Integrative Single-Cell RNA-Seq and ATAC-Seq Analysis of Mouse Corneal Epithelial Cells.

Purpose: Corneal epithelial homeostasis is maintained by coordinated gene expression across distinct cell populations, but the gene regulatory programs underlying this cellular diversity remain to be characterized. Here we applied single-cell multi-omics analysis to delineate the gene regulatory profile of mouse corneal epithelial cells under normal homeostasis. Methods: Single cells isolated from the cornea epithelium (with marginal conjunctiva) of adult mice were subjected to scRNA-seq and scATAC-seq using the 10×Genomics platform. Cell types were clustered by the graph-based visualization method uniform manifold approximation and projection and unbiased computational informatics analysis. The scRNA-seq and scATAC-seq datasets were integrated following the integration pipeline described in ArchR and Seurat. Results: We characterized diverse corneal epithelial cell types based on gene expression signatures and chromatin accessibility. We found that cell type-specific accessibility regions were mainly located at distal regions, suggesting essential roles of distal regulatory elements in determining corneal epithelial cell diversity. Trajectory analyses revealed a continuum of cell state transition and higher coordination between transcription factor (TF) motif accessibility and gene expression during corneal epithelial cell differentiation. By integrating transcriptomic and chromatin accessibility analysis, we identified cell type-specific and shared gene regulation programs. We also uncovered critical TFs driving corneal epithelial cell differentiation, such as nuclear factor I (NFI) family members, Rarg, Elf3. We found that nuclear factor-κB (NF-κB) family members were positive TFs in limbal cells and some superficial cells, but they were involved in regulating distinct biological processes. Conclusions: Our study presents a comprehensive gene regulatory landscape of mouse cornea epithelial cells, and provides valuable foundations for future investigation of corneal epithelial homeostasis in the context of cornea pathologies and regenerative medicine.

The Function of the Mutant p53-R175H in Cancer.

Wild-type p53 is known as "the guardian of the genome" because of its function of inducing DNA repair, cell-cycle arrest, and apoptosis, preventing the accumulation of gene mutations. TP53 is highly mutated in cancer cells and most TP53 hotspot mutations are missense mutations. Mutant p53 proteins, encoded by these hotspot mutations, lose canonical wild-type p53 functions and gain functions that promote cancer development, including promoting cancer cell proliferation, migration, invasion, initiation, metabolic reprogramming, angiogenesis, and conferring drug resistance to cancer cells. Among these hotspot mutations, p53-R175H has the highest occurrence. Although losing the transactivating function of the wild-type p53 and prone to aggregation, p53-R175H gains oncogenic functions by interacting with many proteins. In this review, we summarize the gain of functions of p53-R175H in different cancer types, the interacting proteins of p53-R175H, and the downstream signaling pathways affected by p53-R175H to depict a comprehensive role of p53-R175H in cancer development. We also summarize treatments that target p53-R175H, including reactivating p53-R175H with small molecules that can bind to p53-R175H and alter it into a wild-type-like structure, promoting the degradation of p53-R175H by targeting heat-shock proteins that maintain the stability of p53-R175H, and developing immunotherapies that target the p53-R175H-HLA complex presented by tumor cells.

A fiber-progressive-engagement model to evaluate the composition, microstructure, and nonlinear pseudoelastic behavior of porcine arteries and decellularized derivatives.

The theoretical fiber-progressive-engagement model was proposed to describe the pseudoelastic behavior of an artery pre- and post-decellularization treatments. Native porcine arteries were harvested and decellularized with 0.05% trypsin for 12 h. The uniaxial tensile test data were fitted to the fiber-progressive-engagement model proposed herein. The effects of decellularization on the morphology, structural characteristics, and composition of vessel walls were studied. The experimental stress-strain curve was fitted to the model in the longitudinal and circumferential direction, which demonstrated the adequacy of the proposed model (R2>0.99). The initial and turning strains were similar in the longitudinal and circumferential directions in the aorta, suggesting the occurrence of collagen conjugation in both directions. Discrepancies in the initial and turning strain and initial and stiff modulus in both directions in the coronary artery revealed the anisotropic features of this vessel. Decellularization induced a decrease in the initial and turning strains, a slight change in the initial modulus, and a substantial decrease in the stiffness modulus. The decrease in the initial and turning strain can be attributed to the loss of waviness of collagen bundles because of the considerable decrease in elastin and glycosaminoglycan contents. This simple non-linear model can be used to determine the fiber modulus and waviness degree of vascular tissue. Based on these results, this mechanical test can be used as a screening tool for the selection of an optimized decellularization protocol for arterial tissues. STATEMENT OF SIGNIFICANCE: Decellularized vascular graft has potential in clinical application, such as coronary artery bypass surgery, peripheral artery bypass surgery or microsurgery. An ideal decellularization protocol requires balance in cell removal efficiency and extracellular matrix preserving. Both biochemical and biomechanical properties are crucial to the success of scaffold in cell seeding and animal study. A comprehensive understanding of the composition, microstructure, and mechanical behavior of the arterial wall is the key to the development of decellularized vascular grafts. For this purpose, we proposed this "Fiber-Progressive-Engagement" model to evaluate the microstructure, composition and mechanical properties of porcine coronary artery. The model provides a new perspective regarding the non-linear behavior of arterial tissue and its decellularized derivatives. It can be widely applied to different types of tissues, as demonstrated in the aorta and coronary artery. This model has several advantages; it provides an improved fit of non-linear curves (R2>0.99), can be used to elucidate the pseudoelastic properties of porcine vascular tissues using the concept of fiber engagement, and can estimate an elastic modulus with greater accuracy (compared to the graphical estimation or calculation by simple linear fittings), as well as to plot typical stress-strain curves.

The synthetic flavonoid WYC02-9 inhibits colorectal cancer cell growth through ROS-mediated activation of MAPK14 pathway.

AIM: Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide. In this study, we explored the anti-cancer activity of WYC02-9, a synthetic protoapigenone, on human HCT116 CRC cells. MAIN METHODS: The anti-cancer activity of WYC02-9 and its underlying mechanisms were analyzed using XTT cell proliferation assays, colony formation assays, FACS analysis, annexin V staining, immunoblotting analysis, reactive oxygen species (ROS) generation assays, soft agar assays, a nude mice xenograft study and immunohistochemistry assays. KEY FINDINGS: Data showed that WYC02-9 suppressed CRC cell growth by arresting cells at G2/M and inducing cell death via apoptotic pathways. Further analysis demonstrated that WYC02-9-induced apoptosis was mediated by the activation of a ROS-mediated MAPK14 pathway. An in vivo xenograft study revealed that WYC02-9 enhanced MAP2K3/6 and MAPK14 phosphorylation, induced apoptosis, and suppressed CRC tumor growth. SIGNIFICANCE: WYC02-9 exerts its anti-tumor effect via ROS/MAPK14-induced apoptosis and has the potential to be developed as a chemotherapeutic agent for CRC.