The analysis of human conjunctival epithelium proteome in ocular surface diseases using impression cytology and 2D-DIGE

Soria, Javier Millán; Acera, Arantxa; Durán, Jorge; Boto-de-los-Bueis, Ana; Del-Hierro-Zarzuelo, Almudena; González, Nerea; Reigada, R.; Suárez, Tatiana

doi:10.1016/j.exer.2017.03.006

Cited by 24 publications

(34 citation statements)

References 38 publications

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“…As of January 2019, 936 manuscripts were retrieved in Medline (PubMedNCBI http://ncbi.nlm.nih.gov) using the term "impression cytology" in the title/abstract and only 5 of them were associated also to the word "proteomics". One seminal work on impression cytology and proteomics used 2D-DIGE approach to investigate the ocular dysfunctions in patients with meibomian gland dysfunction and dry eye disease was published (52). In this study, an 8 mm in diameter cellulose disc was applied to the eye on both sides to improve the number of captured cells.…”

Section: Discussionmentioning

confidence: 99%

“…Different materials and filter devices have been tested, ranging from polycarbonate, nitrocellulose and cellulose acetate to commercial devices such as the Eyeprim™ (28,49,50), but no major differences in RNA isolation yields were observed between them (51). Recently, the IC method has been coupled to 2D-DIGE to study protein expression in patients with meibomian gland dysfunction and dry eye disease (52). Barbosa et.al.…”

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confidence: 99%

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CImP: Cellular Imprinting Proteomics applied to ocular disorders elicited by Congenital Zika virus Syndrome

Rosa-Fernandes

Mlb

et al. 2019

Preprint

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IMPORTANCE:Ocular complications in infants with Congenital Zika Syndrome (CZS) have been reported. However, the molecular mechanisms underlying of eye dysfunctions are presently unknown. OBJECTIVE:A method (termed Cellular Imprinting Proteomics, CImP) for the identification and quantification of the ocular surface proteome using a minimally invasive membrane filter device is described. Moreover, The CImP method was applied to profile the molecular alterations in the eyes of infants exposed to Zika virus (ZIKV) infection during gestation. DESIGN, SETTINGS AND PARTICIMPANTS:The CImP method was applied to a cohort divided into three conditions: 1) Ctrl (infants with no infectious diseases, n=5). 2) Zikv (infants exposed to ZIKV gestation, with no microcephaly, n=5). 3) Zikv CZS (infants exposed to ZIKV, with microcephaly, n=3). All conditions were age and sex-matched. An improved impression cytology method was used to capture the outermost ocular surface cells. The number of impression cytology membrane collected was: Ctrl (12), Zikv (14) and Zikv CZS (8). Proteins were extracted and analysed using mass spectrometry-based proteomics technology followed by statistical analysis. Parallel reaction monitoring was performed to validate the expression of specific protein markers. RESULTS:Using the CImP method, 2209 proteins were identified on the membrane-captured conjunctiva epithelial cells. Modulation of neutrophil degranulation, cell death, ocular and neurodevelopment pathways are reported in infants with CZS compared to matched controls.Moreover, the molecular pattern of ocular surface cells retrieved from infants infected during the gestation but with no CZS was different from matched controls. CONCLUSIONS AND PERSPECTIVES: Molecular alterations in the ocular cell surface associatedto ZIKV infection with and without CZS complications are reported for the first time. We predict that this method will be introduced successfully in the study of several neurological diseases with the aim to identify novel diagnostic and therapeutic biomarkers. Introduction:Zika virus (ZIKV) is a positive-strand RNA virus belonging to the Flaviviridae family that is transmitted to humans predominantly by the female Aedes aegypti and Aedes albopictus mosquitos (1). Moreover, sexual contact, blood transfusions, laboratory and healthcare settings Subjects, materials and methods Patient cohortThis study comprises 13 infants with and without CZS referred to the Pediatric Service of the Antonio Pedro University Hospital, Fluminense Federal University, Niteroi, Brazil. This study was approved by Institutional review board and ethics committee of Fluminense Federal University, protocol CAAE number 79890517.6.0000.5243, and followed the guidelines of the Declaration of Helsinki. All samples were collected upon informed and written consent from the parents/legal guardians of each participant. Clinical examination was performed by a multidisciplinary team and all infants included in this study are part of a clinical follow up program currently in pr...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

CImP: Cellular Imprinting Proteomics applied to ocular disorders elicited by Congenital Zika virus Syndrome

