T-cell exhaustion in CAR-T-cell therapy and strategies to overcome it.

Tumour immunotherapy has achieved good therapeutic effects in clinical practice and has received increased attention. Cytotoxic T cells undoubtedly play an important role in tumour immunotherapy. As a revolutionary tumour immunotherapy approach, chimeric antigen receptor T-cell (CAR-T-cell) therapy has made breakthroughs in the treatment of haematological cancers. However, T cells are easily exhausted in vivo, especially after they enter solid tumours. The exhaustion of T cells can lead to poor results of CAR-T-cell therapy in the treatment of solid tumours. Here, we review the reasons for T-cell exhaustion and how T-cell exhaustion develops. We also review and discuss ways to improve CAR-T-cell therapy effects by regulating T-cell exhaustion.

FEN1 inhibitor SC13 promotes CAR-T cells infiltration into solid tumours through cGAS-STING signalling pathway.

It is well known that chimeric antigen receptor T-cell immunotherapy (CAR-T-cell immunotherapy) has excellent therapeutic effect in haematological tumours, but it still faces great challenges in solid tumours, including inefficient T-cell tumour infiltration and poor functional persistence. Flap structure-specific endonuclease 1 (FEN1), highly expressed in a variety of cancer cells, plays an important role in both DNA replication and repair. Previous studies have reported that FEN1 inhibition is an effective strategy for cancer treatment. Therefore, we hypothesized whether FEN1 inhibitors combined with CAR-T-cell immunotherapy would have a stronger killing effect on solid tumours. The results showed that low dose of FEN1 inhibitors SC13 could induce an increase of double-stranded broken DNA (dsDNA) in the cytoplasm. Cytosolic dsDNA can activate the cyclic GMP-AMP synthase-stimulator of interferon gene signalling pathway and increase the secretion of chemokines. In vivo, under the action of FEN1 inhibitor SC13, more chemokines were produced at solid tumour sites, which promoted the infiltration of CAR-T cells and improved anti-tumour immunity. These findings suggest that FEN1 inhibitors could enable CAR-T cells to overcome poor T-cell infiltration and improve the treatment of solid tumours.

Techniques to characterize dynamics in biomaterials microenvironments: XPCS and microrheology of alginate/PEO-PPO-PEO hydrogels.

Many recent studies have highlighted the timescale for stress relaxation of biomaterials on the microscale as an important factor in regulating a number of cell-material interactions, including cell spreading, proliferation, and differentiation. Relevant timescales on the order of 0.1-100 s have been suggested by several studies. While such timescales are accessible through conventional mechanical rheology, several biomaterials have heterogeneous structures, and stress relaxation mechanisms of the bulk material may not correspond to that experienced in the cellular microenvironment. Here we employ X-ray photon correlation spectroscopy (XPCS) to explore the temperature-dependent dynamics, relaxation time, and microrheology of multicomponent hydrogels comprising of commercial poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer F127 and alginate. Previous studies on this system have shown thermoreversible behavior in the bulk oscillatory shear rheology. At physiological temperatures, bulk rheology of these samples shows behavior characteristic of a soft solid, with G' > G'' and no crossover between G' and G'' over the measurable frequency range, indicating a relaxation time >125 s. By contrast, XPCS-based microrheology shows viscoelastic behavior at low frequencies, and XPCS-derived correlation functions show relaxation times ranging from 10-45 s on smaller length scales. Thus, we are able to use XPCS to effectively probe the viscoelasticity and relaxation behavior within the material microenvironments.

Impact of stereochemistry on rheology and nanostructure of PLA-PEO-PLA triblocks: stiff gels at intermediate l/d-lactide ratios.

