TY - JOUR T1 - A protocol to inject ocular drug implants into mouse eyes. JF - STAR Protoc Y1 - 2022 A1 - Lin, Cheng-Hui A1 - Sun, Young Joo A1 - Lee, Soo Hyeon A1 - Mujica, Elena M A1 - Kunchur, Caitlin R A1 - Wu, Man-Ru A1 - Yang, Jing A1 - Jung, Youn Soo A1 - Chiang, Bryce A1 - Wang, Sui A1 - Mahajan, Vinit B KW - Animals KW - Drug Implants KW - Intravitreal Injections KW - Mice KW - Vitreous Body AB -

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' efficacy, toxicity, and pharmacokinetics. For complete details on the use and execution of this protocol, please refer to Sun et al. (2021).

VL - 3 IS - 1 ER - TY - JOUR T1 - Molecular Characterization of a Rare Case of Bilateral Vitreoretinal T Cell Lymphoma through Vitreous Liquid Biopsy. JF - Int J Mol Sci Y1 - 2021 A1 - Cani, Andi K A1 - Toral, Marcus A A1 - Balikov, Daniel A A1 - Betz, Bryan L A1 - Hu, Kevin A1 - Liu, Chia-Jen A1 - Prifti, Matthew V A1 - Chinnaiyan, Arul M A1 - Tomlins, Scott A A1 - Mahajan, Vinit B A1 - Rao, Rajesh C KW - Biomarkers, Tumor KW - DNA (Cytosine-5-)-Methyltransferases KW - DNA Copy Number Variations KW - DNA Methyltransferase 3A KW - Gene Expression Regulation, Neoplastic KW - High-Throughput Nucleotide Sequencing KW - Humans KW - Liquid Biopsy KW - Lymphoma, T-Cell KW - Male KW - Middle Aged KW - Neoplasm Proteins KW - Proto-Oncogene Proteins B-raf KW - Retinal Neoplasms KW - Vitreous Body AB -

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.

VL - 22 IS - 11 ER - TY - JOUR T1 - Proteomic insight into the molecular function of the vitreous. JF - PLoS One Y1 - 2015 A1 - Skeie, Jessica M A1 - Roybal, C Nathaniel A1 - Mahajan, Vinit B KW - Aged KW - Aged, 80 and over KW - Eye Proteins KW - Female KW - Gene Expression Regulation KW - Humans KW - Male KW - Middle Aged KW - Proteome KW - Proteomics KW - Vitreous Body AB -

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.

VL - 10 IS - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26020955?dopt=Abstract ER - TY - JOUR T1 - Proteomic analysis of vitreous biopsy techniques. JF - Retina Y1 - 2012 A1 - Skeie, Jessica M A1 - Brown, Eric N A1 - Martinez, Harryl D A1 - Russell, Stephen R A1 - Birkholz, Emily S A1 - Folk, James C A1 - Boldt, H Culver A1 - Gehrs, Karen M A1 - Stone, Edwin M A1 - Wright, Michael E A1 - Mahajan, Vinit B KW - Adolescent KW - Aged KW - Biological Markers KW - Biopsy KW - Chromatography, Liquid KW - Electrophoresis, Polyacrylamide Gel KW - Eye Diseases KW - Eye Proteins KW - Female KW - Humans KW - Male KW - Middle Aged KW - Proteomics KW - Spectrophotometry, Ultraviolet KW - Tandem Mass Spectrometry KW - Vitrectomy KW - Vitreous Body KW - Young Adult AB -

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.

VL - 32 IS - 10 ER - TY - JOUR T1 - Bilateral intravitreal injection of antivascular endothelial growth factor therapy. JF - Retina (Philadelphia, Pa.) Y1 - 2011 A1 - Mahajan, Vinit B A1 - Elkins, Kori A A1 - Russell, Stephen R A1 - Boldt, H Culver A1 - Gehrs, Karen M A1 - Weingeist, Thomas A A1 - Stone, Edwin M A1 - Abràmoff, Michael D A1 - Liu, Dawei A1 - Folk, James C KW - Aged KW - Aged, 80 and over KW - Antibodies, Monoclonal KW - Case-Control Studies KW - Drug Administration Schedule KW - Drug Combinations KW - Female KW - Follow-Up Studies KW - Humans KW - Incidence KW - Injections, Intraocular KW - Macular Degeneration KW - Male KW - Middle Aged KW - Myocardial Infarction KW - Patient Preference KW - Retrospective Studies KW - Vascular Endothelial Growth Factor A KW - Vitreous Body AB -

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

VL - 31 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21187731?dopt=Abstract ER - TY - JOUR T1 - Dissection of human vitreous body elements for proteomic analysis. JF - Journal of visualized experiments : JoVE Y1 - 2011 A1 - Skeie, Jessica M A1 - Mahajan, Vinit B KW - Dissection KW - Electrophoresis, Polyacrylamide Gel KW - Humans KW - Proteomics KW - Vitreous Body AB -

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.

IS - 47 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21304469?dopt=Abstract ER - TY - JOUR T1 - Evisceration of mouse vitreous and retina for proteomic analyses. JF - Journal of visualized experiments : JoVE Y1 - 2011 A1 - Skeie, Jessica M A1 - Tsang, Stephen H A1 - Mahajan, Vinit B KW - Animals KW - Eye Evisceration KW - Eye Proteins KW - Mice KW - Models, Animal KW - Proteomics KW - Retina KW - Vitreous Body AB -

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.

IS - 50 U1 - http://www.ncbi.nlm.nih.gov/pubmed/21490583?dopt=Abstract ER - TY - JOUR T1 - T-cell infiltration in autosomal dominant neovascular inflammatory vitreoretinopathy. JF - Molecular vision Y1 - 2010 A1 - Mahajan, Vinit B A1 - Vallone, John G A1 - Lin, Jonathan H A1 - Mullins, Robert F A1 - Ko, Audrey C A1 - Folk, James C A1 - Stone, Edwin M KW - Aged, 80 and over KW - Antigens, CD4 KW - Antigens, CD8 KW - Autoantibodies KW - B-Lymphocytes KW - Eye Diseases KW - Female KW - Genes, Dominant KW - Humans KW - Immunoglobulin G KW - Immunologic Techniques KW - Neovascularization, Pathologic KW - Retina KW - Retinal Diseases KW - Staining and Labeling KW - T-Lymphocytes KW - Vitreous Body AB -

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.

VL - 16 U1 - http://www.ncbi.nlm.nih.gov/pubmed/20596252?dopt=Abstract ER - TY - JOUR T1 - A head-tilt test for hypopyon after intravitreal triamcinolone. JF - Retina (Philadelphia, Pa.) Y1 - 2009 A1 - Mahajan, Vinit B A1 - Folk, James C A1 - Boldt, H Culver KW - Adult KW - Anterior Chamber KW - Diagnosis, Differential KW - Diagnostic Techniques, Ophthalmological KW - Endophthalmitis KW - Eye Infections KW - Female KW - Glucocorticoids KW - Head KW - Humans KW - Inflammation KW - Injections KW - Posture KW - Triamcinolone Acetonide KW - Vitreous Body VL - 29 IS - 4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/19092731?dopt=Abstract ER -