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Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks

Receptors, transporters, and ion channels are important targets for therapy development in neurological diseases, but their mechanistic role in pathogenesis is often poorly understood. Gene editing and in vivo imaging approaches will help to identify the molecular and functional role of these target...

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Autores principales: Marciano, Sabina, Ionescu, Tudor M., Saw, Ran Sing, Cheong, Rachel Y., Kirik, Deniz, Maurer, Andreas, Pichler, Bernd J., Herfert, Kristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9546570/
https://www.ncbi.nlm.nih.gov/pubmed/36161926
http://dx.doi.org/10.1073/pnas.2122552119
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author Marciano, Sabina
Ionescu, Tudor M.
Saw, Ran Sing
Cheong, Rachel Y.
Kirik, Deniz
Maurer, Andreas
Pichler, Bernd J.
Herfert, Kristina
author_facet Marciano, Sabina
Ionescu, Tudor M.
Saw, Ran Sing
Cheong, Rachel Y.
Kirik, Deniz
Maurer, Andreas
Pichler, Bernd J.
Herfert, Kristina
author_sort Marciano, Sabina
collection PubMed
description Receptors, transporters, and ion channels are important targets for therapy development in neurological diseases, but their mechanistic role in pathogenesis is often poorly understood. Gene editing and in vivo imaging approaches will help to identify the molecular and functional role of these targets and the consequence of their regional dysfunction on the whole-brain level. We combine CRISPR-Cas9 gene editing with in vivo positron emission tomography (PET) and functional MRI (fMRI) to investigate the direct link between genes, molecules, and the brain connectome. The extensive knowledge of the Slc18a2 gene encoding the vesicular monoamine transporter (VMAT2), involved in the storage and release of dopamine, makes it an excellent target for studying the gene network relationships while structurally preserving neuronal integrity and function. We edited the Slc18a2 in the substantia nigra pars compacta of adult rats and used in vivo molecular imaging besides behavioral, histological, and biochemical assessments to characterize the CRISPR-Cas9–mediated VMAT2 knockdown. Simultaneous PET/fMRI was performed to investigate molecular and functional brain alterations. We found that stage-specific adaptations of brain functional connectivity follow the selective impairment of presynaptic dopamine storage and release. Our study reveals that recruiting different brain networks is an early response to the dopaminergic dysfunction preceding neuronal cell loss. Our combinatorial approach is a tool to investigate the impact of specific genes on brain molecular and functional dynamics, which will help to develop tailored therapies for normalizing brain function.
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spelling pubmed-95465702023-03-26 Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks Marciano, Sabina Ionescu, Tudor M. Saw, Ran Sing Cheong, Rachel Y. Kirik, Deniz Maurer, Andreas Pichler, Bernd J. Herfert, Kristina Proc Natl Acad Sci U S A Biological Sciences Receptors, transporters, and ion channels are important targets for therapy development in neurological diseases, but their mechanistic role in pathogenesis is often poorly understood. Gene editing and in vivo imaging approaches will help to identify the molecular and functional role of these targets and the consequence of their regional dysfunction on the whole-brain level. We combine CRISPR-Cas9 gene editing with in vivo positron emission tomography (PET) and functional MRI (fMRI) to investigate the direct link between genes, molecules, and the brain connectome. The extensive knowledge of the Slc18a2 gene encoding the vesicular monoamine transporter (VMAT2), involved in the storage and release of dopamine, makes it an excellent target for studying the gene network relationships while structurally preserving neuronal integrity and function. We edited the Slc18a2 in the substantia nigra pars compacta of adult rats and used in vivo molecular imaging besides behavioral, histological, and biochemical assessments to characterize the CRISPR-Cas9–mediated VMAT2 knockdown. Simultaneous PET/fMRI was performed to investigate molecular and functional brain alterations. We found that stage-specific adaptations of brain functional connectivity follow the selective impairment of presynaptic dopamine storage and release. Our study reveals that recruiting different brain networks is an early response to the dopaminergic dysfunction preceding neuronal cell loss. Our combinatorial approach is a tool to investigate the impact of specific genes on brain molecular and functional dynamics, which will help to develop tailored therapies for normalizing brain function. National Academy of Sciences 2022-09-26 2022-10-04 /pmc/articles/PMC9546570/ /pubmed/36161926 http://dx.doi.org/10.1073/pnas.2122552119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Marciano, Sabina
Ionescu, Tudor M.
Saw, Ran Sing
Cheong, Rachel Y.
Kirik, Deniz
Maurer, Andreas
Pichler, Bernd J.
Herfert, Kristina
Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title_full Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title_fullStr Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title_full_unstemmed Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title_short Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
title_sort combining crispr-cas9 and brain imaging to study the link from genes to molecules to networks
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9546570/
https://www.ncbi.nlm.nih.gov/pubmed/36161926
http://dx.doi.org/10.1073/pnas.2122552119
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