@article {1901, title = {A protocol to inject ocular drug implants into mouse eyes.}, journal = {STAR Protoc}, volume = {3}, year = {2022}, month = {2022 03 18}, pages = {101143}, abstract = {

Ocular drug implants (ODIs) are beneficial for treating ocular diseases. However, the lack of a robust injection approach for small-eyed model organisms has been a major technical limitation in developing ODIs. Here, we present a cost-effective, minimally invasive protocol to deliver ODIs into the mouse vitreous called Mouse Implant Intravitreal Injection (MI3). MI3 provides two alternative surgical approaches (air-pressure or plunger) to deliver micro-scaled ODIs into milli-scaled eyes, and expands the preclinical platforms to determine ODIs\&$\#$39; efficacy, toxicity, and pharmacokinetics. For complete details on the use and execution of this protocol, please refer to Sun et\ al. (2021).

}, keywords = {Animals, Drug Implants, Intravitreal Injections, Mice, Vitreous Body}, issn = {2666-1667}, doi = {10.1016/j.xpro.2022.101143}, author = {Lin, Cheng-Hui and Sun, Young Joo and Lee, Soo Hyeon and Mujica, Elena M and Kunchur, Caitlin R and Wu, Man-Ru and Yang, Jing and Jung, Youn Soo and Chiang, Bryce and Wang, Sui and Mahajan, Vinit B} } @article {1861, title = {Molecular Characterization of a Rare Case of Bilateral Vitreoretinal T Cell Lymphoma through Vitreous Liquid Biopsy.}, journal = {Int J Mol Sci}, volume = {22}, year = {2021}, month = {2021 Jun 05}, abstract = {

Vitreoretinal lymphoma (VRL) is an uncommon eye malignancy, and VRLs of T cell origin are rare. They are difficult to treat, and their molecular underpinnings, including actionable genomic alterations, remain to be elucidated. At present, vitreous fluid liquid biopsies represent a valuable VRL sample for molecular analysis to study VRLs. In this study, we report the molecular diagnostic workup of a rare case of bilateral T cell VRL and characterize its genomic landscape, including identification of potentially targetable alterations. Using next-generation sequencing of vitreous-derived DNA with a pan-cancer 126-gene panel, we found a copy number gain of and copy number loss of tumor suppressor . To the best of our knowledge, this represents the first exploration of the T cell VRL cancer genome and supports vitreous liquid biopsy as a suitable approach for precision oncology treatments.

}, keywords = {Biomarkers, Tumor, DNA (Cytosine-5-)-Methyltransferases, DNA Copy Number Variations, DNA Methyltransferase 3A, Gene Expression Regulation, Neoplastic, High-Throughput Nucleotide Sequencing, Humans, Liquid Biopsy, Lymphoma, T-Cell, Male, Middle Aged, Neoplasm Proteins, Proto-Oncogene Proteins B-raf, Retinal Neoplasms, Vitreous Body}, issn = {1422-0067}, doi = {10.3390/ijms22116099}, author = {Cani, Andi K and Toral, Marcus A and Balikov, Daniel A and Betz, Bryan L and Hu, Kevin and Liu, Chia-Jen and Prifti, Matthew V and Chinnaiyan, Arul M and Tomlins, Scott A and Mahajan, Vinit B and Rao, Rajesh C} } @article {693, title = {Proteomic insight into the molecular function of the vitreous.}, journal = {PLoS One}, volume = {10}, year = {2015}, month = {2015}, pages = {e0127567}, abstract = {

