## Protocol Section
### Identification Module
**NCT ID:** NCT06427993

**Brief Title:** Urine DNA Methylation Detection for Hematuria Evaluation

**Official Title:** Urine Exfoliated Cell DNA Methylation Detection for Urothelial Carcinomas Diagnosis in Patients With Hematuria--A Prospective, Single-center, Cohort Study

#### Organization Study ID Info
**ID:** CH_Hematuria_Metest
#### Organization
**Class:** OTHER
**Full Name:** Changhai Hospital
### Status Module
#### Completion Date
**Date:** 2025-12-31

**Type:** ESTIMATED

#### Expanded Access Info
#### Last Update Post Date
**Date:** 2024-05-24

**Type:** ACTUAL

**Last Update Submit Date:** 2024-05-20

**Overall Status:** NOT_YET_RECRUITING

#### Primary Completion Date
**Date:** 2025-06-30

**Type:** ESTIMATED

#### Start Date
**Date:** 2024-06-15

**Type:** ESTIMATED

**Status Verified Date:** 2024-05

#### Study First Post Date
**Date:** 2024-05-24
**Type:** ACTUAL
**Study First Submit Date:** 2024-05-20
**Study First Submit QC Date:** 2024-05-20
### Sponsor Collaborators Module

#### Lead Sponsor

**Class:** OTHER

**Name:** Changhai Hospital

#### Responsible Party

**Type:** SPONSOR

### Oversight Module

**Is FDA Regulated Device:** False

**Is FDA Regulated Drug:** False

**Oversight Has DMC:** True

### Description Module

**Brief Summary:** Background: Hematuria, a common symptom of urinary system diseases, can result from various causes including infection, stones, trauma, and tumors. Urothelial carcinoma (UC), the most common malignancy of the urinary system, often presents with hematuria. Current diagnostic methods like urine cytology and cystoscopy have limitations in sensitivity and specificity, and cystoscopy is invasive. DNA methylation biomarkers offer potential for non-invasive UC detection, improving diagnostic accuracy in hematuria patients.

Objective: This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting UC in patients with hematuria.

Methods: This prospective pilot study will involve collecting preoperative urine samples from hematuria patients for DNA methylation testing using MSRE-qPCR. Sample size calculation was based on an assumed 25% prevalence of UC in hematuria patients, resulting in a total of 71 participants after accounting for a 20% dropout rate. Sensitivity, specificity, and diagnostic performance will be assessed using ROC curves.

Conclusion: This study seeks to validate the effectiveness of urine DNA methylation testing for UC detection in hematuria patients, providing a basis for its clinical application and informing the design of larger future studies.

**Detailed Description:** 1. Background Hematuria is a common symptom of urinary system diseases and can be caused by various factors, including infection, stones, trauma, and tumors. Urothelial carcinoma (UC) is the most common malignant tumor of the urinary system, primarily occurring in the bladder, renal pelvis, and ureter. The early symptoms of UC are often not obvious, with hematuria being the most common symptom. However, current diagnostic methods such as urine cytology and cystoscopy have limitations in sensitivity and specificity for diagnosing UC. Additionally, cystoscopy is invasive and can cause discomfort for patients. DNA methylation biomarkers have shown potential in detecting UC, providing a non-invasive method to improve diagnostic sensitivity and specificity, especially in patients with hematuria.
2. Objective This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria. By collecting preoperative urine samples from a small cohort of hematuria patients and performing DNA methylation testing, we aim to explore the feasibility and advantages of this method in clinical applications.
3. Methods

   Study Design:

   This is a prospective pilot study aimed at evaluating the effectiveness of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria.

   Sample Size Calculation:

   Based on our previous observations, the incidence of urothelial carcinoma in patients with hematuria is 20%-30%, slightly higher than reported in other literature. Therefore, we hypothesize that 25% of the patients in the hematuria cohort have UC. The group allocation ratio (R): N-/N+ = 75%/25% = 3.