We report rheology and structural studies of poly(lactide)-poly(ethylene oxide)-poly(lactide) (PLA-PEO-PLA) triblock copolymer gels with various ratios of l-lactide and d-lactide in the PLA blocks. These materials form associative micellar gels in water, and previous work has shown that stereoregular triblocks with a l/d ratio of 100/0 form much stiffer gels than triblocks with a 50/50 l/d ratio. Our systems display an unexpected maximum in the storage modulus, G', of the hydrogels at intermediate l/d ratio. The impact of stereochemistry on the rheology is very striking; gels with an l/d ratio of 85/15 have storage moduli that are ∼1-2 orders of magnitude higher than hydrogels with l/d ratios of 100/0. No stereocomplexation is observed in the gels, although PLLA crystals are found for gels with l/d ratios of 95/5 and 90/10, and SANS results show a decrease in the intermicellar spacing for intermediate l/d ratios. We expect the dominant contribution to the elasticity of the gels to be intermicellar bridging chains and attribute the rheology to a competition between an increase in the time for PLA endblocks to pull out of micelles as the l/d ratio is increased and PLLA crystallization occurs, and a decrease in the number of bridging chains for micelles with crystalline PLA domains, as formation of bridges may be hindered by crowded crystalline PLA domains. These results provide a new strategy for controlling the rheology of PLA-based hydrogels for potential applications in biomaterials, as well as fundamental insights into how intermicellar interactions can be tuned via stereochemistry.

What occupational risk factors significantly affect miners' health: Findings from meta-analysis and association rule mining.

INTRODUCTION: The workplace's health hazard remains a significant concern to workers in the mining industry, where miners are continually exposed to various kinds of exposure sources. METHOD: First, the determinants of miners' health were systematically extracted from 259 publications, comprising chemical, physical, ergonomic, and psychosocial stressors, vulnerability factors, and common health outcomes. Then, 16 meta-analyses were performed to ascertain the epidemiological evidence for associations between four stressors and three health outcomes. The seven top contributing factors affecting miners' health were identified through 166 available prospective studies. Finally, based on the classic and domestic measurement scales, a cross-sectional survey of 559 Chinese miners was conducted to determine the core psychosocial predictors. In addition to the traditional mechanisms, complex interactive networks among the antecedents and consequences and the reversed effects of consequences were also obtained, where 379 strong association rules were yielded via the Apriori algorithm. RESULTS: The results showed that occupational dust, NO2, heavy metals, heat, vibration, awkward posture, and job stress are significant risk factors associated with individuals' health conditions. Psychological capital, coping style, job demand, social support, organizational support, justice, and culture are core psychosocial predictors of miners. CONCLUSIONS: This study presents a case for identifying the most significant occupational risk factors related to individuals' health, which could be extended and applied to other industries, as working populations around the world are suffering from various chemical, physical, ergonomic, and psychosocial stressors. PRACTICAL APPLICATIONS: Identifying the significant occupational risk factors affecting workers' health conditions is essential for comprehensive occupational health risk assessment and management. Therefore, this study could be important for health management in mines and other industries.

Exploring the determinants of health-promoting behaviors among miners: A text mining and meta-analysis.

The health-promoting behaviors of miners are of great significance to their physical and mental well-being. With a focus on enhancing their overall health, this study aimed to explore the determinants and influencing mechanisms of health-promoting behaviors in miners. Initially, the latent Dirichlet allocation (LDA) model was utilized to extract topical keywords from literature over the last 23 years and to categorize the determinants based on integrating the health promotion model and the health belief model. Subsequently, a meta-analysis was performed based on 51 related empirical research to explore the mechanisms between determinants and health-promoting behaviors. The results indicated that (1) the factors influencing miners' health-promoting behaviors can be divided into four dimensions: physical environment, psychosocial environment, individual characteristics, and health beliefs. (2) Noise was negatively related to health-promoting behaviors, while protective equipment, health culture, interpersonal relationships, health literacy, health attitudes, and income were positively related to health-promoting behaviors. (3) Protective equipment and health literacy were positively related to perceived threat, whereas interpersonal relationships were positively related to perceived benefits. This study sheds light on the mechanisms influencing miners' health-promoting behaviors and could inform behavioral interventions in occupational health.

Alloy nanozyme-reinforced hyaluronic acid-based hydrogel with wound environment-responsive properties for synergistically accelerating infectious wound healing.

