Clinical translation of 19F MRI to visualize cancer immunotherapeutic cells

在这一更新应用中，我们建议使用一种新的细胞磁共振成像(MRI)技术来 可视化肿瘤浸润性淋巴细胞(TIL)在头颈癌(HNC)患者中的运输。在 在第一个资金周期中，我们成功地开发并实施了一种颠覆性的临床翻译 基于新型全氟碳(PFC)乳胶探头(CS-1000)的成像技术 19(19F)MRI细胞检测。总体而言，感兴趣的细胞群在培养中使用非细胞内标记 有毒的PFC作为培养基添加剂。在转移到受试者之后，体内的细胞被跟踪 19F磁共振成像。细胞内的氟产生细胞特有的图像，没有背景信号。图像被量化 以测量蓄积部位的表观细胞数量。在试点试验中，免疫治疗性树突状细胞 (DC)用CS-1000标记，并用19F MRI对结直肠癌患者进行纵向检测。建房 在这些努力中，我们的目标是将这项技术用于HNC的TIL成像。HNC是第六大常见癌症 全世界。在美国，HNC占每年诊断的所有癌症的3%，占癌症的2%- 相关死亡，复发后预后差(生存2个月)。免疫疗法正在成为一种关键 抗癌战略，有可能提供针对患者的、毒性较低和更有效的治疗方法。TILs 已被证明在治疗黑色素瘤方面取得了成功，加州大学圣迭戈分校正在进行一项重大努力，以开发这种治疗HNC的方法。 尽管到目前为止，TIL疗法已经在数百名患者中使用，但基本问题仍然是 肿瘤归巢与体内TIL的细胞存活。到目前为止，我们一直对细胞的行为视而不见 给病人输液。重要的是，TIL贩运以及细胞存活率可能是应答者与 对治疗无反应。成像可以提供实时替代标记物来测量TIL肿瘤归巢 每个患者的容量和TIL存活率，这可以更好地为治疗设计和试验后数据提供信息 分析。该提案有三个具体目标：目标1：为TIL贴上PFC标签。(A)我们会发展组织培养 临床规模的临床TIL批次的PFC标记方案(1×109个细胞)。(二)我们将严格评估 PFC标记在体外诱导TIL活性和表型潜在变化的程度。目标2：In 活体啮齿动物研究以评估TIL的生物分布。使用人患者来源的异种移植瘤(PDX) HNC模型，我们将评估CS-1000标记的TIL的肿瘤归巢能力和整体生物分布 (CS-TILs)联合使用PD-1阻滞剂和不联合使用PD-1受体阻滞剂。我们将检验PD-1协同- 给药会增加肿瘤的归巢和蓄积。目的3：HNC患者的临床CS-TIL。 在两个临床试验中，我们将评估使用MRI常规检测(A)的可行性 给予CS-TIL和(B)CS-TIL联合PD-1阻断。19F核磁共振将是 用于纵向测定推测的CS-TIL肿瘤归巢和存活率。总体而言，这项研究将有所帮助 将19F核磁共振的使用扩大到涉及T细胞和其他类型白细胞的广泛临床试验。

In this renewal application, we propose to use a novel cellular magnetic resonance imaging (MRI) technology to visualize the trafficking of tumor infiltrating lymphocytes (TILs) in head and neck cancer (HNC) patients. In the first funding cycle, we successfully developed and implemented ‘first-into-man’ clinical translation of a disruptive imaging technology based on a novel perfluorocarbon (PFC) emulsion probe (CS-1000) that employs fluorine- 19 (19F) MRI cell detection. Overall, cell populations of interest are intracellularly-labeled in culture using non- toxic PFC as an additive to the culture media. Following transfer to the subject, cells are tracked in vivo using 19F MRI. The fluorine inside the cells yields cell-specific images, with no background signal. Images are quantified to measure apparent cell numbers at sites of accumulation. In the pilot trial, immunotherapeutic dendritic cells (DCs) were labeled with CS-1000 and longitudinally detected in colorectal cancer patients using 19F MRI. Building on these efforts, we aim to use this technology for imaging TILs in HNC. HNC is the sixth most common cancer worldwide. In the United States, HNC accounts for 3% of all cancers diagnosed annually and 2% of cancer- related deaths, with poor prognosis (<2 mo survival) after reoccurrence. Immunotherapy is emerging as a key anti-cancer strategy with the potential to provide patient-specific, less toxic and more efficacious treatments. TILs have proven successful in melanoma, and a major effort is underway at UCSD to develop this therapy for HNC. Although TIL therapies have been used in hundreds of patients to date, fundamental questions remain about tumor homing and cell survival of TILs in vivo. Up until now, we have been blind to the behavior of cells after infusion into patients. Importantly, TIL trafficking, as well as cell survival, may be predictive of responders versus non-responders to treatment. Imaging could provide real-time surrogate markers to gauge TIL tumor homing capacity and TIL survival in each patient, which could better inform therapeutic design, as well as post-trial data analysis. The proposal has three Specific Aims: AIM 1: PFC labeling for TILs. (a) We will develop tissue culture protocols for PFC labeling of clinical TIL batches at clinical scale (>1×109 cells). (b) We will rigorously evaluate the degree to which PFC labeling induces potential alterations in TIL viability and phenotype in vitro. AIM 2: In vivo rodent studies to evaluate biodistribution of TILs. Using a human patient-derived xenograft (PDX) tumor model for HNC, we will evaluate the tumor homing ability and overall biodistribution of CS-1000 labeled TILs (CS-TILs) with and without co-administration of PD-1 blockade. We will test the hypothesis that PD-1 co- administration results in increased tumor homing and accumulation. AIM 3: Clinical CS-TILs in HNC patients. In two clinical trial HNC patient cohorts, we will evaluate the feasibility of using MRI to detect (A) conventionally administered CS-TILs and (B) CS-TILs administered in combination with PD-1 blockade. The 19F MRI will be used to assay putative CS-TIL tumor homing and survival in a longitudinal fashion. Overall, this study will help expand the use of 19F MRI to a wide range of clinical trials involving T cells and other types of leukocytes.