Metabolomic profiling of the aqueous humor in patients with pediatric cataract.

Pediatric cataract, including congenital and developmental cataract, is a kind of pediatric vision-threatening disease with extensive phenotypic heterogeneity and multiple mechanisms. We aimed to investigate the metabolite profile of aqueous humor (AH) in patients with pediatric cataracts, and identify underlying mutual correlations between differential metabolites. Metabolomic profiles of AH were analyzed and compared between pediatric cataract patients (n = 33) and age-related cataract patients without metabolic diseases (n = 29), using global untargeted metabolomics with ultra-high-performance liquid chromatography tandem mass spectrometry. Principal component analysis, partial least squares discriminant analysis and heat map were applied. Enriched pathway analysis was conducted using Kyoto Encyclopedia of Genes and Genomes. Receiver-operating characteristic (ROC) analyses were employed to select potential biomarkers. A total of 318 metabolites were identified, of which 54 differential metabolites (25 upregulated and 29 downregulated) were detected in pediatric cataract group compared with controls (variable importance of projection >1.0, fold change ≥1.5 or ≤ 0.667 and P < 0.05). A significant accumulation of N-Acetyl-Dl-glutamic acid was observed in pediatric cataract group. The differential metabolites were mainly enriched in histidine metabolism (increased L-Histidine and decreased 1-Methylhistamine) and the tryptophan metabolism (increased N-Formylkynurenine and L-Kynurenine). 5-Aminosalicylic acid showed strong positive mutual inter-correlation with L-Tyrosinemethylester and N,N-Diethylethanolamine, both of which were down-regulated in pediatric cataract group. The ROC analysis implied 11 metabolites served as potential biomarkers for pediatric cataract patients (all area under the ROC curve ≥0.900). These results illustrated novel potential metabolites and metabolic pathways in pediatric cataract, which provides new insights into the pathophysiology of pediatric cataract.

Effect of Posterior Keratometry on the Accuracy of 10 Intraocular Lens Calculation Formulas: Standard Keratometry versus Total Keratometry.

PURPOSE: To investigate the effect of posterior keratometry (PK) on the accuracy of 10 intraocular lens (IOL) power calculation formulas using standard keratometry (K) and total keratometry (TK). METHODS: This is a retrospective consecutive case-series study. The IOL power was calculated using K and TK measured by IOLMaster 700 in 6 new-generation formulas (Barrett Universal II, Emmetropia Verifying Optical (EVO) 2.0, RBF Calculator 3.0, Hoffer QST, Kane, and Ladas Super Formula) and 4 traditional formulas (Haigis, Hoffer Q, Holladay 1, and SRK/T). The arithmetic prediction error (PE) and mean absolute PE (MAE) were evaluated. The locally-weighted scatterplot smoothing was performed to assess the relationship between PE and PK. RESULTS: A total of 576 patients (576 eyes) who underwent cataract surgery were included. Compared with using K, all formulas using TK showed a hyperopic shift in the whole group. Specifically, for eyes with PK exceeding -5.90 D, all formulas using TK exhibited a hyperopic shift (all P < 0.001), while eyes with PK less than -5.90 D showed a myopic shift (all P < 0.001). The MAE of new-generation formulas calculated with TK and K showed no statistical differences, while the MAE of traditional formulas with TK was larger (TK: 0.34 ~ 0.43 D; K: 0.33 ~ 0.42 D, all P < 0.05). CONCLUSIONS: The prediction bias of formulas with TK increased as PK deviated from -5.90 D. TK did not improve the prediction accuracy of new-generation formulas, and even performed worse in traditional formulas.

Impact of Gold-Standard Label Errors on Evaluating Performance of Deep Learning Models in Diabetic Retinopathy Screening: Nationwide Real-World Validation Study.

