Characterization of extrachromosomal circular DNAs in plasma of patients with clear cell renal cell carcinoma.

BACKGROUND AND PURPOSE: Extrachromosomal circular DNAs (eccDNAs) have been recognized for their significant involvement in numerous biological processes. Nonetheless, the existence and molecular characteristics of eccDNA in the peripheral blood of patients diagnosed with clear cell renal cell carcinoma (ccRCC) have not yet been reported. Our aim was to identify potentially marked plasma eccDNAs in ccRCC patients. METHODS AND MATERIALS: The detection of plasma eccDNA in ccRCC patients and healthy controls was performed using the Tn5-tagmentation and next-generation sequencing (NGS) method. Comparisons were made between ccRCC patients and healthy controls regarding the distribution of length, gene annotation, pattern of junctional nucleotide motif, and expression pattern of plasma eccDNA. RESULTS: We found 8,568 and 8,150 plasma eccDNAs in ccRCC patients and healthy controls, respectively. There were no statistical differences in the length distribution, gene annotation, and motif signature of plasma eccDNAs between the two groups. A total of 701 differentially expressed plasma eccDNAs were identified, and 25 plasma eccDNAs with potential diagnostic value for ccRCC have been successfully screened. These up-regulated plasma eccDNAs also be indicated to originate from the genomic region of the tumor-associated genes. CONCLUSION: This work demonstrates the characterization of plasma eccDNAs in ccRCC and suggests that the up-regulated plasma eccDNAs could be considered as a promising non-invasive biomarker in ccRCC.

DFT Study on the Mechanism of the Palladium-Catalyzed [3 + 2] Annulation of Aromatic Amides with Maleimides via Benzylic and meta-C-H Bond Activation: Role of the External Ligand Ac-Gly-OH.

The mechanism of the Ac-Gly-OH-assisted palladium-catalyzed [3 + 2] annulation of aromatic amides with maleimides is investigated using density functional theory calculations. The results show that the reaction undergoes the sequential steps of N-H bond deprotonation, first benzylic C-H bond activation, maleimide insertion, second meta-C-H bond activation, reductive elimination, and oxidation. The external ligand Ac-Gly-OH acts as the internal base for hydrogen abstraction in the first benzylic C-H bond activation. The maleimide insertion step is found to be the rate-determining step. Based on the nearly same energetic span of the two pathways to generate the enantio products, the computational results are consistent with the experimental observation that the terminal [3 + 2] annulation products are racemic when using an achiral ligand. These calculation results disclose the detailed reaction mechanism and shed light on some experimental ambiguities.

DFT Study on Mechanism of Ni-Al Bimetallic-Catalyzed C-H Cyclization to Construct Tricyclic Imidazoles: Roles of NHC Ligand and AlMe3.

The mechanism of the Ni-Al bimetallic-catalyzed C-H cyclization to construct tricyclic imidazoles is investigated using density functional theory calculations. The calculation result shows that the reaction mechanism involves sequential steps of substrate coordination, ligand-to-ligand hydrogen transfer (LLHT), and C-C reductive elimination to produce the final product tricyclic imidazole. The LLHT step is calculated to be the rate-determining step. The oxidative addition of the benzimidazole C-H bond to the Ni center and the insertion of the alkene into the Ni-H bond occur concertedly in the LLHT step. The effects of N-heterocyclic carbene (NHC) ligands and AlMe3 on the reactivity and regioselectivity were also analyzed. These calculation results shed light on some ambiguous suggestions from experiments.

A Histomorphometric and Computational Investigation of the Stabilizing Role of Pectinate Ligaments in the Aqueous Outflow Pathway.

Murine models are commonly used to study glaucoma, the leading cause of irreversible blindness. Glaucoma is associated with elevated intra-ocular pressure (IOP), which is regulated by the tissues of the aqueous outflow pathway. In particular, pectinate ligaments (PLs) connect the iris and trabecular meshwork (TM) at the anterior chamber angle, with an unknown role in maintenance of the biomechanical stability of the aqueous outflow pathway, thus motivating this study. We conducted histomorphometric analysis and optical coherence tomography-based finite element (FE) modeling on three cohorts of C57BL/6 mice: "young" (2-6 months), "middle-aged" (11-16 months), and "elderly" (25-32 months). We evaluated the age-specific morphology of the outflow pathway tissues. Further, because of the known pressure-dependent Schlemm's canal (SC) narrowing, we assessed the dependence of the SC lumen area on varying IOPs in age-specific FE models over a physiological range of TM/PL stiffness values. We found age-dependent changes in morphology of outflow tissues; notably, the PLs were more developed in older mice compared to younger ones. In addition, FE modeling demonstrated that murine SC patency is highly dependent on the presence of PLs and that increased IOP caused SC collapse only with sufficiently low TM/PL stiffness values. Moreover, the elderly model showed more susceptibility to SC collapse compared to the younger models. In conclusion, our study elucidated the previously unexplored role of PLs in the aqueous outflow pathway, indicating their function in supporting TM and SC under elevated IOP.

