High throughput nanoplasmonic exosome testing (NEXT) of immunotherapies in bladder cancer

挑战一旦发生转移，100名患者中只有5名存活5年。免疫检查点 抑制剂已被证明增加临床牵引力，但常规成像，如CT扫描， 难以准确评估这种治疗背景下的肿瘤反应。可连续访问的 肿瘤和宿主生物标志物可以增加对反应的早期了解， 做出精准免疫治疗的决策。创新：根据竞争前所需 用于EV调查的纳米技术工具，以填补时间和科学空白， 基于免疫疗法的治疗监测，我们的小组开发和验证了磁， 电化学平台，无需EV纯化，可在内部实现96个并行读数 45分钟.该建议利用膀胱在泌尿生殖系统内的私密解剖位置 因此，尿液将为我们提供丰富的膀胱癌EV储存库。我们之前 证明了尿EV检测在肾移植排斥反应中的可行性。通过增加兴奋 通过在金纳米粒子中诱导等离子体共振产生的电荷，我们最近加速了 在我们最新和可扩展的平台内进行电化学反应，以实现12倍信号 EV表面标记物增加。我们假设，推进我们的纳米等离子体EV传感器， 人尿和优化测定方案以测量EV内和表面标志物，可以鉴定 高价值膀胱癌和宿主生物标志物（蛋白质和mRNA），以更好地检查它们的相互作用 随着时间的推移和治疗压力下。我们提出了三个具体目标：目标1：优化我们的 用于高通量尿基分析的纳米等离子体传感器（NEXT）测定和仪器 以及表面和EV内标志物的全面分析。目的2：采用临床前和 用于NEXT分析的生物样本库，以检查性能并提供最佳信息 生物标志物组。目的3：使用NEXT前瞻性监测和描述患者的尿液EV 正在接受基于免疫疗法的膀胱癌治疗。影响：我们的高度互补性 一组有成就的研究人员带来了长期的专业知识和翻译经验 EV生物学、生物工程、系统生物学、生物信息学和临床肿瘤学。如果成功，我们 尿纳米等离子体EV平台将为基于免疫治疗的 晚期膀胱癌的治疗，在其他流行的泌尿生殖系统癌症， 肾脏和前列腺。

Challenges. Once metastatic, only 5 out of 100 patients are alive at five years. Immune checkpoint inhibitors have demonstrated increasing clinical traction yet conventional imaging such as CT scans struggle to accurately assess tumor response in this treatment context. Serially accessible sources of tumor and host biomarkers could add earlier insights into response this informing timely go / no-go decision making to render precision immunotherapy. Innovation: In light of needed pre-competitive nanotechnology tools for EV investigation to fill temporal and scientific gaps precluding accurate immunotherapy-based treatment monitoring, our group developed and validated a magneto- electrochemical platform without need for EV purification and capable of 96 parallel readouts within 45 minutes. This proposal exploits the bladder's intimate anatomical location within the genitourinary system; urine would thus provide us with rich repositories of bladder cancer EVs. We previously demonstrated feasibility of urine EV testing in kidney transplant rejection. Through increased excited charges generated by inducing plasmonic resonance in gold nanoparticles, we recently accelerated electrochemical reactions within our most current and scalable platform to achieve 12-fold signal increase from EV surface markers. We hypothesize that advancing our nanoplasmonic EV sensor for human urine and optimizing assay protocols to measure intra-EV and surface markers, could identify high value bladder cancer and host biomarkers (protein and mRNA) to better examine their interplay over time and under treatment pressures. We propose three specific aim: AIM 1: To optimize our nanoplasmonic sensor (NEXT) assay and instrumentation for high-throughput urine-based analyses and comprehensive profiling of both surface and intra-EV markers. AIM 2: To employ pre-clinical and banked biospecimens for NEXT analyses to examine profiling performance and inform optimal biomarker panel. AIM 3: Use NEXT to prospectively monitor and profile urinary EVs from patients undergoing immunotherapy-based therapies for bladder cancer. Impact: Our highly complementary group of accomplished investigators bring to bear longstanding expertise and translational experience in EV biology, bioengineering, systems biology, bioinformatics, and clinical oncology. If successful, our urinary nanoplasmonic EV platform would add critical actionable insights into immunotherapy-based treatments of advanced bladder cancers with promise in other prevalent genitourinary cancers such as kidney and prostate.