Cargando…

The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis

PURPOSE: Current understanding of local disease pathophysiology in AMD is limited. Analysis of the human disease-affected tissue is most informative, as gene expression, expressed quantitative trait loci, microenvironmental, and epigenetic changes can be tissue, cell type, and location specific. Dev...

Descripción completa

Detalles Bibliográficos
Autores principales: Owen, Leah A., Shakoor, Akbar, Morgan, Denise J., Hejazi, Andre A., McEntire, M. Wade, Brown, Jared J., Farrer, Lindsay A., Kim, Ivana, Vitale, Albert, DeAngelis, Margaret M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Association for Research in Vision and Ophthalmology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6440527/
https://www.ncbi.nlm.nih.gov/pubmed/30924847
http://dx.doi.org/10.1167/iovs.18-24254
_version_ 1783407404826755072
author Owen, Leah A.
Shakoor, Akbar
Morgan, Denise J.
Hejazi, Andre A.
McEntire, M. Wade
Brown, Jared J.
Farrer, Lindsay A.
Kim, Ivana
Vitale, Albert
DeAngelis, Margaret M.
author_facet Owen, Leah A.
Shakoor, Akbar
Morgan, Denise J.
Hejazi, Andre A.
McEntire, M. Wade
Brown, Jared J.
Farrer, Lindsay A.
Kim, Ivana
Vitale, Albert
DeAngelis, Margaret M.
author_sort Owen, Leah A.
collection PubMed
description PURPOSE: Current understanding of local disease pathophysiology in AMD is limited. Analysis of the human disease-affected tissue is most informative, as gene expression, expressed quantitative trait loci, microenvironmental, and epigenetic changes can be tissue, cell type, and location specific. Development of a novel translational treatment and prevention strategies particularly for earlier forms of AMD are needed, although access to human ocular tissue analysis is challenging. We present a standardized protocol to study rapidly processed postmortem donor eyes for molecular biochemical and genomic studies. METHODS: We partnered with the Utah Lions Eye Bank to obtain donor human eyes, blood, and vitreous, within 6 hours postmortem. Phenotypic analysis was performed using spectral-domain optical coherence tomography (SD-OCT) and color fundus photography. Macular and extramacular tissues were immediately isolated, and the neural retina and retinal pigment epithelium/choroid from each specimen were separated and preserved. Ocular disease phenotype was analyzed using clinically relevant grading criteria by a group of four ophthalmologists incorporating data from SD-OCT retinal images, fundus photographs, and medical records. RESULTS: The use of multimodal imaging leads to greater resolution of retinal pathology, allowing greater phenotypic rigor for both interobserver phenotype and known clinical diagnoses. Further, our analysis resulted in excellent quality RNA, which demonstrated appropriate tissue segregation. CONCLUSIONS: The Utah protocol is a standardized methodology for analysis of disease mechanisms in AMD. It uniquely allows for simultaneous rigorous phenotypic, molecular biochemical, and genomic analysis of both systemic and local tissues. This better enables the development of disease biomarkers and therapeutic interventions.
format Online
Article
Text
id pubmed-6440527
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher The Association for Research in Vision and Ophthalmology
record_format MEDLINE/PubMed
spelling pubmed-64405272019-04-03 The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis Owen, Leah A. Shakoor, Akbar Morgan, Denise J. Hejazi, Andre A. McEntire, M. Wade Brown, Jared J. Farrer, Lindsay A. Kim, Ivana Vitale, Albert DeAngelis, Margaret M. Invest Ophthalmol Vis Sci Retina PURPOSE: Current understanding of local disease pathophysiology in AMD is limited. Analysis of the human disease-affected tissue is most informative, as gene expression, expressed quantitative trait loci, microenvironmental, and epigenetic changes can be tissue, cell type, and location specific. Development of a novel translational treatment and prevention strategies particularly for earlier forms of AMD are needed, although access to human ocular tissue analysis is challenging. We present a standardized protocol to study rapidly processed postmortem donor eyes for molecular biochemical and genomic studies. METHODS: We partnered with the Utah Lions Eye Bank to obtain donor human eyes, blood, and vitreous, within 6 hours postmortem. Phenotypic analysis was performed using spectral-domain optical coherence tomography (SD-OCT) and color fundus photography. Macular and extramacular tissues were immediately isolated, and the neural retina and retinal pigment epithelium/choroid from each specimen were separated and preserved. Ocular disease phenotype was analyzed using clinically relevant grading criteria by a group of four ophthalmologists incorporating data from SD-OCT retinal images, fundus photographs, and medical records. RESULTS: The use of multimodal imaging leads to greater resolution of retinal pathology, allowing greater phenotypic rigor for both interobserver phenotype and known clinical diagnoses. Further, our analysis resulted in excellent quality RNA, which demonstrated appropriate tissue segregation. CONCLUSIONS: The Utah protocol is a standardized methodology for analysis of disease mechanisms in AMD. It uniquely allows for simultaneous rigorous phenotypic, molecular biochemical, and genomic analysis of both systemic and local tissues. This better enables the development of disease biomarkers and therapeutic interventions. The Association for Research in Vision and Ophthalmology 2019-03 /pmc/articles/PMC6440527/ /pubmed/30924847 http://dx.doi.org/10.1167/iovs.18-24254 Text en Copyright 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
spellingShingle Retina
Owen, Leah A.
Shakoor, Akbar
Morgan, Denise J.
Hejazi, Andre A.
McEntire, M. Wade
Brown, Jared J.
Farrer, Lindsay A.
Kim, Ivana
Vitale, Albert
DeAngelis, Margaret M.
The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title_full The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title_fullStr The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title_full_unstemmed The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title_short The Utah Protocol for Postmortem Eye Phenotyping and Molecular Biochemical Analysis
title_sort utah protocol for postmortem eye phenotyping and molecular biochemical analysis
topic Retina
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6440527/
https://www.ncbi.nlm.nih.gov/pubmed/30924847
http://dx.doi.org/10.1167/iovs.18-24254
work_keys_str_mv AT owenleaha theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT shakoorakbar theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT morgandenisej theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT hejaziandrea theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT mcentiremwade theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT brownjaredj theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT farrerlindsaya theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT kimivana theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT vitalealbert theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT deangelismargaretm theutahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT owenleaha utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT shakoorakbar utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT morgandenisej utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT hejaziandrea utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT mcentiremwade utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT brownjaredj utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT farrerlindsaya utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT kimivana utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT vitalealbert utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis
AT deangelismargaretm utahprotocolforpostmortemeyephenotypingandmolecularbiochemicalanalysis