Circular RNA
circ-Olfm1 induces progression of Alzheimer’s disease by regulating FOXO3a- Journal of Army Medical University Vol. 47, Issue 1, Pages: 60-70(2025)
- 作者机构：
1.蚌埠医学院第一附属医院神经内科，安徽蚌埠
2.陆军特色医学中心（第三军医大学大坪医院）神经内科，重庆
- 作者简介：
ZHOU Huadong， E-mail： zhouhuad@163.com
- 基金信息：Supported by the General Program of National Natural Science Foundation of China(81571258)
DOI：10.16016/j.2097-0927.202406084
CLC： R394.3;R741.02;R745.7Published：15 January 2025，
Received：25 June 2024，
Revised：14 August 2024，
- 稿件说明：
移动端阅览
杨红岩,廖旗荣,侯明亮,等.环状RNA circ-Olfm1通过FOXO3a调节阿尔茨海默病[J].陆军军医大学学报,2025,47(01):60-70.
circ-Olfm1 induces progression of Alzheimer’s disease by regulating FOXO3a[J].J Army Med Univ,2025,47(1):60-70.
杨红岩,廖旗荣,侯明亮,等.环状RNA circ-Olfm1通过FOXO3a调节阿尔茨海默病[J].陆军军医大学学报,2025,47(01):60-70. DOI：10.16016/j.2097-0927.202406084.
circ-Olfm1 induces progression of Alzheimer’s disease by regulating FOXO3a[J].J Army Med Univ,2025,47(1):60-70. DOI：10.16016/j.2097-0927.202406084.YANG Hongyan,LIAO Qirong,HOU Mingliang,et al.Circular RNA
摘要
目的
2
探讨环状RNAs（circRNAs）在阿尔茨海默病（Alzheimer’s disease，AD）中的作用及其潜在机制。
方法
2
选取6月龄APP/PS1转基因（AD）小鼠和C57BL/6野生型（WT）小鼠，采用随机数字表法将小鼠分为5组（
n
=3）：WT组（野生型小鼠）、WT+敲除
circ-Olfm1
组（敲除
circ-Olfm1
的野生型小鼠）、AD组（APP/PS1转基因小鼠）、AD+敲除
circ-Olfm1
组（敲除
circ-Olfm1
的APP/PS1转基因小鼠）和AD+敲除
FOXO3a
组（敲除
FOXO3a
的APP/PS1转基因小鼠）。采用皮层和海马立体定向注射慢病毒敲除
circ-Olfm1
和
FOXO3a
基因。①提取小鼠脑RNA，检测AD小鼠和WT小鼠中circRNA和mRNA的差异表达，并进行基因本体（gene ontology，GO）分析。②RT‐qPCR检测前10位上调和下调的circRNA、
circ-Olfm1
和miR-330-5p的表达。③制备慢病毒载体并立体定向注射到WT和AD小鼠的皮层或海马，以敲除
circ
-
Olfm1
基因，水迷宫实验评估敲除
circ-Olfm1
基因对小鼠认知功能的影响，免疫荧光检测Aβ斑块沉积。④双荧光素酶基因分析验证
circ-Olfm1
和miR-330-5p之间的相互作用。⑤Western blot检测AMPK和FOXO3a蛋白表达水平。⑥透射电镜检查AD小鼠海马组织的线粒体。⑦ELISA检测炎性因子IL-6、IL-1β和TNF-α的水平。
结果
2
AD小鼠和WT小鼠中共鉴定出52个差异表达的circRNA，其中28个上调，24个下调（fold change
>
1.5，
P
<
0.05）。这些差异表达基因主要涉及信号转导、学习和记忆等功能。
circ-Olfm1
是差异最大circRNA，在神经元中高表达，并在AD小鼠的大脑皮层和海马中上调（fold change
>
1.5，
P
<
0.05）。敲除
circ-Olfm1
基因减少了AD小鼠大脑皮层和海马的β-淀粉样斑块（amyloid β，Aβ）数量（
P
<
0.01）。细胞实验中：通过starBase数据库观察到
circ
-
Olfm1
和miR-330-5p之间存在互补序列。Western Blot结果表明：加入Aβ42后，AMPK和FOXO3a蛋白在神经元细胞中表达明显增加（
P
<
0.01）。沉默
circ-Olfm1
后，AMPK和FOXO3a蛋白在神经元细胞+Aβ42中表达降低（
P
<
0.01）。ELISA检测提示：敲除
FOXO3a
后，炎性因子IL-6、IL-1β和TNF-α均显著增高（
P
<
0.01）。透射电镜检查提示：敲除
FOXO3a
后线粒体损伤显著加重（
P
<
0.01）。
结论
2
circ
-
Olfm1
在AD小鼠脑组织和神经元细胞+Aβ42中表达上调，其导致AD小鼠认知损害的机制可能是通过调节FOXO3a蛋白的表达。
Abstract
Objective To investigate the role of circular RNAs （circRNA） in Alzheimer’s disease （AD） and its potential mechanism. Methods Six-month-old APP/PS1 mouse model of AD and wild type (WT) mice were subjected and then randomly divided into WT group
WT+
circ-Olfm1
knockout group
AD group (transgenic APP/PS1 mice)
AD+
circ-Olfm1
knockout group
AD+FOXO3a knockout group
with 3 mice in each group. ① The total RNA of mouse brain was extracted
and the differential expression of circRNAs and mRNAs between the AD mice and WT mice was detected