BACKGROUND: For medical artificial intelligence (AI) training and validation, human expert labels are considered the gold standard that represents the correct answers or desired outputs for a given data set. These labels serve as a reference or benchmark against which the model's predictions are compared. OBJECTIVE: This study aimed to assess the accuracy of a custom deep learning (DL) algorithm on classifying diabetic retinopathy (DR) and further demonstrate how label errors may contribute to this assessment in a nationwide DR-screening program. METHODS: Fundus photographs from the Lifeline Express, a nationwide DR-screening program, were analyzed to identify the presence of referable DR using both (1) manual grading by National Health Service England-certificated graders and (2) a DL-based DR-screening algorithm with validated good lab performance. To assess the accuracy of labels, a random sample of images with disagreement between the DL algorithm and the labels was adjudicated by ophthalmologists who were masked to the previous grading results. The error rates of labels in this sample were then used to correct the number of negative and positive cases in the entire data set, serving as postcorrection labels. The DL algorithm's performance was evaluated against both pre- and postcorrection labels. RESULTS: The analysis included 736,083 images from 237,824 participants. The DL algorithm exhibited a gap between the real-world performance and the lab-reported performance in this nationwide data set, with a sensitivity increase of 12.5% (from 79.6% to 92.5%, P<.001) and a specificity increase of 6.9% (from 91.6% to 98.5%, P<.001). In the random sample, 63.6% (560/880) of negative images and 5.2% (140/2710) of positive images were misclassified in the precorrection human labels. High myopia was the primary reason for misclassifying non-DR images as referable DR images, while laser spots were predominantly responsible for misclassified referable cases. The estimated label error rate for the entire data set was 1.2%. The label correction was estimated to bring about a 12.5% enhancement in the estimated sensitivity of the DL algorithm (P<.001). CONCLUSIONS: Label errors based on human image grading, although in a small percentage, can significantly affect the performance evaluation of DL algorithms in real-world DR screening.

Biologically Relevant Laminin-511 Moderates the Derivation and Proliferation of Human Lens Epithelial Stem/Progenitor-Like Cells.

Purpose: The role of specific extracellular matrix (ECM) molecules in lens cell development and regeneration is poorly understood, as appropriate cellular models are lacking. Here, a laminin-based lens cell in vitro induction system was developed to study the role of laminin in human lens epithelial stem/progenitor cell (LES/PC) development. Methods: The human embryonic stem cell-based lens induction system followed a three-stage protocol. The expression profile of laminins during lens induction was screened, and laminin-511 (LN511) was tested as a candidate substitute. LN511 induction system cellular and molecular features, including induction efficiency, transcription factor expression related to different lens development stages, ECM alterations, and Hippo/YAP signaling, were evaluated. Results: LAMA5, LAMB1, and LAMC1 were highly expressed around the time of LES/PC derivation. We chose LN511 (product of LAMA5, LAMB1, and LAMC1) and found that it considerably enhanced lens cell induction efficiency, compared to that in Matrigel-coated culture, as more and larger lentoid bodies were detected. Notably, LES/PC induction efficiency improved by promoting lens specification-related transcription factor expression and cell proliferation. Transcriptome analysis revealed that compared to those with Matrigel, ECM accumulation and cell adhesion were downregulated in the LN511 system. Hippo/YAP signaling was hypoactive during LES/P-like cell generation, and small molecule inhibitors of YAP/TAZ activity upregulated LES/PC marker expression and promoted the efficiency of LES/P-like cell derivation. Conclusions: The laminin isoform LN511 is a reliable substitute for the LES/P-like cell induction system, and LN511-YAP acted as efficient modulators of LES/PC derivation; this contributes to knowledge of the role of the ECM in human lens development.

Four-Year Progression of Myopic Maculopathy in Children and Adolescents With High Myopia.

Importance: Individuals with high myopia younger than 18 years are at relatively high risk of progressively worsening myopic maculopathy. Additional studies are needed to investigate the progression of myopic maculopathy in this age group, as well as the risk factors associated with progression. Objective: To investigate the 4-year progression of myopic maculopathy in children and adolescents with high myopia, and to explore potential risk factors. Design, Setting, and Participants: This hospital-based observational study with 4-year follow-up included a total of 548 high myopic eyes (spherical power -6.00 or less diopters) of 274 participants aged 7 to 17 years. Participants underwent comprehensive ophthalmic examination at baseline and 4-year follow-up. Myopic maculopathy was accessed by the International Photographic Classification and Grading System. The data analysis was performed from August 1 to 15, 2023. Main Outcomes and Measures: The progression of myopic maculopathy progression over 4 years and associated risk factors. Results: The 4-year progression of myopic maculopathy was found in 67 of 548 eyes (12.2%) of 274 participants (138 girls [50.4%] at baseline and 4-year follow-up) with 88 lesion changes, including new signs of the tessellated fundus in 16 eyes (18.2%), diffuse atrophy in 12 eyes (13.6%), patchy atrophy in 2 eyes (2.3%), lacquer cracks in 9 eyes (10.2%), and enlargement of diffuse atrophy in 49 eyes (55.7%). By multivariable analysis, worse best-corrected visual acuity (odds ratio [OR], 6.68; 95% CI, 1.15-38.99; P = .04), longer axial length (AL) (OR, 1.73; 95% CI, 1.34-2.24; P < .001), faster AL elongation (OR, 302.83; 95% CI, 28.61-3205.64; P < .001), and more severe myopic maculopathy (diffuse atrophy; OR, 4.52; 95% CI, 1.98-10.30; P < .001 and patchy atrophy; OR, 3.82; 95% CI, 1.66-8.80; P = .002) were associated with myopic maculopathy progression. Conclusions and Relevance: In this observational study, the progression of myopic maculopathy was observed in approximately 12% of pediatric high myopes for 4 years. The major type of progression was the enlargement of diffuse atrophy. Risk factors for myopic maculopathy progression were worse best-corrected visual acuity, longer AL, faster AL elongation, and more severe myopic maculopathy. These findings support consideration of follow-up in these individuals and trying to identify those at higher risk for progression.

