Locoregional delivery of IL-13Rα2-targeting CAR-T cells in recurrent high-grade glioma: a phase 1 trial.

Chimeric antigen receptor T cell (CAR-T) therapy is an emerging strategy to improve treatment outcomes for recurrent high-grade glioma, a cancer that responds poorly to current therapies. Here we report a completed phase I trial evaluating IL-13Rα2-targeted CAR-T cells in 65 patients with recurrent high-grade glioma, the majority being recurrent glioblastoma (rGBM). Primary objectives were safety and feasibility, maximum tolerated dose/maximum feasible dose and a recommended phase 2 dose plan. Secondary objectives included overall survival, disease response, cytokine dynamics and tumor immune contexture biomarkers. This trial evolved to evaluate three routes of locoregional T cell administration (intratumoral (ICT), intraventricular (ICV) and dual ICT/ICV) and two manufacturing platforms, culminating in arm 5, which utilized dual ICT/ICV delivery and an optimized manufacturing process. Locoregional CAR-T cell administration was feasible and well tolerated, and as there were no dose-limiting toxicities across all arms, a maximum tolerated dose was not determined. Probable treatment-related grade 3+ toxicities were one grade 3 encephalopathy and one grade 3 ataxia. A clinical maximum feasible dose of 200 × 106 CAR-T cells per infusion cycle was achieved for arm 5; however, other arms either did not test or achieve this dose due to manufacturing feasibility. A recommended phase 2 dose will be refined in future studies based on data from this trial. Stable disease or better was achieved in 50% (29/58) of patients, with two partial responses, one complete response and a second complete response after additional CAR-T cycles off protocol. For rGBM, median overall survival for all patients was 7.7 months and for arm 5 was 10.2 months. Central nervous system increases in inflammatory cytokines, including IFNγ, CXCL9 and CXCL10, were associated with CAR-T cell administration and bioactivity. Pretreatment intratumoral CD3 T cell levels were positively associated with survival. These findings demonstrate that locoregional IL-13Rα2-targeted CAR-T therapy is safe with promising clinical activity in a subset of patients. ClinicalTrials.gov Identifier: NCT02208362 .

Z-Scheme Modulated Charge Transfer on InVO4 @ZnIn2 S4 for Durable Overall Water Splitting.

The charge transfer within heterojunction is crucial for the efficiency and stability of photocatalyst for overall water splitting (OWS). Herein, InVO4 nanosheets have been employed as a support for the lateral epitaxial growth of ZnIn2 S4 nanosheets to produce hierarchical InVO4 @ZnIn2 S4 (InVZ) heterojunctions. The distinct branching heterostructure facilitates active site exposure and mass transfer, further boosting the participation of ZnIn2 S4 and InVO4 for proton reduction and water oxidation, respectively. The unique Z-scheme modulated charge transfer, visualized by simulation and in situ analysis, has been proved to promote the spatial separation of photoexcited charges and strengthen the anti-photocorrosion capability of InVZ. The optimized InVZ heterojunction presents improved OWS (153.3 µmol h-1  g-1 for H2 and 76.9 µmol h-1  g-1 for O2 ) and competitive H2 production (21090 µmol h-1  g-1 ). Even after 20 times (100 h) of cycle experiment, it still holds more than 88% OWS activity and a complete structure.

3D-organoid culture supports differentiation of human CAR+ iPSCs into highly functional CAR T cells.

Unlimited generation of chimeric antigen receptor (CAR) T cells from human-induced pluripotent stem cells (iPSCs) is an attractive approach for "off-the-shelf" CAR T cell immunotherapy. Approaches to efficiently differentiate iPSCs into canonical αß T cell lineages, while maintaining CAR expression and functionality, however, have been challenging. We report that iPSCs reprogramed from CD62L+ naive and memory T cells followed by CD19-CAR engineering and 3D-organoid system differentiation confers products with conventional CD8αß-positive CAR T cell characteristics. Expanded iPSC CD19-CAR T cells showed comparable antigen-specific activation, degranulation, cytotoxicity, and cytokine secretion compared with conventional CD19-CAR T cells and maintained homogeneous expression of the TCR derived from the initial clone. iPSC CD19-CAR T cells also mediated potent antitumor activity in vivo, prolonging survival of mice with CD19+ human tumor xenografts. Our study establishes feasible methodologies to generate highly functional CAR T cells from iPSCs to support the development of "off-the-shelf" manufacturing strategies.

Defective Patient NK Function Is Reversed by AJ2 Probiotic Bacteria or Addition of Allogeneic Healthy Monocytes.