The human vitreous contains primarily water, but also contains proteins which have yet to be fully characterized. To gain insight into the four vitreous substructures and their potential functions, we isolated and analyzed the vitreous protein profiles of three non-diseased human eyes. The four analyzed substructures were the anterior hyaloid, the vitreous cortex, the vitreous core, and the vitreous base. Proteins were separated by multidimensional liquid chromatography and identified by tandem mass spectrometry. Bioinformatics tools then extracted the expression profiles, signaling pathways, and interactomes unique to each tissue. From each substructure, a mean of 2,062 unique proteins were identified, with many being differentially expressed in a specific substructure: 278 proteins were unique to the anterior hyaloid, 322 to the vitreous cortex, 128 to the vitreous base, and 136 to the vitreous core. When the identified proteins were organized according to relevant functional pathways and networks, key patterns appeared. The blood coagulation pathway and extracellular matrix turnover networks were highly represented. Oxidative stress regulation and energy metabolism proteins were distributed throughout the vitreous. Immune functions were represented by high levels of immunoglobulin, the complement pathway, damage-associated molecular patterns (DAMPs), and evolutionarily conserved antimicrobial proteins. The majority of vitreous proteins detected were intracellular proteins, some of which originate from the retina, including rhodopsin (RHO), phosphodiesterase 6 (PDE6), and glial fibrillary acidic protein (GFAP). This comprehensive analysis uncovers a picture of the vitreous as a biologically active tissue, where proteins localize to distinct substructures to protect the intraocular tissues from infection, oxidative stress, and energy disequilibrium. It also reveals the retina as a potential source of inflammatory mediators. The vitreous proteome catalogues the dynamic interactions between the vitreous and surrounding tissues. It therefore could be an indirect and effective method for surveying vitreoretinal disease for specific biomarkers.

}, keywords = {Aged, Aged, 80 and over, Eye Proteins, Female, Gene Expression Regulation, Humans, Male, Middle Aged, Proteome, Proteomics, Vitreous Body}, issn = {1932-6203}, doi = {10.1371/journal.pone.0127567}, author = {Skeie, Jessica M and Roybal, C Nathaniel and Mahajan, Vinit B} } @article {129, title = {Proteomic analysis of vitreous biopsy techniques.}, journal = {Retina}, volume = {32}, year = {2012}, month = {2012 Nov-Dec}, pages = {2141-9}, abstract = {

PURPOSE: To compare vitreous biopsy methods using analysis platforms used in proteomics biomarker discovery.

METHODS: Vitreous biopsies from 10 eyes were collected sequentially using a 23-gauge needle and a 23-gauge vitreous cutter instrument. Paired specimens were evaluated by UV absorbance spectroscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and liquid chromatography tandem mass spectrometry (LC-MS/MS).

RESULTS: The total protein concentration obtained with a needle and vitrectomy instrument biopsy averaged 1.10 mg/mL (standard error of the mean = 0.35) and 1.13 mg/mL (standard error of the mean = 0.25), respectively. In eight eyes with low or medium viscidity, there was a very high correlation (R = 0.934) between the biopsy methods. When data from 2 eyes with high viscidity vitreous were included, the correlation was reduced (R = 0.704). The molecular weight protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of paired needle and vitreous cutter samples were similar, except for a minority of pairs with single band intensity variance. Using LC-MS/MS, equivalent peptides were identified with similar frequencies (R \≥ 0.90) in paired samples.

CONCLUSION: Proteins and peptides collected from vitreous needle biopsies are nearly equivalent to those obtained from a vitreous cutter instrument. This study suggests both techniques may be used for most proteomic and biomarker discovery studies of vitreoretinal diseases, although a minority of proteins and peptides may differ in concentration.

}, keywords = {Adolescent, Aged, Biological Markers, Biopsy, Chromatography, Liquid, Electrophoresis, Polyacrylamide Gel, Eye Diseases, Eye Proteins, Female, Humans, Male, Middle Aged, Proteomics, Spectrophotometry, Ultraviolet, Tandem Mass Spectrometry, Vitrectomy, Vitreous Body, Young Adult}, author = {Skeie, Jessica M and Brown, Eric N and Martinez, Harryl D and Russell, Stephen R and Birkholz, Emily S and Folk, James C and Boldt, H Culver and Gehrs, Karen M and Stone, Edwin M and Wright, Michael E and Mahajan, Vinit B} } @article {137, title = {Bilateral intravitreal injection of antivascular endothelial growth factor therapy.}, journal = {Retina (Philadelphia, Pa.)}, volume = {31}, year = {2011}, month = {2011 Jan}, pages = {31-5}, abstract = {

The purpose of this study was to review adverse events and patient preference after bilateral intravitreal injection of antibodies to vascular endothelial growth factor.

