Arq Bras Cardiol abc Arquivos Brasileiros de Cardiologia Arq. Bras. Cardiol. 0066-782X 1678-4170 Sociedade Brasileira de Cardiologia - SBC XjbCR6td7GHmJDdjr7qTRZD S0066-782X2022000601134 abc.20201383 10.36660/abc.20201383 Artigo de Revisão Os LncRNAs Estão Envolvidos no Processo de Aterosclerose em Diversos Níveis 0000-0002-9633-1642 Liang Shiyi 1 0000-0001-5095-1697 Xv Weicheng 1 0000-0003-4099-3476 Li Chijian 1 0000-0003-1329-8814 Huang Yuxiang 1 0000-0003-2477-3772 Qian Ge 1 Yan Yuxiang 2 Zou Hequn 3 0000-0002-7572-2353 Li Yongqiang 4 The Third Affiliated Hospital of Southern Medical University Department of Nephrology Guangzhou China The Third Affiliated Hospital of Southern Medical University – Department of Nephrology, Guangzhou – China Capital Medical University Department of Epidemiology and Biostatistics Beijing China Capital Medical University – Department of Epidemiology and Biostatistics, Beijing – China South China Hospital of Shenzhen University Department of Nephrology Beijing China South China Hospital of Shenzhen University, Department of Nephrology, Beijing – China The Third Affiliated Hospital of Southern Medical University General Practice Department Guangzhou China The Third Affiliated Hospital of Southern Medical University – General Practice Department, Guangzhou – China Correspondência: Yongqiang Li • The Third Affiliated Hospital of Southern Medical University – GuangDong GuangZhou 510000 – China. E-mail: liyongqiang851@163.com

Contribuição dos autores

Concepção e desenho da pesquisa: Liang S, Li U; Obtenção de dados: Liang S, Xv W, Li C, Huang Y, Qian G; Obtenção de financiamento: Xv W, Li U; Redação do manuscrito: Liang S; Revisão crítica do manuscrito quanto ao conteúdo intelectual importante: Yan Y, Zou H, Li U.

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 10 06 2022 2022 118 6 1134 1140 15 08 2020 14 04 2021 09 06 2021 This is an open-access article distributed under the terms of the Creative Commons Attribution License Resumo

A aterosclerose é a causa mais comum de doença cardiovascular em todo o mundo, ela está associada a uma alta incidência de eventos clínicos. O acúmulo de evidências elucidou que os RNAs longos não codificantes (LncRNAs) são uma nova classe de transcritos com papéis críticos nos processos fisiopatológicos da aterosclerose. Nesta revisão, resumimos o progresso recente dos LncRNAs no desenvolvimento da aterosclerose. Descrevemos principalmente os diversos mecanismos regulatórios dos LncRNAs nos níveis transcricionais e pós-transcricionais. Este estudo pode fornecer informações úteis sobre os LncRNAs como alvos terapêuticos ou biomarcadores para o tratamento da aterosclerose.

 Palavras-chave LncRNAs Enzimas Inibidores Enzimáticos Inibidores Aterosclerose Doenças Cardiovasculares Células Endoteliais Lipoproteínas VLDL Interferência de RNA “Thirteenth Five-Year Plan” of Guangdong Province Educational Science 2020GXJK441 Traditional Chinese Medicine Bureau of Guangdong Province 20191228 Science and Technology Planning Project of Tianhe District 201704KW011 People’s Hospital of Huadu District 2020C03 O presente estudo foi financiado por “Thirteenth Five-Year Plan” of Guangdong Province Educational Science (No. 2020GXJK441), the Project of Traditional Chinese Medicine Bureau of Guangdong Province (CN) (No. 20191228 to ZY), the Science and Technology Planning Project of Tianhe District, Guangzhou City (No.201704KW011), the Young Plants Project, Hospital scientific research fund of the People’s Hospital of Huadu District, Guangzhou (NO.2020C03) Introdução

As doenças cardiovasculares (DCVs) são consideradas um problema de saúde global, responsável por 17,9 milhões de mortes todos os anos.1 A aterosclerose (AS), a principal impulsionadora das DCV em todo o mundo, é um processo inflamatório crônico conduzido por lipídios com disfunção endotelial, formação de células espumosas e acúmulo final de placa.2 Este processo é acompanhado pela proliferação de células, apoptose e liberação de fatores pró-inflamatórios.3 (Figura 1) Eles podem desencadear a ruptura da placa e a formação de trombose, levando a eventos clínicos agudos, como acidente vascular cerebral e síndrome coronariana aguda.4

A patogênese da aterosclerose.

No genoma de mamífero, os RNAs codificantes de proteínas são apenas <3%.5 Essa fração do gene codificador torna, portanto, difícil explicar o complexo mecanismo regulatório do organismo. Nos últimos anos, estudos têm revelado o importante papel dos RNAs não codificantes de proteínas nos processos fisiopatológicos de várias doenças.6–7 De acordo com o comprimento, os RNAs não codificantes (ncRNAs) podem ser divididos em RNAs não codificantes longos (lncRNA, >200 nucleotídeos) e RNAs não codificantes pequenos (<200 nucleotídeos, como miRNAs, piRNAs e siRNAs).8 Em muitas pesquisas, algumas funções regulatórias e efeitos biológicos de pequenos ncRNAs foram demonstrados.9–11 A função de muitos LncRNAs é desconhecida, mas um número crescente de LncRNAs foi caracterizado.

A biossíntese do lncRNA é semelhante à do mRNA. Os LncRNAs são transcritos pela RNA polimerase II, mas não possuem fases de leitura abertas e estão em uma expressão mais baixa do que os genes codificantes de proteínas.8 Os LncRNAs estão localizados principalmente no núcleo e no citoplasma.12 No citoplasma, os LncRNAs podem se ligar aos ribossomos13 ou originar-se do genoma mitocondrial.14 Os primeiros relatórios mostram que muitos LncRNAs não podem codificar proteínas porque não possuem fases de leitura aberta (ORFs) ou contêm poucas ORFs. Mas evidências emergentes sugerem que alguns LncRNAs contêm pequenas ORFs que codificam pequenas proteínas ou micropeptídeos, que são considerados reguladores-chave em vários processos biológicos.8,15,16 Estudos demonstram que os LncRNAs desempenham papéis críticos na função das células endoteliais e do músculo liso vascular (VSMC), na ativação de macrófagos, no metabolismo lipídico e a resposta inflamatória.17,18 Nesta revisão, discutimos principalmente a regulação dos LncRNAs que estão envolvidos no processo fisiopatológico da aterosclerose em níveis transcricionais e pós-transcricionais.

A patogênese da aterosclerose é acompanhada por disfunção celular, como proliferação, apoptose e migração. O resultado é a formação de células espumosas e o acúmulo de placas.

