during postoperative hospitalization and before discharge. Results Seventy-eight (78/446, 17.4%) patients were diagnosed with POD. The most relevant risk factor for POD was preoperative cognitive dysfunction (odds ratio [OR], 4.37; 95% confidence interval [CI], 1.60 to 11.93; P = 0.004), followed by emergency surgery (OR, 2.70; 95% CI, 1.27 to 5.74; P = 0.01), age (OR, 1.19; 95% CI, 1.13 to 1.26; P < 0.001), and anesthesia time (OR, 1.01; 95% CI 1.00 to 1.01; P = 0.002). Conclusions Preoperative cognitive dysfunction, emergency surgery, age, and anesthesia time were factors that affected POD occurrence after spinal surgery. Patients with such associated factors may be at a higher risk for POD when undergoing spinal surgery, and hence, careful management may be necessary for these patients

IDEA: Integrating Divisive and Ensemble-Agglomerate hierarchical clustering framework for arbitrary shape data

© 2021 IEEE.Hierarchical clustering, a traditional clustering method, has been getting attention again. Among several reasons, a credit goes to a recent paper by Dasgupta in 2016 that proposed a cost function that quantitatively evaluates hierarchical clustering trees. An important question is how to combine this recent advance with existing successful clustering methods. In this paper, we propose a hierarchical clustering method to minimize the cost function of clustering tree by incorporating existing clustering techniques. First, we developed an ensemble tree-search method that finds an integrated tree with reduced cost by integrating multiple existing hierarchical clustering methods. Second, to operate on large and arbitrary shape data, we designed an efficient hierarchical clustering framework, called integrating divisive and ensemble-agglomerate (IDEA) by combining it with advanced clustering techniques such as nearest neighbor graph construction, divisive-agglomerate hybridization, and dynamic cut tree. The IDEA clustering method showed better performance in minimizing Dasgupta&apos;s cost and improving accuracy (adjusted rand index) over existing cost-minimization-based, and density-based hierarchical clustering methods in experiments using arbitrary shape datasets and complex biology-domain datasets.N