Cucurbitacin IIa promotes the immunogenic cell death­inducing effect of doxorubicin and modulates immune microenvironment in liver cancer.

The immunogenic cell death (ICD) has aroused great interest in cancer immunotherapy. Doxorubicin (DOX), which can induce ICD, is a widely used chemotherapeutic drug in liver cancer. However, DOX­induced ICD is not potent enough to initiate a satisfactory immune response. Cucurbitacin IIa (CUIIa), a tetracyclic triterpene, is a biologically active compound present in the Cucurbitaceae family. The present study assessed the effects of the combination of DOX and CUIIa on the viability, colony formation, apoptosis and cell cycle of HepG2 cells. In vivo anticancer effect was performed in mice bearing H22 tumor xenografts. The hallmark expression of ICD was tested using immunofluorescence and an ATP assay kit. The immune microenvironment was analyzed using flow cytometry. The combination of CUIIa and DOX displayed potent apoptosis inducing, cell cycle arresting and in vivo anticancer effects, along with attenuated cardiotoxicity in H22 mice. The combination of DOX and CUIIa also facilitated ICD as manifested by elevated high­mobility group box 1, calreticulin and ATP secretion. This combination provoked a stronger immune response in H22 mice, including dendritic cell activation, increment of cytotoxic T cells and T helper 1 cells. Moreover, the proportion of immunosuppressive cells including myeloid­derived suppressor cells, T regulatory cells and M2­polarized macrophages, decreased. These data suggested that CUIIa is a promising combination partner with DOX for liver cancer treatment, probably via triggering ICD and remolding the immune microenvironment.

Interocular difference in crystalline lens morphology in children and adolescents with unilateral high myopia.

PURPOSE: To investigate the between-eye differences of the crystalline lens in subjects with unilateral high myopia and assess its contribution to the interocular refractive error disparity. METHODS: Children and adolescents with unilateral high myopia, defined as cycloplegic spherical equivalent (SE) ≤ -5D in one eye and ≥ -3D in the other eye, were recruited. Ocular biometric parameters, including axial length (AL) and lens thickness (LT), were measured by IOLMaster 700. Other lens-related parameters, including anterior lens radius of curvature (ALR) and posterior lens radius of curvature (PLR), were measured by CASIA2 swept-source optical coherence tomography. Lens power (LP) was calculated using Bennett's formula. Paired t-test was used to assess the between-eye difference in biometric parameters, and multiple regression analysis was used to assess factors associated with the between-eye SE difference. RESULTS: Ninety-one participants (6-18 years of age; 52.75% girls) were included. The highly myopic eyes showed significantly lower LP (P < 0.001) and smaller ALR (P < 0.001) than the contralateral eyes, while no significant difference was found in central LT. In both eyes, ALR was significantly related to SE (P = 0.001 and P = 0.006, respectively); while LT was not associated with SE (P = 0.051 and P = 0.052, respectively). Paired-eye analysis showed that the between-eye difference in ALR was the only lenticular parameter significantly associated with the between-eye difference in SE (P = 0.005). CONCLUSION: In highly myopic eyes, the crystalline lens reduced total power but morphologically changed to a more curved shape without significant lens thinning, suggesting that the LP loss is mainly achieved by reducing its internal power in high myopes.

Characteristics of myopic maculopathy in Chinese children and adolescents with high myopia.