The recovery of infectious wound tissues presents a significant global health challenge due to the impediments posed by the harsh healing microenvironment, which includes ongoing bacterial invasion, high oxidative stress, inflammatory response, and impaired angiogenesis. To overcome the above issues, we propose a composite hydrogel based on the multiple-crosslinked mechanism involving the covalent network of CC bonds within catechol and maleic-modified HA (CMHA), the self-assembly network of glycyrrhizic acid (GA), and the metal-polyphenol coordination induced by ZHMCe for accelerating infectious wound healing. The resulting CMHA/GA/ZHMCe hydrogels demonstrate enhanced mechanical, adhesive, antioxidative, and antibacterial properties. Importantly, the hydrogel system possesses wound environment-responsive properties that allow it to adapt to the specific therapeutic requirements of different stages by regulating various enzyme activities in the healing of infected wounds. Furthermore, the biocompatible CMHA/GA/ZHMCe shows the ability to promote cell migration and angiogenesis in vitro while reprogramming macrophages toward an anti-inflammatory phenotype due to the effective release of active ingredients. In vivo experiments confirm that the CMHA/GA/ZHMCe hydrogel significantly enhances infectious wound healing by accelerating re-epithelialization, promoting collagen deposition, regulating inflammation, and contributing to vascularization. These findings underscore the therapeutic potential of our hydrogel dressings for the treatment of bacterially infected cutaneous wound healing.

Aging and TNF induce premature senescence of astrocytes after spinal cord injury via regulating YAP expression.

BACKGROUND: Spinal cord injury (SCI) causes chronic functional impairment in patients. In addition, SCI is tormenting more and more older adults, and those who suffer from SCI often have shorter lifespans. Previous studies have confirmed that overexpression of p75 leads to neuroinflammation and motor dysfunction following spinal cord injury in adult mice. METHODS: As TNF-α is upregulated after SCI, targeting TNF-mediated inflammation may be an attractive option to combat trauma, paving the way for new therapeutic insight. In this study, we evaluated behavioral testing, phenotype of senescent cells, reactive oxygen species (ROS), inflammation and mitochondrial damage in adult (2-month-old) and aged (20-month-old) female wild-type (WT) and p75 knockout (KO) mice. RESULTS: Herein, we hypothesized that aged mice were more prone to death after SCI, but p75 deletion could promote motor/sensory function recovery and improve survival in both adult and aged mice. Further exploration of the underlying mechanism revealed that the expression of p-YAP was reduced in vivo and in vitro, and p75 deletion partially rescued aging-induced astrocytes senescence. CONCLUSION: Taken together, our study have identified an unrecognized function of the p75-YAP pathway on preventing astrocytic aging in vitro and in vivo, which may provide further insights and new targets into slowing spinal cord aging and improving dysfunctional remission and longevity.

Biomimetic nanoplatform with anti-inflammation and neuroprotective effects for repairing spinal cord injury in mice.

Regeneration in the therapeutics of spinal cord injury (SCI) remains a challenge caused by the hyperinflammation microenvironment. Nanomaterials-based treatment strategies for diseases with excellent therapeutic efficacy are actively pursued. Here, we develop biodegradable poly (lactic-co-glycolic acid) nanoparticles (PLGA) obtained by loading celastrol (pCel) for SCI thrapy. Cel, as an antioxidant drug, facilitated reactive oxygen species (ROS) scavenging, and decreased the generation of pro-inflammatory cytokines. To facilitate its administration, pCel is formulated into microspheres by oil-in-water (O/W) emulsion/solvent evaporation technique. The constructed pCel can induced polarization of macrophages and obviously improved lipopolysaccharide (LPS) and interferon-γ (IFN-γ)-induced mitochondrial dysfunction, and increased neurite length in PC12 cells and primary neurons. In vivo experiments revealed that pCel regulated the phenotypic polarization of macrophages, prevented the release of pro-inflammatory cytokines, promoted myelin regeneration and inhibited scar tissue formation, and further improve motor function. These findings indicated that the neuroprotective effect of this artificial biodegradable nanoplatform is benefit for the therapy of SCI. This research opens an exciting perspective for the application of SCI treatment and supports the clinical significance of pCel.

Effects of the genetic knockout of the ß-1,3-galactosyltransferase 2 on spatial learning and neurons in the adult mouse hippocampus and somatosensory cortex.

