miR-221/222-Mediated Inhibition of Autophagy Promotes Dexamethasone Resistance in Multiple Myeloma.

Inherent or acquired resistance to chemotherapeutic drugs is still an obstacle for the treatment of multiple myeloma (MM). MicroRNA dysregulation is related to the development of chemoresistance in cancers. However, its role in chemoresistance of MM is largely unknown. Here we demonstrated that miR-221/222 were upregulated in plasma cells from patients with MM, especially those with relapsed or refractory disease. Moreover, expression levels of miR-221/222 were inversely correlated with dexamethasone (Dex) sensitivity of human MM cell lines. Importantly, we found that Dex induced pro-death autophagy in MM cells and the inhibition of autophagy significantly decreased Dex-induced cell death. Mechanistically, autophagy-related gene 12 (ATG12) was identified as a novel target gene of miR-221/222, and miR-221/222 overexpression inhibited autophagy by directly targeting ATG12 and the p27kip (p27)-mammalian target of rapamycin (mTOR) pathway. Indeed, Dex treatment decreased the expression of miR-221/222, thereby activating the ATG12/p27-mTOR autophagy-regulatory axis and inducing cell death in Dex-sensitive MM cells. Furthermore, both in vitro and in vivo results showed that the inhibitions of miR-221/222 increased the expression of ATG12 and p27 and functionally induced extended autophagy and cell death of MM cells. In conclusion, our findings demonstrated the crucial role of the miR-221/222-ATG12/p27-mTOR autophagy-regulatory axis in Dex resistance of MM, and they suggest potential prediction and treatment strategies for glucocorticoid resistance.

MicroRNA-324-5p regulates stemness, pathogenesis and sensitivity to bortezomib in multiple myeloma cells by targeting hedgehog signaling.

Chromosome 17p deletions are present in 10% of patients with newly diagnosed multiple myeloma (MM), and are associated with inferior prognosis. miR-324-5p is located on chromosome 17p, and shows diverse functions in different types of cancers. However, its role in MM is largely unknown. Here we found the expression of miR-324-5p was decreased in MM, especially in del(17p) MM. In contrast, the expression of hedgehog (Hh) signaling components was elevated, indicating a correlation between miR-324-5p and Hh signaling in MM. Hh signaling is important for the pathogenesis of MM and maintenance of MM stem cell compartment. Indeed, overexpression of miR-324-5p significantly decreased Hh signaling components Smo and Gli1, and functionally reduced cell growth, survival as well as stem cell compartment in MM. Moreover, miR-324-5p potentiated the anti-MM efficacy of bortezomib through regulating the activities of multidrug-resistance proteins and the expression of Bcl-2 family genes. Consistent results were obtained in vivo. Finally, miR-324-5p overcame the protective effect of bone marrow stromal cells on MM cells. Taken together, our data demonstrate that miR-324-5p is essential for MM pathogenesis and downregulation of miR-324-5p is a novel mechanism of Hh signaling activation in MM. Therefore, targeting miR-324-5p provides a potential therapeutic strategy for MM.

Hyperreflective Dots on OCT as a Predictor of Treatment Outcome in Diabetic Macular Edema: A Systematic Review.

TOPIC: This review aims to evaluate the role of hyperreflective dots (HRDs), detected using OCT, as a predictor of the treatment outcome in patients with diabetic macular edema (DME). CLINICAL RELEVANCE: The treatment of DME is possible, but its results are often unsatisfactory. Thus, it is important to develop biomarkers that can help to predict the treatment response to optimize the treatment's effect for individual patients. METHODS: PubMed, Embase, Web of science, and Cochrane library were searched (final search date on May 5, 2021). Participants were patients diagnosed with DME and provided with treatment. The predictor was HRDs, detected using OCT, before treatment. The outcomes were best-corrected visual acuity (BCVA) and central macular thickness (CMT), detected using OCT, after treatment. Two reviewers independently screened the titles and abstracts as well as full text. The refined Quality in Prognosis Studies tool was used to assess the risk of bias for each included study. Because of the clinical heterogeneity of the studies, a meta-analysis was not performed. RESULTS: Thirty-six studies were included. The Quality in Prognosis Studies assessment showed that most studies had a low or moderate risk of bias in 6 domains. Six studies could not find any correlation between baseline HRDs (either the presence or absence of HRDs [n = 1] or baseline HRD number [n = 5]) and outcome (BCVA or CMT), whereas 12 studies found a significant correlation between these variables. Eight studies reported that baseline HRDs could predict a poor visual outcome (n = 4 on prescence or abscence of HRD and n = 4 on HRD number), and 4 studies (n = 1 on prescence or abscence of HRD and n = 3 on HRD number) found that HRDs were predictive of visual improvement. Fifteen out of 17 studies found that the HRD number decreased after treatment. CONCLUSION: Based on the current literature, the HRD numbers decrease with treatment, but it is not clear whether HRDs predict the treatment outcome in patients with DME. Future investigations with more uniform approaches are needed to confirm the nature of this biomarker and its effect on DME treatment outcome.