AIMS: To investigate the characteristics of myopic maculopathy among highly myopic Chinese children and adolescents and explore its associated risk factors. METHODS: Children and adolescents aged 7-17 years with spherical equivalent (SE) ≤ -6.00 dioptres (D) were recruited. Myopic maculopathy was categorised based on the International Meta-Analysis of Pathological Myopia Classification. The extent of diffuse choroidal atrophy (DCA) was classified using Early Treatment Diabetic Retinopathy Study grid (ETDRS). The area of DCA was categorised into three classes relative to optic disk area (DA): A1 (≤1 DA), A2 (1 to ≤5 DA) and A3 (5 to ≤10 DA). Logistic regression was used to identify risk factors associated with myopic maculopathy. RESULTS: Of the 425 participants aged 13.66±2.67 years, the proportions of tessellated fundus and DCA were 11.76% and 12.24%, and no more severe fundus lesions or 'plus' lesions. The proportion of DCA was 27.03% in children under 11, significantly higher than the 9.12% observed in those aged 11 and older (p<0.001). The percentages of DCA involving the outer, middle and central circles of the ETDRS grid were 42.31%, 55.77% and 1.92%. Myopic maculopathy was significantly associated with younger age (p<0.001), longer axial length (AL; p<0.001) and larger ß-zone peripapillary atrophy (ß-PPA; p=0.012). CONCLUSION: In highly myopic children and adolescents, myopic maculopathy predominantly manifested as DCA (12.24%), with no cases of worse myopic maculopathy or 'plus' lesions. Younger age, longer AL and larger ß-PPA were risk factors for myopic maculopathy.

Targeting glutamine metabolism improves sarcoma response to radiation therapy in vivo.

Diverse tumor metabolic phenotypes are influenced by the environment and genetic lesions. Whether these phenotypes extend to rhabdomyosarcoma (RMS) and how they might be leveraged to design new therapeutic approaches remains an open question. Thus, we utilized a Pax7Cre-ER-T2/+; NrasLSL-G12D/+; p53fl/fl (P7NP) murine model of sarcoma with mutations that most frequently occur in human embryonal RMS. To study metabolism, we infuse 13C-labeled glucose or glutamine into mice with sarcomas and show that sarcomas consume more glucose and glutamine than healthy muscle tissue. However, we reveal a marked shift from glucose consumption to glutamine metabolism after radiation therapy (RT). In addition, we show that inhibiting glutamine, either through genetic deletion of glutaminase (Gls1) or through pharmacological inhibition of glutaminase, leads to significant radiosensitization in vivo. This causes a significant increase in overall survival for mice with Gls1-deficient compared to Gls1-proficient sarcomas. Finally, Gls1-deficient sarcomas post-RT elevate levels of proteins involved in natural killer cell and interferon alpha/gamma responses, suggesting a possible role of innate immunity in the radiosensitization of Gls1-deficient sarcomas. Thus, our results indicate that glutamine contributes to radiation response in a mouse model of RMS.

Association Between Preoperative Ocular Parameters and Myopic Shift in Children Undergoing Primary Intraocular Lens Implantation.

Purpose: To evaluate the association between preoperative ocular parameters and myopic shift following primary intraocular lens (IOL) implantation in pediatric cataracts. Methods: Eyes from pediatric patients undergoing bilateral cataract surgery with primary IOL implantation were included. Eyes were grouped by age at surgery and subdivided into three axial length (AL) subgroups and three keratometry subgroups. Mixed-effects linear regression was utilized to assess the trend in myopic shift among subgroups. Multivariable analysis was performed to determine factors associated with myopic shift. Results: A total of 222 eyes were included. The median age at surgery was 4.36 years (interquartile range [IQR], 3.16-6.00 years) and the median follow-up was 4.18 years (IQR, 3.48-4.64 years). As preoperative AL increased, a decreased trend was observed in myopic shift and rate of myopic shift (P = 0.008 and P = 0.003, respectively, in the 4 to <6 years old group; P = 0.002 and P < 0.001, respectively, in the ≥6 years old group). Greater myopic shift and rate of myopic shift were associated with younger age at surgery (P = 0.008 and P = 0.008, respectively). Both myopic shift and rate of myopic shift were negatively associated with AL. Conclusions: Age at surgery and preoperative AL were associated with myopic shift in pediatric cataracts following primary IOL implantation. Adjusting the target refraction based on preoperative AL could potentially improve patients' long-term refractive outcome. Translational Relevance: This study may help to guide the selection of postoperative target refraction according to age at surgery and preoperative ocular parameters for pediatric cataracts.

Capsular Tension Ring Implantation for Intraocular Lens Decentration and Tilt in Highly Myopic Eyes: A Randomized Clinical Trial.

