Correlation of gene expression with magnetic resonance imaging features of retinoblastoma: a multi-center radiogenomics validation study.

OBJECTIVES: To validate associations between MRI features and gene expression profiles in retinoblastoma, thereby evaluating the repeatability of radiogenomics in retinoblastoma. METHODS: In this retrospective multicenter cohort study, retinoblastoma patients with gene expression data and MRI were included. MRI features (scored blinded for clinical data) and matched genome-wide gene expression data were used to perform radiogenomic analysis. Expression data from each center were first separately processed and analyzed. The end product normalized expression values from different sites were subsequently merged by their Z-score to permit cross-sites validation analysis. The MRI features were non-parametrically correlated with expression of photoreceptorness (radiogenomic analysis), a gene expression signature informing on disease progression. Outcomes were compared to outcomes in a previous described cohort. RESULTS: Thirty-six retinoblastoma patients were included, 15 were female (42%), and mean age was 24 (SD 18) months. Similar to the prior evaluation, this validation study showed that low photoreceptorness gene expression was associated with advanced stage imaging features. Validated imaging features associated with low photoreceptorness were multifocality, a tumor encompassing the entire retina or entire globe, and a diffuse growth pattern (all p < 0.05). There were a number of radiogenomic associations that were also not validated. CONCLUSIONS: A part of the radiogenomic associations could not be validated, underlining the importance of validation studies. Nevertheless, cross-center validation of imaging features associated with photoreceptorness gene expression highlighted the capability radiogenomics to non-invasively inform on molecular subtypes in retinoblastoma. CLINICAL RELEVANCE STATEMENT: Radiogenomics may serve as a surrogate for molecular subtyping based on histopathology material in an era of eye-sparing retinoblastoma treatment strategies. KEY POINTS: • Since retinoblastoma is increasingly treated using eye-sparing methods, MRI features informing on molecular subtypes that do not rely on histopathology material are important. • A part of the associations between retinoblastoma MRI features and gene expression profiles (radiogenomics) were validated. • Radiogenomics could be a non-invasive technique providing information on the molecular make-up of retinoblastoma.

MRI Features for Identifying MYCN-amplified RB1 Wild-type Retinoblastoma.

Background MYCN-amplified RB1 wild-type (MYCNARB1+/+) retinoblastoma is a rare but clinically important subtype of retinoblastoma due to its aggressive character and relative resistance to typical therapeutic approaches. Because biopsy is not indicated in retinoblastoma, specific MRI features might be valuable to identify children with this genetic subtype. Purpose To define the MRI phenotype of MYCNARB1+/+ retinoblastoma and evaluate the ability of qualitative MRI features to help identify this specific genetic subtype. Materials and Methods In this retrospective, multicenter, case-control study, MRI scans in children with MYCNARB1+/+ retinoblastoma and age-matched children with RB1-/- subtype retinoblastoma were included (case-control ratio, 1:4; scans acquired from June 2001 to February 2021; scans collected from May 2018 to October 2021). Patients with histopathologically confirmed unilateral retinoblastoma, genetic testing (RB1/MYCN status), and MRI scans were included. Associations between radiologist-scored imaging features and diagnosis were assessed with the Fisher exact test or Fisher-Freeman-Halton test, and Bonferroni-corrected P values were calculated. Results A total of 110 patients from 10 retinoblastoma referral centers were included: 22 children with MYCNARB1+/+ retinoblastoma and 88 control children with RB1-/- retinoblastoma. Children in the MYCNARB1+/+ group had a median age of 7.0 months (IQR, 5.0-9.0 months) (13 boys), while children in the RB1-/- group had a median age of 9.0 months (IQR, 4.6-13.4 months) (46 boys). MYCNARB1+/+ retinoblastomas were typically peripherally located (in 10 of 17 children; specificity, 97%; P < .001) and exhibited plaque or pleomorphic shape (in 20 of 22 children; specificity, 51%; P = .011) with irregular margins (in 16 of 22 children; specificity, 70%; P = .008) and extensive retina folding with vitreous enclosure (specificity, 94%; P < .001). MYCNARB1+/+ retinoblastomas showed peritumoral hemorrhage (in 17 of 21 children; specificity, 88%; P < .001), subretinal hemorrhage with a fluid-fluid level (in eight of 22 children; specificity, 95%; P = .005), and strong anterior chamber enhancement (in 13 of 21 children; specificity, 80%; P = .008). Conclusion MYCNARB1+/+ retinoblastomas show distinct MRI features that could enable early identification of these tumors. This may improve patient selection for tailored treatment in the future. © RSNA, 2023 Supplemental material is available for this article. See also the editorial by Rollins in this issue.

