Investigating druggable kinases for targeted therapy in retinoblastoma.

Retinoblastoma (RB) is a childhood retinal neoplasm and commonly treated with cytotoxic chemotherapeutic agents. However, these therapeutic approaches often lead to diverse adverse effects. A precise molecular therapy will alleviate these side effects and offer better treatment outcomes. Over the years, kinases have become potential drug targets in cancer therapy. Hence, we aimed to investigate genetic alterations of putative kinase drug targets in RB. Targeted exome sequencing was performed on 35 RB tumors with paired blood samples using a gene panel consisting of 29 FDA-approved kinase genes. Single nucleotide variants were analyzed for pathogenicity using an in-house pipeline and copy number variations (CNVs) were detected by a depth of coverage and CNVPanelizer. The correlation between genetic changes and clinicopathological features was assessed using GraphPad Prism. Three somatic mutations, two in ERBB4 and one in EGFR were identified. Two of these mutations (ERBB4 c.C3836A & EGFR c.A1196T) were not reported earlier. CNV analysis revealed recurrent gains of ALK, MAP2K2, SRC, STK11, and FGFR3 as well as frequent losses of ATM, PI3KCA and ERBB4. Notably, nonresponsive tumors had a higher incidence of amplifications in clinically actionable genes such as ALK. Moreover, ALK gain and ATM loss were strongly correlated with optic nerve head invasion. In conclusion, our study revealed genetic alterations of druggable kinases in RB, providing preliminary insights for the exploration of kinase-targeted therapy in RB.

COVID-19-associated rhino-orbito-cerebral mucormycosis: a single center prospective study of 264 patients.

PURPOSE: Outbreaks of mucormycosis were reported worldwide throughout the COVID-19 pandemic. We report clinical outcomes of a treatment protocol for COVID-19-associated rhino-orbital-cerebral mucormycosis (ROCM). METHODS: Patients with biopsy-proven mucormycosis and COVID-19 were included. All received intravenous amphotericin B deoxycholate 1 mg/kg and surgical endoscopic sinus debridement (FESS). Those with rhino-orbital or cerebral disease limited to the cavernous sinus were eligible for transcutaneous retrobulbar amphotericin B (TRAMB). Patients were followed with weekly imaging, endoscopic examinations, and serial debridement as necessary. Patients were discharged on oral posaconazole for 6 months. RESULTS: In total, 264 patients were followed for a mean of 2.5 months. On presentation, 163 patients (174 eyes) had eye involvement. Of these, 141 eyes (81.0%) had light perception or worse vision. By the last follow-up, 163 patients (176 eyes) were affected, and of these, 96 eyes (54.5%) had no light perception. Twenty-one patients (8%) died and 3 orbits (0.5%) were exenterated. There was no change in mortality (p = 0.38) or exenteration (p = 0.38) in the 55 patients who received TRAMB compared to patients with rhino-orbital or cerebral disease limited to the cavernous sinus who did not. Asymptomatic COVID-19 was associated with higher mortality than symptomatic COVID-19 (p = 0.025). Uncontrolled diabetes was a risk factor for death (p = 0.022). New diabetes was associated with increased mortality versus pre-existing diabetes (p = 0.005). CONCLUSION: A multidisciplinary approach is crucial to manage COVID-19-ROCM. In our cohort, TRAMB therapy did not increase mortality or exenteration rates. While poor vision on presentation was profound, some vision recovery was noted with treatment. COVID-19 immune dysregulation may predispose patients to ROCM, particularly those with asymptomatic disease.

Focused cancer pathway analysis revealed unique therapeutic targets in retinoblastoma.

Retinoblastoma (RB) is a pediatric cancer of the eye that occurs in 1/15000 live births worldwide. Albeit RB is initiated by the inactivation of RB1 gene, the disease progression relies largely on transcriptional alterations. Therefore, evaluating gene expression is vital to unveil the therapeutic targets in RB management. In this study, we employed an RT2 Profiler™ PCR array for a focused analysis of 84 cancer-specific genes in RB. An interaction network was built with gene expression data to identify the dysregulated pathways in RB. The key transcript alterations identified in 13 tumors by RT2 Profiler™ PCR array was further validated in 15 tumors by independent RT-qPCR. Out of 84 cancer-specific genes, 68 were dysregulated in RB tumors. Among the 68 genes, 23 were chosen for further analysis based on statistical significance and abundance across multiple tumors. Pathway analysis of altered genes showed the frequent perturbations of cell cycle, angiogenesis and apoptotic pathways in RB. Notably, upregulation of MCM2, MKI67, PGF, WEE1, CDC20 and downregulation of COX5A were found in all the tumors. Western blot confirmed the dysregulation of identified targets at protein levels as well. These alterations were more prominent in invasive RB, correlating with the disease pathogenesis. Our molecular analysis thus identified the potential therapeutic targets for improving retinoblastoma treatment. We also suggest that PCR array can be used as a tool for rapid and cost-effective gene expression analysis.