As classificações e mecanismo regulatório de LncRNAs

De acordo com a correlação entre a localização genômica e genes codificadores de proteínas, os LncRNAs podem ser divididos em (1) LncRNAs intergênicos (LincRNAs) que expressam genes codificadores de proteínas como uma unidade independente. (2) LncRNAs intrônicos que derivam dos íntrons de genes codificadores de proteínas. (3) LncRNAs anti-senso transcritos da direção oposta dos genes que codificam proteínas. (4) LncRNAs senso que se sobrepõem aos exons de genes que codificam proteínas na mesma fita. (5) potenciadores que se originam no potenciador de genes que codificam proteínas. (6) LncRNAs bidirecionais que são transcritos a partir dos promotores bidirecionais divergentes.19,20 Os critérios de classificação também incluem as várias funções na regulação do gene local: cis- (regulação da expressão de genes proximais) e trans- (regulação da expressão de genes distantes).21 Além disso, os transcritos de LncRNAs também podem ser categorizados em lineares ou circulares.22

O mecanismo de funcionamento dos LncRNAs não foi completamente elucidado, mas pode ser classificado aproximadamente em vários grupos: 1. a regulação da transcrição está incorporada na interferência da transcrição, remodelação da cromatina e promoção da transcrição; 2. níveis pós-transcricionais se manifestam no controle da tradução da regulação do splicing de mRNAs e até mesmo como esponjas para miRNAs; 3. Outros contêm localização de proteínas, replicação de telômeros e interferência de RNA etc. Além disso, seus mecanismos de direcionamento para regular a expressão de genes são resumidos como o seguinte: sinais, iscas, guias e estruturas.22,23

 Regulação transcricional

Os LncRNAs podem exercer sua regulação transcricional por meio de mecanismos de ação cis e ação trans. (Tabela 1). Os LncRNAs regulam a expressão de genes vizinhos em cis via interferência transcricional ou remodelação da cromatina.24 Os LncRNAs de ação trans podem interagir com RNA polimerases e fatores de alongamento da transcrição ou servir como um arcabouço para complexos de modificação da cromatina para regular os genes distantes.24,25

O papel dos lncRNAs no processo patológico da aterosclerose
LncRNAs Mecanismo Efeito Referências
Função celular Proliferação Apoptose
Células Endoteliais (ECs) MALAT1 MALAT1-Pc2 (CBX4)-E2F1 + 38
GAS5 GAS5 - ceRNA (miR-21) − + 75
HOTTIP TNF-α/PDGFBB-HOTTIP-β-catenina + 47
MALAT1 ceRNA (miR-22-3p) − 60
TUG1 ceRNA (miR-26a) + 71
Macrófagos, Células musculares lisas ANRIL Ligam com CBX7 e SUZ12 + − 32
NEAT1 NEAT1-WDR5-SM-genes específicos + 44
LincRNA-p21 LincRNA-p21-MDM2/ p300-p53 + − 45
HAS2 remodela estructura da cromatina + 49,50
RP11-714G18.1 regula positivamente a expressão de LRP2BP − 53
H19 ceRNA (miR-148b) + − 66
MIAT ceRNA (miR-181b) + − 69
LncRNAs Mecanismo Efeito Referências
Acúmulo de Lípidios MALAT1 captação de lipídios MALAT1-CD36 + 45
NEAT1 captação de lipídios NEAT1-CD36 − 41
MeXis LXR-MeXis-Abca1 − 46
H19 ceRNA (miR-130b) − 65
TUG1 ceRNA (miR-133a) + 72
LncRNAs Mecanismo Efeito Referências
Resposta inflamatória ANRIL TNF-α/NF-kB-ANRIL/YY1-IL6/8 + 34
MALAT1 MALAT1-p65/p50-TNF-α e IL-6 − 40
MALAT1 ceRNA (miR-503 or miR-155) − 61,62
H19 ceRNA (miR-130b) −
NEAT1 ceRNA (miR-342-3p) + 70
TUG1 ceRNA (miR-133a) + 72

representa prompt ou aumento, e

representa prevenir ou diminuir.

O estudo Wellcome Trust Case Control Consortium (WTCCC) e os estudos de associação do genoma completo descobriram que uma região no cromossomo 9p21 (Chr9p21) estava fortemente associada à doença arterial coronariana.26 A região é adjacente a um LincRNA denominado RNA não codificador anti-senso no lócus INK4 (ANRIL, também conhecido como CDKN2BAS).27 Holdt LM et al.,28 revelaram que a expressão de ANRIL estava correlacionada com a gravidade da aterosclerose por afetar a transcrição dos mRNAs, e o ANRIL também foi detectado em placas ateroscleróticas em seu estudo.28

Dois genes codificadores de proteínas, inibidores de quinase dependentes de ciclina (CDKN2A, CDKN2B) e a estrutura de leitura alternativa (ARF) no cromossomo 9p21, estão ligados a ANRIL inextricavelmente, os quais são supressores de tumor.27 O complexo polycomb repressivo-1 (PRC-1) e o complexo polycomb repressivo-2 (PRC-2) são dois tipos de proteínas de grupo polycomb envolvidas na manutenção do estado da cromatina.29 Suas subunidades CBX7 e SUZ12 se ligam ao ANRIL separadamente para silenciar o lócus CDKN2A/B através da trimetilação de H3 lisina27 (K27H3).30,31 Ainda, a repressão de CDKN2A/B pode estar relacionada à proliferação celular e apoptose no processo de aterosclerose.32

Holdt et al.,28 descobriram que ANRIL estava em posição de exercer uma função reguladora na expressão de genes distantes em trans. O elemento Alu, que marca o promotor dos genes trans-regulados de ANRIL, é decisivo para a trans-regulação linear de ANRIL. As proteínas PcG, desencadeadas pela ligação com ANRIL, eram altamente abundantes a jusante dos motivos Alu.33 O recrutamento de proteínas PcG poderia regular a expressão dos genes alvo (TSC22D3, COL3A1) e atenuar funções pró-aterogênicas mediadas por ANRIL, como células adesão, proliferação e apoptose.3,33 Além disso, ANRIL desempenha um papel fundamental nos processos inflamatórios através da via TNF-α/NF-kB-ANRIL/YY1-IL6/8. As proteínas associadas a PRC Yin Yang 1(YY1), um fator de transcrição, formam um complexo funcional com ANRIL.33 O complexo ANRIL-YYI se liga aos loci promotores de IL6/8 e estimula seu recrutamento na sinalização de TNF-α/NF-κB, levando à inflamação vascular.34