   Assumptions:

   Area under the curve (AUC) under H0: 0.5 AUC under H1: 0.8 Power: 0.95 Significance level (Alpha): 0.05 Type of data: Continuous FPR range: 0.00 to 1.00

   Results: Using PASS 15.0 software for sample size calculation to ensure sufficient statistical power. The calculated sample size is:

   N+ (number of patients with UC): 14 N- (number of patients without UC): 42 Total sample size (N): 56

   Considering a 20% dropout rate, the adjusted sample size is:

   N+': 18 N-': 53 Total sample size (N'): 71

   Inclusion and Exclusion Criteria:

   Inclusion Criteria:

   Aged between 18 and 99 years, with gross or microscopic hematuria (\>5/HP). Able to provide 50ml urine for testing before surgery. Consent to participate in the study and sign the informed consent form.

   Exclusion Criteria:

   With history of malignancy or concomitant malignancies other than UC. Severe urinary tract infection leading to sepsis. Patients with indwelling catheters, nephrostomy, or cystostomy. Severe liver or kidney failure or other conditions deemed unsuitable for the study.

   Patients who did not undergo surgical treatment for various reasons. Samples with insufficient DNA content or other quality control failures.

   Sample Collection:

   Clinicians will collect fresh urine samples from enrolled hematuria patients and record their basic information, clinical information, and medical history.

   Samples will be randomly numbered and provided to DNA methylation testing personnel to ensure blinding.

   Testing Method:

   Testing personnel will analyze urine samples using MSRE-qPCR based DNA methylation testing methods to evaluate DNA methylation levels.

   Unblinding and Data Organization:

   After the last sample is successfully enrolled and tested, non-recruiting personnel and testing personnel will unblind the samples and organize clinical and pathological information.

   Data Analysis:

   Using pathological results as the gold standard. Calculate the sensitivity, specificity, positive predictive value, and negative predictive value of DNA methylation biomarkers.

   Use statistical methods (such as ROC curves) to evaluate the diagnostic performance of DNA methylation biomarkers and calculate the AUROC.

   Ethics and Informed Consent:

   This study has been approved by the hospital's ethics committee, and all participants must sign an informed consent form.
4. Conclusion This study aims to validate the effectiveness of urine DNA methylation testing in detecting urothelial carcinoma in patients with hematuria and provide evidence for its clinical application. The results of this preliminary study will offer essential data support and design optimization suggestions for future larger-scale studies.

### Conditions Module

**Conditions:**

- Hematuria

- Urothelial Carcinoma

- Bladder Cancer

- Ureter Cancer

- Renal Pelvis Cancer

**Keywords:**

- Hematuria

- Urothelial Carcinoma

- Urine DNA Methylation

### Design Module

#### Bio Spec

**Description:** Urine sediment DNA.

**Retention:** SAMPLES_WITH_DNA

#### Design Info

**Observational Model:** COHORT

**Time Perspective:** PROSPECTIVE

#### Enrollment Info

**Count:** 71

**Type:** ESTIMATED

**Study Type:** OBSERVATIONAL

### Outcomes Module

#### Primary Outcomes

**Description:** Evaluate the diagnostic performance of DNA methylation biomarkers by calculating the AUROC value.

**Measure:** Diagnostic performance of DNA methylation test

**Time Frame:** 1 year

#### Secondary Outcomes

**Description:** Calculate the sensitivity, specificity, positive predictive value, and negative predictive value of DNA methylation biomarkers.

**Measure:** Sensitivity, specificity, positive predictive value, and negative predictive value of DNA methylation biomarkers

**Time Frame:** 1 year

### Eligibility Module

**Eligibility Criteria:** Inclusion Criteria:

1. Aged between 18 and 99 years, with gross or microscopic hematuria (\>5/HP).
2. Able to provide 50ml urine for testing before surgery.
3. Consent to participate in the study and sign the informed consent form.

Exclusion Criteria:

1. With history of malignancy or concomitant malignancies other than UC.
2. Severe urinary tract infection leading to sepsis.
3. Patients with indwelling catheters, nephrostomy, or cystostomy. 4 Severe liver or kidney failure or other conditions deemed unsuitable for the study.

5. Patients who did not undergo surgical treatment for various reasons. 6. Samples with insufficient DNA content or other quality control failures.

**Maximum Age:** 99 Years

**Minimum Age:** 18 Years

**Sampling Method:** PROBABILITY_SAMPLE

**Sex:** ALL

**Standard Ages:**

- ADULT

- OLDER_ADULT

**Study Population:** This is a prospective pilot study aimed at evaluating the effectiveness of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria.

