Metabolic interventions combined with CTLA-4 and PD-1/PD-L1 blockade for the treatment of tumors: mechanisms and strategies.

Immunotherapies based on immune checkpoint blockade (ICB) have significantly improved patient outcomes and offered new approaches to cancer therapy over the past decade. To date, immune checkpoint inhibitors (ICIs) of CTLA-4 and PD-1/PD-L1 represent the main class of immunotherapy. Blockade of CTLA-4 and PD-1/PD-L1 has shown remarkable efficacy in several specific types of cancers, however, a large subset of refractory patients presents poor responsiveness to ICB therapy; and the underlying mechanism remains elusive. Recently, numerous studies have revealed that metabolic reprogramming of tumor cells restrains immune responses by remodeling the tumor microenvironment (TME) with various products of metabolism, and combination therapies involving metabolic inhibitors and ICIs provide new approaches to cancer therapy. Nevertheless, a systematic summary is lacking regarding the manner by which different targetable metabolic pathways regulate immune checkpoints to overcome ICI resistance. Here, we demonstrate the generalized mechanism of targeting cancer metabolism at three crucial immune checkpoints (CTLA-4, PD-1, and PD-L1) to influence ICB therapy and propose potential combined immunotherapeutic strategies co-targeting tumor metabolic pathways and immune checkpoints.

ATM-ESCO2-SMC3 axis promotes 53BP1 recruitment in response to DNA damage and safeguards genome integrity by stabilizing cohesin complex.

53BP1 is primarily known as a key regulator in DNA double-strand break (DSB) repair. However, the mechanism of DSB-triggered cohesin modification-modulated chromatin structure on the recruitment of 53BP1 remains largely elusive. Here, we identified acetyltransferase ESCO2 as a regulator for DSB-induced cohesin-dependent chromatin structure dynamics, which promotes 53BP1 recruitment. Mechanistically, in response to DNA damage, ATM phosphorylates ESCO2 S196 and T233. MDC1 recognizes phosphorylated ESCO2 and recruits ESCO2 to DSB sites. ESCO2-mediated acetylation of SMC3 stabilizes cohesin complex conformation and regulates the chromatin structure at DSB breaks, which is essential for the recruitment of 53BP1 and the formation of 53BP1 microdomains. Furthermore, depletion of ESCO2 in both colorectal cancer cells and xenografted nude mice sensitizes cancer cells to chemotherapeutic drugs. Collectively, our results reveal a molecular mechanism for the ATM-ESCO2-SMC3 axis in DSB repair and genome integrity maintenance with a vital role in chemotherapy response in colorectal cancer.

SENP1 Decreases RNF168 Phase Separation to Promote DNA Damage Repair and Drug Resistance in Colon Cancer.

The DNA damage response (DDR) is essential for the maintenance of genomic stability. Protein posttranslational modifications play pivotal roles in regulating the DDR process. Here, we found that SUMOylated RNF168 undergoes liquid-liquid phase separation (LLPS), which restricts the recruitment of RNF168 to DNA damage sites, reduces RNF168-catalyzed H2A ubiquitination, restrains 53BP1 in nuclear condensates, and ultimately impairs nonhomologous DNA end joining repair efficiency. Sentrin/SUMO-specific protease 1 (SENP1) was identified as a specific deSUMOylase of RNF168, and it was highly expressed in colorectal adenocarcinoma. In response to DNA damage, SENP1 decreased RNF168 SUMOylation and prevented RNF168 from forming nuclear condensates, thus promoting damage repair efficiency and cancer cell resistance to DNA damaging agents. Moreover, high SENP1 expression correlated with poor prognosis in patients with cancer, and SENP1 depletion sensitized cancer cells to chemotherapy. In summary, these findings reveal DDR is suppressed by SUMOylation-induced LLPS of RNF168 and suggest that SENP1 is a potential target for cancer therapy. SIGNIFICANCE: Sentrin/SUMO-specific protease 1 decreases RNF168 SUMOylation and liquid-liquid phase separation to promote DNA damage repair, safeguarding genomic integrity and driving chemotherapy resistance.