MALAT1, localizado no cromossomo 11q13, é descrito pela primeira vez como lncRNA associado a metástases de tumores de pulmão.35 A expressão de MALAT1 é regulada para baixo em placas ateroscleróticas em comparação com artérias não ateroscleróticas.36 Michalik et al.,37 descobriram que o silenciamento de MALAT1 inibiu a mudança de um estado promigratório para um proliferativo das células endoteliais, resultando na redução do crescimento de vasos.37 E MALAT1 também atua como um andaime molecular para interagir com o Polycomb 2 não metilado (Pc2); a expressão de Pc2 promove a SUMOilação de E2F1 e regula modificações de histonas para aumentar a proliferação celular.38

Em um experimento de controle, Gast et al.,39 observaram que os níveis séricos de TNF, IL-6 e IFN-γ estavam aumentados nos camundongos ApoE -/- deficientes em MALAT1, causando disfunção imunológica e aterosclerose agravada.39 MALAT1 pode estar envolvido na resposta LPS inflamatória induzida via sinalização LPS/TLR4/NF-κB. MALAT1 interage com as subunidades p65/p50 de NF-κB, inibindo a ligação de p65/p50 a promotores alvo, como TNF-α e IL-6, atenuando uma inflamação excessiva.40

No metabolismo lipídico, MALAT1 pode ser regulado positivamente em macrófagos durante a estimulação ox-LDL.41 CD36, um receptor eliminador de classe B, é necessário para a absorção de lipídios de ox-LDL.42 A superexpressão de MALAT1 induz o recrutamento de β-catenina no promotor CD36 para aumentar a transcrição de CD36, promovendo a captação de lipídios em macrófagos e acelerando a formação de células espumosas em placas ateroscleróticas.41

NEAT1, um transcrito adjacente de MALAT1, pode aumentar a formação de paraspeckles em oxLDL-macrófago induzido, que suprime a captação de lipídios ligando-se ao mRNA de CD36 para inibir a expressão de CD36 e estimula a resposta inflamatória por meio da fosforilação de p65 para promover a secreção de TNFα.43 Além disso, ASI et al.,44 descobriram que a expressão de NEAT1 foi regulada positivamente em células do músculo liso vascular (VSMCs) após lesão vascular in vivo e in vitro, levando a um estado de cromatina inativa em genes específicos de SM por meio da ligação com o modificador de cromatina WDR5. A repressão da expressão de genes específicos de SM mudou VSMCs para fenótipo proliferativo, promovendo a proliferação e migração de VSMCs e, assim, a formação de neoíntima.44

A expressão de lincRNA-p21 foi regulada negativamente nas placas ateroscleróticas. O LincRNA-p21 diminuiu a interação MDM2/p53 e aumentou a interação p300/p53 para facilitar a atividade transcricional de p53, levando à repressão da formação neointimal, a inibição da proliferação celular e o aumento da apoptose em VSMCs e células macrófagos mononucleares in vitro e vivo.45

Além disso, alguns outros LncRNAs estão envolvidos no processo AS no nível transcricional, mas as descrições são limitadas. A superexpressão de LncRNA-MeXis em macrófagos pode facilitar a reversão do transporte de colesterol pelos macrófagos através do eixo LXR-MeXis-Abca1, sugerindo que LncRNA-MeXis desempenha um papel protetor no desenvolvimento de aterosclerose.46 A expressão ectópica de LncRNA-HOTTIP, induzida por TNF-α ou fator de crescimento derivado de plaquetas (PDGFBB), aumenta a expressão de marcadores proliferativos ciclina D1 e PCNA através da via Wnt/β-catenina, subsequentemente estimulando a proliferação e migração de células endoteliais.47 A O-GlcNAcilação modula a ativação do promotor HAS2-AS1, o transcrito anti-senso natural HAS2-AS1 pode regular a transcrição de HAS2 em cis através da remodelação da estrutura da cromatina,48 HAS2 pode estar relacionado à proliferação de VSMCs,49,50 recrutamento de macrófagos,50 migração de VSMCs e formação de neoíntima,51,52 e resposta inflamatória.50,52 A expressão de LncRNA RP11-714G18.1 na placa aterosclerótica é baixa. Ainda assim, pode regular positivamente a expressão do gene LRP2BP próximo para prejudicar a migração celular, suprimir a adesão de ECs aos monócitos, reduzir a neoangiogênese, diminuir a apoptose de VSMCs e promover a produção de óxido nítrico. Além disso, o LRP2BP sérico foi positivamente relacionado ao colesterol de lipoproteína de alta densidade.53

HOXC-AS1 pode suprimir o acúmulo de colesterol em macrófagos por meio da promoção da expressão de HOXC6 em níveis de mRNA.54 LEENE pode melhorar a função endotelial aumentando a transcrição inicial do RNA da eNOS.55 Lethe Lin et al.,56 atua como uma isca lncRNA para interagir com a subunidade RelA do NF-κB e inibe a ligação de RelA ao DNA de genes alvo, como IL6, SOD2, IL8, atenuando a resposta inflamatória.56 LncRNA-TSLP induz a transcrição de HOTAIR através da via PI3K/AKT-IRF1, promovendo a proliferação e migração de células endoteliais na aterosclerose.57 Além disso, a TSLP induzida por ox-LDL pode se ligar a células dendríticas (DCs) para ativar a inflamação Th17,58 que está relacionado à gravidade e progressão da AS.59

 Regulação pós-transcricional

Os LncRNAs atuam principalmente como RNAs endógenos competidores (ceRNAs) ou miRNAs “esponja” interagindo com miRNAs no processo de aterosclerose no nível de regulação pós-transcricional. (tabela 1) Além disso, eles também estão envolvidos no controle da tradução, regulação de splicing e o mecanismo de RNA de interferência pequeno (siRNA).24

MALAT1 atua como ceRNA na lesão de células induzidas por ox-LDL e desempenha um papel protetor na doença aterosclerose. MALAT1 poderia competir com miR-22-3p por RNA endógeno e regular positivamente os genes alvo CXCR2 e AKT de miR-22-3p para inibir a apoptose de células endoteliais e promover a migração de ECs e angiogênese.60 Cremer S et al.,61 descobriram que MALAT1 “esponja” o miR-503 para reduzir a liberação de citocinas pró-inflamatórias, atenuando a inflamação da placa.61 Além disso, o supressor de sinalização de citocina 1 (SOCS1) é a proteína alvo do miR-155 que regula negativamente a sinalização do transdutor de sinal da quinase ativada de Janus (JAK) e do ativador da transcrição (STAT). MALAT1 poderia diminuir a regulação de miR-155 e aumentar a expressão de SOCS1 para aliviar a inflamação e apoptose na aterosclerose.62 Assim, MALAT1 pode desempenhar um papel protetor por meio da interação com miRNAs na patogênese da aterosclerose.