PENTOSAN POLYSULFATE AND VISION: Findings from an International Survey of Exposed Individuals.

PURPOSE: To investigate patient-reported visual function among individuals taking pentosan polysulfate (PPS) for interstitial cystitis. METHODS: A 27-item online survey was distributed to an international mailing list of individuals with interstitial cystitis in November 2018. Demographic characteristics, PPS exposure history, subjective visual function, and previous macular diagnoses were queried. The impact of PPS use, grouped by tertile of cumulative exposure, on visual function and macular diagnoses was assessed with multivariate logistic regression. RESULTS: The survey was completed by 912 respondents. Eight hundred and sixty-one (96.4%) were women, and the median age was 55 [interquartile range (IQR), 45-64 years]. Among PPS users, the median exposure was 547.5 g (IQR, 219-1,314 g). Respondents in the highest PPS exposure tertile were more likely to report difficulty with reading small print [adjusted odds ratio 2.29, 95% confidence interval (CI) 1.15-4.57] and to have a diagnosis of macular degeneration and/or pigmentary maculopathy (adjusted odds ratio 2.41, 95% CI 1.44-4.03) than unexposed respondents. CONCLUSION: In this large sample of individuals with interstitial cystitis, those in the highest PPS exposure category were more likely to have difficulties reading small print and to report a previous diagnosis of macular disease. Further study of objective measures of visual function in PPS users is warranted.

Bias Reduction Practices in Underrepresented Groups in Ophthalmology Resident Recruitment.

Importance: Best recruitment practices for increasing diversity are well established, but the adoption and impact of these practices in ophthalmology residency recruitment are unknown. Objective: To describe the adoption of bias reduction practices in groups underrepresented in ophthalmology (URiO) residency recruitment and determine which practices are effective for increasing URiO residents. Design, Setting, and Participants: This cross-sectional survey study used an 18-item questionnaire included in the online survey of the Association of University Professors in Ophthalmology (AUPO) Residency Program Directors. Data collection occurred from July 2022 to December 2022. The data were initially analyzed on January 16, 2023. Participants included residency program directors (PDs) in the AUPO PD listserv database. Main Outcomes and Measures: Descriptive analysis of resident selection committee approaches, evaluation of applicant traits, and use of bias reduction tools. Primary outcome was diversity assessed by presence of at least 1 resident in the last 5 classes who identified as URiO, including those underrepresented in medicine (URiM), lesbian, gay, bisexual, transgender, queer, intersex, and asexual plus, or another disadvantaged background (eg, low socioeconomic status). Multivariate analyses of recruitment practices were conducted to determine which practices were associated with increased URiO and URiM. Results: Among 106 PDs, 65 completed the survey (61.3%). Thirty-nine PDs used an interview rubric (60.0%), 28 used interview standardization (43.0%), 56 provided at least 1 bias reduction tool to their selection committee (86.2%), and 44 used postinterview metrics to assess diversity, equity, and inclusion efforts (67.7%). Application filters, interview standardization, and postinterview metrics were not associated with increased URiO. Multivariate logistic regression analysis showed larger residency class (odds ratio [OR], 1.34; 95% CI, 1.09-1.65; P = .01) and use of multiple selection committee bias reduction tools (OR, 1.47; 95% CI, 1.13-1.92; P = .01) were positively associated with increased URiO, whereas use of interview rubrics (OR, 0.72; 95% CI, 0.59-0.87; P = .001) and placing higher importance of applicant interest in a program (OR, 0.83; 95% CI, 0.75-0.92; P = .02) were negatively associated. URiM analyses showed similar associations. Conclusions and Relevance: Ophthalmology residency interviews are variably standardized. In this study, providing multiple bias reduction tools to selection committees was associated with increased URiO and URiM residents. Prioritizing applicant interest in a program may reduce resident diversity. Interview rubrics, while intended to reduce bias, may inadvertently increase inequity.

