Multiple Signaling Roles of CD3ε and Its Application in CAR-T Cell Therapy.

A T cell receptor (TCR) mediates antigen-induced signaling through its associated CD3ε, δ, γ, and ζ, but the contributions of different CD3 chains remain elusive. Using quantitative mass spectrometry, we simultaneously quantitated the phosphorylation of the immunoreceptor tyrosine-based activation motif (ITAM) of all CD3 chains upon TCR stimulation. A subpopulation of CD3ε ITAMs was mono-phosphorylated, owing to Lck kinase selectivity, and specifically recruited the inhibitory Csk kinase to attenuate TCR signaling, suggesting that TCR is a self-restrained signaling machinery containing both activating and inhibitory motifs. Moreover, we found that incorporation of the CD3ε cytoplasmic domain into a second-generation chimeric antigen receptor (CAR) improved antitumor activity of CAR-T cells. Mechanistically, the Csk-recruiting ITAM of CD3ε reduced CAR-T cytokine production whereas the basic residue rich sequence (BRS) of CD3ε promoted CAR-T persistence via p85 recruitment. Collectively, CD3ε is a built-in multifunctional signal tuner, and increasing CD3 diversity represents a strategy to design next-generation CAR.

The influence of age on prostate cancer screening index.

PURPOSE: This study aimed to identify parameters with a higher diagnostic value for early screening of prostate cancer (PCa) at different ages. MATERIALS AND METHODS: A total of 294 patients were included and divided into two groups according to the age of patients (≤66 and >66 years). Receiver operating characteristic (ROC) curves of total prostate-specific antigen (TPSA), free PSA (FPSA), (F/T)PSA, PSA density (PSAD), PSA-AV score, the ratio of patients' age to prostate volume (AVR) and (F/T)/PSAD were constructed. The area under the ROC curve (AUC) was calculated, and differences in the AUC values among the above-mentioned parameters were compared. RESULTS: There were 121 patients in the ≤66 years age group (benign prostatic hyperplasia BPH, 103 patients; PCa 18 patients) and 173 patients in the >66 years age group (BPH, 100 patients; PCa, 73 patients). In the ≤66 years age group, the AUC value of AVR for PCa diagnosis was the highest; however, there was no statistically significant difference compared with the AUC values of PSAD and (F/T)/PSAD; compared with TPSA, FPSA, (F/T)PSA and PSA-AV, the differences were statistically significant. In the >66 years age group, the AUC values of PSAD and PSA-AV for PCa diagnosis were higher than those of TPSA, FPSA, (F/T)PSA and (F/T)/PSAD, and the difference was statistically significant; however, the difference was not statistically significant when compared with the AUC value of AVR. CONCLUSION: In different age groups, screening indices for PCa diagnosis should be selected according to the age of patients.

The influence of prostate volume on clinical parameters in prostate cancer screening.

PURPOSE: The purpose of the study was to evaluate the diagnostic significance of two new and a few clinical markers for prostate cancer (PCa) at various prostate volumes (PV). METHODS: The study subjects were divided into two groups. Among them, there were 70 cases in the PV ≤30 ml group (benign prostatic hyperplasia [BPH]: 32 cases, PCa: 38 cases) and 372 cases in the PV > 30 ml group (BPH: 277 cases, PCa: 95 cases). SPSS 26.0 and GraphPad Prism 8.0 were used to construct their receiver operating characteristic (ROC) curves for diagnosing PCa and calculating their area under the ROC curve (AUC). RESULTS: In the PV ≤30 ml group, the diagnostic parameters based on prostate-specific antigen (PSA) had a decreased diagnostic significance for PCa. In the PV > 30 ml group, PSAD (AUC = 0.709), AVR (AVR = Age/PV, AUC = 0.742), and A-PSAD (A-PSAD = Age×PSA/PV, AUC = 0.736) exhibited moderate diagnostic significance for PCa, which was better than PSA-AV (AUC = 0.672), free PSA (FPSA, AUC = 0.509), total PSA (TPSA, AUC = 0.563), (F/T) PSA (AUC = 0.540), and (F/T)/PSAD (AUC = 0.663). Compared with AVR, A-PSAD exhibited similar diagnostic significance for PCa, but higher than PSA density (PSAD). CONCLUSIONS: Choosing appropriate indicators for different PVs could contribute to the early screening and diagnosis of PCa. The difference in the diagnostic value of two new indicators (A-PSAD and AVR), and PSAD for PCa may require further validation by increasing the sample size.

Clinical research analysis based on prostate cancer screening diagnosis.

This study aimed to analyse the clinical characteristics and risk factors of patients with positive prostate biopsy at 4-20 ng/mL of prostate-specific antigen (PSA), construct a new parameter based on this characteristics and assess its diagnostic value for prostate cancer (PCa). Logistic regression analysis was used to clarify the risk factors of PCa, and a new parameter based on the results was constructed. Compare the diagnostic value of various diagnostic parameters for PCa. Logistic multivariate regression analysis revealed that age (OR, 5.269; 95%CI, 2.762-10.050), comorbid diabetes (OR, 2.437; 95%CI, 1.162-5.111), PSA (OR, 2.462; 95%CI, 1.198-5.059) and prostate volume (PV) (OR, 0.227; 95%CI, 0.100-0.516) are risk factors for PCa. The age, PSA and PV of patients were combined to construct a new parameter, that is A-PSAD = (age × total PSA [TPSA])/PV]. The area under the receiver-operating characteristic curve(AUC) of A-PSAD (0.728) for PCa diagnosis was higher than the AUCs of TPSA (0.581), free prostate-specific antigen (0.514), (F/T)PSA (0.535) and PSAD (0.696), with significant differences. Age, history of diabetes, TPSA and PV are risk factors for PCa(PSA:4-20ng/mL); in addition, A-PSAD has a moderate diagnostic value for PCa and may become a new indicator for PCa screening.

