The Mechanism of lncRNA NEAT1 in Alleviating Acute Respiratory Distress Syndrome Through miR-27b Regulated Nrf2 Pathway

Inflammation Clinical Trial

Official title:

The Mechanism of lncRNA NEAT1 in Alleviating Acute Respiratory Distress Syndrome Through miR-27b Regulated Nrf2 Pathway

Clinical Trial Details — Status: Enrolling by invitation

Administrative data

• NCT number: NCT04937855
• Other study ID #: 81970067
• Status: Enrolling by invitation
• Start date: July 1, 2021
• Est. completion date: December 31, 2023

Study information

• Verified date: June 2021
• Source: Beijing Anzhen Hospital
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

The acute respiratory distress syndrome, formerly known as the acute lung injury (ARDS/ALI), is a critical illness with high mortality due to the lack of effective treatment. The pathogenesis of ARDS/ALI has not been fully elucidated. Nuclear factor E2-related factor 2 (Nrf2) plays a key role in regulating lung inflammation and oxidative stress which are closely related to lung injury in ARDS/ALI, but its regulatory mechanism remains unclear. The investigator's provious study shown that microRNA-27b (miR-27b) downregulated Nrf2 to aggravate lung inflammation and histological injury. Furthermore, in lipopolysaccharide (LPS)-induced cell (J774A.1) inflammation model, miR-27b was upregulated while the long non-coding RNA (lncRNA) NEAT1 was downregulated, the putative binding sites of lncRNA NEAT1 and miR-27b were successfully predicted by bioinformatics approach. Thus, the investigators propose that NEAT1 plays as a competing endogenous RNA (ceRNA) to adsorb miR-27b and liberate Nrf2, therefore, to attenuate lung inflammation and related lung injury in ARDS/ALI. This project aims to explore the role of the lncRNA NEAT1/ mir-27b /Nrf2 signal axis in the development and treatment of ARDS/ALI in patients, as well as in LPS-induced ALI animal and cell models by using bioinformatics, molecular biology, histomorphology and clinical phenotype approaches, and to clarify the new mechanism in ARDS/ALI development and to provide new therapeutic targets.

Description:

Collect blood and BALF from 400 ARDS patients at different time (at check-in, 24, 48 and 72 h after check-in the hospital) and 25 gender and age matching healthy controls. Use RT-PCR to detect the expression of lncRNA NEAT1、miR-27b and Nrf2 in blood and BALF of ARDS patients and health controls. The expressions of inflammatory and oxidative stress associated factors (NLRP3、NF-κB-P65、 p-P65、IκB、p-IκB、HO-1、NQO1、caspase-1、IL-1β、IL-6、IL-18、TNF-α) will be detected by western blot、ELISA and RT-PCR. Moreover, flow cytometry will be adopted to measure the numbers and kinds of cells in BALF. Then, analyze the differences of the expressions of lncRNA NEAT1、miR-27b and Nrf2 in the groups. To explore the correlation of expressions of lncRNA NEAT1、miR-27b and Nrf2 with inflammation and oxidative stress in the groups. Finally, to declare the relative of lncRNA NEAT1、miR-27b and Nrf2 with the time of mechanical ventilation, severity and mortality in 28 days of ARDS patients.

Recruitment information / eligibility

• Status: Enrolling by invitation
• Enrollment: 425
• Est. completion date: December 31, 2023
• Est. primary completion date: December 31, 2022
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: All
• Age group: 18 Years to 70 Years

Eligibility

Inclusion Criteria:

We included patients with acute respiratory distress according to 2012 ARDS Berlin new definition (Acute Respiratory Distress Syndrome: The Berlin Definition. JAMA, 2012, 307(23):2526).

