Advances in base editing with an emphasis on an AAV-based strategy.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based base editors have been developed for precisely installing point mutations in genomes with high efficiency. Two editing systems of cytosine base editors (CBEs) and adenine base editors (ABEs) have been developed for conversion of C.G-to-T.A and A.T-to-G.C, respectively, showing the prominence in genomic DNA correction and mutation. Here, we summarize recent optimized approaches in improving base editors, including the evolution of Cas proteins, the choice of deamination enzymes, modification on linker length, base-editor expression, and addition of functional domains. Specifically, in this paper we highlight a strategy of split-intein mediated base-editor reconstitution for its adeno-associated virus (AAV) delivery. The purpose of this article is to offer readers with a better understanding of AAV-mediated base editors, and facilitate them to use this tool in in vivo experiments and potential clinical applications.

Direct Evidence for the Critical Role of 5,6-Dihydroxyindole in Polydopamine Deposition and Aggregation.

The definitive role of the intermediate 5,6-dihydroxyindole (DHI) in the formation of polydopamine (PDA) coatings from aqueous dopamine (DA) has not been clearly elucidated and remains highly controversial. Our foray into this debate as reported in this study agrees with some reported assertions that DHI-based coatings are not synonymous with PDA coatings. Our conclusion arises from a systematic comparison of the components and properties of DHI-based coatings and PDA coatings. In addition, through careful copolymerization studies of DA and DHI, our studies reported herein unequivocally suggest that both DA and DHI are partial building blocks for PDA formation. Our results also provide additional evidence of the critical role of DHI in controlling the thickness of PDA coatings, through competitive events between PDA aggregation in solutions and deposition onto substrates. These findings highlight the complex interplay between both DHI and uncyclized DA moieties in the formation of adhesive catechol/amine materials.

Development and Validation of a Novel Ferroptosis-Related LncRNA Signature for Predicting Prognosis and the Immune Landscape Features in Uveal Melanoma.

Background: Ferroptosis is a newly iron-dependent mode of programmed cell death that is involved in a variety of malignancies. But no research has shown a link between ferroptosis-related long non-coding RNAs (FRLs) and uveal melanoma (UM). We aimed to develop a predictive model for UM and explore its potential function in relation to immune cell infiltration. Methods: Identification of FRLs was performed using the Cancer Genome Atlas (TCGA) and FerrDb databases. To develop a prognostic FRLs signature, univariate Cox regression and least absolute shrinkage and selection operator (LASSO) were used in training cohort. Kaplan-Meier (K-M) and receiver operating characteristic (ROC) curve analyses were used to assess the reliability of the risk model. The immunological functions of FRLs signature were determined using gene set enrichment analysis (GSEA). Immunological cell infiltration and immune treatment were studied using the ESTIMATE, CIBERSORT, and ssGSEA algorithms. Finally, in vitro assays were carried out to confirm the biological roles of FRLs with known primer sequences (LINC00963, PPP1R14B.AS1, and ZNF667.AS1). Results: A five-genes novel FRLs signature was identified. The mean risk score generated by this signature was used to create two risk groups. The high-risk score UM patients had a lower overall survival rate. The area under the curve (AUC) of ROC and K-M analysis further validated the strong prediction capacity of the prognostic signature. Immune cells such as memory CD8 T cells, M1 macrophages, monocytes, and B cells showed a substantial difference between the two groups. GSEA enrichment results showed that the FRLs signature was linked to certain immune pathways. Moreover, UM patients with high-risk scores were highly susceptible to several chemotherapy drugs, such as cisplatin, imatinib, bortezomib, and pazopanib. Finally, the experimental validation confirmed that knockdown of three identified lncRNA (LINC00963, PPP1R14B.AS1, and ZNF667.AS1) suppressed the invasive ability of tumor cells in vitro. Conclusion: The five-FRLs (AC104129.1, AC136475.3, LINC00963, PPP1R14B.AS1, and ZNF667.AS1) signature has effects on clinical survival prediction and selection of immunotherapies for UM patients.

Therapeutic implications of nanodrug and tissue engineering for retinal pigment epithelium-related diseases.