Circular RNA HIPK3 facilitates ferroptosis in gestational diabetes mellitus by regulating glutathione peroxidase 4 DNA methylation.

BACKGROUND: Gestational diabetes mellitus (GDM) is the most frequently occurring complication during pregnancy, with a high prevalence rate. Ferroptosis, a type of iron-dependent cell death, is closely associated with GDM nosogenesis. The present study aimed to examine the potential role and mechanism of circHIPK3 in GDM. METHODS: Placental tissues, plasma samples, and HTR-8/SVneo cells were used. A receiver operating characteristic curve was used to analyze the diagnostic value of circHIPK3 in GDM. Actinomycin D and RnaseR were added to identify circHIPK3 characteristics. The expression of circHIPK3, miR-1278, and DNA methyltransferase 1 (DNMT1) was assessed using a quantitative reverse transcriptase-PCR. Cell counting kit-8 and terminal deoxynucleotidyl transferase dUTP nick end labeling assays and specific kits were employed to assess cell viability, apoptosis, reactive oxygen species (ROS), malondialdehyde, iron, glutathione, and glutathione peroxidase 4 (GPX4) levels. RESULTS: The interaction between miR-1278 and circHIPK3 or DNMT1 was validated via luciferase reporter and RNA pull-down assays. circHIPK3 expression was found to be high in GDM placental tissues, plasma, and cells, with a high diagnostic value. In high glucose (HG)-induced HTR-8/SVneo cells, the inhibition of circHIPK3 provoked cell viability and mitigated cell apoptosis, ROS, and iron levels, but it was rescued through the downregulation of miR-1278. Mechanism experiments showed that circHIPK3 bound with miR-1278 targeting DNMT1 in GDM. The elevation in DNMT1 expression abolished the effects of miR-1278 overexpression on ferroptosis in HG-cultured HTR-8/SVneo cells. CONCLUSIONS: Overall, circHIPK3 might facilitate ferroptosis via miR-1278/DNMT1 to regulate GPX4 DNA methylation in HG-cultured HTR-8/SVneo cells. CircHIPK3 could be a therapeutic agent for GDM treatment.

hCINAP regulates the differentiation of embryonic stem cells by regulating NEDD4 liquid-liquid phase-separation-mediated YAP1 activation.

YAP1 functions in lineage differentiation of pluripotent embryonic stem cells (ESCs); however, the detailed mechanisms underlying the regulation of YAP1 activity during ESC differentiation remain elusive. Here, we report that hCINAP serves as a negative regulator of YAP1 during ESC fate decisions. The expression of mCINAP, the murine homolog of hCINAP, is downregulated during the differentiation process of murine ESC (mESC) ectoderm lineage, leading to liquid-liquid phase separation (LLPS) of NEDD4 and activation of YAP1. Mechanistically, hCINAP interacts with and prevents NEDD4 from forming cytoplasmic condensates that compartmentalize YAP1 and its kinase NLK, facilitating YAP1 phosphorylation at Ser128 and promoting YAP1 activation. mCINAP depletion leads to the formation of NEDD4 condensates and YAP1 activation, which impedes endoderm differentiation of mESCs. Our study shows that hCINAP is a vital regulator of YAP1 activity and is essential for stem cell fate decisions, which provides mechanistic insight into early embryogenesis.

Deubiquitinase OTUB2 exacerbates the progression of colorectal cancer by promoting PKM2 activity and glycolysis.