• Acute or progressive dyspnea within 1 week with identify cause;
• Chest radiograph/chest CT showed double lung infiltration, which could not be fully explained by pleural effusion, atelectasis, or nodules;
• Respiratory failure cannot be fully explained by heart failure and fluid overload;
• Hypoxemia, partial pressure of oxygen in arterial blood (PaO2)/oxygen fraction in air (FIO2) <150 mm Hg under PEEP =5 cm H2O, (mild ARDS: 200mmHg<PaO2/FiO2=300mmHg, moderate ARDS: 100mmHg<PaO2/FiO2=200mmHg, severe ARDS: PaO2/FiO2=100mmHg);
• 18~70 years old;
• Agree to participate in the trial, and sign the informed consent.

Exclusion Criteria:

• Age less than 18 years old;
• Time of hospital stay <24 h;
• Pregnancy;
• Using V-V ECOM;
• Cardiac index <1.5L·ml.min-1.m-2;
• Pulmonary resection;
• Pulmonary embolism ;
• Refused to participate in the study.

Related Conditions & MeSH terms

• Acute Lung Injury
• Acute Respiratory Distress Syndrome
• Inflammation
• Respiratory Distress Syndrome, Adult
• Respiratory Distress Syndrome, Newborn
• Syndrome

Intervention

Other:
• no intervention
no intervention

Locations

Country	Name	City	State
China	Department of Respiratory and Critical Care Medicine, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases	Beijing	Beijing

Sponsors (1)

Lead Sponsor	Collaborator
Beijing Anzhen Hospital

Country where clinical trial is conducted

China, 

References & Publications (19)

ARDS Definition Task Force, Ranieri VM, Rubenfeld GD, Thompson BT, Ferguson ND, Caldwell E, Fan E, Camporota L, Slutsky AS. Acute respiratory distress syndrome: the Berlin Definition. JAMA. 2012 Jun 20;307(23):2526-33. doi: 10.1001/jama.2012.5669. — View Citation

Ding L, Ni J, Yang F, Huang L, Deng H, Wu Y, Ding X, Tang J. Promising therapeutic role of miR-27b in tumor. Tumour Biol. 2017 Mar;39(3):1010428317691657. doi: 10.1177/1010428317691657. Review. — View Citation

Fan E, Brodie D, Slutsky AS. Acute Respiratory Distress Syndrome: Advances in Diagnosis and Treatment. JAMA. 2018 Feb 20;319(7):698-710. doi: 10.1001/jama.2017.21907. Review. — View Citation

Huang Y, Huang L, Zhu G, Pei Z, Zhang W. Downregulated microRNA-27b attenuates lipopolysaccharide-induced acute lung injury via activation of NF-E2-related factor 2 and inhibition of nuclear factor ?B signaling pathway. J Cell Physiol. 2019 May;234(5):6023-6032. doi: 10.1002/jcp.27187. Epub 2018 Dec 24. — View Citation

Li T, Cao H, Zhuang J, Wan J, Guan M, Yu B, Li X, Zhang W. Identification of miR-130a, miR-27b and miR-210 as serum biomarkers for atherosclerosis obliterans. Clin Chim Acta. 2011 Jan 14;412(1-2):66-70. doi: 10.1016/j.cca.2010.09.029. Epub 2010 Sep 29. — View Citation

Ling Y, Li ZZ, Zhang JF, Zheng XW, Lei ZQ, Chen RY, Feng JH. MicroRNA-494 inhibition alleviates acute lung injury through Nrf2 signaling pathway via NQO1 in sepsis-associated acute respiratory distress syndrome. Life Sci. 2018 Oct 1;210:1-8. doi: 10.1016/j.lfs.2018.08.037. Epub 2018 Aug 17. — View Citation

Liu Q, Lv H, Wen Z, Ci X, Peng L. Isoliquiritigenin Activates Nuclear Factor Erythroid-2 Related Factor 2 to Suppress the NOD-Like Receptor Protein 3 Inflammasome and Inhibits the NF-?B Pathway in Macrophages and in Acute Lung Injury. Front Immunol. 2017 Nov 9;8:1518. doi: 10.3389/fimmu.2017.01518. eCollection 2017. — View Citation