Joint Segmentation of Multi-Class Hyper-Reflective Foci in Retinal Optical Coherence Tomography Images.

Hyper-reflective foci (HRF) refers to the spot-shaped, block-shaped areas with characteristics of high local contrast and high reflectivity, which is mostly observed in retinal optical coherence tomography (OCT) images of patients with fundus diseases. HRF mainly appears hard exudates (HE) and microglia (MG) clinically. Accurate segmentation of HE and MG is essential to alleviate the harm in retinal diseases. However, it is still a challenge to segment HE and MG simultaneously due to similar pathological features, various shapes and location distribution, blurred boundaries, and small morphology dimensions. To tackle these problems, in this paper, we propose a novel global information fusion and dual decoder collaboration-based network (GD-Net), which can segment HE and MG in OCT images jointly. Specifically, to suppress the interference of similar pathological features, a novel global information fusion (GIF) module is proposed, which can aggregate the global semantic information efficiently. To further improve the segmentation performance, we design a dual decoder collaborative workspace (DDCW) to comprehensively utilize the semantic correlation between HE and MG while enhancing the mutual influence on them by feedback alternately. To further optimize GD-Net, we explore a joint loss function which integrates pixel-level with image-level. The dataset of this study comes from patients diagnosed with diabetic macular edema at the department of ophthalmology, University Medical Center Groningen, The Netherlands. Experimental results show that our proposed method performs better than other state-of-the-art methods, which suggests the effectiveness of the proposed method and provides research ideas for medical applications.

Algorithm for Detection and Quantification of Hyperreflective Dots on Optical Coherence Tomography in Diabetic Macular Edema.

Purpose: To develop an algorithm to detect and quantify hyperreflective dots (HRDs) on optical coherence tomography (OCT) in patients with diabetic macular edema (DME). Materials and Methods: Twenty OCTs (each OCT contains 128 b scans) from 20 patients diagnosed with DME were included in this study. Two types of HRDs, hard exudates and small HRDs (hypothesized to be activated microglia), were identified and labeled independently by two raters. An algorithm using deep learning technology was developed based on input (in total 2,560 OCT b scans) of manual labeling and differentiation of HRDs from rater 1. 4-fold cross-validation was used to train and validate the algorithm. Dice coefficient, intraclass coefficient (ICC), correlation coefficient, and Bland-Altman plot were used to evaluate agreement of the output parameters between two methods (either between two raters or between one rater and proposed algorithm). Results: The Dice coefficients of total HRDs, hard exudates, and small HRDs area of the algorithm were 0.70 ± 0.10, 0.72 ± 0.11, and 0.46 ± 0.06, respectively. The correlations between rater 1 and proposed algorithm (range: 0.95-0.99, all p < 0.001) were stronger than the correlations between the two raters (range: 0.84-0.96, all p < 0.001) for all parameters. The ICCs were higher for all the parameters between rater 1 and proposed algorithm (range: 0.972-0.997) than those between the two raters (range: 0.860-0.953). Conclusions: Our proposed algorithm is a good tool to detect and quantify HRDs and can provide objective and repeatable information of OCT for DME patients in clinical practice and studies.

Whole-genome sequencing suggests a role of MIF in the pathophysiology of TEMPI syndrome.