A expressão de lncRNA H19 foi regulada positivamente em macrófagos tratados com LDL-ox. MiR-130b regula a resposta inflamatória diminuindo os níveis translacionais de TNF-α, Sp1, NF-κB com estimulação lipídica63 e inibe a adipogênese ao direcionar PPAR-g.64 O silenciamento de H19 aumenta significativamente a expressão de miR-130b, que melhora a inflamação e a síntese de lipídios em células Raw264.7 tratadas com LDL-ox.65 O H19 pode acelerar a proliferação e impedir a apoptose em VSMCs estimulados por LDL-ox, suprimindo diretamente a expressão de miR-148b e aumentando a expressão do gene WNT1 do miR-148b alvo.66

O LncRNA-MIAT pode estar envolvido na progressão da placa aterosclerótica. O MIAT é expresso principalmente nos macrófagos de placas ateroscleróticas avançadas. Com o tratamento ox-LDL, a expressão de MIAT é regulada positivamente. A molécula anti-fagocítica CD47, um gene alvo de miR-149-5p, está relacionada à depuração de células apoptóticas e núcleos necróticos67 O MIAT interfere nas vias do miR-149-5p para aumentar o nível de CD47 em macrófagos, promovendo a vulnerabilidade da placa.68 A formação do eixo MIAT/miR-181b/STAT3 desempenha um papel crítico nas células do músculo liso vascular da aorta humana induzidas por ox-LDL (HA-VSMCs) e células mononucleares humanas (U937). O MIAT regula para cima o transdutor de sinal e o ativador do nível da proteína de transcrição 3 (STAT3) por meio do sequestro de miR-181b, promovendo subsequentemente a proliferação, facilitando a parada do ciclo celular e inibindo a apoptose em células HA-VSMCs e U937.69

NEAT1 também estava envolvido no processo aterosclerótico como ceRNA, exceto para a remodelação da cromatina no nível transcricional. Wang et al.,70 descobriram que NEAT1 foi significativamente regulado positivamente na presença de ox-LDL e serviu como uma esponja para reprimir a expressão de miR-342-3p, aumentando o nível sérico de IL-6, IL-1β, COX-2 e colesterol total levando para acelerar o processo de inflamação e a formação de células espumosas.70 LncRNA-TUG1 poderia regular negativamente a expressão de miR-26a e aumentar o mRNA e o nível de proteína de TRPC6 para facilitar a apoptose das células endoteliais.71 Zhang et al.,72 revelou que o TUG1 esponjou miR-133a e regulou positivamente a expressão do fator de crescimento de fibroblastos 1 (FGF1), resultando em aumento da hiperlipidemia e resposta inflamatória excessiva agravou a lesão aterosclerótica.72

Além disso, mais e mais estudos demonstraram que muitos LncRNAs relacionados à aterosclerose desempenham um papel crucial na patogênese da AS, interagindo com miRNAs no nível pós-transcricional. LINC00305 atua como uma esponja endógena para miR-136 e inibe a expressão de miR-136 para suprimir a proliferação de células endoteliais vasculares e aumentar a apoptose.73 LincRNA-p21 funciona como ceRNA para promover a apoptose de ECs e induz a progressão do ciclo celular ao direcionar o miR-130b.74 LncRNA-GAS5 regula negativamente a expressão de miR-21 para aumentar a expressão de morte celular programada 4 (PDCD4), suprimindo a proliferação de ECs e desencadeando a apoptose de ECs.75

 Outras

Os LncRNAs podem funcionar através da localização de proteínas, replicação de telômeros e interferência de RNA em alguns processos,24 tal como localização de partículas de RNP em plantas leguminosas, extensão do telômero durante a replicação do DNA em eucariotos,76 reduzindo o siRNA gerado por Dicer e afetando a expressão de genes regulados por Dicer.77 No entanto seu mecanismo molecular subjacente relacionado ao desenvolvimento de aterosclerose permanece desconhecido.

 Conclusão e Perspectiva

Tomados em conjunto, os LncRNAs podem estar envolvidos em vários processos associados à aterosclerose, incluindo resposta inflamatória, metabolismo lipídico e função celular. Eles regulam a patologia da aterosclerose em níveis epigenéticos, transcricionais e pós-transcricionais, como remodelamento da cromatina, promoção da transcrição e competição endógena por miRNAs. Portanto, os LncRNAs podem servir como novos marcadores diagnósticos e alvos terapêuticos promissores para aterosclerose e doenças vasculares. Além disso, todos esses papéis possíveis nos processos fisiopatológicos abriram espaços para decifrar a função e o mecanismo dos LncRNAs em doenças cardiovasculares e outras doenças, como tumores, doenças renais e nervosas.

Fontes de financiamento

O presente estudo foi financiado por “Thirteenth Five-Year Plan” of Guangdong Province Educational Science (No. 2020GXJK441), the Project of Traditional Chinese Medicine Bureau of Guangdong Province (CN) (No. 20191228 to ZY), the Science and Technology Planning Project of Tianhe District, Guangzhou City (No.201704KW011), the Young Plants Project, Hospital scientific research fund of the People’s Hospital of Huadu District, Guangzhou (NO.2020C03)

Vinculação acadêmica

Não há vinculação deste estudo a programas de pós-graduação.

Aprovação ética e consentimento informado

Este artigo não contém estudos com humanos ou animais realizados por nenhum dos autores.