OBJECTIVE: Glycosyltransferases contribute to the biosynthesis of glycoproteins, proteoglycans and glycolipids and play essential roles in various processes in the brain, such as learning and memory, brain development, neuronal survival and neurodegeneration. ß-1,3-galactosyltransferase 2 (B3galt2) belongs to the ß-1,3-galactosyltransferase gene family and is highly expressed in the brain. Recent studies have indicated that B3galt2 plays a vital role in ischemic stroke through several signaling pathways in a mouse model. However, the function of B3galt2 in the brain remains poorly understood. METHODS: The genotypes of mice were determined by PCR. To verify B3galt2 expression in an adult mouse brain, X-gal staining was performed in 6-month-old B3galt2 heterozygous (B3galt2+/-) mice. Using adult B3galt2 homozygous (B3galt2-/-), heterozygous and wild-type (WT) littermates, spatial learning and memory were determined by the Morris Water Maze test, and neurotoxicity and synaptic plasticity were examined by immunofluorescence. RESULTS: B3galt2 was highly expressed in the adult mouse hippocampus and cortex, especially in the hippocampal dentate gyrus. Compared to that of WT mice, the spatial learning ability of adult B3galt2-/- mice was impaired. B3galt2 mutations also caused neuronal loss and synaptic dysfunction in the hippocampus and somatosensory cortex, and these changes were more obvious in B3galt2-/- mice than in B3galt2+/- mice. CONCLUSIONS: The findings indicate that B3galt2 plays an important role in cognitive function, neuronal maintenance and synaptic plasticity in the adult mouse brain. This study suggests that genetic and/or pharmacological manipulation of glycosyltransferases may be a novel strategy for elucidating the mechanism of and managing various brain disorders.

DC vaccine enhances CAR-T cell antitumor activity by overcoming T cell exhaustion and promoting T cell infiltration in solid tumors.

OBJECTIVE: Great success has been achieved in CAR-T cell immunotherapy in the treatment of hematological tumors. However, it is particularly difficult in solid tumors, because CAR-T is difficult to enter interior and exert long-term stable immune effects. Dendritic cells (DCs) can not only present tumor antigens but also promote the infiltration of T cells. Therefore, CAR-T cells with the help of DC vaccines are a reliable approach to treat solid tumors. METHODS: To test whether DC vaccine could promote CAR-T cell therapy in solid tumors, DC vaccine was co-cultured with MSLN CAR-T cells. The in vitro effects of DC vaccine on CAR-T were assessed by measuring cell proliferation, cell differentiation, and cytokine secretion. Effects of DC vaccine on CAR-T were evaluated using mice with subcutaneous tumors in vivo. The infiltration of CAR-T was analyzed using immunofluorescence. The persistence of CAR-T in mouse blood was analyzed using real-time quantitative PCR. RESULTS: The results showed that DC vaccine significantly enhanced the proliferation potential of MSLN CAR-T cells in vitro. DC vaccines not only promoted the infiltration of CAR-T cells, but also significantly improved the persistence of CAR-T in solid tumors in vivo. CONCLUSION: In conclusion, this study has demonstrated that DC vaccine can promote CAR-T therapy in solid tumors, which provides the possibility of widespread clinical application of CAR-T cells in the future.

Encapsulation of berberine decorated ZnO nano-colloids into injectable hydrogel using for diabetic wound healing.

Chronic wound healing in diabetic patients had been considered a major clinical challenge, so there was an urgent need to establish more effective treatment methods. In this study, we prepared berberine-modified ZnO nano-colloids hydrogel (ZnO-Ber/H) and evaluated its wound healing performance in a diabetic rat. The prepared ZnO-Ber/H had excellent moisturizing, anti-inflammatory and anti-oxidative stress abilities. In vitro, ZnO-Ber/H could effectively up-regulate antioxidant stress factors (Nrf2, HO-1, NQO1) by 4.65-fold, 2.49-fold, 2.56-fold, respectively. In vivo experiments have shown that ZnO-Ber/H could effectively improve the wound healing rate (92.9%) after 15 days of treatment. Meanwhile, the ability of anti-oxidative stress had also been verified in vivo. ZnO-Ber/H down-regulated inflammatory factor (TNF-α, IL-1ß, and IL-6) by 72.8%, 55% and 71% respectively, up-regulated vascular related factors VEGF and CD31 by 3.9-fold and 3.2-fold by Western blot. At the same time, ZnO-Ber/H could promote the expression of EGFR and FGFR, thereby affecting the generation of new epithelial tissue. Based on extensive characterization and biological evaluation, ZnO-Ber/H was expected to be a potential candidate for promoting diabetic wound healing.