Preparation of Yellowing-Resistant Waterborne Polyurethane Modified with Disulfide Bonds.

Waterborne polyurethane, renowned for its lightweight properties, excellent insulation capabilities, and corrosion resistance, has found extensive application in fields such as construction, automotive, leather, and thermal insulation. Nevertheless, during operational usage, waterborne polyurethane materials, akin to other polymeric substances, are susceptible to oxidative aging manifestations like yellowing, cracking, and diminished mechanical performance, significantly curtailing their utility. Consequently, the synthesis of yellowing-resistant polyurethane assumes pivotal significance. This study integrates dynamic reversible reactions into the synthesis process of polyurethane by introducing the dynamic reversible compound 2-hydroxyethyl disulfide as a chain extender, alongside the incorporation of a UV absorber to enhance the polyurethane's resistance to yellowing. When the disulfide bonds absorb heat, they undergo cleavage, yielding thiols that spontaneously recombine into disulfide bonds at ambient temperatures, allowing for the continuous breaking and reformation of disulfide bonds to absorb heat. Concurrently, in collaboration with the UV absorber, the detrimental effects of ultraviolet radiation on the polyurethane material are mitigated, thereby augmenting its resistance to yellowing. This study scrutinizes the positioning of UV absorber addition, the quantity of UV absorber, and the molar ratio of 1,4-butanediol to 2-hydroxyethyl disulfide, characterizing the functional groups of polyurethane through infrared and Raman spectroscopy. It is observed that the successful preparation of yellowing-resistant polyurethane is achieved, and evaluations on the modified polyurethane through color difference, tensile, and centrifugal tests reveal that the optimal yellowing resistance is attained by adding a UV absorber at a mass fraction of 1% to 3% prior to chain extension, resulting in a color change grade of 2, denoting slight discoloration. Simultaneously, the other properties of polyurethane exhibit relative stability. Notably, when the molar ratio of 1,4-butanediol to 2-hydroxyethyl disulfide is 3:2, the overall performance of the polyurethane remains stable, with exceptional yellowing resistance capabilities attaining a color change grade of 2.

Urine-based detection of HPV for cervical cancer screening: Time for standardized tests.

Cervical cancer is preventable because it has an established etiology, mainly attributed to a detectable pathogen, human papillomavirus (HPV). In 2018, the world health organization issued an unprecedented call for global action to eliminate cervical cancer by 2030. The adaptation of regular screening programs is fundamental to achieve the goal of cervical cancer elimination. However, it is still difficult to achieve satisfactory coverage rates of screening in developing countries as well as in developed countries because many women are reluctant to participate in gynecologic examination. HPV detection in urine is a convenient, widely acceptable by women and relatively affordable without the necessity for clinical visits to improve the coverage rates of cervical cancer screening. Unfortunately, the clinical implementation of urine-based tests for HPV detection has been hindered by the lack of standardized tests. Further optimization of protocols and standardization of urinary HPV detection are expected to be realized. With the advantages of urine sampling to overcome cost, personal, and cultural barriers, time has come for the standardized tests to facilitate a wide clinical implementation of urinary HPV detection that will significantly contribute to the WHO's goal, that is, to eliminate the cervical cancer globally.

Vis-NIR Spectroscopy Combined with GAN Data Augmentation for Predicting Soil Nutrients in Degraded Alpine Meadows on the Qinghai-Tibet Plateau.

