Toulouse Purpan Medical School, Toulouse, France.
Department of Neurology, Massachusetts General Hospital, Boston, MA, USA.
Brain Behav. 2021 Apr;11(4):e02048. doi: 10.1002/brb3.2048. Epub 2021 Mar 11.
To identify circular RNAs as candidates for differential expression in the middle temporal (MT) cortex in a well-characterized cohort with contrasting Alzheimer disease (AD) pathology and cognition. Top screen candidates were assessed for proof of circularity and then quantified by qPCR in a larger number of samples.
An initial RNA sequencing screen was performed on n = 20 frozen human tissue samples. Filters were applied to select candidate circular RNAs for further investigation. Frozen human tissue samples were selected for global AD pathology burden and global cognition scores (n = 100). Linear and divergent primers were used to assess circularity using RNaseR digestion. RT-qPCR was performed to quantify relative hsa_circ_0131235 abundance.
Eleven circular RNAs were selected for further investigation. Four candidates produced circular RNA primers with appropriate efficiencies for qPCR. RNaseR treatment and analysis by both basic PCR and qPCR confirmed hsa_circ_0131235 circularity. There was a significant main effect of AD pathology on hsa_circ_0131235 expression.
Elevated hsa_circ_0131235 expression in the MT cortex was significantly associated with AD pathology.
在具有对比性阿尔茨海默病(AD)病理和认知的特征明确队列的颞中(MT)皮层中鉴定出差异表达的环状 RNA 作为候选物。对顶级候选物进行环状 RNA 验证,并通过 qPCR 在更多样本中进行定量。
对 n=20 个冷冻人组织样本进行初始 RNA 测序筛选。应用筛选标准以选择用于进一步研究的环状 RNA 候选物。选择了具有全球 AD 病理负担和全球认知评分的冷冻人组织样本(n=100)。使用 RNaseR 消化来评估环形引物的线性和发散性。采用 RT-qPCR 来定量 hsa_circ_0131235 的相对丰度。
选择了 11 个环状 RNA 进一步研究。4 个候选物生成了适用于 qPCR 的环状 RNA 引物。RNaseR 处理和基本 PCR 和 qPCR 分析均证实了 hsa_circ_0131235 的环状结构。AD 病理对 hsa_circ_0131235 的表达有显著的主效应。
MT 皮层中 hsa_circ_0131235 的表达升高与 AD 病理显著相关。