XPCS Microrheology and Rheology of Sterically Stabilized Nanoparticle Dispersions in Aprotic Solvents.

X-ray photon correlation spectroscopy (XPCS) microrheology and conventional bulk rheology were performed on silica nanoparticle dispersions associated with battery electrolyte applications to probe the properties of these specific complex materials and to explore the utility of XPCS microrheology in characterizing nanoparticle dispersions. Sterically stabilized shear-thickening electrolytes were synthesized by grafting poly(methyl methacrylate) chains onto silica nanoparticles. Coated silica dispersions containing 5-30 wt % nanoparticles dispersed in propylene carbonate were studied. In general, both XPCS microrheology and conventional rheology showed that coated silica dispersions were more viscous at higher concentrations, as expected. The complex viscosity of coated silica dispersions showed shear-thinning behavior over the frequency range probed by XPCS measurements. However, measurements using conventional mechanical rheometry yielded a shear viscosity with weak shear-thickening behavior for dispersions with the highest concentration of 30% particles. Our results indicate that there is a critical concentration needed for shear-thickening behavior, as well as appropriate particle size and surface polymer chain length, for this class of nanoparticle-based electrolytes. The results of this study can provide insights for comparing XPCS microrheology and bulk rheology for related complex fluids and whether XPCS microrheology can capture expected macroscopic rheological properties by probing small-scale particle dynamics.

Hierarchical assembly in PLA-PEO-PLA hydrogels with crystalline domains and effect of block stereochemistry.

Understanding the development of microstructure (e.g., structures with length scales roughly 0.5-500 µm) in hydrogels is crucial for their use in several biomedical applications. We utilize ultra-small-angle neutron scattering (USANS) and confocal microscopy to explore microstructure of poly(lactide)-poly(ethylene oxide)-poly(lactide) (PLA-PEO-PLA) triblock copolymer hydrogels with varying l/d-lactide ratio. We have previously found that these polymers self-assemble on the nanoscale into micelles. Here, we observe large-scale structures with diverse morphologies, including highly porous self-similar networks with characteristic sizes spanning approximately 120 nm-200 µm. These structural features give rise to power-law scattering indicative of fractal structures in USANS. Mass fractal and surface fractal structures are found for gels with l/d ratios of 80/20 and 50/50, respectively. Confocal microscopy shows microscale water-filled channels and pores that are more clearly evident in gels with a higher fraction of l-lactide in the PLA block as compared to the 50/50 hydrogels. Tuning block stereochemistry may provide a means of controlling the self-assembly and structural evolution at both the nanoscale and microscale, impacting application of these materials in tissue engineering and drug delivery.

DC vaccine enhances CAR-T cell antitumor activity by overcoming T cell exhaustion and promoting T cell infiltration in solid tumors

Objective Great success has been achieved in CAR-T cell immunotherapy in the treatment of hematological tumors. However, it is particularly difficult in solid tumors, because CAR-T is difficult to enter interior and exert long-term stable immune effects. Dendritic cells (DCs) can not only present tumor antigens but also promote the infiltration of T cells. Therefore, CAR-T cells with the help of DC vaccines are a reliable approach to treat solid tumors. Methods To test whether DC vaccine could promote CAR-T cell therapy in solid tumors, DC vaccine was co-cultured with MSLN CAR-T cells. The in vitro effects of DC vaccine on CAR-T were assessed by measuring cell proliferation, cell differentiation, and cytokine secretion. Effects of DC vaccine on CAR-T were evaluated using mice with subcutaneous tumors in vivo. The infiltration of CAR-T was analyzed using immunofluorescence. The persistence of CAR-T in mouse blood was analyzed using real-time quantitative PCR. Results The results showed that DC vaccine significantly enhanced the proliferation potential of MSLN CAR-T cells in vitro. DC vaccines not only promoted the infiltration of CAR-T cells, but also significantly improved the persistence of CAR-T in solid tumors in vivo. Conclusion In conclusion, this study has demonstrated that DC vaccine can promote CAR-T therapy in solid tumors, which provides the possibility of widespread clinical application of CAR-T cells in the future (AU)