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

Sponsor

Beijing Anzhen Hospital (Other)

Overall Status

Enrolling by invitation

CT.gov ID

NCT04937855

Collaborator

(none)

425

Enrollment

Location

Anticipated Duration (Months)

14.2

Patients Per Site Per Month

Study Details

Study Description

Brief 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.

Condition or Disease	Intervention/Treatment	Phase
Acute Respiratory Distress Syndrome Inflammation	Other: no intervention

Detailed 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.

Study Design

Study Type:

Observational

Anticipated Enrollment :

425 participants

Observational Model:

Other

Time Perspective:

Prospective

Official Title:

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

Anticipated Study Start Date :

Jul 1, 2021

Anticipated Primary Completion Date :

Dec 31, 2022

Anticipated Study Completion Date :

Dec 31, 2023

Arms and Interventions

Arm	Intervention/Treatment
Control group 25 gender and age matching healthy controls	Other: no intervention no intervention
ARDS group 1 100 ARDS patients at the time of check in hospital	Other: no intervention no intervention
ARDS group 2 100 ARDS patients at the time of 24h after check in hospital	Other: no intervention no intervention
ARDS group 3 100 ARDS patients at the time of 48h after check in hospital	Other: no intervention no intervention
ARDS group 4 100 ARDS patients at the time of 72h after check in hospital	Other: no intervention no intervention

Outcome Measures

Primary Outcome Measures

The expression of lncRNA NEAT1 in blood and BALF in all groups [up to 24 day]
Use RT-PCR to measure the expression of lncRNA NEAT1 in blood and BALF in all groups
The expression of miR-27b in blood and BALF in all groups [up to 3 day]
Use RT-PCR to measure the expression of miR-27b in blood and BALF in all groups
The expression of Nrf2 in blood and BALF in all groups [up to 3 day]
Use RT-PCR and Wsetern blot to measure the expression of Nrf2 in blood and BALF in all groups
The expression of inflammatory factors(IL-1β、IL-6、IL-18、TNF-α) in blood and BALF in all groups [up to 3 day]
Use RT-PCR and ELISA to measure the expression of inflammatory factors(IL-1β、IL-6、IL-18、TNF-α) in blood and BALF in all groups
The expression of oxidative stress associated factors in blood and BALF in all groups [up to 3 day]
Use Western blot to measure the expression of oxidative stress associated factors(NLRP3、NF-κB-P65、 p-P65、IκB、p-IκB、HO-1、NQO1、caspase-1) in blood and BALF in all groups
The numbers and kinds of inflammatory cells in BALF and blood in all groups [up to 3 day]
Use flow cytometry to detect the number of inflammatory cells in BALF and blood in all groups
The kinds of inflammatory cells in BALF and blood in all groups [up to 3 day]
Use flow cytometry to detect the kinds of inflammatory cells(neutrophile、macrophage、 lymphocyte) in BALF and blood in all groups
The time of mechanical ventilation of patients in ARDS groups [up to28 day]
Record the time of mechanical ventilation of patients in ARDS groups
The severity of ARDS patients in ARDS groups [up to 28 day]
Record the severity(PaO2/FiO2、OI、S/F、OSI) of ARDS patients in ARDS groups
the mortality in 28 days of ARDS patients [up to 28 day]
Record the mortality in 28 days of ARDS patients

Secondary Outcome Measures

The differences and correlation of the expressions of lncRNA NEAT1、miR-27b and Nrf2 in the groups [up to 28 day]
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.
The correlation of expressions of lncRNA NEAT1、miR-27b and Nrf2 with inflammation and oxidative stress in the groups. [up to 28 day]
To explore the correlation of expressions of lncRNA NEAT1、miR-27b and Nrf2 with inflammation and oxidative stress in the groups.
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]
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

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years to 70 Years

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Yes

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.

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Department of Respiratory and Critical Care Medicine, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases	Beijing	Beijing	China	100029