How precisely will this be used in humans to get better recovery? WHOM will do the followup research? And create and distribute protocols on this?
Long Noncoding RNA Fos Downstream Transcript Is Developmentally Dispensable but Vital for Shaping the Poststroke Functional Outcome
Abstract
Background and Purpose:
Stroke induces the expression of several long noncoding RNAs in the brain. However, their functional significance in poststroke outcome is poorly understood. We recently observed that a brain-specific long noncoding RNA called Fos downstream transcript (FosDT) is induced rapidly in the rodent brain following focal ischemia. Using FosDT knockout rats, we presently evaluated the role of FosDT in poststroke brain damage.
Methods:
FosDT knockout rats were generated using CRISPR-Cas9 genome editing on a Sprague-Dawley background. Male and female FosDT−/− and FosDT+/+ cohorts were subjected to transient middle cerebral artery occlusion. Postischemic sensorimotor deficits were evaluated between days 1 and 7 and lesion volume on day 7 of reperfusion. The developmental expression profile of FosDT was determined with real-time polymerase chain reaction and mechanistic implications of FosDT in the ischemic brain were conducted with RNA-sequencing analysis and immunostaining of pathological markers.
Results:
FosDT expression is developmentally regulated, with the adult cerebral cortex showing significantly higher FosDT expression than neonates. FosDT−/− rats did not show any anomalies in growth and development, fertility, brain cytoarchitecture, and cerebral vasculature. However, when subjected to transient focal ischemia, FosDT−/− rats of both sexes showed enhanced sensorimotor recovery and reduced brain damage. RNA-sequencing analysis showed that improved poststroke functional outcome in FosDT−/− rats is partially associated with curtailed induction of inflammatory genes, reduced apoptosis, mitochondrial dysfunction, and oxidative stress.
Conclusions:
Our study shows that FosDT is developmentally dispensable, mechanistically important, and a functionally promising target to reduce ischemic brain damage and facilitate neurological recovery.
Introduction
Stroke alters the profiles of various classes of noncoding RNAs (ncRNAs), including microRNAs, long ncRNAs (lncRNAs), circular RNAs, and transcribed ultraconserved regions.1–5 The lncRNAs (200 nucleotides to >100 kb) are the largest class of ncRNAs involved in transcriptional and translational regulation in a developmental-stage and a disease-specific manner.6–8 Many lncRNAs are organ and cell type specific. It was reported that <10% of the lncRNAs in humans ubiquitously express in different cell types, whereas ≈30% are seen in only one cell type.9 Furthermore, ≈40% of the lncRNAs are expressed specifically in the brain.6 Particularly, lncRNAs transcribed within the vicinity of protein-coding genes are preferentially expressed in the brain and share overlapping transcription factor binding sites with protein-coding genes.2,10
We previously showed that induction of a highly conserved brain-enriched lncRNA called Fos downstream transcript (FosDT, MRAK159688) promotes ischemic brain damage by interacting with REST (RE1-silencing transcription factor)-associated chromatin-modifying proteins.2,11FosDT gene is cogenic to the Fos gene, which is a marker of cellular stress.12FosDT and Fos genes are located within a gene desert of ≈240 000 nucleotides on chromosome 6 in rats (chromosome 14 in humans).11 We currently evaluated the functional significance of FosDT in brain development and postischemic outcome by using FosDT−/− rats developed by CRISPR-Cas9 genome editing.
We also evaluated the reproducibility of our data by analyzing the role of FosDT in ischemic brain damage by another lab. We further conducted RNA-sequencing analysis of FosDT−/− and FosDT+/+ rats following transient focal ischemia to understand the mechanistic implications of FosDT in the ischemic brain.
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