Lowering the Incidence of Endophthalmitis Following Intravitreal Anti-VEGF Injection: An Analysis of Aseptic Protocol Adjustment.

BACKGROUND AND OBJECTIVE: The impact of anti-sepsis-anesthesia sequence in intravitreal injection (IVI)-associated endophthalmitis is unknown. We compared outcomes of patients who had 10% topical povidone-iodine before or after 2% topical lidocaine gel during IVIs. PATIENTS AND METHODS: A retrospective study of IVIs in nine clinical sites was undertaken. Group 1 had lidocaine gel applied first. This protocol was changed on March 1, 2020, with Group 2 having povidone-iodine applied first. Visual and micro-biological outcomes were compared. RESULTS: Among 72 cases (0.07%) from 102,908 IVIs, Group 1 had 59 cases from 65,307 IVI (0.09%) and Group 2 had 13 cases from 37,601 IVI (0.03%; P = 0.001). There was no significant difference in the best-corrected visual acuity between groups. Highly virulent bacteria were predominantly isolated in Group 1, but proportions of gram-positive bacterial growth were similar. CONCLUSIONS: Application of povidone-iodine before lidocaine gel, compared to after, significantly decreased rate of IVI endophthalmitis, with no significant changes in visual and microbiological outcomes. [Ophthalmic Surg Lasers Imaging Retina 2023;54:520-525.].

Artificial intelligence in retinal image analysis: Development, advances, and challenges.

Modern advances in diagnostic technologies offer the potential for unprecedented insight into ophthalmic conditions relating to the retina. We discuss the current landscape of artificial intelligence in retina with respect to screening, diagnosis, and monitoring of retinal pathologies such as diabetic retinopathy, diabetic macular edema, central serous chorioretinopathy, and age-related macular degeneration. We review the methods used in these models and evaluate their performance in both research and clinical contexts and discuss potential future directions for investigation, use of multiple imaging modalities in artificial intelligence algorithms, and challenges in the application of artificial intelligence in retinal pathologies.

Human MPox (Monkeypox) Virus Membranous Keratoconjunctivitis With Transient Corneal Hypoesthesia and Late Symblepharon Formation: A Novel Case and Clinical Implications.

PURPOSE: The aim of this study was to describe a case of corneal involvement as an early manifestation of ocular disease in the 2022 human mpox (monkeypox) virus outbreak. METHODS: This is a single case report with longitudinal care. RESULTS: A 47-year-old immunocompetent man presented with viral conjunctivitis before development of skin lesions or systemic symptoms. Subsequently, he developed membranous keratoconjunctivitis and a corneal epithelial defect. Orthopoxvirus-positive polymerase chain reaction test from his ocular surface was positive. The epithelial defect did not heal with conservative treatment but was successfully treated with amniotic membrane transplantation over 8 days. Reduced corneal sensation was noted after epithelial healing, and polymerase chain reaction from the ocular surface remained positive at 17 days from symptom onset, with slowly recovering conjunctivitis at 21 days. Continued membrane formation required repeated removal but significantly improved with topical corticosteroid treatment after epithelial healing by 29 days of symptom onset. Corneal sensation normalized by 87 days from symptom onset at which time symblepharon were noted but PCR testing from the ocular surface was negative. CONCLUSIONS: Early corneal involvement of human monkeypox virus is possible. Transient corneal hypoesthesia may be due to acute inflammation. Chronic inflammatory changes can result in symblepharon. These findings have potential implications in patient care and corneal donation.

Animal Models of Uveal Melanoma.

