Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection

Introduction: Hibernating mammals have evolved physiological strategies to cope with harsh winters by markedly reducing their metabolic rate and core temperature. Interestingly, neural stem cells derived from Arctic ground squirrels (AGS) also possess resilient cell biological processes that preserv...

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Published in:Stroke
Main Authors: Llera, Ted D, Singhal, Neel S, Ma, Dengke
Format: Article in Journal/Newspaper
Language:English
Published: Ovid Technologies (Wolters Kluwer Health) 2022
Subjects:
Online Access:http://dx.doi.org/10.1161/str.53.suppl_1.tp251
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spelling crovidcr:10.1161/str.53.suppl_1.tp251 2023-05-15T15:13:59+02:00 Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection Llera, Ted D Singhal, Neel S Ma, Dengke 2022 http://dx.doi.org/10.1161/str.53.suppl_1.tp251 en eng Ovid Technologies (Wolters Kluwer Health) Stroke volume 53, issue Suppl_1 ISSN 0039-2499 1524-4628 Advanced and Specialized Nursing Cardiology and Cardiovascular Medicine Neurology (clinical) journal-article 2022 crovidcr https://doi.org/10.1161/str.53.suppl_1.tp251 2022-05-29T06:42:01Z Introduction: Hibernating mammals have evolved physiological strategies to cope with harsh winters by markedly reducing their metabolic rate and core temperature. Interestingly, neural stem cells derived from Arctic ground squirrels (AGS) also possess resilient cell biological processes that preserve cell integrity during hypoxia and exposure mitochondrial toxins. We identified AGS mesencephalic astrocyte-derived neurotrophic factor (MANF) as a key cytoprotective gene in diverse survival screens of mouse neural cells expressing an AGS cDNA library. Hypothesis: We hypothesize that adaptively evolved amino acid substitution in AGS MANF confers metabolic stress resilience by modulating actions of MANF including sulfatide-binding, reducing endoplasmic reticulum (ER) stress and altering mitochondrial metabolism. Methodology: CRISPR-Cas9 knock-in (KI) gene editing was utilized to generate mouse neural cells harboring the AGS Manf allele to allow for analysis of the beneficial effects of the AGS amino acid substitutions in cell survival to metabolic stress, mitochondrial metabolism, MANF protein stability, and sulfatide binding.. We also tested AGS compared to human MANF with regard to reducing ER stress under hypoxia, hypothermia, and rotenone. Results: AGS MANF expression reduced ER stress markers compared to human MANF, and demonstrated increased stability following rotenone treatment further pointing to its important role regulating cellular adaptations to metabolic stress and imparting neuroprotection in our in vitro model. We gained key functional insights into how specific amino acid substitutions improve MANF stability to impart cytoprotection to metabolic insults. This detailed dissection of the AGS optimized adaptive stress response pathway can serve as a template for the development of new neuroprotective treatments. Article in Journal/Newspaper Arctic Ovid (via Crossref) Arctic Stroke 53 Suppl_1
institution Open Polar
collection Ovid (via Crossref)
op_collection_id crovidcr
language English
topic Advanced and Specialized Nursing
Cardiology and Cardiovascular Medicine
Neurology (clinical)
spellingShingle Advanced and Specialized Nursing
Cardiology and Cardiovascular Medicine
Neurology (clinical)
Llera, Ted D
Singhal, Neel S
Ma, Dengke
Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
topic_facet Advanced and Specialized Nursing
Cardiology and Cardiovascular Medicine
Neurology (clinical)
description Introduction: Hibernating mammals have evolved physiological strategies to cope with harsh winters by markedly reducing their metabolic rate and core temperature. Interestingly, neural stem cells derived from Arctic ground squirrels (AGS) also possess resilient cell biological processes that preserve cell integrity during hypoxia and exposure mitochondrial toxins. We identified AGS mesencephalic astrocyte-derived neurotrophic factor (MANF) as a key cytoprotective gene in diverse survival screens of mouse neural cells expressing an AGS cDNA library. Hypothesis: We hypothesize that adaptively evolved amino acid substitution in AGS MANF confers metabolic stress resilience by modulating actions of MANF including sulfatide-binding, reducing endoplasmic reticulum (ER) stress and altering mitochondrial metabolism. Methodology: CRISPR-Cas9 knock-in (KI) gene editing was utilized to generate mouse neural cells harboring the AGS Manf allele to allow for analysis of the beneficial effects of the AGS amino acid substitutions in cell survival to metabolic stress, mitochondrial metabolism, MANF protein stability, and sulfatide binding.. We also tested AGS compared to human MANF with regard to reducing ER stress under hypoxia, hypothermia, and rotenone. Results: AGS MANF expression reduced ER stress markers compared to human MANF, and demonstrated increased stability following rotenone treatment further pointing to its important role regulating cellular adaptations to metabolic stress and imparting neuroprotection in our in vitro model. We gained key functional insights into how specific amino acid substitutions improve MANF stability to impart cytoprotection to metabolic insults. This detailed dissection of the AGS optimized adaptive stress response pathway can serve as a template for the development of new neuroprotective treatments.
format Article in Journal/Newspaper
author Llera, Ted D
Singhal, Neel S
Ma, Dengke
author_facet Llera, Ted D
Singhal, Neel S
Ma, Dengke
author_sort Llera, Ted D
title Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
title_short Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
title_full Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
title_fullStr Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
title_full_unstemmed Abstract TP251: CRISPR-engineered Mouse Neural Cells Expressing An Optimized Manf Variant Reveals Mechanisms Of MANF Neuroprotection
title_sort abstract tp251: crispr-engineered mouse neural cells expressing an optimized manf variant reveals mechanisms of manf neuroprotection
publisher Ovid Technologies (Wolters Kluwer Health)
publishDate 2022
url http://dx.doi.org/10.1161/str.53.suppl_1.tp251
geographic Arctic
geographic_facet Arctic
genre Arctic
genre_facet Arctic
op_source Stroke
volume 53, issue Suppl_1
ISSN 0039-2499 1524-4628
op_doi https://doi.org/10.1161/str.53.suppl_1.tp251
container_title Stroke
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