Product for sensitive imaging of cells in vivo

在第二阶段的申请中，我们的目标是继续开发和商业化一种新颖而敏感的 磁共振成像（MRI）探头的细胞治疗工具市场。我们建议实施先进先出- 这项技术的人类临床翻译，以可视化肿瘤浸润淋巴细胞（TILs）的运输， 头颈癌（HNC）患者。Celsense公司，制造成像工具，提供定量 体内细胞运输的评估。本公司的核心产品是基于独特的成像剂 全氟化碳（PFC）纳米乳液组合物。此前，在Celsense与 Ahrens实验室（UCSD），第一代PFC纳米乳液产品的临床使用被用于检测细胞 癌症患者的免疫治疗。在这项工作的基础上，我们建议开发下一代成像PFC 纳米乳液细胞标记物，其结合了新型金属螯合物（“FETRIS”）技术， 提高了在临床试验中检测体内免疫细胞产物的灵敏度。成像起始细胞 生物分布可以提供关于定位、存活、最佳递送途径和 治疗剂量FETRIS纳米乳剂被设计为在培养中和转移后被细胞吸收 对于受试者，使用氟-19（19 F）MRI在体内检测细胞。细胞内的氟产生细胞- 特定图像，没有背景信号。图像易于定量以测量在200 ℃下的表观细胞数。 积累的地方。这些19 F MRI方法已被证明是各种疾病的安全跟踪工具。 干细胞和免疫细胞类型。通过提高使用FETRIS的19 F细胞检测的灵敏度，我们将降低19 F细胞的表达。 将该技术应用于更广泛的细胞治疗应用的障碍。作出重大努力 正在UCSD开发HNC的TIL疗法。关于肿瘤归巢的基本问题仍然存在， TIL在体内的细胞存活。到目前为止，我们一直对细胞输注到患者体内后的行为一无所知。 重要的是，TIL运输以及细胞存活可以预测对肿瘤的应答者与非应答者。 治疗成像可以提供实时替代标记物来测量TIL肿瘤归巢能力和TIL的免疫反应性。 这可以更好地为治疗设计和试验后数据分析提供信息。该提案 有两个特定目的：目的1 - TIL-FETRIS GMP细胞制备。(a)我们将支持制造业数据， 新FETRIS FDA药物主文件（DMF）的方法。≥500 mL规模的其他工程批次 将进行放行检测和加速稳定性研究。(b)从现在的加州大学旧金山分校开始 Diego TIL方案，我们将开发临床规模的TIL-FETRIS批次的组织培养方案 （>1×109个细胞）。我们将为可接受的TIL标签建立放行标准，并严格评估 PFC标记在体外诱导TIL活力和表型的潜在改变的程度。目标2 - CS-TIL在HNC患者中的临床使用。在一个小型HNC患者队列（N=5）中，我们将评估 以及MRI检测TIL-FETRIS的有效性。将使用19 F MRI检测推定的CS-TIL肿瘤 归巢和纵向生存。

In this Phase 2 application we aim to continue development and commercialization of a novel and sensitive magnetic resonance imaging (MRI) probe for the cell therapy tools market. We propose to implement first-into- man clinical translation of this technology to visualize the trafficking of tumor infiltrating lymphocytes (TILs) in head and neck cancer (HNC) patients. Celsense, Inc., manufactures imaging tools that provide quantitative assessment of in vivo cell trafficking. The Company’s core products are imaging agents based on unique perfluorocarbon (PFC) nanoemulsion compositions. Previously, in a collaboration between Celsense and Ahrens lab (UCSD), clinical use of a first-generation PFC nanoemulsion product was used to detect cell immunotherapy in cancer patients. Building on this work, we propose to develop next-generation imaging PFC nanoemulsion cell labels that incorporate a novel metal chelate (“FETRIS”) technology that yields dramatically improved sensitivity to detect immunotherapeutic cell products in vivo in clinical trials. Imaging initial cell biodistribution can provide crucial feedback regarding the localization, survival, optimal routes of delivery and therapeutic doses. FETRIS nanoemulsion is designed to be taken up by cells in culture, and following transfer to the subject, cells are detected in vivo using fluorine-19 (19F) MRI. The fluorine inside the cells yields cell- specific images, with no background signal. Images are readily quantified to measure apparent cell numbers at sites of accumulation. These 19F MRI methods have been demonstrated to be a safe tracking tool for various stem cells and immune cell types. By improving the sensitivity of 19F cell detection using FETRIS, we will lower the barriers for applying this technology to a wider range of cell therapy applications. A major effort is underway at UCSD to develop TIL therapy for HNC. 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 and post-trial data analysis. The proposal has two Specific Aims: Aim 1 - TIL-FETRIS GMP cell preparation. (a) We will bolster manufacturing data and methods for a new FETRIS FDA Drug Master File (DMF). Additional engineering batches at ≥500 mL scale with release testing and accelerated stability studies will be produced. (b) Starting with the current UC San Diego TIL protocol, we will develop tissue culture protocols for TIL-FETRIS batches at clinical scale (>1×109 cells). We will establish a release criteria for acceptable TIL labeling and rigorously evaluate the degree to which PFC labeling induces potential alterations in TIL viability and phenotype in vitro. Aim 2 - Clinical use of CS-TILs in HNC patients. In a small HNC patient cohort (N=5), we will evaluate the safety and efficacy of using MRI to detect TIL-FETRIS. The 19F MRI will be used to assay putative CS-TIL tumor homing and survival longitudinally.