Liu Y, Song M, Zhu G, Xi X, Li K, Wu C, Huang L. Corynoline attenuates LPS-induced acute lung injury in mice by activating Nrf2. Int Immunopharmacol. 2017 Jul;48:96-101. doi: 10.1016/j.intimp.2017.04.029. Epub 2017 May 6. — View Citation

Lu MC, Ji JA, Jiang ZY, You QD. The Keap1-Nrf2-ARE Pathway As a Potential Preventive and Therapeutic Target: An Update. Med Res Rev. 2016 Sep;36(5):924-63. doi: 10.1002/med.21396. Epub 2016 May 18. Review. — View Citation

Rajasekaran S, Pattarayan D, Rajaguru P, Sudhakar Gandhi PS, Thimmulappa RK. MicroRNA Regulation of Acute Lung Injury and Acute Respiratory Distress Syndrome. J Cell Physiol. 2016 Oct;231(10):2097-106. doi: 10.1002/jcp.25316. Epub 2016 Feb 4. Review. — View Citation

Signorelli SS, Volsi GL, Pitruzzella A, Fiore V, Mangiafico M, Vanella L, Parenti R, Rizzo M, Volti GL. Circulating miR-130a, miR-27b, and miR-210 in Patients With Peripheral Artery Disease and Their Potential Relationship With Oxidative Stress. Angiology. 2016 Nov;67(10):945-950. doi: 10.1177/0003319716638242. Epub 2016 Jul 11. — View Citation

Song YX, Sun JX, Zhao JH, Yang YC, Shi JX, Wu ZH, Chen XW, Gao P, Miao ZF, Wang ZN. Non-coding RNAs participate in the regulatory network of CLDN4 via ceRNA mediated miRNA evasion. Nat Commun. 2017 Aug 18;8(1):289. doi: 10.1038/s41467-017-00304-1. Erratum in: Nat Commun. 2021 May 19;12(1):3149. — View Citation

Wang L, Xia JW, Ke ZP, Zhang BH. Blockade of NEAT1 represses inflammation response and lipid uptake via modulating miR-342-3p in human macrophages THP-1 cells. J Cell Physiol. 2019 Apr;234(4):5319-5326. doi: 10.1002/jcp.27340. Epub 2018 Sep 27. — View Citation

Wang Y, Xu Z, Jiang J, Xu C, Kang J, Xiao L, Wu M, Xiong J, Guo X, Liu H. Endogenous miRNA sponge lincRNA-RoR regulates Oct4, Nanog, and Sox2 in human embryonic stem cell self-renewal. Dev Cell. 2013 Apr 15;25(1):69-80. doi: 10.1016/j.devcel.2013.03.002. Epub 2013 Mar 28. — View Citation

Wei J, Chen G, Shi X, Zhou H, Liu M, Chen Y, Feng D, Zhang P, Wu L, Lv X. Nrf2 activation protects against intratracheal LPS induced mouse/murine acute respiratory distress syndrome by regulating macrophage polarization. Biochem Biophys Res Commun. 2018 Jun 7;500(3):790-796. doi: 10.1016/j.bbrc.2018.04.161. Epub 2018 Apr 25. — View Citation

Wu CT, Huang Y, Pei ZY, Xi X, Zhu GF. MicroRNA-326 aggravates acute lung injury in septic shock by mediating the NF-?B signaling pathway. Int J Biochem Cell Biol. 2018 Aug;101:1-11. doi: 10.1016/j.biocel.2018.04.019. Epub 2018 May 1. Erratum in: Int J Biochem Cell Biol. 2021 Mar;132:105922. — View Citation

Xiong DD, Li ZY, Liang L, He RQ, Ma FC, Luo DZ, Hu XH, Chen G. The LncRNA NEAT1 Accelerates Lung Adenocarcinoma Deterioration and Binds to Mir-193a-3p as a Competitive Endogenous RNA. Cell Physiol Biochem. 2018;48(3):905-918. doi: 10.1159/000491958. Epub 2018 Jul 23. Erratum in: Cell Physiol Biochem. 2020;54(3):508-509. — View Citation