The retinal pigment epithelium (RPE), as a single layer of cells that performs multiple functions posteriorly in the eye, is a promising target site for the prevention and treatment of several clinical diseases, including proliferative diabetic retinopathy, age-related macular degeneration, chorionic neovascularization, and retinitis pigmentosa. In recent decades, several nanodrug delivery platforms and tissue-engineered RPE have been widely developed to treat RPE-related diseases. This work summarizes the recent advances in nanoplatforms and tissue engineering scaffolds developed in these fields. The diseases associated with pathological RPE and their common therapy strategies are first introduced. Then, the recent progress made with a variety of drug delivery systems is presented, with an emphasis on the modification strategies of nanomaterials for targeted delivery. Tissue engineering-mediated RPE transplantation for treating these diseases is subsequently described. Finally, the clinical translation challenges in these fields are discussed in depth. This article will offer readers a better understanding of emerging nanotechnology and tissue engineering related to the treatment of RPE-related diseases and could facilitate their widespread use in experiments in vivo and in clinical applications.

Distinct Molecular Subtypes of Diffuse Large B Cell Lymphoma Patients Treated with Rituximab-CHOP Are Associated with Different Clinical Outcomes and Molecular Mechanisms.

OBJECTIVE: Our purpose was to characterize distinct molecular subtypes of diffuse large B cell lymphoma (DLBCL) patients treated with rituximab-CHOP (R-CHOP). METHODS: Two gene expression datasets of R-CHOP-treated DLBCL patients were downloaded from GSE10846 (n = 233, training set) and GSE31312 (n = 470, validation set) datasets. Cluster analysis was presented via the ConsensusClusterPlus package in R. Using the limma package, differential expression analysis was utilized to identify feature genes. Kaplan-Meier survival analysis was presented to compare the differences in the prognosis between distinct molecular subtypes. Correlation between molecular subtypes and clinical features was analyzed. Based on the sets of highly expressed genes, biological functions were explored by gene set enrichment analysis (GSEA). Several feature genes were validated in the molecular subtypes via qRT-PCR and western blot. RESULTS: DLBCL samples were clustered into two molecular subtypes. Samples in subtype I displayed poorer overall survival time in the training set (p < 0.0001). Consistently, patients in subtype I had shorter overall survival (p = 0.0041) and progression-free survival time (p < 0.0001) than those in subtype II. Older age, higher stage, and higher international prognostic index (IPI) were found in subtype I. In subtype I, T cell activation, lymphocyte activation, and immune response were distinctly enriched, while cell adhesion, migration, and motility were significantly enriched in subtype II. T cell exhaustion-related genes including TIM3 (p < 0.001), PD-L1 (p < 0.0001), LAG3 (p < 0.0001), CD160 (p < 0.001), and CD244 (p < 0.001) were significantly highly expressed in subtype I than subtype II. CONCLUSION: Two molecular subtypes were constructed in DLBCL, which were characterized by different clinical outcomes and molecular mechanisms. Our findings may offer a novel insight into risk stratification and prognosis prediction for DLBCL patients.

Smart nano-micro platforms for ophthalmological applications: The state-of-the-art and future perspectives.

Smart nano-micro platforms have been extensively applied for diverse biomedical applications, mostly focusing on cancer therapy. In comparison with conventional nanotechnology, the smart nano-micro matrix can exhibit specific response to exogenous or endogenous triggers, and thus can achieve multiple functions e.g. site-specific drug delivery, bio-imaging and detection of bio-molecules. These intriguing techniques have expanded into ophthalmology in recent years, yet few works have been summarized in this field. In this work, we provide the state-of-the-art of diverse nano-micro platforms based on both the conventional materials (e.g. natural or synthetic polymers, lipid nanomaterials, metal and metal oxide nanoparticles) and emerging nanomaterials (e.g. up-conversion nanoparticles, quantum dots and carbon materials) in ophthalmology, with some smart nano/micro platformers highlighted. The common ocular diseases studied in the field of nano-micro systems are firstly introduced, and their therapeutic method and the related drawback in clinic treatment are presented. The recent progress of different materials for diverse ocular applications is then demonstrated, with the representative nano- and micro-systems highlighted in detail. At last, an in-depth discussion on the clinical translation challenges faced in this field and the future direction are provided. This review would allow the researchers to design more smart nanomedicines in a more rational manner for specific ophthalmology applications.

The Chemistry of Bioinspired Catechol(amine)-Based Coatings.

Surface coatings are widely used for the protection of underlying materials from erosion or contamination by the external environment, with biomimetic organic coatings based on catecholamine chemistry gaining prominence in recent years. Such coatings have found use in the biomedical field, e.g., in diagnostics, implant manufacturing, and biosensing, with coatings based on polydopamine (PDA) being the most popular. This Review aims to summarize the chemistry of catechol(amine) coatings, in particular the adhesion and cohesion properties of catecholamine-based coatings. This will allow for the design and synthesis of new polymers and coating materials in a more rational manner, enabling the selection of parameters and conditions to precisely control the structure of the materials formed. Particular attention is paid to the formation mechanism, structure, and variables affecting the properties of PDA, which is the most widely reported catechol(amine) coating. The use of other catechol(amine) precursors to synthesize biomimetic coatings is also discussed. A summary of the different methods reported in the literature to effect specific chemical properties on catechol(amine) coatings will allow the reader to best choose the technique to tailor coating properties for specific applications.