Mapping prognostic factors for globe survival in panophthalmitis using logistic regression and Cox proportional hazard models: A retrospective study.

Purpose: The aim of this study was to report the outcomes of panophthalmitis and to identify factors significantly affecting globe survival in the disease. Methods: This was a retrospective study on patients with panophthalmitis in a tertiary hospital between January 1, 2017, and December 31, 2019. The demographics, treatment details, culture results, and final outcomes were recorded. Logistic regression and Cox proportional hazards (CPH) were calculated to identify variables associated with globe loss. A P < 0.05 was considered significant. Results: Eighty-five eyes of 85 patients (31 culture positive) were eligible for review. The mean age of the participants was 55.21 ± 20.17 years with a male-to-female ratio of 2.04:1. Corneal ulcer (38.82%; n = 33) and open globe injuries (OGIs) (38.82%; n = 33) were the most common etiologies. Pseudomonas aeruginosa (n = 10; 11.76%) was the most common isolate. The mean duration of hospital stay was 7.58 ± 2.32 days. Overall, 44 (51.76%) globes could be salvaged. The need for evisceration (P = 0.901) and hospital stays (P = 0.095) were similar for culture-positive and -negative cohorts. The unadjusted logistic regression and CPH models showed that culture sterility did not affect globe survival [OR = 1.210 (0.501-2.950), P = 0.668; HR = 1.176 (0.617-2.243), P = 0.623]. The adjusted logistic regression and the CPH models showed that corneal ulcers [OR = 10.900 (2.460-48.200), P = 0.002; HR = 5.393 (1.603-18.140), P = 0.006] and OGI [OR = 7.360 (1.650-32.700), P = 0.009; HR = 4.548 (1.321-15.660), P = 0.016] were significantly associated with globe loss. Conclusion: Corneal ulcer or OGI as the primary etiology is detrimental to globe survival in panophthalmitis.

Frontal osteomyelitis post-COVID-19 associated mucormycosis.

Rhino-orbito-cerebral mucormycosis (ROCM) is the most commonly noted form of mucormycosis, which is the most common secondary fungal infection following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Osteomyelitis is one of the rare sequelae of ROCM, frontal osteomyelitis being the rarest. We present four patients of coronavirus disease 2019 (COVID-19)-associated mucormycosis, who presented with frontal bone osteomyelitis after being treated for ROCM surgically and medically. This is the first case series highlighting this complication in post-COVID-19 mucormycosis patients and needs utmost attention as it can be life-threatening and can cause extreme facial disfiguration. All four patients are alive with salvage of the affected globe and vision being preserved in one patient. If identified early, disfiguration of face and intracranial extension can be avoided.

Orbital tooth - not always a teratoma : a case report.

A 7-year-old girl presented with a painless firm to cystic mass in the infero-temporal quadrant of her right orbit since birth. The mass had recurred with a cutaneous sinus after initial surgery. Right eye vision was affected by mass induced astigmatism. The anterior and posterior segments were normal. Magnetic Resonance Imaging of the orbit suggested a mature teratoma within the orbital bone. Mass excision showed a fully developed molar tooth within a cyst associated with a cutaneous sinus. Histopathological examination reported odontogenic choriostoma. We report this unique case of orbital odontogenic choristoma in an unusual location, associated with a cutaneous sinus, emphasizing the need for complete excision to prevent recurrence.

Searching beyond nevi - A rare case of neurocutaneous ocular syndrome.

Epidermal nevus syndrome is a rare congenital disorder affecting only a few hundred people in the world. It has ophthalmic, dermatological, and neurological manifestations, with varied presentation. Here, we report a case of two-year-old child who presented with epibulbar mass in left eye, pigmented nevi over left side of the body and alopecia over left side of parieto-temporal scalp. Imaging confirmed epibulbar mass and presence of calcification of choroid on ipsilateral side with presence of arachnoid cyst of brain with underlying pachygyria. Neurological examination was normal and dermatologist confirmed presence of verrucous nevi over skin. Excisional biopsy of epibulbar mass revealed a complex choristoma with presence of lacrimal gland tissue. Underlying ocular findings were near normal with normal posterior segment. It is a rare form of epidermal nevus syndrome with near normal ocular findings in the presence of anterior and posterior choristoma, which has not been reported.