}, keywords = {Aged, Aged, 80 and over, Antibodies, Monoclonal, Case-Control Studies, Drug Administration Schedule, Drug Combinations, Female, Follow-Up Studies, Humans, Incidence, Injections, Intraocular, Macular Degeneration, Male, Middle Aged, Myocardial Infarction, Patient Preference, Retrospective Studies, Vascular Endothelial Growth Factor A, Vitreous Body}, author = {Mahajan, Vinit B and Elkins, Kori A and Russell, Stephen R and Boldt, H Culver and Gehrs, Karen M and Weingeist, Thomas A and Stone, Edwin M and Abr{\`a}moff, Michael D and Liu, Dawei and Folk, James C} } @article {139, title = {Dissection of human vitreous body elements for proteomic analysis.}, journal = {Journal of visualized experiments : JoVE}, year = {2011}, month = {2011}, abstract = {

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. (1,2) Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. (1,2) The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

}, keywords = {Dissection, Electrophoresis, Polyacrylamide Gel, Humans, Proteomics, Vitreous Body}, author = {Skeie, Jessica M and Mahajan, Vinit B} } @article {141, title = {Evisceration of mouse vitreous and retina for proteomic analyses.}, journal = {Journal of visualized experiments : JoVE}, year = {2011}, month = {2011}, abstract = {

While the mouse retina has emerged as an important genetic model for inherited retinal disease, the mouse vitreous remains to be explored. The vitreous is a highly aqueous extracellular matrix overlying the retina where intraocular as well as extraocular proteins accumulate during disease.(1-3) Abnormal interactions between vitreous and retina underlie several diseases such as retinal detachment, proliferative diabetic retinopathy, uveitis, and proliferative vitreoretinopathy.(1,4) The relative mouse vitreous volume is significantly smaller than the human vitreous (Figure 1), since the mouse lens occupies nearly 75\% of its eye.(5) This has made biochemical studies of mouse vitreous challenging. In this video article, we present a technique to dissect and isolate the mouse vitreous from the retina, which will allow use of transgenic mouse models to more clearly define the role of this extracellular matrix in the development of vitreoretinal diseases.

}, keywords = {Animals, Eye Evisceration, Eye Proteins, Mice, Models, Animal, Proteomics, Retina, Vitreous Body}, author = {Skeie, Jessica M and Tsang, Stephen H and Mahajan, Vinit B} } @article {165, title = {T-cell infiltration in autosomal dominant neovascular inflammatory vitreoretinopathy.}, journal = {Molecular vision}, volume = {16}, year = {2010}, month = {2010}, pages = {1034-40}, abstract = {

Autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV) is a familial blinding disease of unknown pathophysiology. The eyes and sera from patients with ADNIV were studied to understand the immune response in this condition.

}, keywords = {Aged, 80 and over, Antigens, CD4, Antigens, CD8, Autoantibodies, B-Lymphocytes, Eye Diseases, Female, Genes, Dominant, Humans, Immunoglobulin G, Immunologic Techniques, Neovascularization, Pathologic, Retina, Retinal Diseases, Staining and Labeling, T-Lymphocytes, Vitreous Body}, author = {Mahajan, Vinit B and Vallone, John G and Lin, Jonathan H and Mullins, Robert F and Ko, Audrey C and Folk, James C and Stone, Edwin M} } @article {169, title = {A head-tilt test for hypopyon after intravitreal triamcinolone.}, journal = {Retina (Philadelphia, Pa.)}, volume = {29}, year = {2009}, month = {2009 Apr}, pages = {560-1}, keywords = {Adult, Anterior Chamber, Diagnosis, Differential, Diagnostic Techniques, Ophthalmological, Endophthalmitis, Eye Infections, Female, Glucocorticoids, Head, Humans, Inflammation, Injections, Posture, Triamcinolone Acetonide, Vitreous Body}, author = {Mahajan, Vinit B and Folk, James C and Boldt, H Culver} }