Yan B, Yao J, Liu JY, Li XM, Wang XQ, Li YJ, Tao ZF, Song YC, Chen Q, Jiang Q. lncRNA-MIAT regulates microvascular dysfunction by functioning as a competing endogenous RNA. Circ Res. 2015 Mar 27;116(7):1143-56. doi: 10.1161/CIRCRESAHA.116.305510. Epub 2015 Jan 13. — View Citation

Zhou R, Gong AY, Eischeid AN, Chen XM. miR-27b targets KSRP to coordinate TLR4-mediated epithelial defense against Cryptosporidium parvum infection. PLoS Pathog. 2012;8(5):e1002702. doi: 10.1371/journal.ppat.1002702. Epub 2012 May 17. — View Citation

* Note: There are 19 references in all — Click here to view all references

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	The expression of lncRNA NEAT1 in blood and BALF in all groups	Use RT-PCR to measure the expression of lncRNA NEAT1 in blood and BALF in all groups	up to 24 day
Primary	The expression of miR-27b in blood and BALF in all groups	Use RT-PCR to measure the expression of miR-27b in blood and BALF in all groups	up to 3 day
Primary	The expression of Nrf2 in blood and BALF in all groups	Use RT-PCR and Wsetern blot to measure the expression of Nrf2 in blood and BALF in all groups	up to 3 day
Primary	The expression of inflammatory factors(IL-1ß?IL-6?IL-18?TNF-a) in blood and BALF in all groups	Use RT-PCR and ELISA to measure the expression of inflammatory factors(IL-1ß?IL-6?IL-18?TNF-a) in blood and BALF in all groups	up to 3 day
Primary	The expression of oxidative stress associated factors in blood and BALF in all groups	Use Western blot to measure the expression of oxidative stress associated factors(NLRP3?NF-?B-P65? p-P65?I?B?p-I?B?HO-1?NQO1?caspase-1) in blood and BALF in all groups	up to 3 day
Primary	The numbers and kinds of inflammatory cells in BALF and blood in all groups	Use flow cytometry to detect the number of inflammatory cells in BALF and blood in all groups	up to 3 day
Primary	The kinds of inflammatory cells in BALF and blood in all groups	Use flow cytometry to detect the kinds of inflammatory cells(neutrophile?macrophage? lymphocyte) in BALF and blood in all groups	up to 3 day
Primary	The time of mechanical ventilation of patients in ARDS groups	Record the time of mechanical ventilation of patients in ARDS groups	up to28 day
Primary	The severity of ARDS patients in ARDS groups	Record the severity(PaO2/FiO2?OI?S/F?OSI) of ARDS patients in ARDS groups	up to 28 day
Primary	the mortality in 28 days of ARDS patients	Record the mortality in 28 days of ARDS patients	up to 28 day
Secondary	The differences and correlation of the expressions of lncRNA NEAT1?miR-27b and Nrf2 in the groups	Analyse the differences of the expressions of lncRNA NEAT1?miR-27b and Nrf2 in the groups, and to explore the relations between the three(lncRNA NEAT1?miR-27b and Nrf2) in different groups.	up to 28 day
Secondary	The correlation of expressions of lncRNA NEAT1?miR-27b and Nrf2 with inflammation and oxidative stress in the groups.	To explore the correlation of expressions of lncRNA NEAT1?miR-27b and Nrf2 with inflammation and oxidative stress in the groups.	up to 28 day
Secondary	The relative of lncRNA NEAT1?miR-27b and Nrf2 with the time of mechanical ventilation, severity and mortality in 28 days of ARDS patients	To declare the relative of lncRNA NEAT1?miR-27b and Nrf2 with the time of mechanical ventilation, severity and mortality in 28 days of ARDS patients	up to 28 day