and the obtained circRNAs and mRNAs were analyzed with gene ontology (GO) analysis. ② RT‐qPCR was used to detect the expression of the top 10 up-regulated and down-regulated circRNAs
as well as the expression of
circ-Olfm1
and miR-330-5p. ③ Lentiviral vectors were prepared and stereotaxically injected into the cortex or hippocampus of WT and AD mice to knock out
circ-Olfm1
gene. Water maze test was used to evaluate the effect of
circ-Olfm1
knockout on cognitive function
and immunofluorescence assay was employed to observe the deposition of amyloid β (Aβ) plaque in the brain. ④ The interaction between
circ-Olfm1
and miR-330-5p was verified by double luciferase reporter gene analysis. ⑤ The protein levels of AMPK and FOXO3a were detected by Western blotting. ⑥ Transmission electron microscopy was utilized to observe the mitochondria of the hippocampus. ⑦ The levels of inflammatory factors IL-6
IL-1β and TNF-α were detected by ELISA. Results There were totally 52 differentially expressed circRNAs identified between the AD and WT mice
including 28 up-regulated and 24 down-regulated (fold change
>
1.5
P
<
0.05). These differentially expressed genes are mainly involv
ed in signal transduction
learning and memory and other functions.
circ-Olfm1
was identified as the most significantly differentially expressed circRNA
which is highly expressed in the neurons and up-regulated in the cerebral cortex and hippocampus of the AD mice. Knockout of
circ-Olfm1
reduced the number of Aβ plaques in the cerebral cortex and hippocampus of AD mice (
P
<
0.01). In starBase database
there are complementary sequences observed between
circ-Olfm1
and miR-330-5p. Western blotting showed that the addition of Aβ42 significantly increased the expression of AMPK and FOXO3a in the neuronal cells (
P
<
0.01). And silencing
circ-Olfm1
led to decreased expression of AMPK and FOXO3a in neuronal cells+Aβ42 (
P
<
0.01). ELISA revealed that knockout of FOXO3a significantly increased the levels of inflammatory factors IL-6
IL-1β
and TNF-α (
P
<
0.01). Transmission electron microscopy displayed that knocking FOXO3a out significantly aggravated mitochondrial damage (
P
<
0.01). Conclusion
circ-Olfm1
is up-regulated in the brain tissue and neurons+Aβ42 of AD rats
and the mechanism of cognitive impairment in AD rats may be through its regulating FOXO3a protein.
关键词
阿尔茨海默病环状RNA叉头转录因子
Keywords
Alzheimer’s diseasecircRNAsforkhead transcription factor
references
ARVANITAKIS Z， SHAH R C， BENNETT D A. Diagnosis and management of dementia： review［J］. JAMA， 2019， 322（16）： 1589-1599. DOI： 10.1001/jama.2019.4782http://dx.doi.org/10.1001/jama.2019.4782.
HU J， CHEN Q， ZHU H R， et al. Microglial Piezo1 senses Aβ fibril stiffness to restrict Alzheimer’s disease［J］. Neuron， 2023， 111（1）： 15-29.e8. DOI： 10.1016/j.neuron. 2022.10.021http://dx.doi.org/10.1016/j.neuron.2022.10.021.