Soil nutrients play vital roles in vegetation growth and are a key indicator of land degradation. Accurate, rapid, and non-destructive measurement of the soil nutrient content is important for ecological conservation, degradation monitoring, and precision farming. Currently, visible and near-infrared (Vis-NIR) spectroscopy allows for rapid and non-destructive monitoring of soil nutrients. However, the performance of Vis-NIR inversion models is extremely dependent on the number of samples. Limited samples may lead to low prediction accuracy of the models. Therefore, modeling and prediction based on a small sample size remain a challenge. This study proposes a method for the simultaneous augmentation of soil spectral and nutrient data (total nitrogen (TN), soil organic matter (SOM), total potassium oxide (TK2O), and total phosphorus pentoxide (TP2O5)) using a generative adversarial network (GAN). The sample augmentation range and the level of accuracy improvement were also analyzed. First, 42 soil samples were collected from the pika disturbance area on the QTP. The collected soils were measured in the laboratory for Vis-NIR and TN, SOM, TK2O, and TP2O5 data. A GAN was then used to augment the soil spectral and nutrient data simultaneously. Finally, the effect of adding different numbers of generative samples to the training set on the predictive performance of a convolutional neural network (CNN) was analyzed and compared with another data augmentation method (extended multiplicative signal augmentation, EMSA). The results showed that a GAN can generate data very similar to real data and with better diversity. A total of 15, 30, 60, 120, and 240 generative samples (GAN and EMSA) were randomly selected from 300 generative samples to be included in the real data to train the CNN model. The model performance first improved and then deteriorated, and the GAN was more effective than EMSA. Further shortening the interval for adding GAN data revealed that the optimal ranges were 30-40, 50-60, 30-35, and 25-35 for TK2O, TN, TP2O5, and SOM, respectively, and the validation set accuracy was maximized in these ranges. Therefore, the above method can compensate to some extent for insufficient samples in the hyperspectral prediction of soil nutrients, and can quickly and accurately estimate the content of soil TK2O, TN, TP2O5, and SOM.

Ultrasound-Microwave Combined Extraction of Novel Polysaccharide Fractions from Lycium barbarum Leaves and Their In Vitro Hypoglycemic and Antioxidant Activities.

Ultrasound-microwave combined extraction (UMCE), gradient ethanol precipitation, chemical characterization, and antioxidant and hypoglycemic activities of Lycium barbarum leaf polysaccharides (LLP) were systematically studied. The optimal conditions for UMCE of LLP achieved by response surface method (RSM) were as follows: microwave time of 16 min, ultrasonic time of 20 min, particle size of 100 mesh, and ratio of liquid to solid of 55:1. Three novel polysaccharide fractions (LLP30, LLP50, LLP70) with different molecular weights were obtained by gradient ethanol precipitation. Polysaccharide samples exhibited scavenging capacities against ABTS and DPPH radicals and inhibitory activities against α-glucosidase and α-amylase. Among the three fractions, LLP30 possessed relatively high antioxidant and hypoglycemic activities in vitro, which showed a potential for becoming a nutraceutical or a phytopharmaceutical for prevention and treatment of hyperglycemia or diabetes.

Open-source deep learning-based automatic segmentation of mouse Schlemm's canal in optical coherence tomography images.

The purpose of this study was to develop an automatic deep learning-based approach and corresponding free, open-source software to perform segmentation of the Schlemm's canal (SC) lumen in optical coherence tomography (OCT) scans of living mouse eyes. A novel convolutional neural network (CNN) for semantic segmentation grounded in a U-Net architecture was developed by incorporating a late fusion scheme, multi-scale input image pyramid, dilated residual convolution blocks, and attention-gating. 163 pairs of intensity and speckle variance (SV) OCT B-scans acquired from 32 living mouse eyes were used for training, validation, and testing of this CNN model for segmentation of the SC lumen. The proposed model achieved a mean Dice Similarity Coefficient (DSC) of 0.694 ± 0.256 and median DSC of 0.791, while manual segmentation performed by a second expert grader achieved a mean and median DSC of 0.713 ± 0.209 and 0.763, respectively. This work presents the first automatic method for segmentation of the SC lumen in OCT images of living mouse eyes. The performance of the proposed model is comparable to the performance of a second human grader. Open-source automatic software for segmentation of the SC lumen is expected to accelerate experiments for studying treatment efficacy of new drugs affecting intraocular pressure and related diseases such as glaucoma, which present as changes in the SC area.

A Systematic Review and Meta-Analysis of Metabolic Syndrome Prevalence in Chinese Inpatients with Bipolar Disorder.