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Pan SD Yang X Jia Y Li R Zhao R Microvesicle-shuttled miR-130b reduces fat deposition in recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression J Cell Physiol 2014 229 5 631 639 10.1002/jcp.24486 64 Pan SD, Yang X, Jia Y, Li R, Zhao R. Microvesicle-shuttled miR-130b reduces fat deposition in recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression. J Cell Physiol. 2014;229(5): 631-9. doi: 10.1002/jcp.24486. Han Y Ma J Wang J Wang L Silencing of H19 inhibits the adipogenesis and inflammation response in ox-LDL-treated Raw264.7 cells by up-regulating miR-130b Mol Immunol 2018 93 107 114 10.1016/j.molimm.2017.11.017 65 Han Y, Ma J, Wang J, Wang L Silencing of H19 inhibits the adipogenesis and inflammation response in ox-LDL-treated Raw264.7 cells by up-regulating miR-130b. Mol Immunol.2018;93:107-14. doi: 10.1016/j.molimm.2017.11.017 Zhang L Cheng H Yue Y Li S Zhang D He R H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL -stimulated vascular smooth muscle cells J Biomed Sci 2018 25 1 11 11 2018;1: 11 10.1186/s12929-018-0418-4 66 Zhang L, Cheng H, Yue Y, Li S, Zhang D,He R. H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL -stimulated vascular smooth muscle cells. J Biomed Sci.2018;25(1):11.. 2018; 1: 11. doi: 10.1186/s12929-018-0418-4. Kojima Y Volkmer JP McKenna K Civelek M Lusis AJ Miller CL CD47-blocking antibodies restore phagocytosis and prevent atherosclerosis Nature 2016 536 7614 86 90 10.1038/nature18935 67 Kojima Y, Volkmer JP, McKenna K, Civelek M, Lusis AJ, Miller CL, et al. CD47-blocking antibodies restore phagocytosis and prevent atherosclerosis. Nature. 2016;536(7614):86-90. doi: 10.1038/nature18935. Ye ZM Yang S Xia YP Hu RT Chen S BW LncRNA MIAT sponges miR-149-5p to inhibit efferocytosis in advanced atherosclerosis through CD47 upregulation Cell Death Dis 2019 10 2 138 138 10.1038/s41419-019-1409-4 68 Ye ZM, Yang S, Xia YP, Hu RT, Chen S, BW, et al. LncRNA MIAT sponges miR-149-5p to inhibit efferocytosis in advanced atherosclerosis through CD47 upregulation. Cell Death Dis. 2019;10(2):138. doi: 10.1038/s41419-019-1409-4. Zhong X Ma X Zhang L Li Y He R MIAT promotes proliferation and hinders apoptosis by modulating miR-181b/STAT3 axis in ox-LDL-induced atherosclerosis cell models Biomed Pharmacother Biomed 2018 97 1078 1085 10.1016/j.biopha.2017.11.052 69 Zhong X, Ma X, Zhang L, Li Y, He R. MIAT promotes proliferation and hinders apoptosis by modulating miR-181b/STAT3 axis in ox-LDL-induced atherosclerosis cell models. Biomed Pharmacother Biomed.2018;97: 1078-85. doi: 10.1016/j.biopha.2017.11.052. Wang L Xia JW Ke ZP Zhang H Blockade of NEAT1 represses inflammation response and lipid uptake via modulating miR-342-3p in human macrophages THP-1 cells J Cell Physiol 2019 234 4 5319 5326 10.1002/jcp.27340 70 Wang L, Xia JW, Ke ZP, Zhang H. Blockade of NEAT1 represses inflammation response and lipid uptake via modulating miR-342-3p in human macrophages THP-1 cells. J Cell Physiol.2019;234(4): 5319-26. doi: 10.1002/jcp.27340. Chen C Cheng G Yang X Li C Shi R Zhao N Tanshinol suppresses endothelial cells apoptosis in mice with atherosclerosis via lncRNA TUG1 up-regulating the expression of miR-26a Am J TRanslat Res 2016 8 7 2981 2991 27508018 71 Chen C, Cheng G, Yang X, Li C, Shi R, Zhao N. Tanshinol suppresses endothelial cells apoptosis in mice with atherosclerosis via lncRNA TUG1 up-regulating the expression of miR-26a. Am J TRanslat Res.2016;8(7):2981-91. PMID: 27508018 Zhang L Cheng H Yue Y Li S Zhang D He R TUG1 knockdown ameliorates atherosclerosis via up-regulating the expression of miR-133a target gene FGF1 Cardiovasc Pathol 2018 33 6 15 10.1016/j.carpath.2017.11.004 72 Zhang L, Cheng H, Yue Y, Li S, Zhang D, He R. TUG1 knockdown ameliorates atherosclerosis via up-regulating the expression of miR-133a target gene FGF1. Cardiovasc Pathol. 2018;33: 6-15. doi: 10.1016/j.carpath.2017.11.004. Zhang BY Jin Z Zhao Z Long intergenic non-coding RNA 00305 sponges miR-136 to regulate the hypoxia induced apoptosis of vascular endothelial cells Biomed Pharmacother 2017 94 238 243 10.1016/j.carpath.2017.11.004 73 Zhang BY, Jin Z, Zhao Z. Long intergenic non-coding RNA 00305 sponges miR-136 to regulate the hypoxia induced apoptosis of vascular endothelial cells. Biomed Pharmacother. 2017;94:238-43. doi: 10.1016/j.carpath.2017.11.004 He C Ding JW Li S Wu H Jiang YR Yang W The Role of Long Intergenic Noncoding RNA p21 in Vascular Endothelial Cells DNA Cell Biol 2015 34 11 677 683 10.1089/dna.2015.2966 74 He C, Ding JW, Li S, Wu H, Jiang YR, Yang W, et al. The Role of Long Intergenic Noncoding RNA p21 in Vascular Endothelial Cells. DNA Cell Biol. 2015;34(11):677-83. doi: 10.1089/dna.2015.2966. Shen Z She Q Association Between the Deletion Allele of Ins/Del Polymorphism (Rs145204276) in the Promoter Region of GAS5 with the risk of atherosclerosis Cell Physiol Biochem 2018 49 4 1431 1443 10.1159/000493447 75 Shen Z, She Q. Association Between the Deletion Allele of Ins/Del Polymorphism (Rs145204276) in the Promoter Region of GAS5 with the risk of atherosclerosis. Cell Physiol Biochem. 2018;49(4):1431-43. doi: 10.1159/000493447 Campalans A Kondorosi A Crespi M Enod40, a short open reading frame-containing mRNA, induces cytoplasmic localization of a nuclear RNA binding protein in Medicago truncatula Plant Cell 2004 16 4 1047 1059 10.1105/tpc.019406 76 Campalans A, Kondorosi A, Crespi M. Enod40, a short open reading frame-containing mRNA, induces cytoplasmic localization of a nuclear RNA binding protein in Medicago truncatula. Plant Cell. 2004;16(4): 1047-59. doi: 10.1105/tpc.019406. Hellwig S Bass BL A starvation-induced non-coding RNA modulates expression of Dicer-regulated genes Proc Natl Acad Sci 2008 105 35 1289 1902 10.1073/pnas.0805118105 77 Hellwig S, Bass BL. A starvation-induced non-coding RNA modulates expression of Dicer-regulated genes. Proc Natl Acad Sci. 2008;105(35):1289-902. doi: 10.1073/pnas.0805118105 Review Article LncRNAs are Involved in the Process of Atherosclerosis at Diverse Levels 0000-0002-9633-1642 Liang Shiyi 1 0000-0001-5095-1697 Xv Weicheng 1 0000-0003-4099-3476 Li Chijian 1 0000-0003-1329-8814 Huang Yuxiang 1 0000-0003-2477-3772 Qian Ge 1 Yan Yuxiang 2 Zou Hequn 3 0000-0002-7572-2353 Li Yongqiang 4 Guangzhou China The Third Affiliated Hospital of Southern Medical University – Department of Nephrology, Guangzhou – China Beijing China Capital Medical University – Department of Epidemiology and Biostatistics, Beijing – China Beijing China South China Hospital of Shenzhen University, Department of Nephrology, Beijing – China Guangzhou China The Third Affiliated Hospital of Southern Medical University – General Practice Department, Guangzhou – Chin Mailing Address: Yongqiang Li • The Third Affiliated Hospital of Southern Medical University – GuangDong GuangZhou 510000 – China. E-mail: liyongqiang851@163.com

Author Contributions

Conception and design of the research: Liang S, Li U; Acquisition of data: Liang S, Xv W, Li C, Huang Y, Qian G; Obtaining financing: Xv W, Li U; Writing of the manuscript: Liang S; Critical revision of the manuscript for intellectual contente: Yan Y, Zou H, Li U.