SANTOS R X， CORREIA S C， WANG X L， et al. A synergistic dysfunction of mitochondrial fission/fusion dynamics and mitophagy in Alzheimer’s disease［J］. J Alzheimers Dis， 2010， 20（Suppl 2）： S401-S412. DOI： 10.3233/JAD-2010-100666http://dx.doi.org/10.3233/JAD-2010-100666.
LOPEZ-RODRIGUEZ A B， HENNESSY E， MURRAY C L， et al. Acute systemic inflammation exacerbates neuroin-flammation in Alzheimer’s disease： IL-1β drives amplified responses in primed astrocytes and neuronal network dysfunction［J］. Alzheimers Dement， 2021， 17（10）： 1735-1755. DOI： 10.1002/alz.12341http://dx.doi.org/10.1002/alz.12341.
LIU Y， AO X， DING W， et al. Critical role of FOXO3a in carcinogenesis［J］. Mol Cancer， 2018， 17（1）： 104. DOI： 10.1186/s12943-018-0856-3http://dx.doi.org/10.1186/s12943-018-0856-3.
ZHAO Y， HU X B， LIU Y J， et al. ROS signaling under metabolic stress： cross-talk between AMPK and AKT pathway［J］. Mol Cancer， 2017， 16（1）： 79. DOI： 10.1186/s12943-017-0648-1http://dx.doi.org/10.1186/s12943-017-0648-1.
SONG C H， ZHANG Y F， HUANG W Y， et al. Circular RNA Cwc27 contributes to Alzheimer’s disease pathogenesis by repressing Pur-α activity［J］. Cell Death Differ， 2022， 29（2）： 393-406. DOI： 10.1038/s41418-021-00865-1http://dx.doi.org/10.1038/s41418-021-00865-1.
HENTZE M W， PREISS T. Circular RNAs： splicing’s enigma variations［J］. EMBO J， 2013， 32（7）： 923-925. DOI： 10.1038/emboj.2013.53http://dx.doi.org/10.1038/emboj.2013.53.
ZHOU B， YU J W. A novel identified circular RNA， circRNA_010567， promotes myocardial fibrosis via suppressing miR-141 by targeting TGF-β1［J］. Biochem Biophys Res Commun， 2017， 487（4）： 769-775. DOI： 10.1016/j.bbrc.2017.04.044http://dx.doi.org/10.1016/j.bbrc.2017.04.044.
XIA W J， QIU M T， CHEN R， et al. Circular RNA has_circ_0067934 is upregulated in esophageal squamous cell carcinoma and promoted proliferation［J］. Sci Rep， 2016， 6： 35576. DOI： 10.1038/srep35576http://dx.doi.org/10.1038/srep35576.
QIAN J， CHEN C Y， ZHAO C P， et al. Hsa_circRNA_0084043 promoting tumorigenesis in glioma through miR-577 sponging［J］. Heliyon， 2023， 9（9）： e19219. DOI： 10.1016/j.heliyon.2023.e19219http://dx.doi.org/10.1016/j.heliyon.2023.e19219.
LI Z Y， HUANG C， BAO C， et al. Exon-intron circular RNAs regulate transcription in the nucleus［J］. Nat Struct Mol Biol， 2015， 22（3）： 256-264. DOI： 10.1038/nsmb. 2959http://dx.doi.org/10.1038/nsmb.2959.
PAMUDURTI N R， BARTOK O， JENS M， et al. Translation of circRNAs［J］. Mol Cell， 2017， 66（1）： 9-21.e7. DOI： 10.1016/j.molcel.2017.02.021http://dx.doi.org/10.1016/j.molcel.2017.02.021.
DUBE U， DEL-AGUILA J L， LI Z R， et al. An atlas of cortical circular RNA expression in Alzheimer disease brains demonstrates clinical and pathological associations［J］. Nat Neurosci， 2019， 22（11）： 1903-1912. DOI： 10.1038/s41593-019-0501-5http://dx.doi.org/10.1038/s41593-019-0501-5.
ZHANG S， ZHU D N， LI H， et al. Characterization of circRNA-associated-ceRNA networks in a senescence-accelerated mouse prone 8 brain［J］. Mol Ther， 2017， 25（9）： 2053-2061. DOI： 10.1016/j.ymthe.2017.06.009http://dx.doi.org/10.1016/j.ymthe.2017.06.009.