Bioorthogonal "Labeling after Recognition" Affording an FRET-Based Luminescent Probe for Detecting and Imaging Caspase-3 via Photoluminescence Lifetime Imaging.

Bis-labeling with a luminescent energy donor/acceptor pair onto biological substrates affords probes which give FRET readouts for the detection of interaction partners. However, the covalently bound luminophores bring about steric hindrance and nonspecific interaction, which probably perturb the biological recognition. Herein, we designed a highly sensitive and specific "labeling after recognition" sensing approach, where luminophore labeling occurred after the biological recognition. Taking the cutting enzyme caspase-3 as an example, we demonstrated the detection of its catalytic activity in solution and apoptotic cells using the tetrapeptide motif Asp-Glu-Val-Asp (DEVD) as the cleavable substrate, and an iridium(III) complex and a rhodamine derivative as the energy donor/acceptor pair. The DEVD tetrapeptide was modified with an azide and a GK-norbornylene groups at the amino and carboxyl terminuses, respectively, which allowed donor/acceptor bis-labeling via two independent catalysis-free bioorthogonal reactions. The phosphorescence lifetime of the iridium(III) complex was quenched upon bis-labeling owing to the intracellular FRET to the rhodamine derivative, and significantly elongated upon the peptide being catalytically cleaved by caspase-3. Interestingly, the sensitivity and efficiency of the lifetime responses were much higher in the "labeling after recognition" sensing approach. Molecular docking analysis showed that the steric hindrance and nonspecific interactions partially inhibited the biological recognition of the DEVD substrate by caspase-3. The imaging of the catalytic activity of caspase-3 in apoptotic cells was demonstrated via photoluminescence lifetime imaging microscopy. Lifetime analysis not only confirmed the occurrence of intracellular bioorthogonal bis-labeling and catalytic cleavage, but also showed the extent to which the two dynamic processes occurred.

Radial basis collocation method with parameters optimized for estimating pollutant release history.

The identification of pollutant source release history in rivers is important for emergency response of pollution accidents and formulating remediation strategies. Space-time radial basis collocation method (RBCM), as a meshless method with strong applicability, can directly estimate the release history from the concentration data measured at downstream observation sites. However, the uncertainty of specific parameters in space-time RBCM is the main factor affecting the accuracy of estimation. Therefore, a way to solve the parameters efficiently and accurately is essential. For this purpose, a new model which combines space-time RBCM and differential evolution algorithm (DEA) is established to identify the source release history. First of all, efficient parameter optimizer DEA is introduced to search the parameters that affect the estimation accuracy of space-time RBCM. Then, a new loss function considering the imbalance configuration of RBCM nodes is designed to ensure the rationality of the parameters obtained by DEA. The results of numerical cases and real field case show that the proposed method can accurately estimate the real release history with low time consumption. It is also demonstrated that DEA is more efficient than k-fold cross-validation in searching the optimal parameters for space-time RBCM, and the parameters obtained from the new loss function can make the estimated release history more precise.

Study on the Role and Mechanism of LncRNA ZFasL in Renal Carcinoma.

Background: Renal carcinoma is the 7th most common cancer in the world, with the 7th and 6th highest incidence and mortality rates worldwide. Although great progress has been made in the diagnosis and treatment of renal carcinoma, its prognosis is still unsatisfactory. It is important to study the molecular mechanisms of renal carcinoma occurrence and development and to find potential therapeutic targets. Objective: The main objective is to investigate the effects of long noncoding RNA (lncRNA) ZFAS1 (lncZFAS1) on the proliferation, apoptosis, and migration of renal carcinoma cells and to preliminarily explore its mechanism. Methods: A qRT-PCR method was used to detect the expression of lncZFAS1 in renal carcinoma tissues and renal carcinoma cells. After shRNA interference with lncZFAS1 expression, the effects of lncZFAS1 on cell proliferation, apoptosis, migration, and invasion were detected by CCK-8 method, flow cytometry, scratch test, and Transwell assay. The effect of the knockdown of lncZFAS1 on the growth of transplanted tumors was examined. The expression of lncZFAS1 in renal carcinoma tissues and renal carcinoma cells was significantly higher than that in paracancerous tissues and normal esophageal epithelial cells. Knockdown of lncZFAS1 significantly inhibited the proliferation, migration, and invasive ability of renal carcinoma cells; upregulated miR-150-5P expression and downregulated HMGA2 expression in renal carcinoma cells; and significantly inhibited the growth of transplanted tumors in nude mice. Conclusion: Upregulation of miR-150-5P expression was detected after knockdown of lncZFAS1 in renal carcinoma cells, while both mRNA and protein expression levels of HMGA2 were decreased. lncZFAS1 can promote the proliferation and migration of renal carcinoma cells, and the mechanism may be related to the regulation of the miR-150-5P/HMGA2 molecular axis.

Cholesterol Metabolism in T Cells.

Compartmentalization and spatial control of biochemical reactions is the foundation of cell-based life on earth. The lipid bilayer system employed by eukaryote cells not only keeps them separate from the environment but also provides a platform for key receptors to sense and interact with outside factors. Arguably one of the cell types most reliant on interactions of this kind, immune cells depend on their membrane to keep functioning properly. In this review, the influence of variation in cholesterol levels, a key component of lipid bilayer stability, on T cells will be discussed in detail. In comparison to other cells, T cells must be able to undergo rapid activation followed by proliferation. Furthermore, receptor colocalization is an important mechanism in this activation process. The impact of cholesterol availability on the processes of T cell proliferation and receptor sensitivity, as well as its potential for immunomodulation in disease treatment will be considered.