Aberrant regulation of ubiquitination often leads to metabolic reprogramming in tumor cells. However, the underlying mechanisms are not fully understood. Here we demonstrate that OTUB2, an OTU deubiquitinase, is upregulated in colorectal cancer (CRC) and exacerbates the progression of CRC through modulating the aerobic glycolysis. Mechanistically, OTUB2 directly interacts with pyruvate kinase M2 (PKM2) and inhibits its ubiquitination by blocking the interaction between PKM2 and its ubiquitin E3 ligase Parkin, thereby enhancing PKM2 activity and promoting glycolysis. In response to glucose starvation stress, the effect of OTUB2 on PKM2 is enhanced, which confers metabolic advantage to CRC cells. Moreover, OTUB2 depletion reduces glucose consumption, lactate production, and cellular ATP production. OTUB2-knockout CRC cells exhibit attenuated proliferation and migration, as well as an elevated level of apoptosis and increased sensitivity to chemotherapy drugs. Furthermore, in vivo assays show that knockout of OTUB2 inhibits tumor growth in mice. Taken together, these findings reveal the critical role of OTUB2 in the regulation of glycolysis and illustrate the molecular mechanism underlying its role as a negative regulator of PKM2 ubiquitination in CRC, establishing a bridge between OTUB2-regulated PKM2 ubiquitination and altered metabolic patterns in CRC and suggesting that OTUB2 is a promising target for CRC treatment.

Metagenomic analysis revealed the microbiota and metabolic function during co-composting of food waste and residual sludge for nitrogen and phosphorus transformation.

This paper used bagasse as a composting additive and bulking agent in order to investigate the aerobic composting process of food waste and residual sludge. Accordingly, the variations of nitrogen and phosphorus nutrients, microbiota and metabolic function during the composting process were systematically explored. Three piles with residual sludge, food waste and bagasse mass ratios of 1:1:1, 2:1:1 and 4:1:1 were set. The ammonia nitrogen content in the three compost piles were 3.18 mg/g, 4.68 mg/g and 5.84 mg/g at the end of composting. The final available phosphorus content of the three piles were 3.42 mg/g, 6.70 mg/g and 11.21 mg/g, respectively. X-ray photoelectron spectroscopy (XPS) analysis showed that absorption peaks attributed to amines, amino acids and amides appeared in the 1:1:1 pile. Metagenomic analysis of the glycolysis and ammonia transformation pathways showed that the total relative abundance of key enzyme genes for the conversion of glucose to glucose-6-phosphate in the three plies were 0.326%, 0.213% and 0.248%, respectively. The total relative abundance of 2 glutamate dehydrogenase (GDH2), glud1-2 and E1,4,1,4 dehydrogenases in the three piles was 0.125%, 0.151% and 0.160%, respectively, as the main enzymes for the mutual conversion of ammonia and glutamate.

Effect of excess sludge and food waste feeding ratio on the nutrient fractions, and bacterial and fungal community during aerobic co-composting.

The effect of excess sludge and food waste feeding ratio on the co-composting process was explored using 5% bagasse biochar as an additive and conditioner. Results showed that when the mass ratio was 1:1, nitrogen fixation ability was the strongest and ammonia nitrogen increment in the pile reached 2.31 mg/g. The increase in excess sludge content/food waste ratio during composting was conducive to the accumulation of H2O-P, BD-P, HCl-P, NaOH-P and NaOH85-P. When the ratio of excess sludge to food waste mass was 1:1, the relative abundance of Firmicutes was the largest in the compost, which corresponded to 72.77% at the phylum level. Food waste mass was more beneficial to the growth and reproduction of microorganisms and to the metabolic activities related to membrane transport. Considering the fungal content, Ascomycota and Basidiomycota were maximum, with relative abundance of 69.53% and 20.91%, respectively, at the mass ratio of 1:1.

Similar Repair Effects of Human Placenta, Bone Marrow Mesenchymal Stem Cells, and Their Exosomes for Damaged SVOG Ovarian Granulosa Cells.