MA N N， PAN J， YE X Y， et al. Whole-transcriptome analysis of APP/PS1 mouse brain and identification of circRNA-miRNA-mRNA networks to investigate AD pathogenesis［J］. Mol Ther Nucleic Acids， 2019， 18： 1049-1062. DOI： 10.1016/j.omtn.2019.10.030http://dx.doi.org/10.1016/j.omtn.2019.10.030.
MANSOR N I， NTIMI C M， ABDUL-AZIZ N M， et al. Asymptomatic neurotoxicity of amyloid β-peptides （Aβ1-42 and Aβ25-35） on mouse embryonic stem cell-derived neural cells［J］. Bosn J Basic Med Sci， 2021， 21（1）： 98-110. DOI： 10.17305/bjbms.2020.4639http://dx.doi.org/10.17305/bjbms.2020.4639.
CHEN F， FANG S C， DU Y F， et al. CRISPR/Cas9-mediated CysLT1R deletion reverses synaptic failure， amyloidosis and cognitive impairment in APP/PS1 mice［J］. Aging， 2021， 13（5）： 6634-6661. DOI： 10.18632/aging.202501http://dx.doi.org/10.18632/aging.202501.
CHEN L， WANG C L， SUN H Y， et al. The bioinformatics toolbox for circRNA discovery and analysis［J］. Brief Bioinform， 2021， 22（2）： 1706-1728. DOI： 10.1093/bib/bbaa001http://dx.doi.org/10.1093/bib/bbaa001.
RYBAK-WOLF A， STOTTMEISTER C， GLAŽAR P， et al. Circular RNAs in the mammalian brain are highly abundant， conserved， and dynamically expressed［J］. Mol Cell， 2015， 58（5）： 870-885. DOI： 10.1016/j.molcel. 2015.03.027http://dx.doi.org/10.1016/j.molcel.2015.03.027.
PIWECKA M， GLAŽAR P， HERNANDEZ-MIRANDA L R， et al. Loss of a mammalian circular RNA locus causes miRNA deregulation and affects brain function［J］. Science， 2017， 357（6357）： eaam8526. DOI： 10.1126/science.aam 8526http://dx.doi.org/10.1126/science.aam8526.
ZHAO Y H， ALEXANDROV P N， JABER V， et al. Deficiency in the ubiquitin conjugating enzyme UBE2A in Alzheimer’s disease （AD） is linked to deficits in a natural circular miRNA-7 sponge （circRNA； ciRS-7）［J］. Genes， 2016， 7（12）： 116. DOI： 10.3390/genes7120116http://dx.doi.org/10.3390/genes7120116.
WANG X， XIE J Z， TAN L， et al. N6-methyladenosine-modified circRIMS2 mediates synaptic and memory impairments by activating GluN2B ubiquitination in Alzheimer’s disease［J］. Transl Neurodegener， 2023， 12（1）： 53. DOI： 10.1186/s40035-023-00386-6http://dx.doi.org/10.1186/s40035-023-00386-6.
URDÁNOZ-CASADO A， DE GORDOA J S， ROBLES M， et al. Profile of TREM2-derived circRNA and mRNA variants in the entorhinal cortex of Alzheimer’s disease patients［J］. Int J Mol Sci， 2022， 23（14）： 7682. DOI： 10.3390/ijms23147682http://dx.doi.org/10.3390/ijms23147682.
ZHENG Q P， BAO C Y， GUO W J， et al. Circular RNA profiling reveals an abundant circHIPK3 that regulates cell growth by sponging multiple miRNAs［J］. Nat Commun， 2016， 7： 11215. DOI： 10.1038/ncomms11215http://dx.doi.org/10.1038/ncomms11215.
胡梦婷， 陈俊霞. 环状RNA hsacirc0000231与HnRNPK相互作用对乳腺癌增殖、迁移及凋亡的影响［J］. 陆军军医大学学报， 2022， 44 （12）： 1207-1220. DOI：10.16016/j.2097-0927.202204125http://dx.doi.org/10.16016/j.2097-0927.202204125.