Regurgitation on pressure over the lacrimal sac versus lacrimal irrigation in determining lacrimal obstruction prior to intraocular surgeries.

Purpose: : To determine the diagnostic accuracy of manual regurgitation on pressure over the lacrimal sac (ROPLAS) versus lacrimal irrigation for screening nasolacrimal duct obstruction (NLDO) in adults prior to intraocular surgeries. Methods: This cross-sectional study took place in a tertiary eye care hospital in South Tamil Nadu, India. From January to December 2017 and included consecutive patients who presented for routine cataract surgery. Prospective data collection occurred in 8369 eyes of patients who underwent cataract surgery. All patients underwent ROPLAS testing by an ophthalmologist followed by lacrimal irrigation by trained ophthalmic assistants, rechecked or confirmed in equivocal cases by ophthalmologists who were masked to the ROPLAS status. The primary outcome, the sensitivity, specificity, positive, and negative predictive values to detect lacrimal occlusion by ROPLAS compared with lacrimal irrigation with 95% confidence intervals was estimated. Results: : A total of 8369 eyes underwent cataract surgery during the time periods of the study. ROPLAS and lacrimal irrigation were performed in all eyes. The sensitivity of ROPLAS to diagnose NLDO correctly was 54.5% (95% CI, 44.8%-63.9%) and its specificity was 100% (95% CI, 100%-100%). The positive and negative predictive values were 75.3% (95% CI, 65.6%-83.0%) and 99.4% (95% CI, 99.2%-99.5%), respectively. Conclusion: : We found that ROPLAS when used alone had very low sensitivity and low positive predictive value in detecting NLDO prior to cataract surgery as compared with lacrimal irrigation. Hence, we recommend performing ROPLAS and lacrimal irrigation in every patient as part of the routine preoperative workup prior to cataract surgery.

Relationship Between Cooking Fuel and Lens Opacities in South India: A 15-Year Prospective Cohort Study.

PURPOSE: To determine whether exposure to unclean cooking fuels was associated with subsequent cataract progression as reported in previous cross-sectional studies. DESIGN: Prospective cohort study. METHODS: This is a secondary observational analysis of the community-based Antioxidants in Prevention of Cataracts trial (ClinicalTrials.gov ID NCT01664819). The exposure of interest was cooking fuel type, measured at baseline. Main outcome measures were baseline cataract severity and self-reported cataract surgery at a 15-year visit. RESULTS: Baseline and 15-year follow-up data were available for 798 and 579 participants, respectively. Wood or kerosene was used by 711 of 798 (89.1%) baseline participants, including 539 of 579 (93.1%) participants with complete follow-up. Cooking fuel type was not associated with cataract severity at baseline (P = .443). Of 8334 person-years of follow-up, 90 cataract surgeries were observed over 15 years (1.08 surgeries per 100 person-years; 95% CI = 0.87-1.32). Use of wood or kerosene was not associated with 15-year incidence of cataract surgery relative to use of propane (adjusted P = .154). Cataract surgery was more common in older individuals (HR = 1.1 per year, 95% CI = 1.1-1.2, P < .001), those with baseline myopia (HR = 2.1, 95% CI = 1.2-3.5, P = .009), and women (HR = 3.5, 95% CI = 1.2-10.1, P = .019). CONCLUSIONS: This study found no association between unclean cooking fuels and cataract progression over a 15-year period. No other modifiable risk factors were associated with incident self-reported cataract surgery.

Combined treatment modality including topical 20% fluorescein dye in the management of phthiriasis palpebrarum - A case series.