Potential Conflict of Interest

No potential conflict of interest relevant to this article was reported.

 Abstract

Atherosclerosis is the most common cause of cardiovascular disease globally, associated with a high incidence of clinical events. Accumulating evidence has elucidated that long non-coding RNAs (lncRNAs) as a novel class of transcripts with critical roles in the pathophysiological processes of atherosclerosis. In this review, we summarize the recent progress of lncRNAs in the development of atherosclerosis. We mainly describe the diverse regulatory mechanisms of lncRNAs at the transcriptional and post-transcriptional levels. This study may provide helpful insights about lncRNAs as therapeutic targets or biomarkers for atherosclerosis treatment.

 Keywords LnRNAs Emzymes Enzyme Inhibitors Endothelial Cells Lipoproteins, VLDL RNA Interference “Thirteenth Five-Year Plan” of Guangdong Province Educational Science 2020GXJK441 Traditional Chinese Medicine Bureau of Guangdong Province 20191228 Science and Technology Planning Project of Tianhe District 201704KW011 People's Hospital of Huadu District 2020C03 This study was funded by the “Thirteenth Five-Year Plan” of Guangdong Province Educational Science (No. 2020GXJK441), the Project of Traditional Chinese Medicine Bureau of Guangdong Province (CN) (No. 20191228 to ZY), the Science and Technology Planning Project of Tianhe District, Guangzhou City (No.201704KW011), the Young Plants Project, Hospital scientific research fund of the People's Hospital of Huadu District, Guangzhou (NO.2020C03) Introduction

Cardiovascular diseases (CVDs) are regarded as a global health problem that accounts for 17.9 million deaths every year.1 Atherosclerosis (AS), the principal driver of CVDs worldwide, is a lipid-driven chronic inflammatory process with endothelial dysfunction, foam cells formation and final plaque buildup.2 This process is accompanied by cells proliferation, apoptosis, and the release of pro-inflammatory factors3 (Figure 1). These can trigger plaque rupture and thrombosis formation, leading to acute clinical events, such as stroke and acute coronary syndrome.4

The pathogenesis of atherosclerosis.

In the mammalian genome, the encoded protein RNAs are only < 3%.5 That fraction of the coding gene makes, therefore, hard to explain the complex regulatory mechanism of the organism. In recent years, accumulating studies have revealed the important role of non-coding protein RNAs in the pathophysiological processes of various diseases.6,7 According to the length, the non-coding RNAs (ncRNAs) can be divided into long non-coding RNA (lncRNA, >200 nucleotides) and small non-coding RNA (<200 nucleotides, such as miRNAs, piRNAs and siRNAs).8 In many researches, some small ncRNAs’ regulatory functions and biological effects have been demonstrated.9–11 The function of many lncRNAs is unknown, but an increasing number of lncRNAs have been characterized.

The biosynthesis of lncRNA is similar to that of mRNA. LncRNAs are transcribed by RNA polymerase II but lack open reading frames, and they are in a lower expression than protein-coding genes.8 LncRNAs are mainly located within the nucleus and cytoplasm.12 In the cytoplasm, lncRNAs can bind with ribosomes13 or originate from the mitochondrial genome.14 Early reports show that many lncRNAs can’t encode proteins because they lack open reading frames (ORFs) or contain few ORFs. But emerging evidence suggests that some lncRNAs contain small ORFs encoding small proteins or micropeptides, which are regarded as key regulators in various biological processes.8,15,16 Studies demonstrate that lncRNAs play critical roles in the function of endothelial and vascular smooth muscle cells (VSMC), macrophage activation, lipid metabolism and inflammatory response.17,18 In this review, we mainly discuss the regulation of lncRNAs are involved in the pathophysiologic process of atherosclerosis at transcriptional and post-transcriptional levels.

The pathogenesis of atherosclerosis is accompanied by cell dysfunction, such as proliferation, apoptosis, and migration. The result is foam cells formation and plaque buildup.

The classifications and regulatory mechanism of LncRNAs

According to the correlation between the genomic location and protein-coding genes, lncRNAs can be divided into (1) intergenic lncRNAs (lincRNAs) that express protein-coding genes as an independent unit. (2) intronic lncRNAs that derive from the introns of protein-coding genes. (3) antisense lncRNAs transcribed from the opposite direction of protein-coding genes. (4) sense lncRNAs that overlap with exons of protein-coding genes on the same strand. (5) enhancers that originate in the enhancer of protein-coding genes. (6) bidirectional lncRNAs that are transcribed from the divergent bidirectional promoters.19,20 The criteria of classification also include the various functions in local gene regulation: cis- (regulating proximal genes expression) and trans- (regulating distant genes expression).21 Besides, lncRNAs transcripts can also be categorized into linear or circular.22

The mechanism of lncRNAs functioning has not been completely elucidated, but it can be classified roughly into several groups: 1. transcriptional regulation is embodied in transcriptional interference, chromatin remodeling and promotion of transcription; 2. post-transcriptional levels manifest in mRNAs splicing regulation translational control and even as sponges for miRNAs; 3. Others contain protein localization, telomere replication, and RNA interference, etc. Furthermore, their targeting mechanisms for regulating gene expression are summarized as the following: signals, decoys, guides and scaffolds.22,23

 Transcriptional regulation

LncRNAs can exert their transcriptional regulation through cis-acting and trans-acting mechanisms. (Table 1) LncRNAs regulate neighboring genes expression in cis via transcriptional interference or chromatin remodeling.24 Trans-acting lncRNAs can interact with RNA polymerases and transcription elongation factors or serve as a scaffold for chromatin modification complexes to regulate the distant genes.24,25

The role of lncRNAs in the pathologic process of atherosclerosis
lncRNAs Mechanism Effect References
Cells function Proliferation Apoptosis
Endothelial cells (ECs) MALAT1 MALAT1-Pc2 (CBX4)-E2F1 + 38
GAS5 GAS5 - ceRNA (miR-21) − + 75
HOTTIP TNF-α/PDGFBB-HOTTIP-β-catenin + 47
MALAT1 ceRNA (miR-22-3p) − 60
TUG1 ceRNA (miR-26a) + 71
Macrophages, Smooth muscular cells ANRIL Bind with CBX7 and SUZ12 + − 32
NEAT1 NEAT1-WDR5-SM-specific genes + 44
LincRNA-p21 lincRNA-p21-MDM2/ p300-p53 + − 45
HAS2 remodeling chromatin structure + 49,50
RP11-714G18.1 upregulate LRP2BP expression − 53
H19 ceRNA (miR-148b) + − 66
MIAT ceRNA (miR-181b) + − 69
lncRNAs Mechanism Effect References
Lipid accumulation MALAT1 MALAT1-CD36-lipid uptake + 45
NEAT1 NEAT1-CD36-lipid uptake − 41
MeXis LXR-MeXis-Abca1 − 46
H19 ceRNA (miR-130b) − 65
TUG1 ceRNA (miR-133a) + 72
lncRNAs Mechanism Effect References
Inflammatory response ANRIL TNF-α/NF-kB-ANRIL/YY1-IL6/8 + 34
MALAT1 MALAT1-p65/p50-TNF-α and IL-6 − 40
MALAT1 ceRNA (miR-503 or miR-155) − 61,62
H19 ceRNA (miR-130b) −
NEAT1 ceRNA (miR-342-3p) + 70
TUG1 ceRNA (miR-133a) + 72

represents prompt or increase, and

represents prevent or decrease.