TEMPI syndrome (telangiectasias, elevated erythropoietin level and erythrocytosis, monoclonal gammopathy, perinephric fluid collections, and intrapulmonary shunting) is a newly defined multisystemic disease with its pathophysiology largely unknown. Here, we report the whole-genome sequencing (WGS) analysis on the tumor-normal paired cells from a patient with TEMPI syndrome. WGS revealed somatic nonsynonymous single-nucleotide variants, including SLC7A8, NRP2, and AQP7. Complex structural variants of chromosome 2 were found, particularly within regions where some putative oncogenes reside. Of potential clinical relevance, duplication of 22q11.23 was identified, and the expression of the located gene macrophage migration inhibitory factor (MIF) was significantly upregulated in 3 patients with TEMPI syndrome. Importantly, the level of serum MIF in one patient with TEMPI syndrome was significantly decreased in accordance with the downtrend of plasma cells, M-protein, hemoglobin, and erythropoietin and the improvement of telangiectasias, perinephric fluid collections, and intrapulmonary shunting after treatment with plasma cell-directed therapy. In conclusion, our study provides insights into the genomic landscape and suggests a role of MIF in the pathophysiology of TEMPI syndrome.

A Multitask Deep-Learning System to Classify Diabetic Macular Edema for Different Optical Coherence Tomography Devices: A Multicenter Analysis.

OBJECTIVE: Diabetic macular edema (DME) is the primary cause of vision loss among individuals with diabetes mellitus (DM). We developed, validated, and tested a deep learning (DL) system for classifying DME using images from three common commercially available optical coherence tomography (OCT) devices. RESEARCH DESIGN AND METHODS: We trained and validated two versions of a multitask convolution neural network (CNN) to classify DME (center-involved DME [CI-DME], non-CI-DME, or absence of DME) using three-dimensional (3D) volume scans and 2D B-scans, respectively. For both 3D and 2D CNNs, we used the residual network (ResNet) as the backbone. For the 3D CNN, we used a 3D version of ResNet-34 with the last fully connected layer removed as the feature extraction module. A total of 73,746 OCT images were used for training and primary validation. External testing was performed using 26,981 images across seven independent data sets from Singapore, Hong Kong, the U.S., China, and Australia. RESULTS: In classifying the presence or absence of DME, the DL system achieved area under the receiver operating characteristic curves (AUROCs) of 0.937 (95% CI 0.920-0.954), 0.958 (0.930-0.977), and 0.965 (0.948-0.977) for the primary data set obtained from CIRRUS, SPECTRALIS, and Triton OCTs, respectively, in addition to AUROCs >0.906 for the external data sets. For further classification of the CI-DME and non-CI-DME subgroups, the AUROCs were 0.968 (0.940-0.995), 0.951 (0.898-0.982), and 0.975 (0.947-0.991) for the primary data set and >0.894 for the external data sets. CONCLUSIONS: We demonstrated excellent performance with a DL system for the automated classification of DME, highlighting its potential as a promising second-line screening tool for patients with DM, which may potentially create a more effective triaging mechanism to eye clinics.

A Novel Scoring System for Risk Assessment of Elderly Patients With Cytogenetically Normal Acute Myeloid Leukemia Based on Expression of Three AQP1 DNA Methylation-Associated Genes.

Background: Aquaporin 1 (AQP-1), a transmembrane water channel protein, has been proven to involve in many diseases' progression and prognosis. This research aims to explore the prognostic value of AQP-1 in elderly cytogenetically normal acute myeloid leukemia (CN-AML). Methods: Complete clinical and expression data of 226 elderly patients (aged > 60) with cytogenetically normal acute myeloid leukemia (CN-AML) were downloaded from the databases of The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). We have explored prognostic significance of AQP-1, investigated the underlying mechanism, and developed a novel scoring system for the risk assessment of elderly patients with AML based on AQP1 methylation. Results: In the first and second independent group, AQP1 shows lower expression in CN-AML than normal people, while high AQP1 expression and AQP1 promoter hypomethylation were related to better overall survival (OS; P < 0.05). To understand the underlying mechanisms, we investigated differentially expressed genes (DEGs), miRNA and lncRNA associated with AQP1 methylation. A three-gene prognostic signature based on AQP1 methylation which was highly correlated with OS was established, and the performance was validated by Permutation Test and Leave-one-out Cross Validation method. Furthermore, an independent cohort was used to verify the prognostic value of this model. Conclusions: AQP1 methylation could serve as an independent prognostic biomarker in elderly CN-AML, and may provide new insights for the diagnosis and treatment for elderly CN-AML patients.