Animal models are crucial for the study of tumorigenesis and therapies in oncology research. Though rare, uveal melanoma (UM) is the most common intraocular tumor and remains one of the most lethal cancers. Given the limitations of studying human UM cells in vitro, animal models have emerged as excellent platforms to investigate disease onset, progression, and metastasis. Since Greene's initial studies on hamster UM, researchers have dramatically improved the array of animal models. Animals with spontaneous tumors have largely been replaced by engrafted and genetically engineered models. Inoculation techniques continue to be refined and expanded. Newer methods for directed mutagenesis have formed transgenic models to reliably study primary tumorigenesis. Human UM cell lines have been used to generate rapidly growing xenografts. Most recently, patient-derived xenografts have emerged as models that closely mimic the behavior of human UM. Separate animal models to study metastatic UM have also been established. Despite the advancements, the prognosis has only recently improved for UM patients, especially in patients with metastases. There is a need to identify and evaluate new preclinical models. To accomplish this goal, it is important to understand the origin, methods, advantages, and disadvantages of current animal models. In this review, the authors present current and historic animal models for the experimental study of UM. The strengths and shortcomings of each model are discussed and potential future directions are explored.

Analysis of Toxicity and Clinical Outcomes in Full Versus Reduced Starting Dose Cabozantinib in Metastatic Renal Cell Carcinoma Patients.

BACKGROUND: Full dose cabozantinib for metastatic renal cell carcinoma (mRCC) is 60 mg, but adverse events (AEs) may require dose reductions. Limited data exist comparing efficacy among cabozantinib doses. We compared AEs and clinical outcomes in mRCC patients treated with full vs. reduced starting cabozantinib dose. METHODS: We performed a retrospective analysis of 87 mRCC patients treated with cabozantinib at Winship Cancer Institute from 2016 to 2019. Overall survival (OS), progression-free survival (PFS), and objective response (OR) rate measured clinical outcomes. AEs were collected from clinic notes and the most common were hypertension, mucositis/hand-foot skin reaction (HFSR), or gastrointestinal toxicity. Univariate analysis (UVA) between starting doses and AEs with clinical outcomes was performed using logistic regression model. Multivariable analysis was also performed using Cox proportional hazard model. RESULTS: Most patients were men (71%) with clear-cell RCC (72%). The majority were IMDC intermediate (58%) or poor (35%) risk. One third received first-line cabozantinib and 64% had ≥3 baseline metastatic sites. Most patients (68%) required dose reduction from 60 mg or started at reduced dose without escalation. Reduced dose patients were more likely to have ≥3 distant metastatic sites (70% vs. 58%) and ≥2 prior lines of systemic therapy (50% vs. 40%) compared to full dose patients. UVA revealed a trend towards shorter OS (HR: 1.78, P = .095), PFS (HR: 1.50, P = .107), and lower chance of OR (HR:0.42, P = .149) among reduced dose patients. This trend did not hold in Multivariable analysis (OS HR: 1.20, P = .636; PFS HR: 1.23, P = .4662). Mucositis/HFSR and hypertension were significantly associated with improved outcomes in UVA. CONCLUSIONS: Although we found a trend favoring full dose cabozantinib, this is likely due to worse baseline disease characteristics among patients starting on a reduced dose. Hypertension and mucositis/HFSR may be associated with improved outcomes. Larger studies are warranted to validate these findings.

Clinical and Histopathological Features of Corneal Primary Acquired Melanosis and Melanoma.

AIM: The aim of this study is to describe the clinical and pathologic features of corneal primary acquired melanosis (PAM) and melanoma. METHODS: We describe 3 cases in total: two cases of corneal melanomas and 1 case of corneal PAM. The eyes were processed routinely for histopathological examination. Clinical histories, treatments, and outcomes were reviewed. RESULTS: Corneal melanomas arose from recurrence of conjunctival melanoma or conjunctival PAM at the limbus. One patient had a recurrence after excision of a limbal melanoma, another had a de novo corneal melanoma, and the last patient had corneal PAM in the setting of conjunctival PAM with atypia. All lesions were excised with adjuvant alcohol debridement and cryotherapy with no recurrence ranging from 1 week to 8 years. CONCLUSIONS: Corneal melanomas arise at the limbus from corneal PAM or conjunctival atypia. They can appear after excisional removal of a conjunctival melanoma. Surgical excision with alcohol debridement and adjuvant cryotherapy is successful.

