Comprehensive Analysis of the Key Mutation Spectrum in Bladder Cancer: Establishment and Clinical Validation of a Multiplex Mutation Detection System Based on Nucleic Acid Mass Spectrometry
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 400
- 试验地点
- 1
- 主要终点
- Survival Differences Between Mutated and Non-mutated Groups in Bladder Cancer Patients
研究概览
简要总结
Bladder cancer is a highly heterogeneous malignancy characterized by frequent genetic alterations that are closely associated with disease progression, recurrence risk, and treatment response. However, existing mutation detection approaches are often limited by high cost, complex workflows, or insufficient capacity for multiplex and low-frequency mutation analysis, which restricts their routine clinical application. The purpose of this study is to establish and clinically validate a multiplex mutation detection system for bladder cancer based on nucleic acid mass spectrometry. Using fresh tumor tissue and matched adjacent normal tissue samples collected from patients with bladder cancer, a targeted mutation panel comprising key functional mutations with demonstrated clinical relevance will be constructed. The matched normal tissues serve as germline references to enable accurate identification of somatic mutations. The analytical performance of the system, including sensitivity, specificity, and concordance with whole-genome sequencing, will be systematically evaluated. In addition, the clinical utility of the mutation panel in risk stratification and treatment decision support will be explored by comparing its predictive value with established clinical models and guideline-recommended tools. The ultimate goal is to develop a cost-effective, reproducible, and clinically applicable molecular testing strategy that can support precision diagnosis and individualized management of patients with bladder cancer.
详细描述
Bladder cancer is a highly heterogeneous disease with complex genetic mutations that influence tumor behavior, treatment response, and patient outcomes. Current genetic testing methods often face limitations in simultaneously detecting multiple mutations with high sensitivity and low cost. This study aims to develop and clinically validate a novel multiplex mutation detection system for bladder cancer based on nucleic acid mass spectrometry. The study consists of two phases. In the first phase, a standardized detection panel targeting key bladder cancer-related genes and functional mutation sites will be established, selected based on mutation frequency, clinical significance, survival impact, and evidence from authoritative databases such as The Cancer Genome Atlas (TCGA), OncoKB, and ClinVar. The panel covers critical genes, including Fibroblast Growth Factor Receptor 3 (FGFR3), Tumor Protein P53 (TP53), and others involved in tumor progression, therapeutic response, and prognosis. In the second phase, the clinical utility of this system will be validated using 400 freshly collected bladder cancer tissue samples and paired adjacent normal tissue samples. This detection system offers several advantages: 1. High-throughput multiplexing - simultaneous detection of up to 30 mutation sites in a single run; 2. High sensitivity - capable of detecting low-frequency mutations (as low as 0.1% variant allele frequency); 3. Quantitative analysis - provides allele frequency information to assess tumor burden and monitor treatment response; 4. Cost-effectiveness and simplicity - lower cost and simpler workflow compared to next-generation sequencing, making it suitable for clinical implementation. The clinical value of this system will be rigorously evaluated by: 1. Comparing its risk stratification performance with established clinical tools, such as the European Organisation for Research and Treatment of Cancer (EORTC), European Association of Urology (EAU), and Vesical Imaging-Reporting and Data System (VI-RADS); 2. Assessing its treatment predictive value against current standards, such as the Spanish Bladder Cancer Group (CUETO) and immunohistochemical markers; 3. Validating its accuracy against whole-exome sequencing as the gold standard in paired samples of tumor and adjacent normal tissues. By providing a comprehensive, affordable, and clinically actionable mutation profiling tool, this study aims to improve precision risk stratification, guide individualized treatment decisions, and enable dynamic recurrence monitoring for bladder cancer patients. The ultimate goal is to establish a standardized molecular diagnostic framework that can be integrated into routine clinical practice.
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Histologically confirmed diagnosis of urothelial carcinoma of the bladder (any stage, including non-muscle invasive and muscle invasive).
- •Availability of sufficient tumor tissue specimen (fresh frozen) for DNA extraction and mutation analysis.
- •Age ≥ 18 years at time of diagnosis.
排除标准
- •History of other malignant tumors within the past 5 years, except adequately treated non-melanoma skin cancer or carcinoma in situ of the cervix.
- •Inadequate quality or quantity of tumor tissue DNA for mutation panel analysis (e.g., severe DNA degradation, insufficient DNA yield).
- •Pregnancy or breastfeeding.
- •Serious uncontrolled intercurrent illness that would interfere with study follow-up or compliance, including but not limited to ongoing or active infection, symptomatic congestive heart failure, unstable angina pectoris, cardiac arrhythmia, or psychiatric illness/social situations that would limit compliance with study requirements.
研究组 & 干预措施
NMIBC (Non-Muscle Invasive Bladder Cancer) Study Group
This cohort includes patients diagnosed with non-muscle invasive bladder cancer. Participants are categorized according to post-operative recurrence and progression status, as well as response to intravesical therapy (recurrence versus no recurrence; response versus non-response). Survival differences between mutation-positive and mutation-negative groups will be assessed using Kaplan-Meier survival analysis. Predictive models for post-surgical recurrence, progression, and treatment response will be developed using multivariable Cox proportional hazards regression analysis. Model performance will be validated in 30% of participants and compared with established risk models from the European Association of Urology and the European Organisation for Research and Treatment of Cancer.
干预措施: Multiplex Mutation Detection System for Bladder Cancer (Nucleic Acid Mass Spectrometry) (Genetic)
MIBC (Muscle Invasive Bladder Cancer) Study Group
This cohort includes patients diagnosed with muscle invasive bladder cancer. Participants are categorized according to recurrence after adjuvant therapy and the presence or absence of distant metastasis. Survival differences between mutation-positive and mutation-negative groups will be assessed using Kaplan-Meier survival analysis. A predictive model for recurrence and distant metastasis following adjuvant therapy will be developed using multivariable Cox proportional hazards regression analysis. Model performance will be validated in the remaining 30% of participants. Predictive accuracy will be compared with existing clinical assessment methods, including clinicopathological characteristics, single-gene mutation markers, and immunohistochemical biomarkers.
干预措施: Multiplex Mutation Detection System for Bladder Cancer (Nucleic Acid Mass Spectrometry) (Genetic)
结局指标
主要结局
Survival Differences Between Mutated and Non-mutated Groups in Bladder Cancer Patients
时间窗: Survival will be assessed at post-surgical follow-up at 6 months, 1 year, 2 years, and 3 years, including recurrence, progression, and metastasis-free survival events over a 3-year period.
This outcome measure aims to compare the survival rates between bladder cancer patients with mutations in key bladder cancer-related genes (as determined by the multiplex mutation detection panel) and those without mutations. The mutation status (any gene mutation versus no mutation) will be correlated with clinical outcomes, including recurrence-free survival (RFS), progression-free survival (PFS), and overall survival (OS), using Kaplan-Meier survival analysis. These survival metrics will be assessed to determine whether mutation status influences prognosis and to identify any significant survival differences between mutated and non-mutated groups.
次要结局
- Development of Predictive Models for Post-surgical Recurrence, Progression, and Response to Intravesical Therapy(The predictive model will be developed and evaluated during the 3-year follow-up period post-surgery, with data collected at key intervals: 6 months, 1 year, 2 year, and 3 years post-surgery.)
- Validation of Mutation Panel's Predictive Value in Risk Stratification Using Existing Clinical Models(Validation will occur after 3 years of patient follow-up, at the point of comparing the prediction models for their efficacy in risk stratification and recurrence prediction.)
研究者
Zhilong Dong
Professor of Urology
Lanzhou University Second Hospital