Rosa-Fernandes

Mlb

et al. 2019

Preprint

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“…55 Also, studies have shown that cells on the conjunctival surface of KCS patients increase the expression of genes and proteins related to defense and inflammatory responses. [9][10][11] For example, conjunctival epithelium in KCS eyes expresses beta-defensins which are antimicrobial peptides absent in the normal conjunctival epithelium. 11 The remodeling of chromatin from a more compacted to a relaxed state, as seen in KCS (vs. control), involves epigenetic modifications, such as histone acetylation, which may occur in response to microenvironmental stimuli and has been implicated in the pathogenesis of human autoimmune diseases such as systemic lupus erythematosus and Sj€ ogren's syndrome.…”

Section: F I G U R E 5 Surface Plot Graphsmentioning

confidence: 99%

“…[9][10][11] For example, conjunctival epithelium in KCS eyes expresses beta-defensins which are antimicrobial peptides absent in the normal conjunctival epithelium. 11 The remodeling of chromatin from a more compacted to a relaxed state, as seen in KCS (vs. control), involves epigenetic modifications, such as histone acetylation, which may occur in response to microenvironmental stimuli and has been implicated in the pathogenesis of human autoimmune diseases such as systemic lupus erythematosus and Sj€ ogren's syndrome. 56,57 Sj€ ogren's syndrome is a chronic autoimmune epithelitis that affects salivary and lacrimal glands, resulting in a clinical phenotype of dry eye very similar to that of canine KCS.…”

Section: F I G U R E 5 Surface Plot Graphsmentioning

confidence: 99%

“…3,6,7 With regard to the pathophysiology of KCS, recent advances in the fields of cell biology and genetics suggest that it goes beyond the histopathological manifestations seen in the lacrimal gland and the ocular surface. 2,3,[8][9][10][11][12][13] There is evidence that the metabolism of cells on the surface of the palpebral conjunctiva is also severely altered by the disease. 2 In human patients, KCS has been closely associated with changes in interphase nuclear morphology of palpebral/bulbar conjunctival epithelial cells, and with the appearance of snake-like chromatin, whose clinical and biological significances remain under investigation.…”

Section: Introductionmentioning

confidence: 99%

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Nuclear parameters and chromatin remodeling in epithelial cells and lymphocytes from the palpebral conjunctiva of dogs with keratoconjunctivitis sicca

Santos

Aldrovani

Filezio

et al. 2018

Veterinary Ophthalmology

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Prediction of potential drugs and targets based on meibomian gland dysfunction module classification to guide individualized treatment

Zhao

Gao

et al. 2019

J of Cellular Biochemistry

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Meibomian gland dysfunction (MGD) is an epidemic chronic ocular inflammation. However, little is known about its effective treatment. Here, this study identified important MGD‐related genes, core regulators, and potential drugs and their targets though integrating a series of bioinformational analyses. First, there were 665 differentially expression genes (DEGs) were identified. Then, 56 coexpression modules were exacted based on the expression of DEGs and their interactors. Moreover, core transcription factors (TF) significantly regulated modules were identified, including RELA, HIF1A, SIRT1, and MYC, which related to variety of eye diseases. Finally, the prediction of potential drugs and the identification of their target were performed. The results showed that artenimol, copper, and glutathione may have the remarkable curative effect or the toxicology to MGD. Moreover, their targets module gene LDHA (lactate dehydrogenase A), ENO1 (enolase 1), ALB (albumin), and PKM (pyruvate kinase M) are play important role in eye diseases. It suggests that these potential drugs may be useful for the treatment of MGD by acting on their targets. It provides valuable references for drug redirection and new drug development for drug developers, and provides individualized treatment strategies for tarsal gland dysfunction.

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The analysis of human conjunctival epithelium proteome in ocular surface diseases using impression cytology and 2D-DIGE

Cited by 24 publications

References 38 publications

CImP: Cellular Imprinting Proteomics applied to ocular disorders elicited by Congenital Zika virus Syndrome

CImP: Cellular Imprinting Proteomics applied to ocular disorders elicited by Congenital Zika virus Syndrome

Nuclear parameters and chromatin remodeling in epithelial cells and lymphocytes from the palpebral conjunctiva of dogs with keratoconjunctivitis sicca

Prediction of potential drugs and targets based on meibomian gland dysfunction module classification to guide individualized treatment

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