Importance: Capsular tension rings (CTRs) can support weak zonules and inhibit capsular shrinkage, thus potentially reducing intraocular lens (IOL) decentration and tilt. However, it has been debated whether CTRs can reduce IOL decentration and tilt in highly myopic eyes and whether CTR implantation is necessary for all highly myopic eyes. Objective: To evaluate the influence of CTR implantation on IOL decentration and tilt in highly myopic eyes. Design, Setting, and Participants: This randomized clinical trial was conducted between November 2021 and September 2023 at the Zhongshan Ophthalmic Center, Guangzhou, China. Patients with cataract and an axial length (AL) of 26 mm or longer were enrolled. Interventions: Participants were stratified into 3 groups based on the AL (stratum 1, 26 mm ≤ AL <28 mm; stratum 2, 28 mm ≤ AL <30mm; stratum 3, AL ≥30 mm), and further randomly assigned to the CTR group (a C-loop IOL combined with a CTR) or the control group (only a C-loop IOL) within each stratum. Main Outcomes and Measures: IOL decentration at 3 months after cataract surgery was evaluated using anterior segment optical coherence tomography. Results: A total of 186 eyes of 186 participants (mean [SD] age, 57.3 [10.9] years; 118 female [63.4%]) were randomized into the CTR group (93 [50%]) or control group (93 [50%]), with 87 eyes (93.6%) and 92 eyes (98.9%) completing follow-up at 3 months, respectively. The CTR group showed smaller IOL decentration (0.19 mm vs 0.23 mm; difference, -0.04 mm; 95% CI, -0.07 to -0.01 mm; P = .02) and tilt at 3 months, and lower proportions of clinically significant IOL decentration (≥0.4 mm) and tilt (≥7°) at 3 months compared with the control group. Similar results were only found in eyes with an AL of 30 mm or longer (IOL decentration: 0.20 mm vs 0.28 mm; difference, -0.08 mm; 95% CI, -0.14 to -0.02 mm; P = .01). Additionally, the CTR group showed a smaller change in IOL decentration from 1 week to 3 months, higher prediction accuracy, and better visual quality and patient satisfaction in this stratum. No differences were observed between the CTR and control groups in eyes with an AL less than 30 mm. Conclusions and Relevance: CTR implantation reduced C-loop IOL decentration and tilt, increased position stability, and improved visual quality in eyes with an AL of 30 mm or longer. These findings support use of CTR implantation in eyes with an AL of 30 mm or longer and implanted with C-loop IOLs. Trial Registration: ClinicalTrials.gov Identifier: NCT05161520.

Economic evaluation for medical artificial intelligence: accuracy vs. cost-effectiveness in a diabetic retinopathy screening case.

Artificial intelligence (AI) models have shown great accuracy in health screening. However, for real-world implementation, high accuracy may not guarantee cost-effectiveness. Improving AI's sensitivity finds more high-risk patients but may raise medical costs while increasing specificity reduces unnecessary referrals but may weaken detection capability. To evaluate the trade-off between AI model performance and the long-running cost-effectiveness, we conducted a cost-effectiveness analysis in a nationwide diabetic retinopathy (DR) screening program in China, comprising 251,535 participants with diabetes over 30 years. We tested a validated AI model in 1100 different diagnostic performances (presented as sensitivity/specificity pairs) and modeled annual screening scenarios. The status quo was defined as the scenario with the most accurate AI performance. The incremental cost-effectiveness ratio (ICER) was calculated for other scenarios against the status quo as cost-effectiveness metrics. Compared to the status quo (sensitivity/specificity: 93.3%/87.7%), six scenarios were cost-saving and seven were cost-effective. To achieve cost-saving or cost-effective, the AI model should reach a minimum sensitivity of 88.2% and specificity of 80.4%. The most cost-effective AI model exhibited higher sensitivity (96.3%) and lower specificity (80.4%) than the status quo. In settings with higher DR prevalence and willingness-to-pay levels, the AI needed higher sensitivity for optimal cost-effectiveness. Urban regions and younger patient groups also required higher sensitivity in AI-based screening. In real-world DR screening, the most accurate AI model may not be the most cost-effective. Cost-effectiveness should be independently evaluated, which is most likely to be affected by the AI's sensitivity.

Correction of axial length measurement error by IOLMaster 700 could improve refractive prediction accuracy in silicone oil-filled eyes.

