Expression Analysis of Urinary Exosome in Type 2 Diabetic Nephropathy and Evaluation of Its Clinical Diagnostic Value

Expression Analysis of Urinary Exosome miR-142-3p in Type 2 Diabetic Nephropathy and Evaluation of Its Clinical Diagnostic Value

Expression analysis of urinary exosome miR-142-3p in type 2 diabetic nephropathy and evaluation of its clinical diagnostic value

Study Overview

• Status: Active, not recruiting
• Conditions: Diabetic Nephropathy
• Intervention / Treatment: Other: Grouping based on previous laboratory results, no intervention.

Detailed Description

1. Preliminary screening of miRNAs: Through GEO database and reading related literature, miRNAs differentially expressed in type 2 diabetes and diabetic nephropathy were selected as potential candidate biomarkers for follow-up verification.
2. Collection and treatment of clinical samples: Urine of diabetic patients was collected from the First Affiliated Hospital of Shandong First Medical University in strict accordance with the drainage standard, and the basic information of patients was registered. Meanwhile, the collected samples were divided into type 2 diabetes group and diabetic nephropathy group according to whether the urine was accompanied by UACR≥30/g. Subsequently, the collected samples were centrifuged and retained for supernatant.
3. Isolation and identification of urinary exosomes: Ultrafast centrifugation method was used to separate the treated urine samples of exosomes, and then identified by transmission electron microscopy, nanoparticle analysis and Western blot respectively.
4. Real-time PCR was used to detect the changes in urinary exosomal miRNA expression levels of the two groups of patients, and statistical analysis and correlation analysis of clinical indicators of the differentially expressed mirnas were performed.

Target gene prediction and enrichment pathway analysis of differentially expressed mirnas were performed to screen out pathways that might be related to the occurrence and development of DKD, and then the relevant pathways were verified by immunofluorescence, immunocoprecipitate, Western blot, Real-time PCR and other methods.

• Study Type: Observational
• Enrollment (Estimated): 44

Contacts and Locations

Study Locations

• China
  • Shandong
    • Jinan, Shandong, China, 250000
      • Qianfoshan Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

• Sampling Method: Non-Probability Sample

Study Population

Patients with a clinically confirmed diagnosis of type 2 diabetes during hospitalization in the Department of nephrology and Endocrinology from December 2020 to June 2022.

Description

Inclusion Criteria:

1. Age ≥18 years old;
2. Previously diagnosed with type 2 diabetes;
3. Typical diabetic symptoms (polydipsia, polydipsia, polyuria and weight loss) plus random blood glucose ≥11.1mmol/l;
4. Fasting blood glucose ≥7.0mmol/l, fasting is defined as at least 8 hours without intake of calories;
5. Oral glucose tolerance test (OGTT) 2-hour blood glucose ≥11.1mmol/l, using a glucose load equivalent to 75 grams of anhydrous glucose dissolved in water (venous blood was drawn in all patients;
6. With or without UACR≥30mg/g.

Exclusion Criteria:

(1) Refused to enroll patients; （2）Type 1 diabetic nephropathy and other special types of diabetic nephropathy; （3）Patients with kidney stones and urinary system infection; （4）Patients with autoimmune system diseases, malignant tumors, and blood system diseases; （5）Patients with severe chronic cardiopulmonary disease, chronic liver and kidney disease; （6）Patients with severe infectious diseases; （7）Use of glucocorticoids, immunosuppressants or cytotoxic drugs.

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Type 2 diabetes group; The group was based on the clinically confirmed diagnosis of type 2 diabetes	Other: Grouping based on previous laboratory results, no intervention.; The patients were grouped according to the results of previous inpatient tests, and no intervention measures were taken
Diabetic nephropathy group; The group was based on the clinically confirmed diagnosis of type 2 diabetes.Patients were re-screened strictly according to the following admission criteria: persistent albuminuria and/or decreased eGFR, while other causes of chronic kidney disease (CKD) were excluded. When diabetes mellitus is identified as the cause of kidney damage and other primary and secondary glomerular diseases and systemic diseases are excluded, at least one of the following conditions can be diagnosed as DKD: (1) Urinary Albumin/Creatinine Ratio (UACR)≥30 mg/g or Urinary albumin excretion rate (UAER)≥30 mg/24 h, The UACR or UAER was checked again within 3 to 6 months, and 2 out of 3 times reached or exceeded the critical value; Eliminate other interfering factors such as infection; (2) eGFR< 60 ml·min-1· (1.73 m2) -1 for more than 3 months; (3) Renal biopsy was consistent with DKD pathological changes	Other: Grouping based on previous laboratory results, no intervention.; The patients were grouped according to the results of previous inpatient tests, and no intervention measures were taken

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Preliminary screening of miRNA	Through GEO database and reading related literature, mirnas that are differentially expressed in type 2 diabetes and diabetic nephropathy were selected as potential candidate biomarkers for follow-up verification.	2020-2024
Clinical sample collection and processing	(2) The urine of diabetic patients was collected from the First Affiliated Hospital of Shandong First Medical University in strict accordance with the drainage standard, and the basic information of patients was registered. Meanwhile, the collected samples were divided into type 2 diabetes group and diabetic nephropathy group according to whether the urine was accompanied by UACR≥30/g. Subsequently, the collected samples were centrifuged and retained for supernatant.	2020-2024
Statistical analysis and target gene prediction of misexpressed mirnas and enrichment pathway analysis	Statistical analysis and correlation analysis of clinical indicators of differentially expressed mirnas were performed to further evaluate their clinical diagnostic value. Subsequently, the target genes of the mirnas differentially expressed in urinary exosomes of the two groups were analyzed by miRTarBase database and R software package, and the relevant pathways were screened out through KEGG enrichment pathway analysis. Subsequently, the relevant pathways were verified by immunofluorescence, immunocoprecipitation, Western blot, Real-time PCR and other methods.	2020-2024

Collaborators and Investigators

• Sponsor: Yipeng Liu
• Investigators: Study Chair: Yipeng Liu, Qianfoshan Hospital

Study record dates

Study Major Dates

• Study Start (Actual): June 1, 2020
• Primary Completion (Estimated): December 30, 2023
• Study Completion (Estimated): January 31, 2024

Study Registration Dates

• First Submitted: October 18, 2023
• First Submitted That Met QC Criteria: October 18, 2023
• First Posted (Actual): October 24, 2023

Study Record Updates

• Last Update Posted (Actual): October 26, 2023
• Last Update Submitted That Met QC Criteria: October 24, 2023
• Last Verified: October 1, 2023

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Urologic Diseases; Endocrine System Diseases; Diabetes Complications; Diabetes Mellitus; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Male Urogenital Diseases; Kidney Diseases; Diabetic Nephropathies
• Other Study ID Numbers: YXLL-KY-2023(106)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No