Exploring the therapeutic potential of precision T-Cell Receptors (TCRs) in targeting KRAS G12D cancer through in vitro development

Objectives:The Kirsten rat sarcoma virus(KRAS)G12D oncogenic mutation poses a significant challenge in treating solid tumors due to the lack of specific and effective therapeutic interventions.This study aims to explore innovative approaches in T cell receptor(TCR)engineering and characterization to target the KRAS G12D7-16 mutation,providing potential strategies for overcoming this therapeutic challenge.Methods:In this innovative study,we engineered and characterized two T cell receptors(TCRs),KDA11-01 and KDA11-02 with high affinity for the KRAS G12D7-16 mutation.These TCRs were isolated from tumor-infiltrating lymphocytes(TILs)derived from tumor tissues of patients with the KRAS G12D mutation.We assessed their specificity and anti-tumor activity in vitro using various cancer cell lines.Results:KDA11-01 and KDA11-02 demonstrated exceptional specificity for the HLA-A*11:01-restricted KRAS G12D7-16 epitope,significantly inducing IFN-γrelease and eliminating tumor cells without cross-reactivity or alloreactivity.Conclusions:The successful development of KDA11-01 and KDA11-02 introduces a novel and precise TCR-based therapeutic strategy against KRAS G12D mutation,showing potential for significant advancements in cancer immunotherapy.

Effects of probiotics and its fermented milk on constipation: a systematic review

This study reviewed clinical evidence of probiotics-mediated effects on constipation.Four databases were used in the literature searches,namely Cochrane Library,Embase,Pubmed,and Web of Science.The last retrieval time was March 2021.Collected data were analyzed by Review Manager(version 5.3)and Stata(version 14).The primary search retrieved 8418 articles after removing duplicates.Eventually,26 studies were included in the meta-analysis.Administration of probiotics signifi cantly increased stool frequency(MD:1.15 times/week;95%CI:0.59 to 1.70;P<0.0001),improved bloating(MD:-0.48;95%CI:-0.89 to-0.07;P=0.02),reduced abdominal pain(MD:-0.71;95%CI:-1.25 to-0.16;P=0.01),and improved quality of life(MD:-0.73;95%CI:-1.37 to-0.10;P=0.02).However,the clinical effect of intake of probiotics on stool consistency was non-signifi cant(MD:0.07;95%CI:-0.34 to 0.48;P=0.73).Based on our analysis,probiotic supplements were effective in relieving constipation through symptom improvement.The effectiveness of dosage forms of intervention ranked:capsules>others(tablets,powder)>fermented milk.

Construction of an alternative NADPH regeneration pathway improves ethanol production in Saccharomyces cerevisiae with xylose metabolic pathway

Full conversion of glucose and xylose from lignocellulosic hydrolysates is required for obtaining a high ethanol yield.However,glucose and xylose share flux in the pentose phosphate pathway(PPP)and glycolysis pathway(EMP),with glucose having a competitive advantage in the shared metabolic pathways.In this work,we knocked down ZWF1 to preclude glucose from entering the PPP.This reduced the[NADPH]level and disturbed growth on both glucose or xylose,confirming that the oxidative PPP,which begins with Zwf1p and ultimately leads to CO_(2) production,is the primary source of NADPH in both glucose and xylose.Upon glucose depletion,gluconeogenesis is necessary to generate glucose-6-phosphate,the substrate of Zwf1p.We re-established the NADPH regeneration pathway by replacing the endogenous NAD^(+)-dependent glyceraldehyde-3-phosphate dehydrogenase(GAPDH)gene TDH3 with heterogenous NADP^(+)-GAPDH genes GDH,gapB,and GDP1.Among the resulting strains,the strain BZP1(zwf1Δ,tdh3::GDP1)exhibited a similar xylose consumption rate before glucose depletion,but a 1.6-fold increased xylose consumption rate following glucose depletion compared to the original strain BSGX001,and the ethanol yield for total consumed sugars of BZP1 was 13.5%higher than BSGX001.This suggested that using the EMP instead of PPP to generate NADPH reduces the wasteful metabolic cycle and excess CO_(2)^(++) release from oxidative PPP.Furthermore,we used a copper-repressing promoter to modulate the expression of ZWF1 and optimize the timing of turning off the ZWF1,therefore,to determine the competitive equilibrium between glucose-xylose co-metabolism.This strategy allowed fast growth in the early stage of fermentation and low waste in the following stages of fermentation.