### Contacts Locations Module

#### Locations

**Location 1:**

**City:** Shanghai

**Contacts:**

***Contact 1:***

- **Email:** chenqing1002@163.com

- **Name:** Qing Chen, Dr

- **Phone:** +86 18301921727

- **Role:** CONTACT

***Contact 2:***

- **Name:** Chuanliang Xu, Dr

- **Role:** PRINCIPAL_INVESTIGATOR

**Country:** China

**Facility:** Changhai Hospital

**State:** Shanghai

## Derived Section

### Condition Browse Module - Ancestors

- ID: D000009375

- Term: Neoplasms, Glandular and Epithelial

- ID: D000009370

- Term: Neoplasms by Histologic Type

- ID: D000009369

- Term: Neoplasms

- ID: D000014571

- Term: Urologic Neoplasms

- ID: D000014565

- Term: Urogenital Neoplasms

- ID: D000009371

- Term: Neoplasms by Site

- ID: D000052776

- Term: Female Urogenital Diseases

- ID: D000005261

- Term: Female Urogenital Diseases and Pregnancy Complications

- ID: D000091642

- Term: Urogenital Diseases

- ID: D000014570

- Term: Urologic Diseases

- ID: D000052801

- Term: Male Urogenital Diseases

- ID: D000014555

- Term: Urination Disorders

- ID: D000006470

- Term: Hemorrhage

- ID: D000010335

- Term: Pathologic Processes

- ID: D000014515

- Term: Ureteral Diseases

### Condition Browse Module - Browse Branches

- Abbrev: BC04

- Name: Neoplasms

- Abbrev: All

- Name: All Conditions

- Abbrev: BXS

- Name: Urinary Tract, Sexual Organs, and Pregnancy Conditions

- Abbrev: BC23

- Name: Symptoms and General Pathology

- Abbrev: Rare

- Name: Rare Diseases

### Condition Browse Module - Browse Leaves

- ID: M5534

- Name: Carcinoma

- Relevance: HIGH

- As Found: Carcinoma

- ID: M5030

- Name: Urinary Bladder Neoplasms

- Relevance: LOW

- As Found: Unknown

- ID: M5551

- Name: Carcinoma, Transitional Cell

- Relevance: HIGH

- As Found: Urothelial Carcinoma

- ID: M9504

- Name: Hematuria

- Relevance: HIGH

- As Found: Hematuria

- ID: M17266

- Name: Ureteral Neoplasms

- Relevance: HIGH

- As Found: Ureter Cancer

- ID: M13297

- Name: Pelvic Neoplasms

- Relevance: HIGH

- As Found: Pelvis Cancer

- ID: M12320

- Name: Neoplasms, Glandular and Epithelial

- Relevance: LOW

- As Found: Unknown

- ID: M12315

- Name: Neoplasms by Histologic Type

- Relevance: LOW

- As Found: Unknown

- ID: M17320

- Name: Urologic Neoplasms

- Relevance: LOW

- As Found: Unknown

- ID: M17315

- Name: Urogenital Neoplasms

- Relevance: LOW

- As Found: Unknown

- ID: M2875

- Name: Urogenital Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M27093

- Name: Female Urogenital Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M14127

- Name: Pregnancy Complications

- Relevance: LOW

- As Found: Unknown

- ID: M8399

- Name: Female Urogenital Diseases and Pregnancy Complications

- Relevance: LOW

- As Found: Unknown

- ID: M17319

- Name: Urologic Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M27095

- Name: Male Urogenital Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M17305

- Name: Urination Disorders

- Relevance: LOW

- As Found: Unknown

- ID: M9556

- Name: Hemorrhage

- Relevance: LOW

- As Found: Unknown

- ID: M17265

- Name: Ureteral Diseases

- Relevance: LOW

- As Found: Unknown

- ID: T5693

- Name: Transitional Cell Carcinoma

- Relevance: HIGH

- As Found: Urothelial Carcinoma

### Condition Browse Module - Meshes

- ID: D000002277

- Term: Carcinoma

- ID: D000002295

- Term: Carcinoma, Transitional Cell

- ID: D000014516

- Term: Ureteral Neoplasms

- ID: D000010386

- Term: Pelvic Neoplasms

- ID: D000006417

- Term: Hematuria

### Misc Info Module

- Version Holder: 2024-05-31