PURPOSE: To determine whether correcting the axial length (AL) measurement error of the IOLMaster 700 could improve the refractive prediction accuracy in silicone oil-filled eyes. METHODS: This study included 265 cataract patients (265 eyes) with silicone oil tamponade who were scheduled for phacoemulsification with intraocular lens (IOL) implantation. The performances of various formulas, including Barrett Universal II, Emmetropia Verifying Optical, Hoffer-QST, Kane, Ladas Super Formula, Pearl-DGS, Radial Basis Function and traditional formulas (Haigis, Hoffer Q, Holladay 1 and SRK/T), were evaluated. The refractive prediction errors (PE) calculated with measured AL (ALmeas) and corrected AL with silicone oil adjustment (SOAL) were compared. Subgroup analysis was performed based on the ALmeas (<23 mm; 23-26 mm; ≥26 mm). RESULTS: Using SOAL significantly reduced the hyperopic PE of formulas when compared to ALmeas (-0.05 to 0.17 D vs 0.15 to 0.38 D, p < 0.001). After applying AL correction, all formulas showed a lower mean absolute PE (0.47-0.57 D vs 0.50-0.69 D). The percentage of eyes within ±1.0 D of PE increased from 84.91%-88.68% to 89.81%-91.32% for new formulas and from 78.11%-83.40% to 85.66%-88.68% for traditional formulas, with the use of SOAL. Subgroup analysis showed that the majority of formulas with SOAL in prediction accuracy for eyes with an AL ≥26 mm (p < 0.05). CONCLUSIONS: The refractive prediction accuracy in silicone oil-filled eyes was improved by correcting the AL measurement error of the IOLMaster 700, especially for long eyes.

Accuracy of 11 intraocular lens calculation formulas in shallow anterior chamber eyes.

PURPOSE: To evaluate the performance of intraocular lens (IOL) calculation formulas and the effect of anterior chamber depth (ACD), axial length (AL) and lens thickness (LT) on the prediction accuracy in shallow ACD eyes. METHODS: This retrospective, consecutive case-series study included 648 eyes of 648 patients with an ACD < 3.0 mm who underwent phacoemulsification and IOL implantation. Eleven formulas were evaluated: Barrett Universal II (BUII), Emmetropia Verifying Optical (EVO) 2.0, Hill-Radial Basis Function (RBF) 3.0, Hoffer QST, Kane, Olsen, Pearl-DGS and traditional formulas (Haigis, Hoffer Q, Holladay 1 and SRK/T). Subgroup analysis was performed based on ACD, AL and LT. RESULTS: Overall, the Hoffer QST and Kane showed no systematic bias. The Kane, EVO 2.0, Hill-RBF 3.0 and Hoffer QST had relatively lower mean absolute error and higher percentages of prediction error within ±0.5 D. For the ACD of 2.5-3.0 mm and AL < 22.0 mm subgroup, the Pearl-DGS exhibited the lowest MAE (0.45 D) and MedAE (0.41 D). Most formulas had a significant myopic bias (-0.43 to -0.18 D, p < 0.05) in the LT < 4.3 mm subgroup and a significant hyperopic bias (0.09-0.29 D, p < 0.05) in the LT ≥ 5.1 mm subgroup. CONCLUSION: The Kane and Hoffer QST were recommended for shallow ACD eyes. In eyes with an ACD between 2.5 and 3.0 mm and a short AL, the Pearl-DGS showed excellent performance. Clinicians need to fine-tune the target refraction according to LT in shallow ACD eyes.

Spontaneous expression of the CIC::DUX4 fusion oncoprotein from a conditional allele potently drives sarcoma formation in genetically engineered mice.

CIC::DUX4 sarcoma (CDS) is a rare but highly aggressive undifferentiated small round cell sarcoma driven by a fusion between the tumor suppressor Capicua (CIC) and DUX4. Currently, there are no effective treatments and efforts to identify and translate better therapies are limited by the scarcity of patient tumor samples and cell lines. To address this limitation, we generated three genetically engineered mouse models of CDS (Ch7CDS, Ai9CDS, and TOPCDS). Remarkably, chimeric mice from all three conditional models developed spontaneous soft tissue tumors and disseminated disease in the absence of Cre-recombinase. The penetrance of spontaneous (Cre-independent) tumor formation was complete irrespective of bi-allelic Cic function and the distance between adjacent loxP sites. Characterization of soft tissue and presumed metastatic tumors showed that they consistently expressed the CIC::DUX4 fusion protein and many downstream markers of the disease credentialing the models as CDS. In addition, tumor-derived cell lines were generated and ChIP-seq was preformed to map fusion-gene specific binding using an N-terminal HA epitope tag. These datasets, along with paired H3K27ac ChIP-sequencing maps, validate CIC::DUX4 as a neomorphic transcriptional activator. Moreover, they are consistent with a model where ETS family transcription factors are cooperative and redundant drivers of the core regulatory circuitry in CDS.