The aim of the work was to study the prevalence of metabolic syndrome in Chinese patients with bipolar disorder. We searched Chinese literature related to the study in prevalence of metabolic syndrome in bipolar disorder in Chinese language, among which results such as comments, letters, reviews and case reports were excluded. The prevalence of metabolic syndrome in bipolar disorder was researched and discussed. A total of 1562 subjects were included in 11 studies. The prevalence of MetS in bipolar disorder was 33% (95% CI=0.29-0.37), which was higher significantly than normal control (10.82%), but similar to schizophrenia (31.59%). The 41.41% prevalence of MetS in male patients was higher significantly than that in females (26.83%).The prevalence of MetS in BD treated by AAP was 47.54%, by MS was 19.19%, by MS+AAP was 40%.The prevalence of MetS in BD treated by carbamazepine was 28.21%, by lithium was 30%, by valproate was 21.71%, by clozepine was 51.43%, by olanzapine was 39.84%, by quetiapine was 39.44%, and by risperidone was 35%. The prevalence of MetS in bipolar disorder was 33% (95% CI=0.29-0.37), which was higher significantly than normal control (10.82%), but similar to schizophrenia (31.59%). AAP and MS were the main one risks of MetS in BD.

Association of METS-IR with incident hypertension in non-overweight adults based on a cohort study in Northeastern China.

BACKGROUND: Insulin resistance (IR) plays an important role in the progression of hypertension (HTN); therefore, early identification of IR is clinically important for preventing HTN. Our study aims to explore the relationship between the metabolic score for IR (METS-IR) and HTN in Chinese population who maintained non-overweight. METHODS: A total of 4678 adults who underwent annual health check-up in our institution from 2010 to 2017, did not have HTN at the first check-up and maintained non-overweight at follow-up were selected as subjects. The baseline METS-IR was calculated and the outcome was incident HTN. Cox proportional hazards regression models were used to evaluate hazards ratios of HTN for METS-IR. Additionally, sensitive analyses and stratification analyses were used to deeply verify the relationship of METS-IR with HTN. The dose-response association between METS-IR and HTN risk was investigated using restricted the cubic spline analysis fitted for the Cox proportional hazards model. RESULTS: Compared with the first quartiles of METS-IR, the risk of incident HTN was increased by 58% [hazard ratio (HR) 1.58, 95% confidence interval (CI) 1.12-2.22] and 96% (HR 1.96, 95% CI 1.40-2.76) in the Q3 group and the Q4 group, respectively. The results remained consistent when analyses were restricted to people without abnormal high-density lipoprotein cholesterol, triglyceride or fasting plasma glucose level at baseline. A linear dose-response relationship between METS-IR and HTN risk was identified (HR 1.08, 95% CI 1.04-1.12). CONCLUSIONS: The risk of incident HTN was associated with elevated METS-IR levels in non-overweight individuals. METS-IR could help predict the risk of HTN in non-overweight individuals.

In vivo measurement of trabecular meshwork stiffness in a corticosteroid-induced ocular hypertensive mouse model.

Ocular corticosteroids are commonly used clinically. Unfortunately, their administration frequently leads to ocular hypertension, i.e., elevated intraocular pressure (IOP), which, in turn, can progress to a form of glaucoma known as steroid-induced glaucoma. The pathophysiology of this condition is poorly understood yet shares similarities with the most common form of glaucoma. Using nanotechnology, we created a mouse model of corticosteroid-induced ocular hypertension. This model functionally and morphologically resembles human ocular hypertension, having titratable, robust, and sustained IOPs caused by increased resistance to aqueous humor outflow. Using this model, we then interrogated the biomechanical properties of the trabecular meshwork (TM), including the inner wall of Schlemm's canal (SC), tissues known to strongly influence IOP and to be altered in other forms of glaucoma. Specifically, using spectral domain optical coherence tomography, we observed that SC in corticosteroid-treated mice was more resistant to collapse at elevated IOPs, reflecting increased TM stiffness determined by inverse finite element modeling. Our noninvasive approach to monitoring TM stiffness in vivo is applicable to other forms of glaucoma and has significant potential to monitor TM function and thus positively affect the clinical care of glaucoma, the leading cause of irreversible blindness worldwide.

Hepatoprotective Effects of Phloridzin against Isoniazid-Rifampicin Induced Liver Injury by Regulating CYP450 and Nrf2/HO-1 Pathway in Mice.

The protective effect of phloridzin (PHL) and its potential mechanism were examined in mice with liver injury induced by isoniazid (INH) and rifampicin (RFP). The mice were randomly divided into normal control group, model group, low (80 mg/kg), medium (160 mg/kg) and high (320 mg/kg) phloridzin-treated groups. After 28 d treatment, blood and liver tissue were collected and analysed. The results revealed that PHL regulated liver function related indicators and reduced the pathological tissue damage, indicating that PHL significantly alleviated the liver injury. Furthermore, the level of CYP450 enzyme, the expression of CYP3A4, CYP2E1, heme oxygenase-1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA and protein were inhibited by PHL. These results indicated that PHL exerts a protecting effect against liver injury induced by combination of RFP and INH. The potential mechanisms may be concerned with the activation of Nrf2/HO-1 signaling pathway containing its key antioxidant enzymes and regulation of CYP3A4 and CYP2E1.