In this paper, we present the role of autologous and allogeneic monocytes from healthy individuals and those of the cancer patients, with a number of distinct cancers, in activating the function of natural killer (NK) cells, in particular, in induction of IFN-γ secretion by the NK cells and the functional capability of secreted IFN-γ in driving differentiation of the tumor cells. In addition, we compared the roles of CD16 signaling as well as sonicated probiotic bacteria AJ2 (sAJ2)-mediated induction and function of IFN-γ-mediated differentiation in tumor cells. We found that monocytes from cancer patients had lower capability to induce functional IFN-γ secretion by the autologous CD16 mAb-treated NK cells in comparison to those from healthy individuals. In addition, when patient monocytes were cultured with NK cells from healthy individuals, they had lower capability to induce functional IFN-γ secretion by the NK cells when compared to those from autologous monocyte/NK cultures from healthy individuals. Activation by sAJ2 or addition of monocytes from healthy individuals to patient NK cells increased the secretion of functional IFN-γ by the NK cells and elevated its functional capability to differentiate tumors. Monocytes from cancer patients were found to express lower CD16 receptors, providing a potential mechanism for their lack of ability to trigger secretion of functional IFN-γ. In addition to in vitro studies, we also conducted in vivo studies in which cancer patients were given oral supplementation of AJ2 and the function of NK cells were studied. Oral ingestion of AJ2 improved the secretion of IFN-γ by patient derived NK cells and resulted in the better functioning of NK cells in cancer patients. Thus, our studies indicate that for successful NK cell immunotherapy, not only the defect in NK cells but also those in monocytes should be corrected. In this regard, AJ2 probiotic bacteria may serve to provide a potential adjunct treatment strategy.

[Impact of a Sewage Treatment Plant on the Accumulation of Microplastics in Freshwater Organisms in the Lijiang River of the Guilin Urban Section].

Microplastics (MPs, particle size<5 mm), as a new pollutant, have attracted wide attention in recent years. The distributions of MPs in effluent of a sewage treatment plant (STP) were examined. Surface water, sediment, and freshwater organism samples were taken from the STP discharge outlet in the Lijiang River tributary (S1), the confluence of tributaries and main streams in the Lijiang River (S2), and downstream locations in the Lijiang River (S3). The impact of STP discharge effluent on the characteristics and spatial distribution of MPs pollution in freshwater organisms was studied. The results showed that the freshwater organisms had a probability of uptake of MPs by 94.2%. The mean abundance of MPs in S1 (2.7 n·ind-1) was significantly higher than that of S3 (1.9 n·ind-1, P<0.05). The MPs found in S1 and S3 were mainly <0.10 mm, accounting for 46.0% and 30.5%, respectively. The fiber type of MPs was observed in the body of freshwater organisms. Polyethylene terephthalate was the major polymer form in S1 organisms, while polypropylene was the major polymer form in S3. The effluent discharged from the STP led to the accumulation of MPs in freshwater organisms.

Performance and functional microbial communities of denitrification process of a novel MFC-granular sludge coupling system.

The performance, microbial communities and functional gene metabolism of the novel microbial fuel cell (MFC)-granular sludge coupling system was investigated. The results showed that COD and nitrogen removal can be up to 1.3-2.0 kg COD/L, 20-30 mg NO2--N/L, and 60-70 mg NO3--N/L, respectively. Proteobacteria, Chloroflexi, and Firmicutes were the dominant bacterial phyla, and the denitrification process was mainly consisted of the dominant denitrifying bacteria: Thauera (26.21%) and Pseudomonas (14.79%) in the first compartment, combining with denitrifying anaerobic methane oxidation bacteria: NC10 phylum of 0.072% (the first compartment) and 0.089% (the fourth compartment), Candidatus Methylomirabilis oxyfera of 0.044% (the first compartment) and 0.048% (the fourth compartment). According to functional gene classification for Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, metabolism was the main cluster for the whole sequence in the KEGG (7.17-11.41%), indicating that the dominant metabolic pathway played an important role in the degradation of pollutants.

Natural Killer Cells: Diverse Functions in Tumor Immunity and Defects in Pre-neoplastic and Neoplastic Stages of Tumorigenesis.

Natural killer (NK) cells are the key immune effectors with the ability to mediate selection and differentiation of a number of different cancer stem cells/undifferentiated tumors via lysis, and secreted or membrane-bound interferon (IFN)-γ and tumor necrosis factor (TNF)-α, respectively, leading to curtailment of tumor growth and metastasis. In this review, we present an overview of our recent findings on the biology and significance of NK cells in selection and differentiation of stem-like tumors using in vitro and in vivo studies conducted in humanized-BLT mice and in cancer patients. In addition, we present current advances in NK cell expansion and therapeutic delivery, and discuss the utility of allogeneic supercharged NK cells in the treatment of cancer patients. Moreover, we discuss the potential loss of NK cell numbers and function at the neoplastic and pre-neoplastic stages of tumorigenesis in induction and progression of pancreatic cancer. Therefore, because of their indispensable role in targeting cancer stem-like/undifferentiated tumors, NK cells should be placed high in the armamentarium of tumor immunotherapy. A combination of allogeneic supercharged NK cells with other immunotherapeutic strategies such as oncolytic viruses, antibody-dependent cellular cytotoxicity (ADCC)-inducing antibodies, checkpoint inhibitors, chimeric antigen receptor (CAR) T cells, CAR NK cells, and chemotherapeutic and radiotherapeutic strategies can be used for the ultimate goal of tumor eradication.