BACKGROUND: This study is aimed at investigating the repairing effect of mesenchymal stem cells and their exosomes from different sources on ovarian granulosa cells damaged by chemotherapy drugs-phosphoramide mustard (PM). METHODS: In this study, we choose bone marrow mesenchymal stem cells (BMSCs) and human placental mesenchymal stem cells (HPMSCs) for research. Then, they were cocultured with human ovarian granulosa cells (SVOG) injured by phosphoramide mustard (PM), respectively. ß-Galactosidase staining, flow cytometry, and Western blot were used to detect the changes in the senescence and apoptosis of SVOG cells before and after their coculture with the above two types of MSCs. Subsequently, exosomes from these two types of MSCs were extracted and added to the culture medium of SVOG cells after PM injury to test whether these two types of exosomes played a role similar to that of MSCs in repairing damaged SVOG cells. RESULTS: PM treatment-induced apoptotic SVOG cells were significantly decreased after HPMSCs and BMSCs as compared with control group. After coculturing with these two types of MSCs, PM-treated SVOG cells showed significantly reduced senescence and apoptosis proportions as well as cleaved-Caspase 3 expression, and HPMSCs played a slightly stronger role than BMSCs in repairing SVOG cells in terms of the above three indicators. In addition, the ratios of senescent and apoptotic SVOG cells were also significantly reduced by the two types of exosomes, which played a role similar to that of MSCs in repairing cell damages. CONCLUSIONS: The results indicated that BMSCs, HPMSCs, and their exosomes all exerted a certain repair effect on SVOG cells damaged by PM, and consistent repair effect was observed between exosomes and MSCs. The repair effect of exosomes secreted from BMSCs and HPMSCs on the SVOG cells was studied for the first time, and the results fully demonstrated that exosomes are the key carriers for MSCs to play their role.

Cellular redox sensor HSCARG negatively regulates the translesion synthesis pathway and exacerbates mammary tumorigenesis.

The translesion synthesis (TLS) pathway is a double-edged sword in terms of genome integrity. Deficiency in TLS leads to generation of DNA double strand break (DSB) during replication stress, while excessive activation of the TLS pathway increases the risk of point mutation. Here we demonstrate that HSCARG, a cellular redox sensor, directly interacts with the key protein PCNA in the TLS pathway. HSCARG enhances the interaction between PCNA and the deubiquitinase complex USP1/UAF1 and inhibits the monoubiquitination of PCNA, thereby impairing the recruitment of Y-family polymerases and increasing cell sensitivity to stimuli that trigger replication fork blockades. In response to oxidative stress, disaggregation of HSCARG dimers into monomers and the nuclear transport of HSCARG activate the regulatory function of HSCARG in the TLS pathway. Moreover, HSCARG, which is highly expressed in breast carcinoma, promotes the accumulation of DSBs and mutations. HSCARG knockout PyMT transgenic mice exhibit delayed mammary tumorigenesis compared with that in HSCARG wild-type or heterozygous PyMT mice. Taken together, these findings expand our understanding of TLS regulatory mechanisms and establish a link between the cellular redox status and the DNA damage response (DDR).

[Effects of Magnetic Fe3O4 Nanoparticles on the Characteristics of Anaerobic Granular Sludge and Its Interior Microbial Community].

In this study, the effects of magnetic Fe3O4 nanoparticles (Fe3O4 NPs) on soluble microbial products (SMP), loosely bound extracellular polymeric substances (LB-EPS), and tightly bound extracellular polymeric substances (TB-EPS) in anaerobic granular sludge were examined. In addition, the anaerobic granular sludge interior microbial community dynamics were investigated using high-throughput sequencing. The results demonstrated that the removal rate of COD was 83.6% after long-term exposure in the experimental reactor, namely, the anaerobic reactor containing Fe3O4 NPs. It was reduced by 5.7% in comparison with the removal rate in the control reactor. The total amount of TB-EPS in anaerobic granular sludge in the experimental and control reactors was 178.20 mg·g-1 and 138.24 mg·g-1, respectively, while the total amount of SMP in anaerobic granular sludge was 34.88 mg·L-1 and 27.44 mg·L-1, respectively. With regard to the LB-EPS in anaerobic granular sludge in the experimental reactor, the peak of humic acid disappeared and the peak intensity of coenzyme F420 decreased slightly using excitation-emission matrix (EEM) fluorescence spectra. In terms of the microbial community dynamics in the experimental reactor, the abundance of Methanobacterium was greatly augmented from 76.15% to 86.76%; whereas, the abundance of Methanothrix decreased from 17.1% to 7.51%. This indicated that Methanothrix was more sensitive to Fe3O4 NPs. Moreover, the changes in bacterial communities were evident:①the abundance of Proteobacteria dropped from 66.44% to 47.16%; ② the abundance of Actinobacteria grew from 8.97% to 17.33%; and ③ the abundance of Bacteroidetes increased from 8.07% to 17.74%. The increasing abundance of Actinobacteria and Bacteroidetes plays a positive role in the anaerobic hydrolysis of organic matter.