Repeatability, interocular correlation and agreement of quantitative swept-source optical coherence tomography angiography macular metrics in healthy subjects.

PURPOSE: To investigate the repeatability, interocular correlation and agreement of quantitative swept-source optical coherence tomography angiography (SS-OCTA) metrics in healthy subjects. METHODS: Thirty-three healthy normal subjects were enrolled. The macula was scanned four times by an SS-OCTA system using the 3 mm×3 mm mode. The superficial capillary map images were analysed using a MATLAB program. A series of parameters were measured: foveal avascular zone (FAZ) area, FAZ perimeter, FAZ circularity, parafoveal vessel density, fractal dimension and vessel diameter index (VDI). The repeatability of four scans was determined by intraclass correlation coefficient (ICC). Then the averaged results were analysed for intereye difference, correlation and agreement using paired t-test, Pearson's correlation coefficient (r), ICC and Bland-Altman plot. RESULTS: The repeatability assessment of the macular metrics exported high ICC values (ranged from 0.853 to 0.996). There is no statistically significant difference in the OCTA metrics between the two eyes. FAZ area (ICC=0.961, r=0.929) and FAZ perimeter (ICC=0.884, r=0.802) showed excellent binocular correlation. Fractal dimension (ICC=0.732, r=0.578) and VDI (ICC=0.707, r=0.547) showed moderate binocular correlation, while parafoveal vessel density had poor binocular correlation. Bland-Altman plots showed the range of agreement was from -0.0763 to 0.0954 mm2 for FAZ area and from -0.0491 to 0.1136 for parafoveal vessel density. CONCLUSIONS: The macular metrics obtained using SS-OCTA showed excellent repeatability in healthy subjects. We showed high intereye correlation in FAZ area and perimeter, moderate correlation in fractal dimension and VDI, while vessel density had poor correlation in normal healthy subjects.

Macular Pigment Optical Density Measured by a Single Wavelength Reflection Photometry with and without Mydriasis.

PURPOSE: 1. To evaluate the repeatability of macular pigment optical density (MPOD) measurements in non-mydriatic and mydriatic conditions with a single wavelength reflection photometry. 2. To compare the non-mydriatic measurement with the mydriatic measurement of MPOD values. METHODS: Fifty-one healthy young subjects were recruited in this prospective study. MPOD parameters including maximum optical intensity (OD), mean OD, volume and area, were measured using a single 460-nm wavelength reflection method (Visucam 200, Carl Zeiss Meditec) twice both before and after pupil dilation. Paired t test, Pearson's correlation, intraclass correlation coefficient (ICC) and Bland-Altman plots were used to evaluate the agreement and correlation of MPOD parameters before and after mydriasis. RESULTS: The differences of maximum OD, mean OD and volume between the two measurements without mydriasis was statistically significant for all parameters, but not significant between the two measurements with mydriasis or between non-mydriatic and mydriatic measures. The ICCs were higher for the two measurements with mydriasis (range: 0.687-0.840) than those without mydriasis (range: 0.448-0.695) or non-mydriatic and mydriatic measures (range: 0.473-0.769). The 95% limit of agreement (LOA) of measurements between the two measurements after mydriasis was the smallest among all the other comparisons. The coefficient of repeatability was lower for the measurements with mydriasis than those without mydriasis. CONCLUSIONS: MPOD measurement by Visucam 200 with mydriasis has higher repeatability than the measurement without mydriasis. The non-mydriatic and mydriatic measures had less agreement compared to the two measure after mydriasis. Therefore, mydriasis is recommended.

MicroRNA-324-5p suppresses the migration and invasion of MM cells by inhibiting the SCFß-TrCP E3 ligase.