A goal-programming based optimal port docking scheme under COVID-19.

The Covid-19 epidemic, has caused a large-scale congestion in many ports around the world. This increases the cost of port docking, as well as delays the loading and unloading of goods, which affects the price and timely supply of many products. Although scholars have carried out in-depth discussion and analysis on the port congestion problem from different perspectives, there is still no appropriate model and algorithm for the large-scale comprehensive port docking problem. This paper presents a new mixed integer programming model for optimal docking of ships in ports that is comprehensive enough to include four essential objectives. It discusses the generalization and application of the model from the perspectives of the shortest overall waiting time of ships, the balance of tasks at each berth, completion of all docking tasks as soon as possible and meeting the expected berthing time of ships. We demonstrate the results of our models using relevant examples and show that our model can obtain the optimal docking scheme based on different perspectives and relevant objectives. We also show that the scale of the exact solution can reach tens of thousands of decision variables and more than a million constraints. This fully reflects the possibility that the model can be put into use in any real life scenario. This model can not only effectively improve the docking efficiency of the port, but is also suitable for the complex queuing problem of multi window and the same type of service.

In vivo estimation of murine iris stiffness using finite element modeling.

The iris plays an important role in certain types of glaucoma, including primary angle-closure glaucoma and pigmentary glaucoma. Iris mechanics are also important in influencing trabecular meshwork deformation in response to intraocular pressure changes in some animal species. Although mice are widely used to study ocular disease, including glaucoma, the in vivo biomechanical properties of the murine iris are unknown. Thus, the primary objective of this study was to estimate murine iris biomechanical stiffness. We used optical coherence tomography (OCT) images of the anterior segment of living mice (n = 13, age = 7.3 ± 3.2 [mean ± SD] months) at sequentially increasing IOP levels, observing IOP-dependent iris deformations. We then used an inverse finite element model to predict iris deformations under the same conditions, estimating iris stiffness by maximizing agreement between OCT data and numerical simulations. Our results show an in vivo murine iris stiffness of 96.1 ± 54.7 kPa (mean ± SD), which did not correlate with age but was dependent on gender. Our results further showed strong evidence of reverse pupillary block, with mean posterior chamber pressure remaining at approximately 12 mmHg even as anterior chamber pressure was set to much higher levels. Our approach to monitoring iris stiffness in vivo is applicable to study potential changes of iris stiffness in various pathophysiological conditions and thus has significant potential for clinical care of ocular disease involving iris biomechanics.

Integral role for lysyl oxidase-like-1 in conventional outflow tissue function and behavior.

Lysyl oxidase-like-1 (LOXL1), a vital crosslinking enzyme in elastin fiber maintenance, is essential for the stability and strength of elastic vessels and tissues. Variants in the LOXL1 locus associate with a dramatic increase in risk of exfoliation syndrome (XFS), a systemic fibrillopathy, which often presents with ocular hypertension and exfoliation glaucoma (XFG). We examined the role of LOXL1 in conventional outflow function, the prime regulator of intraocular pressure (IOP). Using Loxl1-/- , Loxl1+/- , and Loxl1+/+ mice, we observed an inverse relationship between LOXL1 expression and IOP, which worsened with age. Elevated IOP in Loxl1-/- mice was associated with a larger globe, decreased ocular compliance, increased outflow facility, extracellular matrix (ECM) abnormalities, and dilated intrascleral veins, yet, no dilation of arteries or capillaries. Interestingly, in living Loxl1-/- mouse eyes, Schlemm's canal (SC) was less susceptible to collapse when challenged with acute elevations in IOP, suggesting elevated episcleral venous pressure (EVP). Thus, LOXL1 expression is required for normal IOP control, while ablation results in altered ECM repair/homeostasis and conventional outflow physiology. Dilation of SC and distal veins, but not arteries, is consistent with key structural and functional roles for elastin in low-pressure vessels subjected to cyclical mechanical stress.