Estimation of the timing of BAP1 mutation in uveal melanoma progression.

Uveal melanoma is the most common primary intraocular malignancy. A vast majority of metastasizing tumors have mutations in the BAP1 gene. Here, we investigate the spatiotemporal timing of these mutations. The size of 177 uveal melanomas and 8.3 million individual tumor cells was measured. BAP1 sequencing results and BAP1 IHC were available and for 76 (43%) and 101 (57%) of these, respectively. Tumors with a BAP1 mutation had significantly larger volume (2109 vs. 1552 mm3, p = 0.025). Similarly, tumor cells with loss of BAP1 protein expression had significantly larger volume (2657 vs. 1593 µm3, p = 0.027). Using observations of the time elapsed between mitoses, the BAP1 mutation was calculated to occur when the primary tumor had a size of a few malignant cells to 6 mm3, 0.5 to 4.6 years after tumor initiation and at least 9 years before diagnosis. We conclude that BAP1 mutations occur early in the growth of uveal melanoma, well before the average tumor is diagnosed. Its timing coincides with the seeding of micrometastases.

Reactivation of Retinopathy of Prematurity in Adults and Adolescents.

PURPOSE: To review the clinical features, treatment outcomes, and prevalence within our clinic population of adolescents and adults with previously regressed retinopathy of prematurity (ROP) who demonstrate late-onset exudation and vasoproliferative changes. DESIGN: Retrospective review of consecutive patients at a single center. PARTICIPANTS: Five patients (5 eyes) with a history of ROP who showed new exudates or worsening fibrovascular proliferation diagnosed after 10 years of age. METHODS: Patients were identified by a computerized search of the Emory Eye Center billing records. Data extracted from charts included baseline ROP information, visual acuity and other examination findings, imaging, and treatments. MAIN OUTCOME MEASURES: Status of exudation and vasoproliferation. RESULTS: Among 138 patients older than 10 years with ROP seen at our tertiary referral center from 2000 through 2018, 5 (3.6%) demonstrated late-onset exudation or vasoproliferation. Three patients were female and 3 underwent ROP treatment as neonates. Mean age at onset of late reactivation was 25.6 years (range, 13-43 years). Previous treatments for neonatal ROP included peripheral laser ablation (n = 3), scleral buckle (n = 2), pars plicata vitrectomy (n = 2), and no treatment (n = 2). Management strategies for late reactivation included observation (n = 1), intravitreal anti-vascular endothelial growth factor agents (n = 4), vitrectomy (n = 2), and cryotherapy (n = 1). With mean follow-up of 4.8 years (range, 1-7 years), outcomes were resolution of exudation or proliferation with return to baseline vision (n = 2), stable mild exudation (n = 1), and progressive vasoproliferation with traction leading to phthisis (n = 2). CONCLUSIONS: Late-onset exudation and fibrovascular proliferation in adolescents and adults with ROP can occur rarely with previously regressed ROP. Two of 5 patients were refractory to all treatments and demonstrated phthisis bulbi. One patient showed reactivation in the form of a reactive retinal astrocytic tumor. Our findings highlight the importance continued monitoring with regular fundus examination in adolescents and adults with regressed ROP.

Ophthalmology Education in COVID-19: A Remote Elective for Medical Students.

