Evaluation of Pre-Analytical Factors of Urine Samples for Urine Cancer Cell Cultures (UCCC) --A Non-Invasive Biomarker – in Monitoring Response and Recurrence of Bladder Cancer

尿癌细胞培养 (UCCC) 尿液样本分析前因素的评估——一种非侵入性生物标志物 — 用于监测膀胱癌的反​​应和复发

• 批准号: 10640606
• 负责人: Xuefeng Liu
• 金额: $ 36.03万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-06 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640606
• 关键词: Address; Affect; Antibiotics; Antibody-drug conjugates; Biological Markers; Bladder; Cancer Center; Cancer Detection; Cancer Patient; Cell Count; Cell Culture System; Cell Culture Techniques; Cell Survival; Cell model; Cells; Clinic; Clinical; Clinical Treatment; Clinical Trials; Coculture Techniques; Collection; Conditioned Culture Media; Cystoscopy; Data; Development; Diagnosis; Early Diagnosis; Epithelial Cell Proliferation; European; Evaluation; FDA approved; Freezing; Funding; Future; General Anesthesia; Goals; Health Personnel; Hypoxia; Immunotherapy; Intravesical Instillation; Laboratories; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Medical; Methods; Modeling; Monitor; Monitoring for Recurrence; Needle biopsy procedure; Non-Invasive Detection; Operative Surgical Procedures; Patients; Persons; Pharmacotherapy; Procedures; Prognosis; Proteins; Protocols documentation; Recurrence; Recurrent Malignant Neoplasm; Recurrent tumor; Relapse; Sampling; Specimen; Specimen Handling; Standardization; System; Technology; Temperature; Testing; Therapeutic Agents; Time; Tissues; Transportation; Transurethral Resection; Treatment outcome; Urine; Urology; Variant; Work; X-Ray Computed Tomography; cancer cell; cancer recurrence; cancer type; chemotherapy; clinical application; clinical assay development; clinically relevant; contextual factors; cost; drinking water; drug sensitivity; evidence base; high risk; improved; invention; liquid biopsy; member; mortality; patient response; predicting response; protein kinase inhibitor; response; rho; sample collection; success; targeted treatment; tumor progression

PROJECT SUMMARY Bladder cancer is one of the top 10 most frequent cancers, yet the most underfunded cancer by NCI based on funding dollars and ratio of funding/mortality. There has been no significant improvement in overall survival and prognosis over the last thirty years except for the recent development of immunotherapy. After initial diagnosis and treatment, over 60% of the BC patients will relapse within two years and progression into advanced stages in up to 25% of patients. Therefore, almost all patients will need long-term expensive cystoscopy which makes bladder cancer the costliest cancer (per case) of all cancer types. If a sensitive but less expensive method to detect cancer were available, it would improve the treatment outcomes and decrease the cost. Dr. Xuefeng Liu (PI) and his team recently invented a robust method of conditional reprogramming (CR) technology to establish patient-derived cell models from cystoscopy or needle biopsies and urine samples of BC patients. The overall success rate was near 100%. These urine cancer cell cultures (UCCC) provide a simple, non-invasive, comfortable, reliable method to detect BC recurrence, and represent a living biomarker to predict patients’ response to chemotherapy and targeted therapy. Several cancer centers and laboratories including PI’s lab are actively involved in clinical trials to validate clinical utility of the UCCC in BC clinics. Since a variety of patient- context and collection, storage and transportation of urine samples to laboratories for cell cultures affect enrichment and viability of cancer cells in urine samples, it is urgent needed to evaluate pre-analytical factors of urine samples for UCCC and optimize these non-Invasive and living liquid biopsies as a routine approach in monitoring recurrence and prediction of response of BC patients. In this application, we propose to address the gaps by extending the work done by our team members on evaluating effects of sample processing protocols on UCCC analysis, to further investigate effect of patient-specific context. In this application, we will first further optimization of UCCC system to establish a culture kit for clinical applications, specifically we will optimize and simplify conditions with co-culture kits, conditioned medium (CM), hypoxic condition for maximal efficiency and robustness. Second, we will establish a Standard of Procedure - SOP- for urine collection and transportation in clinics: we will test pH, temperatures, minimal cell number, a cocktail with anti-biotics and Y-27632 affect UCCC cultures and success rate. Last, we will determine patient-context factors that affect UCCC cultures: we will evaluate how urine timing (days before surgery, and after surgery, early morning, noon, night, drinking water before urine collection, etc), urine volume affect cell viability and UCCC. Last, we will optimize a self- sampling procedures for BC patients. The overall goal is to expedite UCCC clinical assay development through evidence-based standardization of urine collection and handling practices, potentially approved by FDA as a standard method for bladder clinics.

项目摘要 膀胱癌是十大最常见的癌症之一，但基于NCI资金最多的癌症 资金美元和资金/死亡率的比率。总体生存没有显着改善， 在过去三十年中的预后，除了最近的免疫疗法发展。初始诊断后 和治疗，超过60％的卑诗省患者将在两年内中继并发展为高级阶段 多达25％的患者。因此，几乎所有患者都需要长期昂贵的膀胱镜检查，这使得 膀胱癌是所有癌症类型的最昂贵的癌症（每病例）。如果是一种敏感但较便宜的方法 可检测癌症，它将改善治疗结果并降低成本。 Xuefeng Liu博士 （PI）和他的团队最近发明了一种有条件重编程（CR）技术的强大方法来建立 来自膀胱镜检查或针头活检的患者衍生细胞模型以及卑诗省患者的尿液样本。总体 成功率接近100％。这些尿液癌细胞培养物（UCCC）提供了简单的，无创的， 舒适，可靠的方法检测BC复发，并代表生物标志物来预测患者 对化学疗法和靶向治疗的反应。包括Pi的实验室在内的几个癌症中心和实验室 积极参与临床试验，以验证BC诊所中UCCC的临床实用性。由于多种患者 - 将尿液样本到实验室进行细胞培养的背景和收集，储存和运输影响 癌细胞在尿液样品中的富集和生存力，迫切需要评估分析前因子 用于UCCC的尿液样本，并优化这些非侵入性和活的液体活检作为常规方法 监测卑诗省患者反应的复发和预测。在此应用程序中，我们建议解决 通过扩展我们的团队成员在评估样本处理协议效果方面完成的工作来差距 关于UCCC分析，以进一步研究患者特定环境的影响。在此应用程序中，我们将首先进一步 优化UCCC系统以建立用于临床应用的文化套件，特别是我们将优化和 用共培养试剂盒，条件培养基（CM），低氧条件来简化条件，以最大程度的效率和 鲁棒性。其次，我们将建立一个程序标准-SOP-用于尿液收集和运输 诊所：我们将测试pH值，温度，最小细胞数，抗生素和Y-27632的鸡尾酒 UCCC文化和成功率。最后，我们将确定影响UCCC培养物的患者可能性因素：我们 将评估尿液计时（手术前几天，手术后，清晨，中午，晚上，喝酒 尿液收集前的水等），尿量会影响细胞活力和UCCC。最后，我们将优化一个自我 卑诗省患者的抽样程序。总体目标是通过 基于证据的尿液收集和处理方法的标准化，有可能被FDA批准为 膀胱诊所的标准方法。