Thiol functionalized covalent organic framework for highly selective enrichment and detection of mercury by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

A spherical thiol-functionalized covalent organic framework (COF-SH) was designed via a facile thiol-yne click reaction of a alkynyl-terminated COF and pentaerythritol tetra(3-mercaptopropionate). The COF-SH was explored as a new adsorbent for the selective enrichment of Hg2+. The as-prepared COF-SH exhibited a uniform mesoporous structure, a high abundance of binding sites, and good chemical stability, which endow it with great performance for the adsorption of Hg2+ and its corresponding maximum adsorption capacity was up to 617.3 mg g-1. Furthermore, the adsorption behavior of Hg2+ on the COF-SH wasin good agreement with the Langmuir and pseudo-second-order models. The influences of adsorbent dosage, pH, selectivity, and reusability of the COF-SH on Hg2+ adsorption were also investigated. Besides this, the COF-SH showed high selectivity towards Hg2+ even in the presence of a high concentration of K+, Na+, Ca2+, Mg2+ and Zn2+ metal ions. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), the corresponding limit of detection (LOD) of Hg2+ was determined at very low concentrations of 80 pg mL-1 (equal to 396 amoL µL-1). In addition, the COF-SH was successfully applied to rapidly enrich and sensitively detect Hg2+ in industrial sewage, with recoveries in the range of 101.8-103.4%, demonstrating the promising potential of COF-SH as an effective adsorbent for use in environmental sample pretreatment.

Coumarin Sulfonamides and Amides Derivatives: Design, Synthesis, and Antitumor Activity In Vitro.

Coumarins possesses immeasurable antitumor potential with minimum side effects depending on the substitutions on the basic nucleus, which exhibits great prospects for antitumor drug development. In an attempt to develop novel antitumor candidates, a series of coumarin sulfonamides and amides derivatives were designed and synthetized. The majority of these derivatives showed good cytotoxic activity against MDA-MB-231 and KB cell lines, among which compound 9c was the most potent against MDA-MB-231 cells, with IC50 value of 9.33 µM, comparable to 5-fluorouracil. Further investigation revealed that compound 9c had versatile properties against tumors, including inhibition of cell migration and invasion as well as inducing apoptosis. Reactive oxygen species (ROS) assay and western blotting analysis suggested that compound 9c promoted cancer cell apoptosis by increasing ROS levels and upregulating the expression of caspase-3 in MDA-MB-231 cells. These results indicated that compound 9c could be promising lead compound for further antitumor drug research.

A tumor suppressor DLC1: The functions and signal pathways.

Deleted in liver cancer-1 (DLC1), a potential tumor suppressor, acts as a GTPase-activating protein for Rho family members. In many human cancers, the DLC1 expression is frequently downregulated or inactivated, which allows cancer cells to proliferate and disseminate. In this review, we describe the characteristics and other members of the DLC1 family and delineate the signal pathways DLC1 involved in regulating cancer cell growth, colony formation, apoptosis, senescence, autophagy, migration and invasion. In addition, we explore the clinical data of DLC1 and the mechanisms that natural products upregulate the DLC1 expression to inhibit cancer. Despite these insights, many important unanswered questions remain about the exact mechanisms of DLC1-mediated cancer suppression.

Targeted Delivery of Cell Softening Micelles to Schlemm's Canal Endothelial Cells for Treatment of Glaucoma.

Increased stiffness of the Schlemm's canal (SC) endothelium in the aqueous humor outflow pathways has been associated with elevated intraocular pressure (IOP) in glaucoma. Novel treatments that relax this endothelium, such as actin depolymerizers and rho kinase inhibitors, are in development. Unfortunately, these treatments have undesirable off-target effects and a lower than desired potency. To address these issues, a targeted PEG-b-PPS micelle loaded with actin depolymerizer latrunculin A (tLatA-MC) is developed. Targeting of SC cells is achieved by modifying the micelle surface with a high affinity peptide that binds the VEGFR3/FLT4 receptor, a lymphatic lineage marker found to be highly expressed by SC cells relative to other ocular cells. During in vitro optimization, increasing the peptide surface density increased micellar uptake in SC cells while unexpectedly decreasing uptake by human umbilical vein endothelial cells (HUVEC). The functional efficacy of tLatA-MC, as measured by decreased SC cell stiffness compared to non-targeted micelles (ntLatA-MC) or targeted blank micelles (tBL-MC), is verified using atomic force microscopy. tLatA-MC reduced IOP in an in vivo mouse model by 30-50%. The results validate the use of a cell-softening nanotherapy to selectively modulate stiffness of SC cells for therapeutic reduction of IOP and treatment of glaucoma.