HU M T， CHEN J X. Effect of circular RNA hsa_circ_0000231 interacting with HnRNPK on proliferation， migration， and apoptosis in breast cancer［J］.J Amry Med Univ，2022，44（12）：1207-1220. DOI：10.16016/j.2097-0927.202204125http://dx.doi.org/10.16016/j.2097-0927.202204125.
LU Y J， TAN L， WANG X. Circular HDAC9/microRNA-138/sirtuin-1 pathway mediates synaptic and amyloid precursor protein processing deficits in Alzheimer’s disease［J］. Neurosci Bull， 2019， 35（5）： 877-888. DOI： 10.1007/s12264-019-00361-0http://dx.doi.org/10.1007/s12264-019-00361-0.
CHEN R X， CHEN X， XIA L P， et al. N6-methyladenosine modification of circNSUN2 facilitates cytoplasmic export and stabilizes HMGA2 to promote colorectal liver metastasis［J］. Nat Commun， 2019， 10（1）： 4695. DOI： 10.1038/s41467-019-12651-2http://dx.doi.org/10.1038/s41467-019-12651-2.
ZHAO Y， ZHAO R R， WU J T， et al. Melatonin protects against Aβ-induced neurotoxicity in primary neurons via miR-132/PTEN/AKT/FOXO3a pathway［J］. Biofactors， 2018， 44（6）： 609-618. DOI： 10.1002/biof.1411http://dx.doi.org/10.1002/biof.1411.
SAWIKR Y， YARLA N S， PELUSO I， et al. Neuroinflammation in Alzheimer’s disease： the preventive and therapeutic potential of polyphenolic nutraceuticals［J］. Adv Protein Chem Struct Biol， 2017， 108： 33-57. DOI： 10.1016/bs.apcsb.2017.02.001http://dx.doi.org/10.1016/bs.apcsb.2017.02.001.
KINNEY J W， BEMILLER S M， MURTISHAW A S， et al. Inflammation as a central mechanism in Alzheimer’s disease［J］. Alzheimers Dement， 2018， 4： 575-590. DOI： 10.1016/j.trci.2018.06.014http://dx.doi.org/10.1016/j.trci.2018.06.014.
廖旗荣，杨红岩，卢静，等.胰淀素对阿尔茨海默病小鼠学习和记忆能力以及Akt信号通路的影响［J］.陆军军医大学学报，2024，46（21）：2467-2474.DOI：10.16016/j.2097-0927.202404087http://dx.doi.org/10.16016/j.2097-0927.202404087.
LIAO Q R，YANG H Y，LU J， et al.Effect of amylin on learning and memory abilities and Akt signaling pathway in mice with Alzheimer’s disease［J］.J Amry Med Univ，2024，46（21）：2467-2474.DOI：10.16016/j.2097-0927.202404087http://dx.doi.org/10.16016/j.2097-0927.202404087.
VARNUM M M， IKEZU T. The classification of microglial activation phenotypes on neurodegeneration and regeneration in Alzheimer’s disease brain［J］. Arch Immunol Ther Exp， 2012， 60（4）： 251-266. DOI： 10.1007/s00005-012-0181-2http://dx.doi.org/10.1007/s00005-012-0181-2.
PARK J， KIM S H， KIM Y J， et al. Elevation of phospholipase C-β1 expression by amyloid-β facilitates calcium overload in neuronal cells［J］. Brain Res， 2022， 1788： 147924. DOI： 10.1016/j.brainres.2022.147924http://dx.doi.org/10.1016/j.brainres.2022.147924.
QIN Y R， MA C Q， WANG D P， et al. Bilobalide alleviates neuroinflammation and promotes autophagy in Alzheimer’s disease by upregulating lincRNA-p21［J］. Am J Transl Res， 2021， 13（4）： 2021-2040.
OMATA Y， LIM Y M， AKAO Y， et al. Age-induced reduction of autophagy-related gene expression is associated with onset of Alzheimer’s disease［J］. Am J Neurodegener Dis， 2014， 3（3）： 134-142.
0
Views
0
下载量
0
CSCD
- L-PDFDownload
- WORDDownload
- Meta-XMLDownload
- BibTeXDownload
- EndnoteDownload
- RefMan Download
- NoteExpressDownload
- NoteFirstDownload
Publicity Resources
Related Articles
Related Author
Related Institution
- Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19
京公网安备11010802024621 - It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
- Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.