Akt Inhibitor Perifosine Prevents Epileptogenesis in a Rat Model of Temporal Lobe Epilepsy

Accumulating data have revealed that abnormal activity of the mTOR(mammalian target of rapamycin)pathway plays an important role in epileptogenesis triggered by various factors. We previously reported that pretreatment with perifosine, an inhibitor of Akt(also called protein kinase B), abolishes the rapamycin-induced paradoxical increase of S6 phosphorylation in a rat model induced by kainic acid(KA). Since Akt is an upstream target in the mTOR signaling pathway, we set out to determine whether perifosine has a preventive effect on epileptogenesis. Here, we explored the effect of perifosine on the model of temporal epilepsy induced by KA in rats and found that pretreatment with perifosine had no effect on the severity or duration of the KA-induced status epilepticus. However, perifosine almost completely inhibited the activation of p-Akt and p-S6 both acutely and chronically following the KA-induced status epilepticus.Perifosine pretreatment suppressed the KA-induced neuronal death and mossy fiber sprouting. The frequency of spontaneous seizures was markedly decreased in rats pretreated with perifosine. Accordingly, rats pretreated with perifosine showed mild impairment in cognitive functions. Collectively, this study provides novel evidence in a KA seizure model that perifosine may be a potential drug for use in anti-epileptogenic therapy.

Engineering of Saccharomyces cerevisiae for co-fermentation of glucose and xylose:Current state and perspectives

The use of non-food lignocellulosic biomass to produce ethanol fits into the strategy of a global circular economy with low dependence on fossil energy resources.Xylose is the second most abundant sugar in lignocellulosic hydrolysate,and its utilization in fermentation is a key issue in making the full use of raw plant materials for ethanol production and reduce production costs.Saccharomyces cerevisiae is the best ethanol producer but the organism is not a native xylose user.In recent years,great efforts have been made in the construction of xy-lose utilizing S.cerevisiae strains by metabolic and evolutionary engineering approaches.In addition,managing global transcriptional regulation works provides an effective means to increase the xylose utilization capacity of recombinant strains.Here we review the common strategies and research advances in the research field in order to facilitate the researches in xylose metabolism and xylose-based fermentation.

1D/2D composite subnanometer channels for ion transport:The role of confined water

As a mass transport media,water is an alternative of organic solvent applied in rechargeable batteries,due to its unique properties,including fast ionic migration,easy-processibility,economic/environmental friendliness,and flame retardancy.However,due to the high activity of water molecules in aqueous electrolytes,the corrosion of metal anode,side reactions,and inferior metal electrodeposition behavior leads to unstable cycling performance,poor Coulombic efficiency(CE),and early-staged failure of batteries.Despite several attempts to regulate the activity of water,migration of ions is sacrificed,due to the limited methods to control the water states.Herein,we developed a subnanoscale confinement strategy based on a nacre-like structure to modulate the activity of water in the solid electrolytes.By tuning the ratio between the two-dimensional(2D)vermiculite and one-dimensional(1D)cellulose nanofibers(CNFs),the capillary size in the 1D/2D structure is altered to achieve a fast Zn^(2+)transport.Our dielectric relaxation and molecular dynamics studies indicate that the enhanced Zn^(2+)conductivity is attributed to the fast water relaxation in the precisely defined 1D/2D capillary.Taking advantage of the regulated activity of the confined water in 2D capillary,the composite vermiculite membrane can suppress the corrosion and side reactions between Zn electrode and water molecular,endowing a reversible Zn^(2+)stripping/plating behavior and a stable cycling performance for 900 h.Based on our confinement strategy to control the water states by 1D/2D structures,this work will open an avenue toward aqueous energy storage devices with excellent reversibility,high safety,and long-term stability.

Recurring Real-Time Monitoring of Inflammations in Living Mice with a Chemiluminescent Probe for Hypochlorous Acid

Probes for in vivo imaging of hypochlorous acid(HOCl),one of the most important reactive oxygen species in innate immunity,are urgently needed to understand the pathogenesis of autoimmune and neuroinflammatory disorders.As a strong oxidant,HOCl could bleach near-infrared sensors and inactivate luciferase readily,making in vivo imaging overwhelmingly challenging.Via fine-tuning of a selective HOCl sensing moiety,HOCl stable spacer,and bright chemiluminescent scaffold,we have developed HOCl-CL-510 as a highly selective and sensitive probe for HOCl detection both in vitro and in vivo.In particular,we achieved recurring real-time monitoring of HOCl in both acute and chronic inflammation models in living mice,providing a new chemical tool for dynamic monitoring of disease development with reduced usage of experimental animals.