Multiple myeloma (MM) is a cytogenetically heterogeneous malignancy of plasma cells in bone marrow. Among the cytogenetic abnormalities of MM, del(17p) is a well-recognized high-risk genetic lesion associated with the late stage and progression of the disease. MicroRNA (miR)-324-5p, located at 17p13.1, was identified to be involved in the dysregulation of a number of types of malignant disease. However, whether miR-324-5p is associated with the development and progression of MM remains unknown. In the present study, the expression status of miR-324-5p in MM, and its effect on the migratory and invasive ability of MM cells were investigated. Using ubiquitination pathway polymerase chain reaction array, the inhibitory effect of miR-324-5p on the ubiquitinated proteins was investigated. It was identified that miR-324-5p levels were decreased in samples from patients with MM and MM cell lines. Increased expression of miR-324-5p by transfection of miR-324-5p mimic suppressed the proliferative, migratory and invasive abilities of MM.1R cells. Furthermore, increased expression of miR-324-5p in MM.1R cells inhibited the ubiquitination pathway and decreased the levels of ubiquitination-associated proteins, particularly the Skp1-Cullin1-F-box ß-transducin repeat-containing protein (SCFß-TrCP) E3 ligase. In addition, the results of the present study demonstrated that the SCFß-TrCP E3 ligase may contribute to the suppression of MM cell motility by inhibiting the expression of metastasis-associated genes, including metastasis suppressor 1. In conclusion, the results of the present study suggested that miR-324-5p may act as a tumor suppressor by impairing the motility of MM cells by suppressing the ubiquitination pathway.

Change of Optical Intensity during Healing Process of Corneal Wound on Anterior Segment Optical Coherence Tomography.

The purpose of this study is to investigate the process of corneal wound healing after penetrating injury with the change in optical intensity on anterior segment optical coherence tomography (AS-OCT) and to investigate factors associated with severity of corneal scar. Forty-seven eyes from 47 patients with repaired corneal laceration were included. AS-OCT was performed on 1day, 1week, 1, 3 and 6 months after primary repair. Internal aberrations of wound edges were observed on AS-OCT images. Parameters including height of steps, width of gaps, maximal corneal thickness, area and optical intensity of corneal wound/scar were measured. The relationship between the parameters at day 1 and the optical intensity at 6 months were analyzed. The results showed that optical intensity of corneal wound/scar increased from 124.1 ± 18.8 on day 1 postoperatively to 129.3 ± 18.7, 134.2 ± 23.4, 139.7 ± 26.5, 148.2 ± 26.4 at 1 week, 1 month, 3 months and 6 months postoperatively. Height of steps at 1 day after surgery was the only factor identified as correlated with optical intensity of corneal scar at 6 months (beta = 0.34, p = 0.024). The increase of optical intensity represents the process of fibrosis of corneal wound healing. Higher step after suturing is associated with more severity of corneal scar at last.

Two novel compound heterozygous mutations associated with types I and II protein C deficiency with unusual phenotypes.

INTRODUCTION: We diagnosed two Chinese hereditary PC deficiency families and identified two novel compound heterozygous mutations (p.Arg194Cys/Gly324Ser and p.Glu274X/Asp297His) in the protein C (PROC) gene. The probands were classified as types I and II PC deficiency. The aim of this article is to access the influence of the mutations on PC activity, antigen and protein structure, and to evaluate whether there is abnormal PC localization. MATERIALS AND METHODS: Genomic DNA of all family members was extracted, PCR amplified, and sequenced. The mutant PC expression plasmids were constructed. Expression assays, intracellular localization, and molecular modeling were performed. RESULTS: Proband 1, a type II PC defect, harbored a compound heterozygous mutation, p.Arg194Cys/Gly324Ser in the PROC gene, underwent two thromboembolic events. Expression assays indicated that the p.Arg194Cys mutant lead to decreased PC activity and normal PC Ag levels. Intracellular localization showed that both p.Arg194Cys and p.Gly324Ser co-localized with the endoplasmic reticuli and the Golgi apparatus. Molecular modeling suggested that the p.Gly324Ser mutation disturbed the interaction between the heavy and light chains of the PC protein. Proband 2, a type I PC defect, harbored a compound heterozygous PROC gene mutation, p.Glu274X/Asp297His, presented with recurrent spontaneous abortion and right popliteal vein thrombosis. Expression results were in accordance with the PC changes of the patient, and existed in defective PC transport. Structural model suggested p.Glu274X lead to disulfide bond between heavy and light chain cannot form. CONCLUSIONS: Our results confirm that two novel compound heterozygous PROC gene mutations are causative on the two PC deficiency families.