Effect of circulation and micro-aeration on sludge characteristics and microbial community in an ABR for treating traditional Chinese medicine wastewater.

The effects of circulation reflux and micro-aeration on the performance of a modified anaerobic baffled reactor (ABR) for treatment of traditional Chinese medicine (TCM) wastewater were evaluated. The characteristics of anaerobic sludge and microbial community structure in the modified ABR were also investigated. The results indicated that with conditions of reflux ratio of 1, reflux ratio of 2, reflux ratio of 2 with micro-aeration, and reflux ratio of 3, the modified ABR achieved an average COD removal efficiency of 90%, 87.7%, 87.8%, and 88.4%, respectively. In addition, the NH3-N average removal efficiency was 45.1%, 50%, 55.9%, and 55.4%, respectively. The analysis of excitation-emission matrix (EEM) fluorescence spectra of soluble microbial products (SMP) and extracellular polymeric substances (EPS) showed that there were tyrosine-like, aromatic protein-like, and coenzyme F420 substances in the sludge. The EPS were analysed by the Fourier transform infrared spectroscopy (FTIR), which showed that aromatic compounds were partially degraded, while the protein and polysaccharide compounds increased in each compartment of the modified ABR. Interestingly, the microbial community of anaerobic sludge analysis results showed that Chloroflexi was the dominant in the first, third and fourth compartments. Meanwhile, Levilinea and Methanothrix were the dominant species in the first and third compartments at the genus level.

Performance of a novel ABR-bioelectricity-Fenton coupling reactor for treating traditional Chinese medicine wastewater containing catechol.

In this study, a novel anaerobic baffled reactor-bioelectricity-Fenton (ABR-BEF) coupling reactor, combining an ABR, microbial fuel cell (MFC), and Fenton system, was used to treat traditional Chinese medicine (TCM) wastewater containing catechol. The bio-electrochemical degradation of the catechol reached 99.7% after 8 h at dissolved oxygen (DO) concentration of 4 mg/L in the cathodic chamber. The removal rates of chemical oxygen demand (COD) reached 91.7%, when the ratio rate was 1 and the DO concentration was 4 mg/L. Moreover, the maximum open-circuit voltage and power density of the coupling reactor reached 424.9 mV and 77.1 mW/m3, respectively. According to the PICRUSt analysis, carbohydrate metabolism took up the most abundant function of metabolism and the enrichment of membrane transporters may relieve TCM wastewater toxicity. These results suggest that the ABR-BEF coupling reactor could be applied as an efficient approach to treat TCM wastewater.

Influence of amoxicillin after pre-treatment on the extracellular polymeric substances and microbial community of anaerobic granular sludge.

A combined process coupling a Fe3O4 nanoparticles (Fe3O4 NPs) heterogeneous Fenton-like reaction and an anaerobic biological technology was investigated in order to effectively treat amoxicillin-containing wastewater. With the increase in the pretreatment degree, the average COD removal rate correspondingly increased from 84.8% to 92.4% using the anaerobic biological treatment, and the biodegradability and COD removal efficiency was improved by the pretreatment processes. During the process of amoxicillin degradation, hydroxyl free radicals tended to attack the lactamide, amide and pentacyclic rings of amoxicillin. In the excitation-emission matrix (EEM) spectra of soluble microbial products (SMPs), the absorption peak of humic acid gradually decreased with application of the pretreatment. The pretreatment products were more beneficial to the characteristics of anaerobic granular sludge. For the microbial community structure, the proportion of Methanothrix and Clostridia increased with addition the heterogeneous Fenton-like pretreatment, which favored conversion of organic contaminants to volatile fatty acids and biogas.

On the predictability of the orientation of protein domains joined by a spanning alpha-helical linker.

Connecting proteins together in prescribed geometric arrangements is an important element in new areas of biomolecular design. In this study, we characterize the degree of three-dimensional orientational control that can be achieved when two protein domains that have alpha-helical termini are joined using an alpha-helical linker. A fusion between naturally oligomeric protein domains was designed in this fashion with the intent of creating a self-assembling 12-subunit tetrahedral protein cage. While the designed fusion protein failed to assemble into a tetrahedral cage in high yield, a series of crystal structures showed that the two fused components were indeed bridged by an intact alpha helix, although the fusion protein was distorted from the intended ideal configuration by bending of the helix, ranging from 7 to 35°. That range of deviation in orientation creates challenges for designing large, perfectly symmetric protein assemblies, although it should offer useful outcomes for other less geometrically demanding applications in synthetic biology.