Performance and microbial community of CIC anaerobic reactor treating food waste under different grease contents and inner circulation ratio.

High concentrations of grease easily inhibit anaerobic digestion. The stability of the process and microbial responses in the controlling internal circulation (CIC) reactor used for treating food waste were investigated under different grease contents and inner circulation ratios. Results showed that at the grease content of 1 g/L, the removal rates of 94% and 86-93% were achieved for chemical oxygen demand (COD) and NH3-N, respectively. In contrast, when the grease content increased to 7 g/L, removal rates for COD and NH3-N significantly decreased to 42.8 and 10%, respectively. In the three-dimensional excitation and emission matrix (3D-EEM) spectra of LB-EPS (loosely bound extracellular polymeric substances), the fluorescence intensity of coenzyme F420 was weakened in the granular sludge, and the fluorescence peak of aromatic protein disappeared in the TB-EPS (tightly bound EPS). The activity and stability of the granular sludge deteriorated with increasing grease content, in this case at 7 g/L. However, when the inner cycle ratio was increased to 4, the removal rate of COD and NH3-N increased to about 70 and 76%, respectively. The adverse effects of grease could be decreased by increasing the inner cycle ratio. When the grease content increased from 1 to 7 g/L, the abundance of Methanofollis increased from 9.93 to 46.41%, while Methanothrix abundance was reduced from 18.4 to 3.07%. It could indicate that Methanothrix was sensitive to high grease content.

Lasing from lead halide perovskite semiconductor microcavity system.

Organic-inorganic halide perovskite semiconductors are ideal gain media for fabricating laser and photonic devices due to high absorption, photoluminescence (PL) efficiency and low nonradiative recombination losses. Herein, organic-inorganic halide perovskite CH3NH3PbI3 is embedded in the Fabry-Perot (FP) microcavity, and a wavelength-tunable excitonic lasing with a threshold of 12.9 µJ cm-2 and the spectral coherence of 0.76 nm are realized. The lasing threshold decreases and the spectral coherence enhances as the temperature decreases; these results are ascribed to the suppression of exciton irradiative recombination caused by thermal fluctuation. Moreover, both lasing and light emission below threshold from the perovskite microcavity (PM) system demonstrate a redshift with the decreasing temperature. These results provide a feasible platform based on the PM system for the study of light-matter interaction for quantum optics and the development of optoelectronic devices such as polariton lasers.

Exciton-polariton light-emitting diode based on a ZnO microwire.

Room temperature electrically pumped exciton-polariton light-emitting diode (LED) based on the n-ZnO microwire/MgO/p-GaN heterojunction was fabricated. With the injection current of 1.5 mA, the ultraviolet electroluminescence centered at wavelength of 400 nm is obtained. The whispering gallery cavity model combined with the coupling oscillator model is used to describe the emission intensity modulation, from which the strong coupling regime of the system is proved to be surviving at room temperature.

Exciton-Polariton Fano Resonance Driven by Second Harmonic Generation.

Angle-resolved second harmonic generation (SHG) spectra of ZnO microwires show characteristic Fano resonances in the spectral vicinity of exciton-polariton modes. We observe a resonant peak followed by a strong dip in SHG originating from the constructive and destructive interference of the nonresonant SHG and the resonant contribution of the polariton mode. It is demonstrated that the Fano line shape, and thus the Fano asymmetry parameter q, can be tuned by the phase shift of the two channels. We develop a model to calculate the phase-dependent q as a function of the radial angle in the microwire and achieve a good agreement with the experimental results. The deduced phase-to-q relation unveils the crucial information about the dynamics of the system and offers a tool for control on the line shape of the SHG spectra in the vicinity of exciton-polariton modes.