The Wellcome Trust Case Control Consortium (WTCCC) study and the genome-wide association studies found that a region on chromosome 9p21 (Chr9p21) was strongly associated with coronary artery disease strongly.26 The region is adjacent to a lincRNA named antisense non-coding RNA in the INK4 locus (ANRIL, also known as CDKN2BAS).27 Holdt LM et al.28 had revealed that ANRIL expression was correlated with atherosclerosis severity by affecting mRNAs’ transcription, and the ANRIL was also detected in atherosclerotic plaques in their study.28

Two protein-coding genes, cyclin-dependent kinase inhibitors(CDKN2A, CDKN2B) and the alternative reading frame (ARF) on chromosome 9p21, are tied to ANRIL inextricably, which are tumor suppressors.27 The polycomb repressive complex-1 (PRC-1) and polycomb repressive complex-2 (PRC-2) are two kinds of polycomb group proteins involved in maintaining chromatin state.29 Their subunits CBX7 and SUZ12 bind ANRIL separately to silence CDKN2A/B locus through H3 lysine27 (K27H3) trimethylation.30,31 Yet, the repression of CDKN2A/B may be related to cell proliferation and apoptosis in the atherosclerosis process.32

Holdt et al.28 found that ANRIL was in a position to exert a regulatory function in distant gene expression in trans. Alu element, marking the promoter of the ANRIL trans-regulated genes, is decisive for linear ANRIL trans-regulation. PcG proteins, triggered by binding with ANRIL, were highly abundant downstream of the Alu motifs.33 The recruitment of PcG proteins could regulate the expression of the target genes (TSC22D3, COL3A1) and attenuate ANRIL-mediated pro-atherogenic functions, such as cell adhesion, proliferation, and apoptosis.3,33 Furthermore, ANRIL plays a pivotal role in the inflammatory processes through TNF-α/NF-kB-ANRIL/YY1-IL6/8 pathway. PRC-associated proteins Yin Yang 1 (YY1), a transcriptional factor, form a functional complex with ANRIL.33 ANRIL-YYI complex binds to IL6/8 promoter loci and stimulates their recruitment in the TNF-α/NF-κB signaling, leading to vascular inflammation.34

MALAT1, located on chromosome 11q13, is first described as lncRNA associated with metastasis of lung tumors.35 MALAT1 expression is downregulated in atherosclerotic plaques in comparison to non-atherosclerotic arteries.36 Michalik et al.37 found that silencing of MALAT1 inhibited a switch from a promigratory to a proliferative state of the endothelial cells, resulting in the reduction of vessel growth.37 And MALAT1 also acts as a molecular scaffold to interact with unmethylated Polycomb 2 (Pc2); the expression of Pc2 promotes E2F1 SUMOylation and regulates histone modifications to increase cell proliferation.38

In a control experiment, Gast et al.39 observed that the serum levels of TNF, IL-6, and IFN-γ were increased in the MALAT1-deficient ApoE-/- mice, causing immune dysfunction and aggravated atherosclerosis.39 MALAT1 may be involved in the LPS‐induced inflammatory response via LPS/TLR4/NF-κB signaling. MALAT1 interacts with NF-κB subunits p65/p50, inhibiting p65/p50 binding to target promoters such as TNF-α and IL-6, then attenuating an excessive inflammation.40

In lipid metabolism, MALAT1 may be upregulated in macrophages during ox-LDL stimulation.41 CD36, a class B scavenger receptor, is required for lipid uptake of ox-LDL.42 MALAT1 overexpression induces the recruitment of β-catenin on the CD36 promoter to enhance CD36 transcription, promoting lipid uptake in macrophages and accelerating the foam cell formation in atherosclerotic plaques.41

NEAT1, an adjacent transcript of MALAT1, can enhance the paraspeckles formation in oxLDL-induced macrophage, which suppresses lipid uptake by binding CD36 mRNA to inhibit CD36 expression and stimulates inflammatory response via phosphorylating p65 to promote TNF-α secretion.43 Besides, Ahmed ASI et al.44 found that NEAT1 expression was upregulated in vascular smooth muscle cells (VSMCs) after vascular injury in vivo and in vitro, leading to an inactive chromatin state in SM-specific genes through binding with the chromatin modifier WDR5. The repression of SM-specific genes expression switched VSMCs to proliferative phenotype, promoting VSMCs proliferation and migration and thereby neointima formation.44

The expression of lincRNA-p21 was downregulated in the atherosclerotic plaques. LincRNA-p21 decreased MDM2/p53 interaction and increased p300/p53 interaction to facilitate the transcriptional activity of p53, leading to the repression of neointimal formation, the inhibition of cell proliferation and the enhancement of apoptosis in VSMCs and mononuclear macrophage cells in vitro and vivo.45

Also, some other lncRNAs are involved in the AS process at the transcriptional level, but the descriptions are limited. The overexpression of lncRNA-MeXis in macrophages may facilitate macrophage reversing cholesterol transport via the LXR-MeXis-Abca1 axis, suggesting that lncRNA-MeXis plays a protective role in the development of atherosclerosis.46 Ectopic expression of lncRNA-HOTTIP, induced by TNF-α or platelet-derived growth factor (PDGFBB), increases proliferative markers cyclin D1 and PCNA expression through the Wnt/β-catenin pathway, subsequently prompting the endothelial cell proliferation and migration.47 The O-GlcNAcylation modulates HAS2-AS1 promoter activation, HAS2-AS1 natural antisense transcript can regulate HAS2 transcription in cis through remodeling chromatin structure,48 HAS2 may be related to VSMCs proliferation,49,50 macrophages recruitment,50 VSMCs migration and neointima formation,51,52 inflammatory response.50,52 The expression of lncRNA RP11-714G18.1 in atherosclerotic plaque is low. Still, it can upregulate nearby gene LRP2BP expression to impair cell migration, suppress the adhesion of ECs to monocytes, reduce the neoangiogenesis, decrease VSMCs apoptosis and promote nitric oxide production. Furthermore, the serum LRP2BP was positively related to high-density lipoprotein cholesterol.53