The LISA-PPV Formula: An Ensemble Artificial Intelligence-Based Thick Intraocular Lens Calculation Formula for Vitrectomized Eyes.

PURPOSE: To investigate the relationship between effective lens position (ELP) and patient characteristics, and to further develop a new intraocular lens (IOL) calculation formula for cataract patients with previous pars plana vitrectomy (PPV). DESIGN: Cross-sectional study. METHODS: A total of 2793 age-related cataract patients (group 1) and 915 post-PPV cataract patients (group 2) who underwent phacoemulsification with IOL implantation were included. The ELP of 2 groups was compared and the association between ELP and patient characteristics was further evaluated using standardized multivariate regression coefficients. An ensemble artificial intelligence-based ELP prediction model was developed using a training set of 810 vitrectomized eyes, and a thick-lens IOL formula (LISA-PPV) was constructed and compared with 7 existing formulas on an external multi-center testing set of 105 eyes. RESULTS: Compared to eyes with age-related cataract, vitrectomized eyes showed a similar ELP distribution (P = .19), but different standardized coefficients of preoperative biometry for ELP. The standardized coefficients also varied with the type of vitreous tamponade, history of scleral buckling, and ciliary sulcus IOL implantation. The LISA-PPV formula showed the lowest mean and median absolute prediction error (MAE: 0.63 D; MedAE: 0.44 D), and the highest percentages of eyes within ±0.5 D of prediction error (57.14%) in the testing dataset. CONCLUSIONS: The ELP prediction required optimization specifically for vitrectomized eyes based on their biometric and surgical characteristics. The LISA-PPV formula is a useful and accurate tool for determining IOL power in cataract patients with previous PPV (available at http://ppv-iolcalculator.com/).

Predicting the risk of glaucoma-related adverse events following secondary intraocular lens implantation in paediatric eyes: a 3-year study.

AIMS: To establish and evaluate predictive models for glaucoma-related adverse events (GRAEs) following secondary intraocular lens (IOL) implantation in paediatric eyes. METHODS: 205 children (356 aphakic eyes) receiving secondary IOL implantation at Zhongshan Ophthalmic Center with a 3-year follow-up were enrolled. Cox proportional hazard model was used to identify predictors of GRAEs and developed nomograms. Model performance was evaluated with time-dependent receiver operating characteristic (ROC) curves, decision curve analysis, Kaplan-Meier curves and validated internally through C-statistics and calibration plot of the bootstrap samples. RESULTS: Older age at secondary IOL implantation (HR=1.5, 95% CI: 1.03 to 2.19), transient intraocular hypertension (HR=9.06, 95% CI: 2.97 to 27.67) and ciliary sulcus implantation (HR=14.55, 95% CI: 2.11 to 100.57) were identified as risk factors for GRAEs (all p<0.05). Two nomograms were established. At postoperatively 1, 2 and 3 years, model 1 achieved area under the ROC curves (AUCs) of 0.747 (95% CI: 0.776 to 0.935), 0.765 (95% CI: 0.804 to 0.936) and 0.748 (95% CI: 0.736 to 0.918), and the AUCs of model 2 were 0.881 (95% CI: 0.836 to 0.926), 0.895 (95% CI: 0.852 to 0.938) and 0.848 (95% CI: 0.752 to 0.945). Both models demonstrated fine clinical net benefit and performance in the interval validation. The Kaplan-Meier curves showing two distinct risk groups were well discriminated and robust in both models. An online risk calculator was constructed. CONCLUSION: Two nomograms could sensitively and accurately identify children at high risk of GRAEs after secondary IOL implantation to help early identification and timely intervention.

Intraocular lens tilt and decentration in secondary ciliary sulcus implantation in paediatric eyes: A 3-year prospective study.