BACKGROUND: The novel coronavirus disease (COVID-19) has created obstacles for medical student education, as clinical rotations were temporarily halted. Recent literature shows online electives may provide an alternative learning platform. We developed a tele-ophthalmology student elective for rising third-year (MS3) and fourth-year (MS4) medical students to continue teaching and exposure to the field. METHODS: A 4-week remote elective was approved by Emory University School of Medicine and offered between April 18, 2020 to May 15, 2020 for rising MS3 and MS4 students. The curriculum consisted of online self-study materials, student presentations, chart review assignments, case-based discussions with faculty, and telehealth experiences. All students were surveyed and tested with questions from USMLE World (UWorld) test bank at the end of the course. RESULTS: A total of 18 students enrolled, with 66.7% MS3 and 33.3% MS4 participance. The mean rating of fulfillment of course learning objectives was 8.1/10 (range, 6.7-8.8), with mean ratings of 8.2 for MS3s and 7.7 for MS4s. There was a significant increase in self-reported knowledge in ophthalmology, with an increase from 4.6 to 8.1 for MS3s (p=0.002) and 6.7 to 8.0 for MS4s (p=0.04). Students also reported higher interest in the field, with an increase from 4.9 to 7.8 for MS3s (p=0.01) and 7.5 to 8.7 for MS4s (p=0.1). The students performed significantly higher on the post-course test (94.8%) than UWorld question bank users (74.1%) (p<0.001). CONCLUSION: Our novel ophthalmology elective significantly enhanced self-reported medical student knowledge and interest in the field during a crisis that required transition to remote learning. Further study of student telehealth experience and objective assessment is needed to improve online learning in ophthalmology.

Visual and Anatomic Outcomes of Premacular Hemorrhage in Non-Accidental Trauma Infants Managed With Observation or Vitrectomy.

BACKGROUND AND OBJECTIVE: The management of premacular hemorrhage secondary to non-accidental trauma (NAT) is unclear. The authors describe the outcomes of NAT infants referred for surgical evaluation of premacular hemorrhage. PATIENTS AND METHODS: Retrospective institutional review board-approved case series between 2000 and 2019 of vision-threatening premacular hemorrhage (sub-hyaloid or sub-internal limiting membrane hemorrhage without vitreous hemorrhage) in NAT infants. Time to hemorrhage resolution, vision, and comorbidities were collected. RESULTS: Thirty-six patients (62 eyes) with mean age of 5.4 months (range: 2-10 months) were included. Nine eyes (14.5%) underwent vitrectomy. Median time to hemorrhage resolution by observation was 75 days (interquartile range [IQR]: 60-120 days), and time to vitrectomy was 54.5 days (IQR: 47.8-58.5 days). Eight eyes (12.9%) had amblyopia, which was not significantly different between groups. Despite hemorrhage clearance, a higher proportion of eyes in the vitrectomy group had pigmentary changes (P = .04) and strabismus (P = .002) at follow-up. CONCLUSIONS: Most cases of NAT-related premacular hemorrhage resolve within 3 months without surgical intervention. Comorbidities may limit visual prognosis. [Ophthalmic Surg Lasers Imaging Retina. 2020;51:715-722.].

SAP97 regulates behavior and expression of schizophrenia risk enriched gene sets in mouse hippocampus.

Synapse associated protein of 97KDa (SAP97) belongs to a family of scaffolding proteins, the membrane-associated guanylate kinases (MAGUKs), that are highly enriched in the postsynaptic density of synapses and play an important role in organizing protein complexes necessary for synaptic development and plasticity. The Dlg-MAGUK family of proteins are structurally very similar, and an effort has been made to parse apart the unique function of each Dlg-MAGUK protein by characterization of knockout mice. Knockout mice have been generated and characterized for PSD-95, PSD-93, and SAP102, however SAP97 knockout mice have been impossible to study because the SAP97 null mice die soon after birth due to a craniofacial defect. We studied the transcriptomic and behavioral consequences of a brain-specific conditional knockout of SAP97 (SAP97-cKO). RNA sequencing from hippocampi from control and SAP97-cKO male animals identified 67 SAP97 regulated transcripts. As large-scale genetic studies have implicated MAGUKs in neuropsychiatric disorders such as intellectual disability, autism spectrum disorders, and schizophrenia (SCZ), we analyzed our differentially expressed gene (DEG) set for enrichment of disease risk-associated genes, and found our DEG set to be specifically enriched for SCZ-related genes. Subjecting SAP97-cKO mice to a battery of behavioral tests revealed a subtle male-specific cognitive deficit and female-specific motor deficit, while other behaviors were largely unaffected. These data suggest that loss of SAP97 may have a modest contribution to organismal behavior. The SAP97-cKO mouse serves as a stepping stone for understanding the unique role of SAP97 in biology.