Purpose: To study the clinical presentations and outcomes of phthiriasis palpebrarum (PP) managed with combined treatment modality comprising of 20% fluorescein dye, mechanical removal of the ectoparasites, and topical application of antibiotic eye ointment with paraffin base. Methods: This is a prospective interventional noncomparative hospital-based series of 13 patients of PP. All the patients underwent local application of 20% fluorescein dye over the eyelashes and eyebrows of both the eyes followed by the mechanical removal of all the ectoparasites and trimming of the eyelashes from the base followed by application of ophthalmic ointment. Results: Mean age of the patients was 28 ± 22 years (range 3-60 Years). Out of the total of 13 patients, 11 patients (85%) were having bilateral involvement. The mean duration of symptoms in 11 patients (rest 2 were accidental findings) was 4 ± 3 weeks (range 1-10 weeks). Three patients (23%) had a history of sexual contact with possible parasite-infested partners. Four patients (31%) had additional parasites in the pubic area or torso. All the patients were completely parasite free at the end of 1 month. There was no infestation of the treating medical personnel from the patient. The average follow-up was 8 ± 5 months (range 3-21 months). Conclusion: Itching and irritation of the eyes were the most common presentations of PP. Combined treatment modality comprising of 20% fluorescein dye, mechanical removal of ectoparasites, and topical application of antibiotic eye ointment with paraffin base is effective in the management of PP.

Parental age and retinoblastoma-a retrospective study of demographic data and genetic analysis.

OBJECTIVE: To determine the association between the parental age gap and the absolute parental age with the risk of retinoblastoma (RB) development in an offspring. METHODS: RB individuals diagnosed between March 2013 and December 2019 in a single tertiary eye care centre were included. We recorded the demographic data, parental age and RB1 gene mutation status in the patient's tumour, blood and the parental blood. We categorised RB1 mutation inheritance as sporadic RB with somatic mutations (only present in tumour), heritable RB with de novo (present in patient's blood) and familial (present in patient and parents' blood) germline mutations. The statistical significance was confirmed by Fisher's exact/Chi-square test. RESULTS: Out of 259 RB patients, 247 were included in our study. Heritable RB with de novo germline mutations was significantly less common (p value: 0.0387; 95% CI: 0.2676-0.9329) and sporadic RB with somatic mutations was more common (p value: 0.0545; 95% CI: 1.025-3.39), if the parental age gap was <10 years. There were increased odds of a heritable RB with de novo germline mutation with an increase in paternal age and this was more intensified when combined with parental age gap of more than ≥10 years. The heritable RB with de novo germline mutations significantly increased as maternal age progressed, only when it was adjusted to ≥10 years parental age gap (p value: 0.0262; 95% CI: 1.26-17.91). CONCLUSIONS: An increased parental age gap and increased paternal age are independent risk factors for the development of heritable RB with de novo germline mutation.

A Rare Case of Acute Hemorrhagic Retinopathy Following Intravitreal Melphalan Injection for Persistent Vitreous Seeds in Retinoblastoma.

Purpose: This report describes a case of acute occlusive hemorrhagic complication after intravitreal melphalan for vitreous seeds in retinoblastoma. Methods: A case report is presented. Results: Intravitreal melphalan has been used extensively for vitreous seeds in retinoblastoma. Although melphalan is relatively safe at optimal doses, it can sometimes cause inadvertent complications like hemorrhagic events if the drug is administered close to the retina or in more pigmented eyes. We report a case of a 5-month-old patient with bilateral retinoblastoma who underwent enucleation of the right eye and 2 intravitreal melphalan injections in the left eye (20 µg/0.02 mL) at a 1-month interval for persistent vitreous seeds. After the second injection, there was a sudden decrease in the child's visual acuity in the left eye, and the retina showed multiple intraretinal hemorrhages and diffuse chorioretinal atrophy. Conclusion: Intravitreal melphalan may cause acute hemorrhagic complications after intravitreal use for retinoblastoma seeds, especially in pigmented eyes.

Human papillomavirus in retinoblastoma: A tertiary eye care center study from South India.

Purpose: This study is aimed to investigate the presence of Human papillomavirus (HPV) DNA in tumors obtained from sporadic retinoblastoma patients. Methods: One hundred six tumor tissues obtained from sporadic RB patients were analyzed for HPV infection by use of both seminested PCR and real-time quantitative PCR. Results: Of 106 RB patients, 55 were male and 51 were female. The mean age at diagnosis was 26.77 ± 15.36 (mean ± Std. dev) months. Almost all patients presented with leukocoria. Molecular investigation by different methods revealed no HPV positivity in any tumor genome. Conclusion: Our study demonstrates no association between HPV and RB, postulating HPV may not be a major risk factor in the etiology of RB.