PURPOSE: The purpose of this study was to compare the tilt and decentration of one-piece anti-vaulting haptic intraocular lenses (IOL) and three-piece C-loop haptic IOLs in paediatric eyes undergoing secondary IOL implantation into the ciliary sulcus. METHODS: Paediatric aphakic patients receiving either one-piece anti-vaulting haptic or three-piece C-loop haptic IOL implants into the ciliary sulcus were enrolled in this prospective non-randomized interventional study and followed up for 3 years. IOL decentration and tilt were measured using Scheimpflug images. Preoperative and postoperative information, including demographic data and ocular biometric parameters and complications, were collected and analysed. RESULTS: Among 123 eyes of 79 paediatric patients, there were 72 eyes (58.54%) in the anti-vaulting haptic IOL group and 51 eyes (41.46%) in the C-loop haptic group. The anti-vaulting haptic IOL group had a lower incidence of clinically significant vertical IOL decentration than the C-loop haptic IOL group (23.88% vs. 43.14%, p = 0.037). No intergroup differences were observed in vertical or horizontal tilt or in horizontal decentration (all p > 0.05). One-piece anti-vaulting haptic IOL implantation was associated with a lower risk of clinically significant vertical decentration than three-piece C-loop haptic IOL implantation (odds ratio: 0.42, p = 0.037). There was a higher incidence of IOL dislocation in the C-loop haptic IOL group (15.22% vs. 4.17%, p = 0.046). CONCLUSIONS: In paediatric aphakic eyes undergoing secondary IOL implantation into the ciliary sulcus, one-piece anti-vaulting haptic IOLs can reduce the risk of clinically significant vertical IOL decentration compared with three-piece C-loop haptic IOLs and may favour long-term IOL positional stability.

p53 promotes revival stem cells in the regenerating intestine after severe radiation injury.

Ionizing radiation induces cell death in the gastrointestinal (GI) epithelium by activating p53. However, p53 also prevents animal lethality caused by radiation-induced acute GI syndrome. Through single-cell RNA-sequencing of the irradiated mouse small intestine, we find that p53 target genes are specifically enriched in regenerating epithelial cells that undergo fetal-like reversion, including revival stem cells (revSCs) that promote animal survival after severe damage of the GI tract. Accordingly, in mice with p53 deleted specifically in the GI epithelium, ionizing radiation fails to induce fetal-like revSCs. Using intestinal organoids, we show that transient p53 expression is required for the induction of revival stem cells and is controlled by an Mdm2-mediated negative feedback loop. Together, our findings reveal that p53 suppresses severe radiation-induced GI injury by promoting fetal-like reprogramming of irradiated intestinal epithelial cells.

Preclinical Evaluation of the ATR Inhibitor BAY 1895344 as a Radiosensitizer for Head and Neck Squamous Cell Carcinoma.

PURPOSE: Despite aggressive multimodal treatment that typically includes definitive or adjuvant radiation therapy (RT), locoregional recurrence rates approach 50% for patients with locally advanced human papillomavirus (HPV)-negative head and neck squamous cell carcinoma (HNSCC). Thus, more effective therapeutics are needed to improve patient outcomes. We evaluated the radiosensitizing effects of ataxia telangiectasia and RAD3-related (ATR) inhibitor (ATRi) BAY 1895344 in preclinical models of HNSCC. METHODS AND MATERIALS: Murine and human HPV-negative HNSCC cells (MOC2, MOC1, JHU-012) were treated with vehicle or ATRi with or without 4 Gy. Checkpoint kinase 1 phosphorylation and DNA damage (γH2AX) were evaluated by Western blot, and ATRi half-maximal inhibitory concentration was determined by MTT assay for HNSCC cells and immortalized murine oral keratinocytes. In vitro radiosensitization was tested by clonogenic assay. Cell cycle distribution and mitotic catastrophe were evaluated by flow cytometry. Mitotic aberrations were quantified by fluorescent microscopy. Tumor growth delay and survival were assessed in mice bearing MOC2 or JHU-012 transplant tumors treated with vehicle, ATRi, RT (10 Gy × 1 or 8 Gy × 3), or combined ATRi + RT. RESULTS: ATRi caused dose-dependent reduction in checkpoint kinase 1 phosphorylation at 1 hour post-RT (4 Gy) and dose-dependent increase in γH2AX at 18 hours post-RT. Addition of RT to ATRi led to decreased BAY 1895344 half-maximal inhibitory concentration in HNSCC cell lines but not in normal tissue surrogate immortalized murine oral keratinocytes. Clonogenic assays demonstrated radiosensitization in the HNSCC cell lines. ATRi abrogated the RT-induced G2/M checkpoint, leading to mitosis with unrepaired DNA damage and increased mitotic aberrations (multinucleated cells, micronuclei, nuclear buds, nucleoplasmic bridges). ATRi and RT significantly delayed tumor growth in MOC2 and JHU-012 in vivo models, with improved overall survival in the MOC2 model. CONCLUSIONS: These findings demonstrated that BAY 1895344 increased in vitro and in vivo radiosensitivity in HPV-negative HNSCC preclinical models, suggesting therapeutic potential warranting evaluation in clinical trials for patients with locally advanced or recurrent HPV-negative HNSCC.