In situ insights into the nanoscale deposition of 5,6-dihydroxyindole-based coatings and the implications on the underwater adhesion mechanism of polydopamine coatings.

The biomimetic coating polydopamine (PDA) has emerged as a promising coating material for various applications. However, the mechanism of PDA deposition onto surfaces is not fully understood, and the coating components of PDA and its relation to the putative intermediate 5,6-dihydroxyindole (DHI) are still controversial. This investigation discloses the deposition mechanisms of dopamine (DA)-based coatings and DHI-based coatings onto silicon surfaces by monitoring the nanoscale deposition of both coatings in situ using high-precision ellipsometry. We posit that the rapid and instantaneous nano-deposition of PDA coatings onto silicon surface in the initial stages critically involves the oxidation of DHI and/or its related oligomers. Our studies also show that the slow conversion of DA to DHI in PDA solution and the coupling between DA and DHI-derived precursors could be crucial for subsequent PDA coating growth. These findings elucidate the critical role of DHI, acting as an 'initiator' and a 'cross linker', in the PDA coating formation. Overall, our study provides important information on the early stage nano-deposition behavior in the construction of PDA coatings and DHI-based coatings.

A one step method for the functional and property modification of DOPA based nanocoatings.

Biomimetic poly(catecholamine) coatings have gained much attention in recent years due to their versatility as functional materials. Despite this, only limited methods are available to modify the function and property of poly(catecholamine) coatings, primarily through post-modification methods. Our approach reported herein provides a simple approach to the fabrication of novel functionalized poly(catecholamine) coatings. The strategy employs the copolymerization of N-Ac-3,4-dihydroxyphenylalanine methyl ester (NADOPAMe) with nucleophilic additives, giving rise to nano-coatings on various surfaces including plastic, metal, glass and polymers. With the appropriate choice of nucleophilic additives, coatings with desired properties can be achieved. This is demonstrated through the fabrication of a redox responsive coating based on NADOPAMe with cysteamine as additive, which shows a concentration-dependent glutathione (GSH) responsive behavior. The ability to utilize this as a controlled release system is also demonstrated.

Postoperative Regulatory T-Cells and Natural Killer Cells in Stage I Nonsmall Cell Lung Cancer Underwent Video-assisted Thoracoscopic Lobectomy or Thoracotomy.

BACKGROUND: Regulatory T-cells (Treg) play key roles in suppressing cell-mediated immunity in cancer patients. Little is known about perioperative Treg fluctuations in nonsmall cell lung cancer (NSCLC). Video-assisted thoracoscopic (VATS) lobectomy, as a minimal invasive procedure for treating NSCLC, may have relatively less impact on the patient's immune system. This study aimed to observe perioperative dynamics of circulating Treg and natural killer (NK) cell levels in NSCLC patients who underwent major lobectomy by VATS or thoracotomy. METHODS: Totally, 98 consecutive patients with stage I NSCLC were recruited and assigned into VATS or thoracotomy groups. Peripheral blood samples were taken on 1-day prior to operation, postoperative days (PODs) 1, 3, 7, 30, and 90. Circulating Treg and NK cell counts were assayed by flow cytometry, defined as CD4 + CD25 + CD127 low cells in CD4 + lymphocytes and CD56 + 16 + CD3- cells within CD45 + leukocytes respectively. With SPSS software version 21.0 (SPSS Inc., USA), differences between VATS and thoracotomy groups were determined by one-way analysis of variance (ANOVA), and differences between preoperative baseline and PODs in each group were evaluated by one-way ANOVA Dunnett t-test. RESULTS: In both groups, postoperative Treg percentages were lower than preoperative status. No statistical difference was found between VATS and thoracotomy groups on PODs 1, 3, 7, and 30. On POD 90, Treg percentage in VATS group was significantly lower than in thoracotomy group (5.26 ± 2.75 vs. 6.99 ± 3.60, P = 0.012). However, a higher level of NK was found on all PODs except on POD 90 in VATS group, comparing to thoracotomy group. CONCLUSIONS: Lower Treg level on POD 90 and higher NK levels on PODs 1, 3, 7, 30 in VATS group might imply better preserved cell-mediated immune function in NSCLC patients, than those in thoracotomy group.