Epidemiology, clinical profile, management, and outcome of COVID-19-associated rhino-orbital-cerebral mucormycosis in 2826 patients in India - Collaborative OPAI-IJO Study on Mucormycosis in COVID-19 (COSMIC), Report 1.

Purpose: COVID-19-associated rhino-orbital-cerebral mucormycosis (ROCM) has reached epidemic proportion during India's second wave of COVID-19 pandemic, with several risk factors being implicated in its pathogenesis. This study aimed to determine the patient demographics, risk factors including comorbidities, and medications used to treat COVID-19, presenting symptoms and signs, and the outcome of management. Methods: This was a retrospective, observational study of patients with COVID-19-associated ROCM managed or co-managed by ophthalmologists in India from January 1, 2020 to May 26, 2021. Results: Of the 2826 patients, the states of Gujarat (22%) and Maharashtra (21%) reported the highest number of ROCM. The mean age of patients was 51.9 years with a male preponderance (71%). While 57% of the patients needed oxygen support for COVID-19 infection, 87% of the patients were treated with corticosteroids, (21% for > 10 days). Diabetes mellitus (DM) was present in 78% of all patients. Most of the cases showed onset of symptoms of ROCM between day 10 and day 15 from the diagnosis of COVID-19, 56% developed within 14 days after COVID-19 diagnosis, while 44% had delayed onset beyond 14 days. Orbit was involved in 72% of patients, with stage 3c forming the bulk (27%). Overall treatment included intravenous amphotericin B in 73%, functional endoscopic sinus surgery (FESS)/paranasal sinus (PNS) debridement in 56%, orbital exenteration in 15%, and both FESS/PNS debridement and orbital exenteration in 17%. Intraorbital injection of amphotericin B was administered in 22%. At final follow-up, mortality was 14%. Disease stage >3b had poorer prognosis. Paranasal sinus debridement and orbital exenteration reduced the mortality rate from 52% to 39% in patients with stage 4 disease with intracranial extension (p < 0.05). Conclusion: : Corticosteroids and DM are the most important predisposing factors in the development of COVID-19-associated ROCM. COVID-19 patients must be followed up beyond recovery. Awareness of red flag symptoms and signs, high index of clinical suspicion, prompt diagnosis, and early initiation of treatment with amphotericin B, aggressive surgical debridement of the PNS, and orbital exenteration, where indicated, are essential for successful outcome.

Emerging role of tumor microenvironment derived exosomes in therapeutic resistance and metastasis through epithelial-to-mesenchymal transition.

The tumor microenvironment (TME) constitutes multiple cell types including cancerous and non-cancerous cells. The intercellular communication between these cells through TME derived exosomes may either enhance or suppress the tumorigenic processes. The tumor-derived exosomes could convert an anti-tumor environment into a pro-tumor environment by inducing the differentiation of stromal cells into tumor-associated cells. The exosomes from tumor-associated stromal cells reciprocally trigger epithelial-to-mesenchymal transition (EMT) in tumor cells, which impose therapeutic resistance and metastasis. It is well known that these exosomes contain the signals of EMT, but how these signals execute chemoresistance and metastasis in tumors remains elusive. Understanding the significance and molecular signatures of exosomes transmitting EMT signals would aid in developing appropriate methods of inhibiting them. In this review, we focus on molecular signatures of exosomes that shuttle between cancer cells and their stromal populations in TME to explicate their impact on therapeutic resistance and metastasis through EMT. Especially Wnt signaling is found to be involved in multiple ways of exosomal transport and hence we decipher the biomolecules of Wnt signaling trafficked through exosomes and their potential in serving as therapeutic targets.

Ocular oncology practice guidelines during COVID-19 pandemic-An expert consensus.

The outbreak of rapidly spreading COVID-19 pandemic in December 2019 has witnessed a major transformation in the health care system worldwide. This has led to the re-organization of the specialty services for the effective utilization of available resources and ensuring the safety of patients and healthcare workers. Suspension of oncology services will have major implications on cancer care due to delayed diagnosis and treatment leading to irreversible adverse consequences. Therefore various oncology organizations have called for a continuation of cancer care during this crisis with diligence. The COVID-19 pandemic has forced the clinicians to transform the components of care from screening to outpatient care and primary management. The purpose of this article is to establish guidelines and recommendations for ocular oncology in the management of ocular tumors set by a multidisciplinary team of experts including ocular, medical and radiation oncologists, and pathologists. As the pandemic is evolving fast, it will require constant updates and reformation of health strategies and guidelines for safe and quality health care.