HOXC-AS1 may suppress the cholesterol accumulation in macrophages via promoting HOXC6 expression at mRNA levels.54 LEENE can improve endothelial function by enhancing eNOS initial RNA transcription.55 Lethe Lin et al.56 acts as a decoy lncRNA to interact with the NF-κB subunit RelA and inhibits RelA binding to target genes DNA, such as IL6, SOD2, IL8, attenuating the inflammatory response.56 LncRNA-TSLP induces HOTAIR transcription through PI3K/AKT-IRF1 pathway, promoting endothelial cell proliferation and migration in atherosclerosis.57 Besides, ox-LDL induced TSLP may bind to dendritic cells (DCs) to activate the Th17 inflammation,58 which is related to the severity and progression of AS.59

 Post-transcriptional regulation

LncRNAs mainly act as competing endogenous RNAs (ceRNAs) or miRNAs “sponge” interacting with miRNAs in the process of atherosclerosis at the post-transcriptional regulation level. (Table 1) Furthermore, they are also involved in translational control, splicing regulation and small interfering RNA (siRNA) mechanism.24

MALAT1 acts as ceRNA in ox-LDL-induced cells injury and plays a protective role in atherosclerosis disease. MALAT1 could compete with miR-22-3p for endogenous RNA and upregulate the target genes CXCR2 and AKT of miR-22-3p to inhibit endothelial cells apoptosis and promote the ECs migration and angiogenesis.60 Cremer S et al.61 found that MALAT1 “sponged” miR-503 to reduce the release of pro-inflammatory cytokines, attenuating plaque inflammation.61 Besides, the suppressor of cytokine signaling 1 (SOCS1) is the target protein of miR-155 that negatively regulates Janus activated kinase (JAK)-signal transducer and activator of transcription (STAT) signaling. MALAT1 could downregulate miR-155 and increase the expression of SOCS1 to alleviate the inflammation and apoptosis in atherosclerosis.62 Thus, MALAT1 may play a protective role via interacting with miRNAs in the pathogenesis of atherosclerosis.

The expression of lncRNA H19 was up-regulated in ox-LDL treated macrophages. MiR-130b regulates the inflammatory response by decreasing the translational levels of TNF-α, Sp1, NF-κB with lipid stimulation63 and inhibits adipogenesis by targeting PPAR-g.64 Silencing of H19 significantly increases the expression of miR-130b, which ameliorates inflammation and lipid synthesis in ox-LDL-treated Raw264.7 cells.65 H19 can accelerate proliferation and impede apoptosis in ox-LDL-stimulated VSMCs by directly suppressing miR-148b expression and enhancing miR-148b target gene WNT1 expression.66

LncRNA-MIAT may be involved in atherosclerotic plaque progression. MIAT is mainly expressed in the macrophages of advanced atherosclerotic plaques. With the ox-LDL treatment, the expression of MIAT is upregulated. Anti-phagocytic molecule CD47, a target gene of miR-149-5p, is related to apoptotic cell clearance and necrotic cores.67 MIAT interferes with miR-149-5p pathways to increase the CD47 level in macrophages, promoting plaque vulnerability.68 The formation of the MIAT/miR-181b/STAT3 axis plays a critical role in ox-LDL induced human aorta vascular smooth muscle cells (HA-VSMCs) and human mononuclear cells (U937). MIAT up-regulates signal transducer and activator of transcription 3 (STAT3) protein level through sequestering miR-181b, subsequently promoting proliferation, facilitating cell cycle arrest and inhibiting apoptosis in HA-VSMCs and U937 cells.69

NEAT1 was also involved in the atherosclerotic process as ceRNA except for remodeling chromatin at the transcriptional level. Lei Wang et al.70 found that NEAT1 was significantly upregulated in the presence of ox-LDL and served as a sponge to repress the expression of miR-342-3p, increasing the serum level of IL-6, IL-1β, COX-2, and total cholesterol leading to accelerating inflammation process and the formation of foam cells.70 LncRNA-TUG1 could down-regulate the expression of miR-26a and increase the mRNA and protein level of TRPC6 to facilitate the endothelial cells apoptosis.71 Lei Zhang et al.72 revealed that TUG1 sponged miR-133a and up-regulated fibroblast growth factor 1 (FGF1) expression, resulting in increased hyperlipidemia and excessive inflammatory response aggravated atherosclerotic lesion.72

In addition, more and more studies have demonstrated that plenty of atherosclerosis-related lncRNAs plays a crucial role in the pathogenesis of AS by interacting with miRNAs at the post-transcriptional level. LINC00305 acts as an endogenous sponge for miR-136 and inhibits miR-136 expression to suppress the vascular endothelial cells proliferation and enhance apoptosis.73 LincRNA-p21 functions as ceRNA to promote ECs apoptosis and induces cell cycle progression by targeting the miR-130b.74 LncRNA-GAS5 negatively regulates miR-21 expression to enhance programmed cell death 4 (PDCD4) expression, suppressing ECs proliferation and triggering ECs apoptosis.75

 Others

LncRNAs may function through protein localization, telomere replication and RNA interference in some processes,24 such as localizing RNP particles in legume plants, extending telomere during DNA replication in eukaryote,76 reducing Dicer-generated siRNA and affecting the expression of Dicer-regulated genes.77 While their underlying molecular mechanism related to the development of atherosclerosis remains unknown.

 Conclusion and Perspective

Taken together, lncRNAs can be involved in several processes associated with atherosclerosis, including inflammatory response, lipid metabolism and cells function. They regulate the pathology of atherosclerosis at epigenetic, transcriptional and post-transcriptional levels, such as chromatin remodeling, promotion of transcription and competing endogenous for miRNAs. Therefore, lncRNAs may serve as promising novel diagnostic markers and therapeutic targets for atherosclerosis and vascular diseases. Moreover, all of these possible roles in physiopathologic processes have opened venues to decipher the function and mechanism of lncRNAs in cardiovascular diseases and other diseases, such as tumors, renal diseases and nervous diseases.

Sources of Funding

This study was funded by the “Thirteenth Five-Year Plan” of Guangdong Province Educational Science (No. 2020GXJK441), the Project of Traditional Chinese Medicine Bureau of Guangdong Province (CN) (No. 20191228 to ZY), the Science and Technology Planning Project of Tianhe District, Guangzhou City (No.201704KW011), the Young Plants Project, Hospital scientific research fund of the People's Hospital of Huadu District, Guangzhou (NO.2020C03)

Study Association

This study is not associated with any thesis or dissertation work.

Ethics approval and consent to participate

This article does not contain any studies with human participants or animals performed by any of the authors.